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Editor-in-Chief
Prof. VK Joshi
Professor and Head,
Fermentation Technology Lab, Department of Food Science and Technology
Dr YS Parmar University of Horticulture and Forestry
Nauni, Solan, Himachal Pradesh, India
E Mail:vkjoshipht@rediffmail.com
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EDITORIAL BOARD
Editor-In-chief
Prof. V.K. Joshi
Professor and Head,
Fermentation Technology Lab,
Department of Food Science and Technology,
Dr YS Parmar University of Horticulture and
Forestry, Nauni, Solan, Himachal Pradesh, India
vkjoshipht@rediffmail.com
editorijfft@gmail.com
Dr Neerja S. Rana
Department of Basic Sciences
Dr. Y.S. Parmar University of Horticulture and
Forestry, Nauni-Solan, Himachal Pradesh, India
drneerjauhf24@yahoo.co.in
Dr Shashi Bhushan
Division of Biotechnology,
Institute of Himalayan Bio-resource Technology (CSIR),
Palampur,DisttKangra, Himachal Pradesh, India
sbhushan@ihbt.res.in,
Dr Satish Kumar Sharma
G B Pant University of Agriculture & Technology, Hill campus,
Ranichauri, Distt Tehri-Garhwal,
Uttarakhand, India
drsatish10@yahoo.com
Dr Om Prakash Chauhan
Scientist
Fruits and Vegetables Technology Division
Defence Food Research Laboratory
Siddarthanagar, Mysore, India
opchauhan@gmail.com
Dr PS Panesar
Associate Professor
Biotechnology Research Laboratory
Department of Food Engineering and Technology
SL Institute of Engineering and Technology,
Longowal, Punjab,India
pspanesarrr@yahoo.com
Contents
International Journal of Food and Fermentation Technology
Vol. 2 No. 1, June 2012
Review paper
A Panorama of lactic acid bacterial fermentation of vegetables
V.K. Joshi and Somesh Sharma
1-12
Research Paper
Application of thermostable -amylase in Streptomyces erumpens liquefaction of cassava bagasse for production of lactic acid 13-18
Ramesh C. Ray and Shaktimay Kar
Changes in phytochemicals during fermentation of wine grapes
A.K. Sharma, S.V. Navale, S.N. Aute, G. S. Karibasappa, D. P. Oulkar and P. G. Adsule
19-25
Cloning and expression of rec-pediocin CP2 in Escherichia coli using OmpA and TAP gene fusion approach
Balvir Kumar, P. P. Balgir and B. Kaur
27-36
37-41
Design of cascade membrane filtration process for clarification of whey proteins and membrane fouling
Pranav Kaushik Pidatala and Senthil R. Kumar
43-48
Microbiological and biochemical characterization of Seera: A traditional fermented food of Himachal Pradesh
Savitri, N. Thakur, D. Kumar and T.C. Bhalla
49-56
57-61
-amylase production from Endomyces fibuliger an indigenous yeast isolate of Western Himalayas
Keshani Bhushan, Anamika Jain, O.P. Sharma, B. Singh and S.S. Kanwar
63-69
Statistical screening of media components for the production of arginine deiminase by Weissella confusa GR7
Baljinder Kaur and Rajinder Kaur
71-79
Effect of nitrogen source on the fermentation behaviour of musts and quality of wine from two Cvs. of pineapple
B.L. Attri
81-86
Research Note
Effect of Solid state fermentation and yeast species on composition of apple pomace: Application of PCA
V.K. Joshi and D.K. Sandhu
87-91
Processing potential of newly introduced Amla cultivars grown in lower Himalayan Region of Himachal Pradesh
Manisha Kaushal and Shashi K. Sharma
Evaluation of the stability of plum anthocyanin powder in RTS based model solution
M. Preema Devi, V.K. Joshi and Y. Indrani Devi
Book Review
93-97
99-101
103
V.K. Joshi
Conceptual Editorial
enzymes and microorganisms, or packaging in order to curtail the loss of perishables. Utilizing improved postharvest practices
thus often results in reduced food losses, improved overall quality and food safety, and higher profits for growers and marketers.
The most important goals of post-harvesting handling are to keep the product cool, to avoid moisture loss and slow down
undesirable chemical changes, and avoiding physical damage such as bruising, to delay spoilage. The way the grains are stored
after harvest becomes the major cause of contamination with fungi some of which are producer of toxin like aflatoxin. The fruit
like apple after contamination has a toxin called patulin, if not stored properly. Sanitation is also an important factor, to reduce the
possibility of pathogens that could be carried by fresh produce, for example, as residue from contaminated washing water.
Consumption of such food results in the diseases of gastro-intestinal track which sometimes prove to be fatal.
Postharvest loss reduction technology encompasses the usage of optimum harvest factors, reduction of losses in handling,
packaging, transportation and storage with modern infrastructure machinery, processing into a wide variety of products, home
scale preservation with low cost technology. Use of thermal processing, low temperature storage drying chemical and biological
reactions coupled with other preservation techniques are applied to enhance the storability of the perishables.
There are different constraints for postharvest management like: large number of small and marginal farmers with primitive
system of cultivation, poor infrastructure in terms of handling, transport, storage, processing and marketing, lack of adequate quality
systems and procedure like grading and sorting, hot and humid climates, cost of installation of post-harvest treatment facilities is
expensive, lack of awareness training for the rural farmers and inconsistency in supply due to seasonality of the produce.
Adoption of latest techniques could make available a large quantity of food by avoiding losses and provide quality food and
nutrition, more raw materials for processing. Postharvest technology has the capability to meet food requirement of growing
population. By adopting improper and inefficient methods of storage we are losing a substantial portion of production. If better
methods of processing and storage are adopted, the losses could be reduced to a large extent. The developing countries need
proper infrastructure for this and a strong will to adopt modern postharvest technology to save the wastage. Thus, it could
certainty prove to be a panacea for the problems faced by the developing countries.
V K Joshi
Review paper
Department of Food Science and Technology, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal
Pradesh,India
2
Department of Biotechnology, School of Bioengineering and Food Technology, Shoolini University, Bajhol, Solan, Himachal
Pradesh,India
*Email: vkjoshipht@rediffmail.com
Paper no: 31
Received: 14 February,2011
Abstract
Vegetables occupy a prominent position in our diet and form the most nourishing diet.Preservation of
vegetables is carried out to prevent their postharvest losses of functional quality whilst providing safe
and stable products. In general, preservation process consists of a combination of mild heat stress and
low concentration of chemical preservatives to control the food spoilage and the growth of pathogenic
spore- forming bacteria. But, consumer demand today is for natural and minimally processed foods, with
a fresh like appearance and taste, easy-to-eat and with high level of safety.It is also dependent on a wide
range of technologies to ensure that food is maintained at an acceptable level of quality from the time of
manufacture till the time of consumption. As a result, research and development of new products is
leading to the reduction or even replacement of heat treatments and encouraging the use of traditional
preservatives as these are capable of assuring the sensory and nutritional properties of the product
without reducing food safety. Non-thermal preservation methods are thus gaining interest as alternative
treatments in the food industry due to their capability of assuring the quality and safety of foods.
Among them, use of natural antimicrobials from plants (essential oil) and microorganisms (LAB and
Bacteriocins) have gained wide range of popularity as bio-preservatives. Traditionally, microorganisms
have been used in fermentation from ages but now modern large scale production of foods exploits use
of defined strains to ensure consistency and quality in the final product. The biopreservation offer
potential application in food preservation and helps in reducing the addition of chemical preservatives as
well as the intensity of heat treatment, resulting in foods which are more naturally preserved and richer in
sensory and nutritional qualities. Beside this, biopreservation also provides health benefits such as
reduction in cardiovascular diseases, improving gastro intestinal microorganisms and immune system,
helps in curing diarrhea and lowering cholesterol in the consumers of lactic acid fermented foods. The
preservation is accomplished through the production of antimicrobial substances such as lactic acid,
diacetyl, hydrogen peroxide, ethanol reutrin and bacteriocin. The bacteria from the genera Lactobacillus,
Leuconostoc, Pediococcus and Streptococcus are the main species involved in the lactic acid fermentation.
Thus,the lactic acid fermentation of vegetables is an important method of bio-preservation and providing
healthful foods to the consumers.
The Chinese were the first to ferment the vegetables and there
is evidence to show that they had prepared such vegetables
at the time of building the great wall of China (Pederson, 1971).
Today, because of the development of efficient heat sterilizing
and refrigeration systems, lactic acid fermentation, to some
extent has lost its significance as a food preservation method
in industrialized countries, but is being used as a taste
diversification tool. It is extensively used in developing
countries, but is gaining importance more in developed
countries as a food with therapeutic value. It is true especially
for vegetables, where either natural fermentation or
fermentation by pure culture of lactic acid bacteria is practiced
even in the developed countries (Pederson, 1971; Fleming and
Mcfeeters, 1981; Anderson et al., 1990).
Role and advantages of Vegetables Fermentation
Fermented foods including fruits and Vegetables play an
important role in providing food security, enhancing livelihoods
and improving the nutrition and social well being of millions of
people around the world, particularly the marginalized and
vulnerable groups (Table 1). This is achieved through improved
food preservation that combines salting to selectively control
the microorganisms and fermentation to stabilize the treated
2
Many vegetables contain naturally occurring toxins and antinutritional compounds. These can be removed or detoxified
by the action of micro-organisms during fermentation. For
instance the fermentation process that produces the Sudanese
product Kawal removes the toxins from the leaves of Cassia
obtusifolia and fermentation is an important step in ensuring
that cassava is safe to eat. The production of fermented
vegetable products provides income and employment to
millions of people around the world.
The optimum health and nutrition of individuals is dependent
upon a regular supply of food and a balanced diet. When diets
are sub-optimal, the individuals capacity for work and
achievements are greatly reduced. The most vulnerable groups
are women, children and weaning infants. Availability of food,
dietary restrictions and taboos, misconceptions, limited time
available for feeding or eating contribute to create a group of
individuals who are nutritionally disadvantaged.
Facultative homofermenter
Obligate heterofermenter
Enterococcus faecium
Enterococcus faecalis
Lactobacillus acidophilus
Lactobacillus lactis
Lactobacillus delbrueckii
Lactobacillus salivarius
Pediococcus pentocacus
Streptococcus thermophilus
Pediococcus acidilactici
Pedicoccus damnosus
Lactobacillus bavaricus
Lactobacillus casei
Lactobacillus coryniformis
Lactobacillus curvatus
Lactobacillus plantarum
Lactobacillus sake
Lactobacillus brevis
Lactobacillus buchneri
Lactobacillus cellobiosus
Lactobacillus confuses
Lactobacillus coprophilus
Lactobacillus fermentatum
Lactobacillus sanfrancisco
Leuconostoc dextranicum
Leuconostoc mesenteroides
Leuconostoc paramesenteroides
Microorganisms
As described earlier ,lactic acid bacteria (LAB) responsible for
vegetable fermentation. Micro-organisms vary in their optimal
pH requirements for growth. Most bacteria favour conditions
with a near neutral pH . The varied pH requirements of different
groups of micro-organisms is used to effect fermented foods
where successions of micro-organisms take over from each
other as the pH of the environment changes. Certain bacteria
are acid tolerant and will survive at reduced pH levels. Notable
acid-tolerant bacteria include the Lactobacillus and
Streptococcus species, which play a role in the fermentation
of vegetable products. Oxygen requirements vary from species
to species. Many researchers have studied the changing
pattern of microflora involved in the fermentation of kimchi
along with the taste and odour produced during fermentation.
Major microflora isolated from kimchi were; Lactobacillus
plantarum, Lactobacillus brevis, Streptococcus faecalis,
Leuconostoc mesentroides, Pediococcus cerevisae and
Achromobacter. Recently, most of the fermented vegetable
juices are manufactured by lactoferment Process through
the use of pure starter cultures (Buckenhuskes, 1993)
Water activity
Temperature
Additives
Oxygen availability
Salt concentration
Type of salting
2-3
3
5-8
4-7
1.5-32.52.5
Dry salt
Dry salt
Brine
Brine
BrineDry saltDry salt
2-5
5.3
7.5-8.5
2.2
2.5
Brine
Brine
Brine
Dry salt
Dry salt
CabbagesSauerkraut
Korean Kimchi
Cucumbers
Green olives
Carrots Raddish
Black olives
Over-night Dill pickles
Genuine Dill pickles
Sauerrruben (turnip)
Pickled leafy vegetables
References
Fleming, 1982
Steinkraus et al., 1983
Fleming, 1982
Fleming, 1982
Niketic-Aleksic et al., 1973
Joshi, et. al., 2008
Sharma and Joshi, 2007
Battcock and Azam-Ali, 1998
Frazier and Westhoff, 1998
Frazier and Westhoff, 1998
Frazier and Westhoff, 1998
Frazier and Westhoff, 1998
Microorganisms involved
Reference
Lactobacillus casei
Lactobacillus strains
Lactobacillus brevis,Pediococcus
cerevisiaeLactobacillus plantarum
Lactobacillus plantarum,Pediococcus
pentosaceus
Lactobacillus plantarum,Lactobacillus
brevis,Streptococcus faecalis,Leuconostoc
mesenteroides,Pediococcus pentosaceus
Lactobacillus plantarum,Lactobacillus
brevis,Bacillus coagulans
Pediococcus pentosaceus
Bacillus subtilis,Lactobacillus plantarum
Gundruk
Bossam kimchi (Pickled cabbage kimchi)
Karki, 1986
Joshi et al., 2003
Makayama, 1957
Table 5: Metabolites of lactic bacteria which may be inhibitory to other pathogenic and food spoilage organisms
Product
Organic acids
Lactic acid
Putrefactive and Gram-negative bacteria, some fungi
Acetic acid
Putrefactive bacteria, clostridia, some yeasts and some fungi
Hydrogen peroxide
Pathogens and spoilage organisms, especially in protein rich foods
Enzymes
Lactoperoxidase system
Pathogens and spoilage causing bacteria (milk and diary products) with hydrogen peroxide
Lysozyme
(by recombinant DNA)
Undesired Gram-positive bacteria
Low molecular weight metabolites
Reuterin
Wide spectrum of bacteria, yeasts and molds
Diacetyl
Gram-negative bacteria
Fatty acids
Different bacteria
Nisin
Some LAB and Gram-positive bacteria, notably endospore-formers
Other
Gram-positive bacteria, inhibitory spectrum according to producer strain and bacteriocin type
Source: Breidt & Fleming, 1997, Joshi et al., 2006
(Bhunia et al., 1998). The use of pure starter culture (LAB) has
been reported for the inhibition of spoilage organism
Pseudomonas fluorescens and Staphylococcus typhimurium
(Raccach et al., 1979). The growth of Staphylococcus aureus
is also inhibited when grown with Pediococcus cerevisiae and
Lactobacillus plantarum. A wide variety of raw foods are
preserved by LAB fermentation including milk, meat, fruit and
vegetables (Daeschel, 1989). Reduction of pH and removal of
a large amount of carbohydrates by fermentation are the primary
preserving actions that these bacteria provide, though, LAB
are capable of producing inhibitory substances that are
antagonistic towards other microorganisms as described earlier
(Joshi and Thakur, 2000; Battcock and Azam-Ali, 1998).
Bacteriocin or bacteriocin
like substances
Lactobacillus plantarum
Plantacin B, Plantaricin C,
Plantacin N
Nisin, Lacticin, Lactococcin
G
Diplococcin, Lactococcin
Pediocin AcH, Pediocin
P-A-1
Mesenterocin 5
Lactocidin, Acidophilin
Caseicin
Bacteriocin
Inhibitory Spectrum
Bacillus subtilis
Pediococcus acidilactici
Lactobacillus sake
Leuconostoc mesenteroides UL5
Propionibacterium shermanni
Lactobacillus sakei
Future Trends
Hypercholesteremic effect
Hepner et al. (1979) reported hypercholesteremic effect of
yoghurt in human subjects receiving a one-week dietary
supplement. Studies on supplementation of infant formulation
with Lactobacillus acidophilus showed that the serum
cholesterol in infants was reduced from 147 mg/ml to 119 mg/
100 ml (Harrison and Peat, 1975). The ability of yoghurt to
lower the cholesterol in serum by controlled human trials had
also been reported (Poppel and Schafsma 1996). Possible role
of LAB in lowering cholesterol concentration and various
mechanisms by which it may be possible has also been
discussed (Haberer et al., 1997).
Anticarcinogenic effect
Apart from this, there are interesting data on anticarcinogenic
effect of fermented foods showing a potential role of lactobacilli
in reducing or eliminating procarcinogens and carcinogens
from the alimentary canal (Shahani, 1983; Mital and Garg, 1995).
Mutagenicity
The fermentation of foods is also reported to reduce the
mutagenicity of foods by degrading the mutagenic substances
during the process. Lactic acid bacteria isolated from dadih, a
traditional Indonesian fermented milk, were found to be able
to bind mutagens and inhibit mutagenic nitrosamines. Milk
fermented with Lactobacillus acidophilus LA-2 was
demonstrated to suppress faecal mutagenicity in the human
intestine.
Immune system
Some LAB present in fermented milk products, are found to
play an important role in the immune system of the host after
colonisation in the gut (De Simone, 1986). The mechanism of
this effect is not clearly known, but it is speculated that the
10
References
Adams, M.R 1990. Topical aspects of fermented foods. Trends in
Food Sci. Technol., 1:141-144.
Adams, M.R. and Moss, M.O. 1996. Fermented and microbial foods.
In: Food Microbiology. New Age International (P) Ltd.
Publishers, New Delhi. pp. 252-302.
Anand, J.C. and Johar, D.S. 1957. Effect of condiments on control of
Aspergillus niger in mango pickle. J. Sci. Indus. Res., 16A:
370.
Anderson, R.E., Eriksson, C.E., Salmonsson, A.C. and Theander, D.
1990. Lactic acid fermentation of fresh and stored carrot chemical, microbial and sensory evluation of of products.
Lebensm-Wiss-Technol., 23:34-40.
Arthur, R.A.J., 1986. Tribal Recipe may help to Feed the World,
London Press Service 060416, UK
Axelsson, L., 1998. Lactic Acid Bacteria: Classification and
Physiology. In: Lactic Acid Bacteria, Microbiology and
functional Aspects. Ed. S Salminen and A von Wright, Marcel
Decker Inc, New York, USA.
Axelsson, L., Chaung, T.C., Dobrogosz, W.J. and Lindgren, S.
1998. Discovery of a broadspectrum antimicrobial substances
provided by Lactobacillus reuterii. Unpublished manuscript.
North Carolina State University, Releigh.
Barnby-Smith, F.M. 1992. Bacteriocins: Applications in Food
Preservation. Trends Food Sci. Technol., 3(6):133.
Battcock, M. and Azmi-ali, S. 1998. Fermented fruits and vegetables.
In: Fermented Fruits and Vegetables. A Global Perspective.
FAO Agricultural services bulletin No 134, Food and
Agriculture Organization of the United Nations, Italy, Rome.
Beuchat, L. R., 1995. Application of biotechnology to fermented foods.
Food Technology, 97-99.
Bhunia, A.K., Johnson, M.C. and Ray, B. (1988). Purification,
characterization and antimicrobial spectrum of a bacteriocin
produced by Pediococcus acidilacti H. J. Appl. Bacteriol.,
65: 261-268.
Boonlong, N. 1986. Traditional technologies of Thailand : Traditional
Fermented Food Products. In: Traditional Foods: Some
Products and Technologies. Central Food Technological
Research Institute, Mysore, India.
Breidt, F. and Fleming, H.P. 1997. Using lactic acid bacteria to improve
the safety of minimally processed fruits and vegetables. Food
Technol., 51:44-46, 48,51.
Broughton, J.D. 1990. Nisin and its uses as a food preservative.
Food Technol., 44(5):110-117.
Buckenhuskes, H.J. (1993). Selection criteria for lactic acid bacteria
to be used as starter cultures for various food commodities.
FEMS Microbiol. Rev., 12:253-272.
Buckenhuskes, H.J. 2001. Fermented vegetables. In: Food
Microbiology : Fundamentals and Frontiers, 2nd edn. M.P.
11
Research Paper
Microbiology Laboratory, Regional Center of Central Tuber Crops Research Institute, Bhubaneswar, India
*Email: rc_rayctcri@rediffmail.com
Paper no: 32
Abstract
Application of thermostable - amylase (crude and purified) produced by Streptomyces erumpens on
liquefaction of cassava bagasse into sugars for further conversion to lactic acid (LA), was compared
with that of a commercial enzyme Termamyl (Novozyme, Denmark). By applying crude S. erumpens
amylase (10%, v/v), 21.89 g LA was achieved per 100g bagasse at 96h of incubation, which was 44.01%
less than LA yield obtained with Termamyl (2%, v/v). However, when partially purified S. erumpens
amylase was applied at 5% (v/v) concentration or Termamyl at 2% (v/v) concentration to liquefy
cassava bagasse, the LA yield was similar, i.e., 38.45 LA/100g bagasse. The purified form of S. erumpens
amylase was found as effective as commercial Termamyl enzyme in liquefying cassava bagasse to
sugars for further conversion to LA.
2012 New Delhi Publishers. All rights reserved
Commercial enzymes
Termamyl is a thermostable (>90C) - amylase enzyme and
Dextrozyme (>45C) is a fungal amyloglucosidase
(glucoamylase) enzyme. The enzymes were procured from Arun
and Co. Mumbai, India.
S. erumpens enzyme
Crude -amylase was prepared from S. erumpens as follows;.
Sterile SB medium (50 ml) taken in 250 ml Erlenmeyer flasks (in
triplicate) were inoculated with 2% inoculum and agitated at
120 rpm in an incubator shaker (Remi Pvt Ltd, Mumbai, India)
at 50C. After 36h, the culture broths from the flasks were
pooled and centrifuged at 8000 rpm in a refrigerated centrifuge
(Model C-24, Remi Pvt. Ltd, Mumbai, India) for 20 min at 4C.
The clear cell free supernatant was used as the crude enzyme.
Cassava bagasse
LA production
The starch slurry was prepared in 500 ml Erlenmeyer flask by
mixing cassava bagasse (25g) with 250 ml of distilled water. It
14
Application of thermostable -amylase in Streptomyces erumpens liquefaction of cassava bagasse for production of lactic acid
Figure 1: Flow chart for lactic acid production from cassava bagasse
15
Statistical analysis
Total
volume (ml)
Total enzyme
activity (Units)
Total
protein (mg)
Yield
(%)
Specific activity
(Units/mg protein)
Fold of
purification
100
15
16
34000
6747
8241.6
250
24.8
20.2
100
19.8
24.24
136
272
408
0
2.0
3.0
16
Application of thermostable -amylase in Streptomyces erumpens liquefaction of cassava bagasse for production of lactic acid
Reference
Amerine ,M.A. and Ough, C. 1984. Methods for Analysis of Musts
and Wines, New York, Wiley- Inter Science Publication, John
Wiley and Sons, pp. 447.
Anuradha, R., Suresh, A.K. and Venkatesh, K.V. 1999. Simultaneous
saccharification and fermentation of starch to lactic acid.
Process Biochem. 35: 367375.
Edison, S., Anantharaman, M. and Srinivas, T. 2006. Status of cassava
in India an overall view. Tech Bull Ser, 46, Pp, 79, Central
Tuber Crops Research Institute, Thiruvanathapuram, India,
Fitzpatrick, J.J. and O_Keeffe, U. 2001. Influence of whey protein
hydrolyzate addition to whey permeate batch fermentations
for producing lactic acid. Process Biochem. 37: 183186.
Gupta, R., Gigras, P., Mohapatra, H., Goswami, V.K. and Chauhan,
B. 2003. Microbial - amylases: a biotechnological
perspective. Process Biochem. 38: 1599-1616.
Haki ,G.D. and Rakshit, S.K. (2003). Developments in industrially
important thermostable enzymes. Biores. Technol. 89: 17-34.
Hassan, K.S., Moldes, B., Richard, G.K. and Richard, J.S. 2001.
Lactic acid production by simultaneous saccharification and
fermentation of alfalfa fiber. J. Biosci. Bioeng. 92: 518-523.
John, R.P., Nampoothiri ,K.M. and Pandey, A. 2006. Solid state
fermentation for L- lactic acid production from agro wastes
Conclusion
It was concluded that application of partially purified S.
erumpens -amylase was more effective than crude amylase
17
18
Research Paper
Email: sharmadrajay@gmail.com
Paper no: 33
Abstract
Changes in various phytoemicals during fermentation of grape cultivars for red wine (Cabernet
Sauvignon and Zinfandel) and white wine (Sauvignon Blanc and Charark-4; a cross of Chardonnay
and Arkavati) were determined. The fermentation was performed in food grade plastic vessels, using
a commercial wine yeast strain Premier Cuveeat 201C. The data were recorded on total phenols,
anthocyanin,Ferric ion reducing antioxidant power (FRAP), DPPH assay and individual phenolic
content and amino acids. At the initial stage of fermentation, total phenols were higher in Zinfandel
than in Cabernet Sauvignon, but increased to 4-folds at the completion of fermentation. Higher levels
of total phenols and antioxidant activities were noted in red wines than in white ones. Free radical
scavenging activity (DPPH) was higher in Cabernet Sauvignon than in Zinfandel. However,resveratrol
content was more than twice in Zinfandel wine than in Cabernet Sauvignon wine. White wines
showed low levels of phenolic compounds and some phenolics viz.; epicatechingallate, resveratrol
and kaempferol were found absent. During the process of fermentation quantities of various amino
acids were changed.Alanine, serine, lysine and vaniline amino acids were found absent at beginning
but on the completion of fermentation presence of these amino acids was confirmed and quantified.
2012 New Delhi Publishers. All rights reserved
Sharma et al.
Physico-chemical analysis
The collected samples were analyzed following standard
operating procedures. Total phenols were quantified by
following the procedure of Singleton and Rossi (1965).
Content of anthocyanins in samples was estimated by using
method of Fulekiand Francis (1968). Antioxidant activities were
measured by Ferric ion reducing antioxidant power (FRAP),
and Free radical scavenging activity (DPPH assay). In
measuring of FRAP method of Benzie and Strain (1996) was
adopted and in case of DPPH assay method of Arnouset al.
(2001) was followed. LC-MS/MS (Agilent Technologies with
series hyphenated to API 4000 Qtrap (ABS Sciex) mass
spectrometer equipped with electrospray ionization (ESI+)
probe 1200) was used for quantification of individual phenolic
content in samples. For amino acids, the HPLC (Perkin Elmer
200 series) of column G8 (50 x 4.6 mm ID x 1.8 m) was used,
with mobile phase A: 0.1 % pentadecaflurooctonoic acid and
0.1 % formic acid in Water: Methanol (90:10), B: 0.1% formic
acid in water: methanol (10:90). Calibration curves were
obtained by plotting the peak areas against different
concentration of amino acids and poly phenolic compounds.
Table 1: Changes in total phenols and anthocyanin content at different intervals during fermentation process of wine grapes.
Days
Initial
2
3
6
11
SEM
CD at1%
950
1770
1990
3880
3760
5.362
16.16
Anthocyanin (mg/L)
White
Red
330
260
230
230
220
2.828
8.52
652
680
545
393
370
6.251
18.84
20
White
CabernetSauvignon
Zinfandel
1070
8330
14330
19080
15030
2.236
6.74
3400
7940
10210
12350
8340
3.162
9.53
Figure 1: Total ion chromatograph of 100 ng/mL calibration standard (A), red wine (B), white wine (C)
21
Sharma et al.
Table 3: Changes in antioxidant property during fermentation process at the interval of days
FRAP (mg/l)
Red
White
CabernetSauvignon
Initial
2
3
6
11
SEM
CD at1%
DPPH (mM)
Red
290
280
340
320
660
5.477
16.51
130
380
510
600
570
10.624
32.02
0.33
0.32
0.19
0.28
0.26
0.010
0.03
CabernetSauvignon
1.7
2.3
1.5
1.1
1.3
0.028
0.09
White
Zinfandel Sauvignon Blanc
1.24
1.51
1.46
2.10
2.29
0.054
0.16
1.19
1.23
1.60
1.76
1.90
0.053
0.16
0.39
0.42
0.35
1.13
0.36
0.019
0.06
Charark-4
0.60
0.74
0.61
0.52
0.63
0.079
NS
Epicatechingallate
Caffeicacid
Epicatechin
p-Coumaricacid
Quicetrin
Syringicacid
Vanillicacid
Quercetin
Resvaretrol
Catechin
Kaempferol
7.87
7.42
7.14
7.86
8.24
7.29
7.21
9.63
9.24
6.84
7.46
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
[M+H]+
443
181
291
165
449
199
169
303.0
229.0
291.0
287.0
Quantifier
Qualifier
Q3
DP
CE
CXP
Q3
CE
CXP
123.0
89.1
123.0
147.0
303.2
155.0
125
153.0
135
165.0
165.0
51
26
50
56
45
56
55
55
29
38
53
19
43
23
16
15
14
14
45
22
17
40
5
6
5
6
9
7
5
5
6
7
2
273
135
139,165
119 , 91
129, 85
123, 77
93
137, 69
107, 119
139,123
121
12
27
20 ,18
25, 35
22, 35
18,38
21
45,80
30,55
22
50
5
8
6
5,3
6
5,2
3
5,1
4,5
7, 2
5
* Q1- Precursor ion, Q3- product ion, DP- declusturing potential, CE- collision energy, CXP- collision exit potential
22
Concentration (mg/L)
Red
Epicatechingallate
Caffeic acid
Epicatechin
p- Coumaric acid
Quicetrin hydrate
Rutin hydrate
Syringic acid
Quercetin
Resveratrol
Catechin
Kaempfero
Total Phenolics
White
Cabernet Sauvignon
Zinfandel
Sauvignon Blanc
Charark-4
1.10
2.14
59.80
0.52
9.10
7.98
5.27
7.08
0.50
117.00
0.316
4160.00
0.39
2.47
59.30
2.65
1.65
1.97
5.29
2.71
1.08
71.40
0.14
1050.00
0.00
0.35
0.11
0.00
0.60
0.24
0.16
0.11
0.00
0.07
0.00
20.00
0.00
0.33
1.54
0.81
4.07
0.22
0.29
0.18
0.00
3.51
0.00
50.00
Table 5: Changes in quantity of amino acids (mg/l) during fermentation of wine grapes
Amino acid
Arginine
Aspartic Acid
Glutamic Acid
Alanine
Histidine
Hydroxyproline
Leucine
Lysine
Methionine
Ornithine
Phenylalanine
Proline
Serine
Tryptophan
Tyrosine
Vaniline
th
st
Sauvignon Blanc
th
th
Charak-4
1 day
12 day
1 day
12 day
1 day
12 day
1 day
12th day
113
0
0
0
5.08
15.5
1.36
9.53
1.53
4.17
0
448
0
0
1.37
0
79.9
5.7
23
94.5
36.9
32.4
32.7
30.2
0
31.3
9.06
1150
13.5
0.285
11.4
10.5
702
21.8
70.9
242
35.4
5.33
15.1
0
1.58
9.37
26.9
421
81.3
6.19
25
30
80.1
26.1
41.6
54.5
12.9
8.22
31.9
33.4
12.5
8.19
13.1
462
19.2
0
24.2
21.8
34.4
0.274
0.105
0.94
2.97
10.3
0.367
0
0
0.831
0
940
4.39
0
1.05
0.646
80.4
29
30.1
58.3
8.74
20.2
25.8
29.2
11.4
10.9
9.22
928
21.1
0.946
20.1
18.3
37.3
2.49
0.203
15.3
3.39
9.93
0
0
1.55
2.61
0.156
200
0
0.063
3.37
0
126
69.4
67
189
16.5
18.2
107
64.3
29.3
52.7
26.8
733
39.2
0
55.4
55.3
st
st
23
Sharma et al.
24
Ough, C.S. 2009. Acids and Amino Acid in Grapes and Wines.Modern
Methods of Plant Analysis Wine Analysis, pp 98-121
Linskens, H. F., and Jackson, J. F. (Eds.) Springer.
Pakhale, S.S., Karibasappa, G.S., Ramchandani, A.G., Bhushan, B.,
and Sharma A 2007. Scavenging effect of Indian grape
polyphenols on 2,2-diphenyl-1-picrylhydrazyl (DPPH)
radical by electron spin resonance spectrometry. Indian Jour
Experi Biol., 45: 968-973.
Price, S.F., Breen, P.J., Valladao, M. and Watson, B.T. 1995. Cluster
sun exposure and quercetin in Pinot noir grapes and wines.
Amer. J. Enol. Vitic., 46:187194.
Schmandke, H. 2002. Resveratrol and piceid in grapes and soybeans
and products made from them. Ernhrungs-Umsch 49: 349
352.
Sims, C.A. and Bates, R.P. 1994. Effects of skin fermentation time
on the phenols, anthocyanins, ellagic acid sediment, and
sensory characteristics of a red Vitisrotundifoliawine.
American Journal Enol Vitic 45:56-62.
25
Research paper
Accepted: 19 May,2012
Abstract
Study was aimed at cloning, expression and characterization of rec-pediocin CP2 in Escherichia coli
BL21(DE3) using ompA and tap gene fusion expression approach. pET32(b)-pedA vector containing
an inframe fusion of sequences encoding E. coli ompA secretion signal, two tandem affinity purification
tags and rec-pediocin synthetic gene was introduced into a pediocin CP2 resistant strain of E. coli
BL21(DE3). Rec-protein was accumulated in periplasmic fraction as well as inclusion bodies of
transformed E. coli cultures and was extracted by cell lysis. Crude fusion protein did not show any
biological activity. Purification of rec-pediocin involved two tandem affinity chromatographic steps
based on 6XHis-tag and Strep-tag. In between these, enterokinase cleavage was carried out. After
digestion, rec-pediocin displayed a very strong bactericidal activity against Listeria monocytogenes.
The process based on T7-driven pET expression system and tandem affinity chromatography resulted
in approximately 10 times higher yield of rec-pediocin (in terms of specific activity) as compared to the
native P. acidilactici MTCC5101.
2012 New Delhi Publishers. All rights reserved
Kumar et al.
Figure 1: Comparison of sequence features of rec-pediocin fusion protein and native pediocin CP2
28
Cloning and expression of rec-pediocin CP2 in Escherichia coli using OmpA and TAP gene fusion approach
29
Kumar et al.
30
Cloning and expression of rec-pediocin CP2 in Escherichia coli using OmpA and TAP gene fusion approach
31
Kumar et al.
32
Cloning and expression of rec-pediocin CP2 in Escherichia coli using OmpA and TAP gene fusion approach
Total Volume
(ml)
Total Protein
(mg)
Total Bacteriocin
Activity
(AU)
Specific
Activity
(AU/mg)
Folds of
Purity
% Recovery
400
10
392
40
1,20,00,000
38,00,000
30,612 a
95,000 a
3.103
100
31.66
400
10
ND
90
Nil
8,10,00,000
ND
9,00,000 a
29.4
675
Kumar et al.
Cloning and expression of rec-pediocin CP2 in Escherichia coli using OmpA and TAP gene fusion approach
Beaulieu, L., Groleau, D., Miguez, C.B., Jette, J.F., Aomari, H. and
Subirade, M. 1999. Production of pediocin PA-1 in the
methylotrophic yeast Pichia pastoris reveals unexpected
inhibition of its biological activity due to the presence of
collagen-like material. Protein Expr. Purif., 43(2): 111-125.
Bhunia, A.K., Johnson, M.C. and Ray, B. 1988. Purification,
characterization and antimicrobial spectrum of a bacteriocin
produced by Pediococcus acidilactici. J. Appl. Bacteriol., 65:
261-268.
Bhunia, A.K., Johnson, M.C., Ray, B. and Belden, E.L. 1990.
Antigenic property of pediocin AcH produced by
Pediococcus acidilactici H. J. Appl. Bacteriol., 69: 211-215.
Coderre, P.E. and Somkuti, G.A. 1999. Cloning and expression of the
pediocin operon in Streptococcus thermophilus and other lactic
fermentation bacteria. Curr. Microbiol., 39: 295-301.
Daba, H., Lacroix, C., Huang, J., Simard, R.E. and Lemieux, L. 1994.
Simple method of purification and sequencing of a bacteriocin
produced by Pediococcus acidilatici UL5. J. Appl. Bacteriol.,
77: 682-688.
de Bernardez Clark, E. 2001. Protein refolding for industrial processes.
Curr. Opin. Biotechnol., 12(2): 202-207.
Eisenmesser, E.Z., Kapust, R.B., Nawrocki, J.P., Mazzulla, M.J.,
Pannell, L.K., Waugh, D.S. and Byrd, R.A. 2000. Expression,
purification, refolding and characterization of recombinant
human interleukin-13: utilization of intracellular processing.
Protein Expr. Purifi., 20(2): 186-195.
Elegado, F.B., Kim, W.J. and Kwon, D.Y. 1997. Rapid purification,
partial characterization and antimicrobial spectrum of the
bacteriocin, pediocin AcM, from Pediococcus acidilactici M.
Int. J. Food Microbiol., 37: 1-11.
Fickett, J.W. 1982. Recognition of protein coding regions in DNA
sequences. Nucl. Acids Res., 10(17): 5303-5318.
Gillor, O., Etzion, A. and Riley, M.A. 2008. The dual role of bacteriocins
as anti- and probiotics. Appl. Microbiol. Biotechnol., 81(4):591606. Doi: 10.1007/s00253-008-1726-5.
Grodberg, J. and Dunn, J.J. 1988. OmpT encodes the Escherichia
coli outer membrane protease that cleaves T7 RNA
polymerase during purification. J. Bacteriol., 170: 1245-1253.
Guerra, N.P., Rua, M.L. and Pastrana, L. 2001. Nutritional factors
affecting production of two bacteriocins from lactic acid
bacteria on whey. Int. J. Food Microbiol., 70: 267-281.
Halami, P.M. and Chandrashekar, A. 2007. Heterologous expression,
purification and refolding of an anti-listerial peptide produced
by Pediococcus acidilactici K7. Electron J. Biotechnol., 10(4).
Doi:10.2225.
Halami, S. and Prakash, M. 2008. Cloning of pediocin PA-1 and its
immunity genes from Pediococcus acidilactici K7 using pAMJ
shuttle vector into Lactococcus lactis MG1363. Ind. J.
Biotechnol., 7(4): 550-553.
Horn, N., Martinez, M.I., Martinez, J.M., Hernandez, P.E., Gasson,
M.J., Rodrguez, J.M. and Dodd, H.M. 1998. Production of
pediocin PA-1 by Lactococcus lactis using the lactococcin A
secretory apparatus. Appl. Environ. Microbiol., 64: 818-823.
Horn, N., Martinez, M.I., Martinez, J.M., Hernandez, P.E., Gasson,
M.J., Rodriguez, J.M. and Dodd, H.M. 1999. Enhanced
production of pediocin PA-1 and co-production of nisin and
pediocin PA-1 by Lactococcus lactis. Appl. Environ.
35
Kumar et al.
36
Research paper
1*
Email: vrtembhurkar@gmail.com.
Paper no: 35
Abstract
Influence of few growth conditions and media components on alpha amylase production by Bacillus
thuringiensis in lab scale batch process were determined. B. thuringiensis produced maximum amylase
after 72hrs of incubation. The optimum temperature and pH for amylase production were found to be
31oC and 7, respectively. The optimized media composed of [g/l] 10.0g Rice flour; 5.0 Peptone; CaSO4
4.0; MgSO4 2.0; NaCl 1.0; FeSO4 0.5 and pH 7. The alpha amylase produced was working optimally at
60oC, pH 5, [S] 20 mg/mL, [E] 2.0 mL. Many of these parameters are in compliance with the insecticidal
crystal protein production by B. thuringiensis. Therefore, the process can be used for simultaneous
production of alpha amylase and bioinsecticide in a single fermentation cycle.
2012 New Delhi Publishers. All rights reserved
Tembhurkar et al.
24
48
72
96
120
22.77
15.92
112.14
88.96
16.66
4
5
6
7
8
9
7.40
11.11
14.81
122.2
14.81
14.81
Starch (Pure)
Rice flour
Wheat Flour
Corn Flour
Potato
90.72
104.79
102.67
103.12
101.61
Yeast Extract
Tryptone
Beef extract
Peptone
Urea
Ammonium Nitrate
Ammonium Sulfate
66.66
96.29
77.77
107.40
155.55
66.66
48.14
Mg++
Ca++
Fe++
Concentration (%)
0.05
0.1
0.2
0.4
0.05
0.1
0.2
0.4
0.05
0.1
0.2
0.4
0.05
0.1
92
116
72
48
80
76
100
48
72
140
124
176
116
68
39
Tembhurkar et al.
40
25th min. (48.14 U/mL) after that activity reduced slightly and
remained stable up to 60min. Lee et al., characterized alpha
amylase from 19 serotypes of B. thuringiensis (Lee et al., 1988).
They found amylase worked optimally in temperature range 55
to 60 oC and optimum pH range was 6.7 to 7.2. Kinetic constants
for Bt alpha amylase were determined by plotting Lineweaver
Burk double reciprocal plot (Figure 6). The values obtained for
Vmax and Km were 666.67 U/ml and 3.49 mg/mL respectively.
These values are fairly high than reported by Elif Demirkan
(2011). It was found Km and Vmax of B. subtilis amylase were
1.08 mg/ml and 100 U/ml, respectively. After random
mutagenesis he could isolate a strain that produced amylase
with higher Km (1.43 mg/ml) and Vmax (151 U/ml).
Conclusion
Bacillus thuringiensis ATCC 10752 is potential producer of alpha
amylase. Studies of lab scale batch process indicate optimum
fermentation time was 72hrs and optimum temperature was 31oC.
The optimized media composed of [g/l] 10.0g Rice flour; 5.0 Peptone;
CaSO4 4.0; MgSO4 2.0; NaCl 1.0; FeSO4 0.5; pH 7. The alpha amylase
produced was partially characterized. The enzyme worked efficiently
at 60 oC, pH 5. The alpha amylase activity was maximum when
substrate concentration was kept 20 mg/mL. With increase in enzyme
concentration upto 2.0mL the enzyme activity linearly increased.
The optimum reaction time was 25 min. The values of Vmax and Km
were 666.67 U/ml and 3.49 mg/mL respectively. Most of the optimum
production parameters matched with the production parameters of
ICP by B. thuringiensis previously reported by other workers. Thus
B. thuringiensis can be used for production of alpha amylase and
ICP simultaneously in single fermentation cycle.
Acknowledgement
We express our gratitude to MGMs Institute of Biosciences and
Technology, Aurangabad for providing all the necessary financial and
technical support.
References
Abdul-Hameed, A., Carlberg, G. and El-Tayeb, O.M. 1991. Studies
on the Bacillus thuringiensis H-14 strains isolated in EgyptIV Characterization of fermentation conditions for endotoxin
prodtucion. World. J. Microbiol. Biotechnology. 7: 231-236.
Ajayi, A.O. and Fagade, O.E. 2006. Growth pattern and structural
nature of amylases produced by some Bacillus species in
starchy substrates. Afr. J. Biotechnol. 5(5): 440-444.
Beegle, C.C. and Yamamoto, T. 1992. History of Bacillus
thuringiensis Berliner research and development. Can.
Entomol. 124:587616.
Bernfeld, P. 1955. Amylases, alpha and beta in: Methods in
Enzymology. Academic Press, USA, pp 49-158.
Dey, G.; Singh, B. and Banerjee, R. 2003. Immobilization of -amylase
produced by Bacillus circulans GRS 313. Braz. Arch. Biol.
Technol. 46(2): 167-176.
Dharani Aiyer, P.V. 2004. Effect of C:N ratio on alpha amylase
production by Bacillus licheniformis SPT 27. Afr. J.
41
Research paper
Downstream Processing Lab, Department of Biotechnology, SASTRA University, Thanjavur, Tamil Nadu, India
*Email: pranavkaushik.p@gmail.com
Paper no: 36
Abstract
Whey protein isolates are processed by membrane separation techniques - Micro-filtration, Nanofiltration and Ultra-filtration. Proteins, Glyco-macropeptide, Lacto-albumin, Lacto-globulin and Bovine
Serum Albumin are collected by employment of 10, 30, 50 and 100 kilo-Daltons (kDa) ultra-filtration
membranes, respectively based mainly on protein molecular weights. Pressure drop, trans-membrane
pressure, flux and feed volume were optimized for lab scale. Volume and concentration values for all
streams were calculated for micro-filtration, nano-filtration and 4 ultra-filtration membranes and
satisfactory yield values were obtained. Through scale-up criteria, a cascade design has been proposed
for pilot scale whose pressure values fall within the range of standard pressure values. Fouling
studies were carried out in lab scale by calculating fouling and membrane resistances for all membranes
and effect of anti-fouling chemical agents SDS and NaOH on the membrane cleaning has been
analyzed. The study has given on overall idea about the design corresponding to 6 membranes
arranged in series to clarify 4 proteins from milk.
2012 New Delhi Publishers. All rights reserved
Cascade Mode: MF
NF 1kDa
UF 10kDa UF 30kDa
UF 50kDa UF 100kDa
44
Design of cascade membrane filtration process for clarification of whey proteins and membrane fouling
C. Fouling of Membranes
B. Optimization
1.
2.
3.
Volumes used
Amount (ml)
Comments
1,000
738
22
563
32
25
608
Whey (Cascade)
Time(sec)
Flux(lt/sq.mt-hr)
1
MF
166
67.77
2
NF 1 kDa
1328
8.47
3
UF 10 kDa
744
18.25
4
UF 30 kDa
667
18.78
5
UF 50 kDa
621
18.12
6
UF 100 kDa
133
52.05
C. Design of Cascade Membrane Separation Process
45
Pin
Pout
TMP
Pressure Drop
2.347
2.443
2.312
2.320
2.250
2.250
1.297
1.313
1.122
1.130
1.020
0.980
0.822
0.878
0.782
0.725
0.635
0.615
1.05
1.13
1.06
1.19
1.23
1.27
Initially, pure water was sent into the membranes to note the
flux values. Then whey was sent into the membranes to check
flux values. Again, water was sent and flux values were noted.
Water flux values were decreased when both experiments were
compared. Due to the continuous purification process, fouling
phenomena occurred. Hence, the flux value decreased. The
fouling phenomena were measured in terms of fouling and
membrane resistance values. Membrane cleaning was
performed to observe alteration in flux values and reduction of
3.
Table 3: Volume & Concentration values for feed, retentate & permeate of all 6 membranes
Type
MF-F
MF-R
MF-P / NF-F
NF-P
UF 10-P
204
1.25
198.8
1.50
Volume (ml)
608
103.9
504.1
101.3
402.8
Concentration(g/l)
0.75
0.50
0.75
0.75
0.75
MF- Microfiltration, NF- Nanofiltration, UF- Ultrafiltration, F- Feed, P- Permeate, R- Retentate
Type
Volume (ml)
Concentration (g/l)
UF 30-P
UF 30R/UF 50-F
UF 50-P
UF 50R/UF 100-F
UF 100-P
UF 100-R
121
1.625
77.8
2.75
47.2
3.00
30.6
0.75
26.7
0.50
3.9
0.25
Basis for pilot plant scale was taken as 500 liters as for linear scale up, there should be an initial basis
value as per the real time industry requirement.
Table 4: Permeate Recovery and Yield values
Membrane
Flu
x(lt/sq.mt-hr)
TMP
(bar)
Pressure
Drop(bar)
Permeate
Recovery
Yield
Permeate
Lab Scale
Volume-ml
Pilot Scale
Volume-lt
MF(0.1um)
NF 1kDa
UF 10kDa
UF 30kDa
UF 50kDa
UF 100kDa
67.77
8.47
18.25
18.78
18.12
52.05
0.822
0.878
0.782
0.725
0.635
0.615
1.05
1.13
1.06
1.19
1.23
1.27
79.76
79.91
50.746
60.865
60.668
87.25
79.76
79.91
84.57
65.73
73.054
58.166
608
504.1
402.8
198.8
77.8
30.6
500
414.550
331.250
163.486
63.980
25.164
Parameters
MF
NF
UF 10
UF 30
UF 50
UF100
1.32
89.18
3.15
1.32
11.14
3.39
1.32
24.01
3.18
1.32
24.7
3.57
1.32
23.84
3.69
2.14
68.48
3.8
46
Design of cascade membrane filtration process for clarification of whey proteins and membrane fouling
Table 6: Membrane Fouling studies Flux and resistance values for whey and water
a) Before Fouling with i).Pure water and ii).Whey
Sl. no
Pure water
MF
NF
UF 10
UF 30
UF 50
UF 100
40
281.25
1121
10.04
618
18.2
596
18.88
468
24.04
101
68.54
679
16.57
668
16.84
623
18.06
141
49.09
After whey filtration is done for about 3 hours, the membranes were tested for fouling phenomena.
b) After Fouling with i).Pure water and ii).Whey
1.
Time for 5 ml permeate rise(sec)
51
1206
635
2.
Flux (lt/sq.mt-hr)
220.59
9.33
17.72
743
15.14
529
21.27
138
50.16
692
16.26
648
17.36
189
36.63
1.
2.
After cleaning with water for 20 minutes, whey is sent into the filtration cartridges.
1.
Time for 5 ml permeate rise(sec)
158
1305
2.
Flux (lt/sq.mt-hr)
71.2
8.62
1425
7.89
685
16.43
the flux values near to the initial values indicating that the
membranes were washed properly and pores were cleared.
These flux values are inversely proportional to total resistance
of the membranes. So, membrane resistance and fouling
resistance for each membrane were found out as other
parameters were known on lab scale. Using the correlations,
the pilot plant fouling resistance for each membrane was found
out. Usually pilot plant scale operations either go for WaterBack Washing or Air Flushing for the reduction of fouling as
use of reagents is very expensive and these reagents cannot
be reusable on pilot scale.
Conclusion
Whey proteins have a high nutritional value. They are
Whey
MF
NF
UF 10
UF 30
UF 50
UF 100
1.01095
11.44768
10,09,416
1.01095
10.109716
8,72,687.9
1.01825
4.64071
2,75,888
1.02373
4.20543
1,31,987
1.0438
3.38443
1,14,394
1.0073
1.24776
4,29,859
NF
UF 10
UF 30
UF 50
UF 100
672
16.74
502
22.41
124
55.83
601
18.72
483
23.29
112
61.81
Water
MF
47
48
Research paper
Department of Biotechnology, Himachal Pradesh University, Summer hill, Shimla, Himachal Pradesh, India
Shoolini University of Biotechnology and Management Sciences, Bhajol, Solan, Himachal Pradesh, India
*Email: savvy2000in@yahoo.com
Paper no: 37
Abstract
Seera is a nutritious, easily digestible traditional fermented food made from wheat grains in Bilaspur,
Kangra, Hamirpur, Mandi and Kullu districts of Himachal Pradesh, India. Samples during seera
fermentation were analysed for various microbiological and biochemical parameters. The microflora
isolated from seera mainly comprised of Saccharomyces cerevisiae, Cryptococcus laurentii and
Torulospora delbrueckii among yeasts and Lactobacillus amylovorus, Cellulomonas sp.,
Staphylococcus sciuri, Weisella cibaria, Bacillus sp., Leuconostoc sp. and Enterobacter sakazakii
among bacteria. Protein content decreased from 14.9% on day 1 to 8.2% on 5th day of fermentation.
However, final product obtained had higher protein content i.e. 10.4%. Total sugars decreased with
the time of fermentation.Seera had 11.9 mg reducing sugars/g dry matter. Starch content decreased
initially from 72.1 to 57.0 % (w/w) on dry weight basis from first to fourth day. After steeping and
drying of seera, the starch content increased to 87.4% (w/w) on dry weight basis. Amylase and
protease activity increased with the fermentation up to 4th day, then it started decreasing and very low
amylase activity was recorded in the final product. A significant increase in thiamin, riboflavin, nicotinic
acid and cyanocobalamin was observed during fermentation of seera. The level of essential amino
acids especially methionine, phenylalanine, threonine, lysine and leucine also increased during seera
fermentation.
2012 New Delhi Publishers. All rights reserved
Savitri et al.
Biochemical analysis
Samples during seera fermentation were analysed for various
biochemical parameters viz., total acidity by Amerine et al.
(1980) while pH was measured by using digital pH meter (MAC).
Total proteins were estimated by using the methods of Lowry
et al. (1951) and total carbohydrates were estimated by phenol
sulphuric acid method (Dubois et al., 1956). Reducing sugars
were estimated by DNSA method given by Miller (1959). Starch
was estimated according to Hedge and Hofreiter (1962). The
amount of different enzymes during the course of seera
fermentation was also studied. Protease activity was assayed
by the method given by Manachini et al. (1988) and amylase
activity of the samples during seera fermentation was assayed
according to Bernfield (1955).
Preparation of seera
The details about seera and the traditional method of seera
preparation were studied by visiting various places in Himachal
Pradesh and by active interaction with local people.. To make
seera under laboratory conditions, 200g of wheat grains were
cleaned thoroughly and soaked in 300ml tap water without the
50
Figure 1: Changes in pH, titrable acidity and total count during seera
fermentation
51
Savitri et al.
Values*
pH
Titrable acidity (% of dry matter)
Total count (log cfu/g dry matter)
Protein (mg/g of dry matter)
Total sugars (% of dry matter)
Reducing sugars (mg/g of dry matter)
Starch (% of dry matter)
Amylase** (U/g of dry matter)
Protease***(U/g of dry matter)
3.450.055
0.440.006
5.39
10.40.20
89.00.43
11.90.53
87.41.51
3.60.36
1.020.05
52
active at low pH i.e. below 5.5. After 3rd day, protease activity
started decreasing and finally the activity was 1.02 U/g on dry
weight basis (Table 2). Decrease in activity may be due to the
disappearance of microorganisms responsible for the
production of protease. The degree of proteolysis and the
amount of acids and volatile compounds formed depend both
on the kinds of microorganisms present in the starter and on
the process parameters. Lactic acid bacteria possessing
proteolytic and amylolytic properties have been considered
to be most effective in delaying staling in bread (Corsetti et
al., 1998).
53
Savitri et al.
Table 3: Vitamin and amino acid content in wheat grains and seera
*
Wheat
0.620.03
0.0030.0002
0.0490.003
ND
0.0060.0008
2.70.26
2.80.43
1.70.062
1.10.08
4.10.09
Seera
3.70.17
0.1290.005
0.560.04
0.0790.004
0.330.04
8.40.3
12.21.2
2.40.26
1.40.026
4.20.36
Conclusion
The present study has demonstrated for the first time that
during soaking of wheat grains, a mixed microbial fermentation
of complex ecology develops. This fermentation adds quality
to seera by way of enhancing its protein content, vitamin and
essential amino acids as compared to the control.While the
Western world can afford to enrich its foods with synthetic
vitamins, the developing world must rely upon biological
Figure 5: GC chromatogram of amino acids content of 1) wheat grains and 2) Seera (Peaks 1- chloroform, 2,3- leucine, lysine and threonine,
4- methionine and 5-phenylalanine)
54
Acknowledgement
One the author (Savitri) gratefully acknowledges Council of
Scientific and Industrial Research (CSIR), New Delhi, India for
financial support in the form of Junior Research Fellowship
and Senior Research Fellowship.
References
Abegaz, K., Beyene, F., Langsrud, T. and Narvhus, J.A. 2002.
Parameters of processing and microbial changes during
fermentation of borde, a traditional Ethiopian fermented
beverage. J Food Technol Afr.,7: 85-92.
Akolkar, P.N.., 1977. Studies on soyidli fermentation.Ph.D. Thesis,
M.S. University of Baroda, Baroda, India.
Amerine, M.A., Berg, H.W.,Kunkee, R.E.,Ough, E.S., Singleton, V.L.
and Webb AD. 1980. The technology of wine making. AVI
Publishing Co Inc, Westport.
Battcock, M. and Azam-Ali, S.A. 1998. Fermented fruits and
vegetables: a global perspective. FAO Agricultural Services
Bulletin No. 134.
Bernfield P. 1955. Amylases, and . In:Methods of Enzymology(Eds.
S Colowick and NO Kaplan), Academic Press, New York,
USA, pp. 149-158.
Beuchat, L.R. 1983. Indigenous fermented foods. In: Biotechnology:
Food and feed production with microorganisms (Ed. G Reed),
vol. 5, VerlagChemie, Weinheim, pp. 477-528.
Corgan, T.M., Bresford, T.P., Steele, J., Broadbent, J., Shah, N.P.
and Ustunol, Z. 2007.Advances in starter cultures.J Dairy
55
Savitri et al.
Law, S.V., Bakar, A.F., Hashim, M.D. and Hamid , A.A. 2011. Popular
fermented foods and beverages in Southeast Asia. Int Food
Res J., 18: 475-484.
Lowry, O.H., Rosenbrough, N.J., Farr, A.L. and Randall, R.J. 1951.
Protein measurement with folin phenol reagent. J Biol
Chem.,193: 256.
Manachini, P.L.,Fortina, M.G. and Parinic, C. 1988.Thermostable
alkaline protease produced by Bacillus thermoruber-a new
species of Bacillus. ApplMicrobiolBiotechnol.,28: 409-413.
Mensah, P. 1997.Fermentation- the key food safety assurance in
Africa?Food Cont, 8:271-278.
Miller, G.L. 1959. Use of dinitrosalicyclic acid reagent for the
determination of reducing sugars.Anal Chem., 31: 426-428.
Mosha, A.C. and Svanberg, U. 1983.Preparation of weaning foods
with high nutrient density using flour of germinated
cereals.Food Nutr Bull., 5: 10-14.
Narashimahan, S. and Rajalakshmi, D. 1999.Sensory evaluation of
fermented foods. In: Biotechnology: Food Fermentation,
Microbiology, Biochemistry and Technology(Eds. VK Joshi
and A Pandey), Vol II, Educational Publishers and
Distributors, New Delhi, pp. 345-382.
Niba, L. 2003. The relevance of biotechnology in the development of
functional foods for improved nutritional and health quality
in developing countries. Afr J Biotechnol., 2: 631-635.
Oyewole, O.B. 1990. Optimization of cassava fermentation for fufu
production: effect of single starter cultures. J. Appl Bacteriol.,
68: 49-54.
Parvez, S., Malik, K.A., Ah Kang, S. and Kim, H.Y. 2006. Probiotic
and their fermented food products are beneficial for health. J
ApplMicrobiol., 100:1171-1185.
Rajalakshmi, R. and Vanaja, K. 1967. Chemical and biological
evaluation of the effects of fermentation on the nutritive value
of food prepared from rice and gram. Brit J Nutr., 21:467473.
Sankaran, R. 1998. Fermented foods of Indian subcontinent. In:
Microbiology of Fermented Foods(Ed. BJB Wood),vol 2,
Blackie Academic and Professional, New York, pp. 753-789.
56
Research paper
Email: priti_june10@rediffmail.com
Paper no: 38
Abstract
Mahua is used for preparation of distilled liquors since time immemorial. Vermouth was prepared from
young mahua wine after its fortification and flavouring with traditional Indian herbs, viz. black pepper,
cinnamon, clove, cumin, fenugreek, large cardamom, nutmeg, fennel and Indian Cassia. Initially, mahua
wine had 6.00 B TSS, 0.63 % acidity, 2.11 mg/L 100 ml ascorbic acid, 0.09 % tannins, 0.19 % reducing
sugars and 10.5 % alcohol. The fortified mahua vermouth after one year of ageing had 6.40 B TSS, 0.57
% acidity, 1.19 mg/L 100 ml ascorbic acid, 0.11 % tannins, 0.27 % reducing sugars and 18.4% alcohol.
The vermouth was organoleptically more acceptable than the mahua wine. Phenolics, viz. gallic acid,
caffeic acid, pcoumaric acid and kaempferol as well as the flavanols, (+)-catechins and (-)-epi-catechin
were detected in both the mahua wine and vermouth.
2012 New Delhi Publishers. All rights reserved
Yadav et al.
Juice extraction
Statistical Analysis
Yeast culture
Sensory Analysis
Mahua vermouth
Physicochemical Analysis
Vermouth was analyzed for various biochemical and sensory
parameters at 3 months interval. Total soluble solids (TSS)
content of vermouth were recorded using hand refractometer
(Erma, Japan). The titratable acidity, tannins, ascorbic acid,
non enzymatic browning was determined using the methods
described by Ranganna (1986). The reducing sugar was
determined using the methods of AOAC (1985). Ethanol
Table 1: Spices and herbs used in mahua vermouth preparation
Common name
Black pepper
Cinnamon
Clove
Cumin
Fenugreek
Large cardamom
Nutmeg
Fennel
Indian Cassia
15.5
9.2
7.0
12.8
6.0
6.5
8.2
50
8
58
Botanical name
Part used
Piper nigrum L.
Cinnamonum zeylanicum Beryn.
Syzygium aromaticum L.
Cumis cyminum L.
Zingiber officinale Rosc.
Amomum subulatum Roxb.
Myristica fragrans Houth.
Foeniculum vulgare
Cinnamomum tamala
Fruit
Bark
Fruit
Seeds
Dried roots
Seeds
Seeds
Seed
Leafs
Mahua juice
T.S.S (0B)
Acidity (%)
Ascorbic Acid (mg/100ml)
Tannins (%)
Reducing sugar (g %)
13.0
0.11
3.15
0.11
1.04
Parameters
Mahua wine
T.S.S (0B)
Acidity (%)
Volatile acidity (%)
Ascorbic acid (mg/100ml)
Tannins (%)
Reducing Sugar (%)
Alcohol (%)
NEB
6.0
0.63
0.07
2.11
0.09
0.19
10.50
0.021
SO2
(ppm)
107
105
103
100
97
TSSb
(0B)
6.2
6.2
6.4
6.4
6.4
Acidityc
Ascorbic
(%) acidd (mg/100g)
0.59
0.58
0.58
0.57
0.57
1.26
1.24
1.23
1.20
1.19
Tanninse
sugarg (g %)
Reducing
sugarg (%)
Total
sugarf(g %)
Alcoholh
(%)
Non-enzymatici
browning
0.32
0.29
0.28
0.20
0.11
0.16
0.17
0.19
0.23
0.27
0.39
0.37
0.35
0.33
0.33
18.40
18.40
18.40
18.40
18.40
0.020
0.029
0.035
0.038
0.040
Data followed by different letters for each column are significantly different by statistical (p > 0.01).
Statistical analysis due to storage periods: a 0.001; b 0.001; c non-significant; d 0.001; e 0.001; f non-significant; g 0.001; h non-significant; i
0.002
59
Yadav et al.
494.5410.28a
475.3119.23a
436.2039.11a
417.1719.03a
409.457.72a
Gallic acid
(+)-Catechin
(-)-Epi-catechin
Caffeic acid
p-coumaric acid
122.5925.76b
112.4513.76ab
107.3818.67ab
99.1914.98ab
92.8724.74ab
35.31 1.45b
32.561.65b
30.851.78b
30.192.98b
28.752.57a
39.516.98a
35.266.34ab
32.834.96ab
29.744.77ab
29.184.29b
10.763.06a
11.982.95b
13.112.25b
16.871.98b
18.351.59b
4.180.95a
5.351.45b
5.871.34b
5.991.75b
6.181.11b
Data followed by different letters for each column are significantly different by LSD
Values are expressed as mg/100gram standard error (n=33).
kaempferol
282.1649.4a
281.1946.4b
280.9753.1b
280.9151.9b
280.5749.6b
test (p<00.1).
60
61
-amylase
from
Endomyces
fibuliger an indigenous yeast isolate of Western Himalayas
Intl. J. of Food. Ferment.
Technol.production
2(1): 63-69,
June,
2012
Research paper
Department of Microbiology, Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India
Regional Station, Indian Veterinary Research Institute, Palampur, Himachal Pradesh, India
* Department of Microbiology, Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India
2
Email: sskanwar1956@gmail.com
Paper no: 39
Abstract
Amylase production, its purification, characterization and optimization have been studied in Endomyces
fibuliger. The enzyme was purified to about 13-fold by using gel permeation chromatography (Sephadex
G-25, Sephadex G-100). The purity of enzyme was checked by the use of Native-PAGE and SDS-PAGE.
The molecular weight of enzyme was 55KD with the presence of two polypeptide units. Enzyme was
most active at pH 7.0, temperature 30 C with optimum incubation time of 30 minutes. The maximum
activity of the enzyme was at 2 % (w/v) soluble starch concentration. The enzyme was para-constitutive
in nature. The parameters for amylase production were optimized by Response Surface Methodology
(RSM) . The best carbon source was soluble starch, the optimum pH values were 5.0 and 8.0, and the
optimum temperatures were 30 C and 37 C. Ca+ and Mg+ had enhancing effect on its production.
2012 New Delhi Publishers. All rights reserved
63
Bhushan et al.
64
-amylase production from Endomyces fibuliger an indigenous yeast isolate of Western Himalayas
-2
2
25
4
4
-1
4
30
5
6
0
5
31
6
8
+1
6
37
8
12
(1)
+2
8
43
10
16
Starch
Temperature
concentration(%) (A)
(C) (B)
4
2
2
8
5
8
8
8
5
6
2
2
8
2
5
6
5
5
5
8
8
5
5
5
5
5
5
2
2
5
25
37
25
25
30
37
37
37
43
30
25
37
37
37
25
31
30
31
31
25
25
37
30
37
31
31
31
37
25
30
pH
(C)
CaCl2
(mg) (D)
8.0
3.0
4.0
4.0
5.0
8.0
5.0
8.0
6.0
6.0
4.0
4.0
4.0
4.0
6.0
6.0
6.0
10
6.0
8.0
8.0
8.0
6.0
4.0
6.0
6.0
4.0
8.0
8.0
6.0
12
4.0
12
12
12
12
16
4.0
8.0
8.0
12
4.0
12
12
8.0
8.0
6.0
8.0
12
12
4.0
4.0
12
4.0
8.0
16
12
12
4.0
8.0
65
Predicted
Activity(U/mg)
Experimental
0.1
0.56
0.006
-0.067
0.53
0.91
1.13
1.01
-0.033
0.67
0.006
-0.014
0.098
-0.007
0.33
0.7
0.7
0.027
0.74
0.47
0.56
0.76
0.71
-0.037
0.67
0.99
0.21
0.36
0.29
6.64
0.039
0.366
0.0
0.0
1.133
0.9228
1.1263
0.9684
0.0
0.8043
0.0
0.0
0.0
0.0
0.0
0.8043
0.803
0.0
0.796
0.497
0.544
0.92
0.788
0.0
0.0
0.8
0.0
0.437
0.489
0.8
Bhushan et al.
Volume of
extract (ml)
Protein
(mg)
Total activity
(units)
Specific activity
(U/mg)
Recovery
(%)
Purification fold
Crude extract
Sephadex G-25
Sephadex G-100
100
40
25
159
2.8
1.03
95
18.47
7.8
.8545
6.59
7.57
100
19.2
8.2
1
11.04
13
Sum of square
Degree of freedom
Mean square
F-value
P-value
Model
Error
Total
3.9
0
5.1
14
1
29
0.28
0
3.48
0.0112
R=0.7645
Adjusted R=0.5447
Predicted R=0.3713
Adequate precision=5.996
-amylase production from Endomyces fibuliger an indigenous yeast isolate of Western Himalayas
Figure 3: 3-D surface diagram of amylase production shows on zaxis against the effect of temperature and pH plottted on x-axis and yaxis, respectively.
67
Bhushan et al.
68
-amylase production from Endomyces fibuliger an indigenous yeast isolate of Western Himalayas
the predicted one and the prediction error was calculated. The
observed and the predicted values obtained were quite close
(Table 2) and therefore, showed good predictability of the
model.
Conclusion
Endomyces fibuliger is an excellent producer of alpha-amylase
enzyme, which is para-constitutive in nature. Organism showed
enzyme production in both alkaline and acidic conditions.
Enzyme is active at a wide range of temperature. So, it can play
an important role in various industrial processes viz. brewing,
baking, distillery etc. The indigenous yeast isolate can also be
directly supplied to the local people by co-immobilizing it with
alcohol producing strains for cereal based beverages.
References
Banerjee, M., Debnath, S. and Majumdar, S.K. 1988. Production of
alcohol from starch by direct fermentation. Biotechnology
Bioengineering, 32:831-834.
De Mot, R. and Verachtert, H. 1985. Purification and characterization
of the extracellular amylolytic enzymes from the yeast
Filobasidium capsuligenum. Appl Environmental Microbiol,
50: 1474-1482.
Galdino, A.S., Silva, R.N, Lottermann, M.T., Alvares, A.C.M.,
Moraes LMP, Torres FAG, de Freitas SM, and Ulhoa CJ.
2011. Biochemical and Structural Characterization of Amy1:
An Alpha-Amylase from Cryptococcus flavus Expressed in
Saccharomyces cerevisiae. Enzyme Research, doi:10.4061/
2011/157294.
Gigras, P., Sahai, V. and Gupta, R. 2002. Statistical media optimization
and production of its alpha-amylase from Aspergillus oryzae
in a bioreactor. Curr Microbiol, 45: 203-208.
Haaland, P.D. 1989. Statistical problem solving. In Experimental
Design in Biotechnology ed. Haaland, PD pp. 1-18. New
York and Basel: Marcel Dekker, Inc.
He, G.Q., Kong, Q. and Ding, L.X. 2004. Response surface
methodology for optimizing the fermentation medium of
Clostridium butyricum. Lett Applied Microbiol, 39: 363-368.
69
Research paper
Email: baljinderbt@hotmail.com
Paper no: 40
Abstract
A lactic acid bacterial strain capable of metabolizing arginine via arginine deiminase pathway was
isolated from lassi, an Indian fermented food. On the basis of biochemical analysis it was identified as
Weissella confusa. Response surface methodology (RSM) was used to optimize media components
such as carbon and nitrogen sources, minerals, inducers and metal ions for enhancing its specific
arginine deiminase activity. A Placket-Burman design was applied to identify significant factors affecting
enzyme production in Weissella confusa GR7. Central composite rotatable design was applied further
to optimize the concentrations of sucrose, copper, magnesium, manganese and arginine for maximizing
arginine deiminase activity. Statistical design has suggested a Two Factor Interaction model which
was validated experimentally where it showed 10 fold increases in specific arginine deiminase activity
over basal medium. By solving the regression equation and analyzing the response surface cartons,
optimal concentration of various components was determined as 2.5g/l sucrose, 5g/l fructose, 10g/l
tryptone, 5mM MgSO4, 5mM MnSO4, 25mM arginine, 5mM Copper, 50mg/ml CTAB and 50mM Tween
80. The model was found satisfactory as the coefficient of determination was 0.83. Results indicate an
improved arginine deiminase production in statistically optimized medium using response surface
methodology.
2012 New Delhi Publishers. All rights reserved
Keywords: Arginine deiminase, Weissella confusa, Central composite rotatable design, Response
surface methodology, Lactic acid, Bacteria
Statistical screening of media components for the production of arginine deiminase by Weissella confusa GR7
Y = +0.68+0.12*A-0.15*B-0.011*C-0.11
*D+0.11*E-0.17*A*B+0.046*A*C0.18*A*D+0.12*A*E+0.051*B*C+0.19*B*D
-0.11*B*E-0.010*C*D+0.064*C*E-0.18*D*E........................(3)
Where, Y is the specific enzyme activity and A, B, C, D, E is
coded values of sucrose, copper, magnesium, manganese, and
arginine respectively.
Table 3 shows ANOVA results for the RSM Two Factor
Interaction (2FI) for response Y. According to the present model
A, B, D, E, AB, AD, AE, BD, BE and DE are significant model
terms.2FI model was found to be the best fit model for the
specific enzyme activity with the highest F-value when
compared to other models (Table 4). ANOVA for ADI specific
activity (Y, IU/mg) indicated the F-value to be 11.10, which
implies the model to be significant. Model terms having values
of Prob > F less than 0.0500 indicate model terms are
significant, whereas those greater than 0.10 are insignificant.
The Lack of Fit F-value of 138.35 implies the Lack of Fit is
significant. There is only a 0.01% chance that a Lack of Fit Fvalue this large could occur due to noise. ANOVA indicated
the R2 value of 0.8305 for response Y. This ensured a good
73
Fructose)
(g/l
0
5
0
5
5
0
0
5
0
5
5
0
Run
1
2
3
4
5
6
7
8
9
10
11
12
5
0
5
0
5
0
0
5
0
0
5
5
0
0
10
10
0
0
0
0
10
10
10
10
10
0
10
10
0
0
10
10
0
10
0
0
10
10
0
10
0
0
0
10
10
0
0
10
0
0
0
0
10
0
10
10
10
10
0
10
MgSO
(mM)
4
10
10
0
0
0
0
10
0
0
10
10
10
MnSO
(mM)
4
50
50
50
0
50
0
0
0
50
50
0
0
CTAB
(mg/ml)
50
0
0
50
50
0
50
0
50
0
50
0
0
50
50
0
0
0
50
50
50
0
50
0
Tween-80 Arginine
(mM)
(mM)
0
5
5
5
5
0
5
0
0
0
0
5
%
CO2
0.43
1.62
0.4
0.27
2.04
-0.01
0.58
0.38
0.44
0.15
0.2
0.25
Specific
activity
Experimental
value
0.57
0.57
0.57
0.57
0.57
0.57
0.57
0.57
0.57
0.57
0.57
0.57
ADI
(IU/mg)
Predicted
value
74
Statistical screening of media components for the production of arginine deiminase by Weissella confusa GR7
Sucrose
(g/l)
CuSO4
(mM)
MgSO4
(mM)
MnSO4
(mM)
Arginine
(mM)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
5
5
0
2.5
5
8.4
2.5
2.5
5
0
2.5
2.5
2.5
0
5
5
0
0
0
-3.4
0
5
2.5
5
5
0
5
0
5
2.5
5
0
5
2.5
0
0
0
2.5
5
0
2.5
2.5
2.5
5
2.5
2.5
2.5
0
5
0
10
0
10
5
0
5
-6.9
5
10
10
5
5
5
0
10
10
10
10
0
5
10
0
5
0
0
0
0
0
0
5
0
0
10
5
10
10
0
5
10
10
5
5
5
10
5
16.9
5
0
10
0
0
0
10
5
10
5
5
-6.9
10
0
5
5
5
10
10
0
0
0
0
5
10
0
5
10
0
10
10
10
0
16.9
10
0
0
5
10
10
0
5
0
0
5
5
5
10
5
5
5
0
10
10
0
0
10
5
0
5
5
5
0
0
5
-6.9
5
0
10
10
0
10
10
5
0
10
5
0
10
0
10
10
0
5
10
0
10
5
0
10
10
5
0
10
16.9
5
5
0
5
5
5
0
10
10
0
50
50
25
50
25
25
25
50
50
84.5
25
25
50
0
0
0
0
50
25
50
0
25
0
50
0
0
0
0
25
50
50
50
25
0
0
0
25
50
50
25
25
25
0
-34.5
25
25
0
50
50
75
0.68
0.28
0.97
0.56
1.76
1.03
1.04
0.68
0.71
2.17
2.05
0.44
0.17
0.48
0.31
1.05
0.45
1.16
0.57
0.95
0.39
0.24
0.9
069
0.31
0.39
0.41
0.4
0.68
0.58
0.33
0.68
0.71
1.02
0.21
0.68
0.32
0.83
0.42
0.43
0.68
1.21
0.44
0.25
0.96
0.68
0.32
0.65
0.93
0.68
Sum of squares
Model
A-Sucrose
B-Copper
C-Magnesium
D-Manganese
E-Arginine
AB
AC
AD
AE
BC
BD
BE
CD
CE
DE
Residual
Lack of Fit
Pure Error
Cor Total
7.87
0.61
1.02
5.42E-03
0.56
0.48
0.91
0.067
1.03
0.47
0.082
1.14
0.37
3.39E-03
0.13
0.99
1.61
1.6
3.01E-03
9.48
Degree of freedom
15
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
34
27
7
49
Mean squares
F- value
p-value
prob > F
0.52
0.61
1.02
5.42E-03
0.56
0.48
0.91
0.067
1.03
0.47
0.082
1.14
0.37
3.39E-03
0.13
0.99
0.047
0.059
4.29E-04
11.1
12.87
21.51
0.11
11.9
10.18
19.25
1.43
21.71
9.88
1.74
24.2
7.89
0.072
2.8
21
< 0.0001
0.001
< 0.0001
0.737
0.0015
0.003
0.0001
0.2407
< 0.0001
0.0035
0.1964
< 0.0001
0.0082
0.7903
0.1033
< 0.0001
significant
138.35
< 0.0001
significant
Sum of Squares
Mean vs Total
Linear vs Mean
2FI vs Linear
Quadratic vs 2FI
Cubic vs Quadratic
Residual
Total
23.07
2.67
5.2
0.13
1.34
0.14
32.54
Degree of freedom
1
5
10
5
15
14
50
Mean Square
23.07
0.53
0.52
0.026
0.089
9.93E-03
0.65
F-value
p-value
Prob > F
3.46
11
0.5
9
0.0101
< 0.0001
0.7734
< 0.0001
Linear vs Mean
Suggested
Aliased
Statistical screening of media components for the production of arginine deiminase by Weissella confusa GR7
(a)
Figure1: Three-dimensional response cartons showing effects and interaction of (a) arginine (mM) and sucrose (g/l), (b) copper sulphate (mM)
and sucrose (g/l), (c) manganese sulphate (mM) and sucrose (g/l), (d) arginine (mM) and manganese sulphate (mM) and (e) arginine (mM) and
copper sulphate (mM) on specific ADI activity of W. confusa GR7.
77
78
Statistical screening of media components for the production of arginine deiminase by Weissella confusa GR7
soli sp. nov., a lactic acid bacterium isolated from soil. Int. J.
Syst. Evol. Microbiol., 52:831-834.
Milbourne, K. 1983. Thermal tolerance of Lactobacillus viridescens
in ham. Meat Sci. 9: 113-119.
Mira de Ordua, R., Liu, S.Q., Patchett, M.L. and Pilone, G.J. 2000.
Ethyl carbamate precursor citrulline formation from arginine
degradation by malolactic wine lactic acid bacteria. FEMS
Microbiol. Lett., 183:31-35.
Montel, M.C. and Champomier, M.C. 1987. Arginine catabolism in
Lactobacillus sake isolated from meat. Appl. Environ.
Microbiol., 53:2683-2685.
Niven, C.F., JR., and Evans, J.B. 1957. Lactobacillus viridescens
nov. sp., a heterofermentative species that produces a green
discolouration of cured meat pigments. J. Bacteriol., 73:758759.
79
Research paper
Central Institute of Temperate Horticulture, Regional Station, Mukteshwar - 263 138 (Uttarakhand), India
*Email: attribl_cith@rediffmail.com
Paper no: 41
Abstract
The composition of the musts prepared from the pineapple fruits cvs. Kew and Queen by amelioration
of TSS by addition of KMS (100ppm), with and without DAHP (0.1%) as nitrogen source was found
to vary significantly. The fermentation rate of the pineapple musts having nitrogen source, carried out
by the yeast Saccharomyces cerevisiae var. ellipsoideus was comparatively faster than those without
DAHP and over a period of 10 days. After six months of storage the physico-chemical characteristics
of the pineapple wines revealed significantly higher alcohol (10.56 and 11.44 %) in the treatments
having nitrogen source with better fermentation efficiency of yeast (48 and 52%). The acidity and
volatile acidity of these treatments was significantly higher. The pineapple wines having more alcohol
were found to have lower aldehydes (112.78 and 117.47 mg/litre) whereas total esters (94.65 and 104.77
mg/litre) and phenolic contents (148.70 and 156.54 mg/litre) were found to be higher. From the sensory
quality point of view, the wine prepared from cv. Queen was preferred to cv Kew. The costs of the final
products were Rs. 35.38 and Rs. Rs. 31.82 per 650 ml bottle in cv. Kew and Queen, respectively.
2012 New Delhi Publishers. All rights reserved
Keywords: Keywords: Pineapple nitrogen source, must, fermentation, wine, phenolic content,
Saccharomyces cerevisiae, sensory
Introduction
The pineapple (Ananas comosus (L.) Merr. Syn. Ananas sativus
Schult.f) originated from Paraguay belongs to family
Bromeliaceae (Bertoni 1919). It is one of the most important
and wanted tropical fruits because of good source of vitamin
A, B, C and calcium. Apart from its various products viz., slices,
juice, squash, jam, and mixed fruit jam, other by-products are
alcohol, calcium citrate, citric acid and vinegar. The dried waste
after juice extraction is valuable cattle feed. It is being cultivated
in different countries like Hawaiian Islands, Philippines,
Malaysia, Thailand, Brazil, Ghanna, Kenya, Mexico, Taiwan,
S. Africa, Australia and India. It was introduced in India in
Attri
82
Effect of nitrogen source on the fermentation behaviour of musts and quality of wine
yeast cells which has also been reported earlier by Joshi et al.
(1991). The fermentation rate however declined towards the
completion of fermentation which may be due to more alcohol
production inhibiting the fermentation efficiency of the yeast
(Attri et al 1994). The fermentation efficiency of the yeast was
found 48 and 52 per cent in T1 and T-3 (nitrogen source) as
compared to 45 and 50 per cent in T2 and T-4 (without nitrogen
source) which has been depicted in Fig. 3. The higher efficiency
may be attributed to the addition of nitrogen source for the
yeast during fermentation.
Table 1: Composition of different cultivars of pineapple
Sensory analysis
Characters
Length (cm)
Breadth (cm)
Weight (g)
Volume (ml)
Specific gravity (w/v)
Juice (%)
Waste (%)
T.S.S. (0B)
Acidity (% MA)
Brix acid ratio
Ascorbic acid (mg/100g)
Carotenoids (g/100g)
Moisture (%)
Dry matter (%)
Statistical analysis
The available data for physico-chemical characteristics of
different pineapple musts and wines were analysed in a
Completely Randomized Block Design (CRBD) as described
by Panse and Sukhatme (1989).
Cost of production of pineapple wine
The cost of production of pineapple wines from both the cvs.
Kew and Queen was also estimated keeping in view the present
rates of various items used in the fermentation and thereafter.
The overhead charges as well as additional charges on
fermentation were also added along with sales tax and profit of
the producer.
\Cultivars
Kew
Queen
15.700.040
12.500.040
1060.008.16
1099.007.55
0.9640.0008
56.000.489
44.000.489
15.500.081
0.9380.0016
16.520.057
39.130.257
38.990.608
78.830.048
21.170.048
10.500.092
9.800.092
680.004.08
707.001.53
0.9620.0002
52.000.408
48.000.408
12.000.081
1.0450.0008
11.480.069
45.000.326
58.000.347
76.9800.122
23.100.122
83
Attri
Treatments
Kew
Characteristics of musts
TSS (oB)
Acidity (%MA)
22.0
22.0
22.0
22.0
NS
0.850
0.875
1.010
1.014
0.004
T1
T2
T3
T4
-
Queen
CD=0.05
34.35
35.50
41.20
42.45
0.465
27.65
29.57
43.87
45.28
0.254
T1=Must with 0.1% DAHP, T2= Must without DAHP, T3= Must with 0.1% DAHP, T4 =Must without DAHP
Kew
Queen
CD=0.05
T1
T2
T3
T4
Characteristics of wines
TSS
(o B)
Acidity
(%MA)
4.8
5.0
4.6
5.0
0.399
1.012
0.975
1.045
1.020
0.0023
pH
Ascorbic acid
(mg/100g)
(% v/v)
4.00
4.06
3.89
3.94
0.113
18.70
15.40
21.76
19.50
0.118
84
Alcohol
acidity
(%AA)
Volatile
(mg/litre)
Aldehydes
(mg/ litre)
10.56
9.90
11.44
11.00
0.143
0.009
0.007
0.010
0.008
0.012
112.78
119.45
117.47
123.86
0.330
Total esters
Total
(mg/litre) phenols
94.65
86.97
104.77
101.20
0.173
148.70
141.63
156.54
151.87
0.131
Effect of nitrogen source on the fermentation behaviour of musts and quality of wine
Kew
Quantity
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
Queen
Rate (Rs.)
Amount (Rs.)
Quantity
Rate (Rs.)
Amount (Rs.)
Pineapple fruit
100kg
Transportation
Juice extraction
2 mandays
Sugar
4.0 kg
Potassium metabisulphite @ 100 ppm
DAHP @ 0.1%
65.0g
Glass bottles (650 ml)
100
Crown corks
100
Total cost
Overhead charges @20%
Additional charges @ 15% on fermentation
Excise duty
Grand total
Sales tax @20%
Profit @ 20%
-
10.00/kg
60.00/day
18.00/kg
6.5g
3.00/bottle
0.25/cork
3.00/bottle
-
100 kg
2 mandays
5.0 kg
15.00
65.0g
100
100
240.00
-
8.00/kg
60.00/day
18.00/kg
6.5g
3.00/bottle
0.25/cork
3.00/bottle
-
Sale price/bottle
1000.00
50.00
120.00
72.00
15.00
300.00
25.00
1597.00
320.00
300.00
2457.00
491.00
590.00
3538.00
35.38
800.00
50.00
120.00
90.00
15.00
15.00
300.00
25.00
1415.00
283.00
212.00
300.00
2210.00
442.00
530.00
3182.00
31.82
Conclusion
The wine made by the addition of nitrogen sources had better
fermentability than those without it. The physico-chemical and
sensory qualities of both the wines were acceptable but the
product from cv. Queen was preferred. The cost of production
of pineapple wine was also preferable. It can be concluded
that there is an ample scope for the utilization of pineapple
fruits for production of wine which are good in nutrients and
available throughout the year in the islands by the method
described here.
References
Amerine MA and Ough CS. 1979. Wine and Must Analysis, 2nd Edn.
John Wiley and Sons, New York.
Anonymous. 1985. Home Scale Processing and Preservation of Fruits
and Vegetables. 7th Edn. CFTRI, Mysore. Pp: 28-29.
AOAC. 1975. Official Methods of Analysis. Association of Official
Analytical Chemists. 12th Edn. Washington, D.C.
Attri BL, Lal BB and Joshi VK. 1994. Technology for the preparation
of sand pear vermouth. Indian Food Packer, 48(1):39-45.
Attri BL. 2009. Effect of initial sugar concentration on the physicochemical characteristics and sensory qualities of cashew apple
wine. Natural Product Radiance, 8(4):374-379.
Bertoni MS. 1919. Ancient Paraguay, Series II, 4: 250-332.
Caputi A, Ucda Jr M and Brown T. 1968. Spectrophotometric
determination of ethanol in wine. Am. J. Enol. Vitic., 19:160165.
Joshi VK, Attri BL, Gupta JK and Chopra SK. 1990. Comparative
fermentation behaviour, physico-chemical characteristics and
85
Attri
86
Research note
Department of Food Science and Technology Dr YS Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal
Pradesh,India
2
Department of Microbiology,Guru Nanak Dev University,Amritsar,Punjab,India
*
Email: v.k.joshipht@rediffmail.com
Paper no: 42
Abstract
Apple pomace (AP) fermented powders were made after solid state fermentation with different yeasts
(Saccharomyces cerevisiae,Candida utilis and Torula utilis) and after removal of ethanol was made
into powders. Application of Principal Component Analysis (PCA) to the compositional data clearly
separated the unfermented apple powders from the fermented apple fomace powders. The PCA separated
the fermented apple pomace from unfermented based upon the parameters selected viz. total sugars,
TSS, vitamin C, crude proteins, moisture, crude fat and soluble protein. The summary of Eigenanalysis
clearly shows that the first two PCs accounted for most of the variations found in the composition of
samples. Out of the two PCs, the PC 1 was defined by TSS, vitamin-C, crude fat and titratable acidity
while PC 2 contrasted the treatment based upon the total sugar and crude proteins. However, yellow
colour and moisture content were less related as shown by their intensities. The unfermented AP was
distinct for total sugar and TSS while fermented apple pomace samples were specific in vitamin-C,
crude protein, crude fat and soluble protein. Based on the mineral analysis, the PCA failed to differentiate
between unfermented and fermented APP when treatment and attributes (vector loadings) were plotted
together. Ferrous defined the PC 1 strongly while as shown by their intensities K, Cu, Na and Ca, were
weekly correlated. The PC-2 was defined by Zn and Mn contents.
2012 New Delhi Publishers. All rights reserved
Keywords: Solid state fermentation, PCA, Apple pomace, Protein, Minerals, Yeast
88
Effect of Solid state fermentation and yeast species on composition of apple pomace: Application of PCA
Figure 1: Projection of physico-chemical analysis data of apple pomace powders obtained with or without fermentation by different yeasts
in planes defined by principle component 1 and 2 (1) = unfermented apple pomace, Apple pomace fermented by (2)= Sacchamyces,(3)=Candida,
(4)= Torula
89
Table 1: Principal Components Analysis output of data of physico-chemical parameters of apple pomace fermented by different yeasts
Eigen values
(a) All parameters except minerals (12 attributes)
1 = 3.486
2 = 0.502
3 = 0.011
4 = 0.001
(b) All minerals (8 attributes)
1 = 3.998
2 = 0.001
3 = 0.00
4 = 0.00
87.2
12.5
0.3
0.0
87.2
99.7
100.0
100.0
100.00
0.00
0.00
0.00
100.0
100.0
100.0
100.0
Figure 2: Projection of mineral analysis data of apple pomace powders obtained with or withot fermentation by different yeasts in planes
defined by principal component 1 and 2 (1) Unfermeted apple pomace, Apple pomace fermented by (2) = Saccharomyces, (3) = Candida,
(4) = Torula
90
Effect of Solid state fermentation and yeast species on composition of apple pomace: Application of PCA
Joshi, V.K. and Sandhu, D.K. 1994. Solid state fermentation of apple
pomace for production of ethanol and animal feed. In Solid
State Fermentation. Ashok Pandey, Eastern Wiley Ltd., New
Delhi, pp. 93-98.
Joshi, V.K.and Sandhu, D.K. 1996. Composition of the distillates,
from the solid state fermentation of apple pomace by different
yeasts. Nat. Acad. Sci. letter,49(1),1-13
Joshi, V.K. and Sandhu, D.K. 1996. Preparation and evaluation
of animal feed using solid state fermentation of apple
pomace. Bioresource Technol., 56:251-255.
Joshi, V.K. and Rana S. Neerja. 2009. Microbial Technology for the
Production of Value Added Products from Apple Pomace,
Agriculturally Impotant Microorganisms (Vol.II),
(Eds.George, G.K., D.K. Arora, T.P.R., A.K.Srivatava
Academic World (13) pp.271-286.
Kaur, K. 1989. Microbial transformation of apple pomace to recover
industrial Products. M.Sc. (Hons.) Thesis, Punjab University,
Chandigarh.
Keunedy, M.J. 1994. Apple pomace and Kiwifruit : Processing
options. Australasian Biotechnol. 4:43-49.
Lee, C.Y. and Mattick, L.R. 1989. Composition and Nutritive value
of apple; products. In: Processed Products. (C.D.L.
Downing, ed). AVI Novanstrand Reinhold, New York, pp.
303-322.
Lammel, S.A., Heimsch, R.C. and Edwards, L.L., 1979. Optimizing
the continous production of Candidta utilis and
Saccharomyces fibuliger on potato processing waste water.
Appl. Environ. Microbiol, 37:227-232.
Ngadi, M.O. and Corrota, L.R. 1992. Solid state fermentation of
apple pomace as affected by moisture and bioreactor mixing
speed. J. Fd. Sci., 57(3):667-670.
Ranganna, S. 1986. Hand book of Analysis and Quality Control for
Fruit and Vegetable Products. 2nd Edn. Tata McGraw Hill
Publishing Co., Ltd. New Delhi.
Ratledge, C. 1989. Microbiol Technology of Lipids : Lipid Technol.
1:34-39.
References
AOAC 1980. Association of Official Analytical Chemists. Official
Methods of Analysis. ed. Hortwitz. W. 13th edn, Washington.
D.C.
Downing, D.L. 1989. Processed Apple Products. AVI Van Nostrand
Reinhold, Newyork. p. 433.
Fontenot, J.P., Bovard K.P., Oltzen, R.R., Rumsey T.S. and Driode,
B.M. 1977. Supplementation of apple pomace with nonprotein nitrogen for gestating cows. J. Anim. Sci.,
46:513-522.
Ghildyal, N.P., Lonsane, B.L., Sreekantiah, K.R. and
Sreenivasamurthy, V. 1985. Economics of submerged and
solids state fermentation for the production of
amyloglucosidase. J. Food Sci. Technol. 22:171-176.
Gupta, L.K., Pathak, G. and Tiwari, R.P. 1990. Effect of nutrition
variables on solid state alcoholic fermentation of apple pomace
by yeasts. J. Sci. Food. Agric. 50:55-62.
Hang, Y.D. and Walter, R.H. 1989. Treatment and utilization of
apple processing waste.In; Processed Apple Products, ed.
Downing D.L. AVI Van Nostrand Reinhold, New York,
365-376.
Hang, Y.D., Lee, C.Y. and Woodams, E.F. 1982. A solid state
fermentation system for production of ethanol from apple
pomace. J. Food Sci. 47:1851-1852.
Hang, Y.D. and Woodams, E.F. 1984. Apple pomace : A potential
substrate for citric acid production by Aspergillus niger.
Biotechnol Lett. 6:763-764.
Hang, Y.D. 1988. Improvement of the nutritional value of apple
pomace by fermentation. Nutrition Reports International,
38(1): 207-209.
Holdsworth, J.E. and Ratledge, C. 1991. Triglyceride synthesis in
the oleaginous yeast Candida curvata D. Lipids, 26(2):111118.
Joshi, C. and Joshi, V.K. 1990. Food Processing Waste Management
Technology, Need for an integrated approach. Indian Food
Packer, 44: 56-67.
91
Research note
1*
Department of Food Science and Technology, Nauni, Dr Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan,
Himachal Pradesh,India
2
Institute of Biotechnology and Environmental Science, Neri, Hamirpur, Dr YS Parmar University of Horticulture and Forestry,
Nauni, Solan, Himachal Pradesh,India
*
Email: manishapht@gmail.com
Paper no: 43
Abstract
Varietal performance studies of wild and introduced variety of Amla were conducted at Regional
Horticultural and Forestry Research Station, Bhota, Himachal Pradesh. The selections of amla (NA-6,
NA-7 and NA-10) were made from Faizabad (UP) and introduced at Research station Bhota, Hamirpur
and at farmers field during the year 2002 to evaluate their performance. It was inferred that cultivar
NA-7 and NA-10 have outperformed the traditional local strain for almost all the characteristics studied.
These cultivars were having higher survival rate, shorter juvenile phase, better tree canopy and yield.
The fruit of Local (wild) variety was found smaller than the introduced variety like NA-7 and NA-10. In
terms of chemical characteristics, the berries of local cultivars had comparatively higher soluble solids
(11.36oB) and ascorbic acid (275.7 mg/100g) than NA-7 and NA-10 varieties. However, on the basis of
antioxidant potential NA-7 had higher phenol content (21.02 g/100g) as compared to the local and NA10 cultivars. The fruits of local variety were used to prepare pickle while NA-7 and NA-10 were
employed to prepare preserve.
2012 New Delhi Publishers. All rights reserved
Amla pickle
Figure 2: Flow diagram of amla/aonla pickle preparation
94
Processing potential of newly introduced Amla cultivars grown in lower Himalayan Region of Himachal Pradesh
Performance studies
The performance data (pooled averages) of different amla
cultivars presented in Table 1 revealed that survival per cent
was recorded highest for local strains though statistically it
was at par with cultivar NA-10 (71%). The length of juvenile
period (years) was observed almost similar in all the cultivars.
As far as tree growth was concerned the tree height was
recorded highest for NA-7 which was significantly greater than
the local strains. The height of NA-10 was statistically at par
with NA-7. Tree spread was almost similar among the different
cultivars. Per tree yield data indicates that highest yield was
obtained for cultivar NA -7 which was significantly higher
than that of local. From the results presented it can easily be
inferred that cultivar NA-7 and NA-10 have out-performed the
95
Table 1: Performance of introduced amla cultivars under low hill region of Himachal Pradesh (Pooled data for tree size and yield for two
years)
Cultivar
NA-7
NA-10
Local
CD(0.05)
Survival
(%)
Juvenile
Period (years)
Height
Yield
(kg)
5
71
72
7.2
3
3
5
NS
4.75
4.40
3.3
1.33
4.92
4.63
3.90
NS
92
83
52
18.7
Physical characteristics
The comparative study of three cultivars of amla viz., NA-7,
NA 10 and Local showed that the fruits were round, ribbed
and pale green. The average fruit weight and seed weight varies
from 6.58 to 33.8g and 0.94 to 2.70g respectively (Plate 1). The
different components of amla fruit accounted for 6.07-14.26
per cent peel, 69.77-85.49 per cent pulp and 7.10- 15.12 per cent
seed. The maximum and minimum fruit weight was attained by
NA-10 (33.8g) and Local (6.58g), while the maximum seed weight
by NA-7 (2.70g) with a minimum of 0.94 g by Local variety. Out
of the three varieties, the skin of NA-7 cultivar was glossy and
shiny with colour parameters of Yellow Green 144C as read
from Horticulture Colour Charts, while the other two cultivars
were having Yellow Green 145A colour. The size parameters of
three cultivars showed that the Local, NA-7 and NA-10 had a
length (mm) of 21.8, 36.44 and 33.47 with a diameter measuring
22.19, 38.08 and 38.47 mm respectively. Thus, the average size
of NA-7 aonla cultivar was found larger than the other two (
Table 2). The data were in conformity with those obtained by
Goyal et al (2008), Mishra (2009) and Kalra (1988) for different
amla cultivars.
(Emblica officinalis)
Local
NA-7
NA-10
Moisture (%)
Total solids (%)
TSS (oB)
Titratable acidity
(% citric acid)
Brix/acid ratio
Sugars (%)
Reducing
Total
Ascorbic acid (mg/100g)
Total phenols (g/100g)
Fibre (%)
Juice recovery (%)
84.6
15.4
11.36
2.73
82.2
17.8
9.8
2.97
86.18
13.82
9.15
2.17
4.16
3.29
4.21
4.76
5.54
275.7
20.63
2.70
40.0
3.17
4.54
205.0
21.02
3.83
42.0
3.21
5.12
200.3
20.99
3.41
44.0
Chemical characteristics
Data pertaining to the chemical composition of amla cultivars
are presented in Table 3 indicate that moisture content in amla
berries (Local, NA-7 and NA-10) ranged between 82.2 to 86.18
per cent with a maximum in NA-10 and minimum in NA-7
cultivars. In all the three cultivars of amla, the total soluble
solids (oB) and titratable acidity (% citric acid) lied between
9.15 to 11.36 and 2.17 to 2.97 respectively, where Local and
NA-7 cultivar had highest TSS and acidity respectively. The
brix/acid ratio was maximum in NA-10 (4.21) and minimum in
NA-7 (3.29) cultivars. As reported earlier, the amla fruit is
regarded as a rich source of ascorbic acid besides other vital
nutrients. The level of vitamin C in amla berries varied from
200.3 to 275.7mg/100g with the maximum in Local cultivar.
Besides vitamins, the amla fruit also contained good proportion
of total phenols, in different cultivars; their respective values
were recorded as 20.63, 21.09 and 20.99 g/100g for Local, NA7 and NA-10, respectively. The fibre content in the amla
cultivars was found maximum in NA-7 (3.83%) with a minimum
in Local (2.70%). Thus, the berries of amla were regarded as a
fruit containing good proportion of vitamin C and phenols
Parameters
NA-10
33.8
33.47
38.47
0.86
10.89
77.32
11.77
1:9.2
Yellow
Green 145C
27.0
96
Processing potential of newly introduced Amla cultivars grown in lower Himalayan Region of Himachal Pradesh
References
AOAC. 1995. Official Methods of Analysis 16th edn, Association of
Official Analytical Chemists, Washington, DC.
Goyal, R.K., Patil, R.T., Kingsly, A.R.P., Walia, H., and Kumar, P.
2008. Status of postharvest technology of aonla in India- a
review. American Journal of Food Technology. 3 (1):13-23
Jamwal, K.S., Sharma, I.P.,and Chopra, L. 1959. Pharmacological
investigations on the fruits of Emblica officinalis. J. Sci.
Ind.Res. 18: 180-181.
Jayshri, S, and Jolly, C.I. 1993. Phytochemical , antibacterial and
pharmacological investigations on Monordica chiranlia and
Emblica officinalis. Ind. J. Pharm. Sci. 1: 6-13.
Kalra, C.L. 1988.The chemistry and technology of Amla (Phyllanthus
emblica) - a resume. Indian Food Packer. 42(4): 67-82.
Kumar, G. S., Nayaka, H., Dharmesh, S. M. and Salimath, P. V. 2006.
Free and bound phenolic antioxidants in amla (Emblica
officinalis) and turmeric (Curcuma longa). Journal of Food
Composition and Analysis. 19: 446-452.
Lal, G, Siddappa, G.S. and Tandon, G.L. 1959. Preservation of Fruits
and Vegetables, Indian Council of Agriculture Research, New
Delhi, p 483.
Mishra, P., Srivastava, V., Verma, D., Chauhan, O. P. and Rai, G. K.
2009. Physico-chemical properties of Chakiya variety of
Amla (Emblica officinalis) and effect of different dehydration
methods on quality of powder. African Journal of Food
Science. 3(10): 303-306.
Pathak, R. K. 2003. Status Report on Genetic Resources of Indian
Gooseberry - Aonla (Emblica officinalis Gaertn.) in South
and Southeast Asia. IPGRI Office for South Asia National
Agriculture Science Centre (NASC). 99p.
Patil, S. R., Suryawanshi, A. B., and Phad, G. N. 2010. Performance
of some aonla (Emblica officinalis Gaertn) cultivars under
Vidarbha condition of Maharashtra. International Journal of
Plant Sciences, 5 (1): 36-37.
Rao, K. D. and Subramanyam, K. 2009. Growth and yield performance
of aonla varieties under scarce rainfall zone. Agric. Sci. Digest,
29 (2): 45-47.
Ranganna, S. 1986. Handbook of Analysis and Quality Control for
Fruit and Vegetable Products, 2nd edn, Tata McGraw Hill
Pub. Co. Ltd. New Delhi. p 1110.
Rao, T.S., Kumari, K.K., Netaji, B., and Subhokta, P.K. 1985.
Ayurveda Siddha. J Res. 6:213-224.
Rastogi, R.P. 1993. Compendium of Indian Medicinal plants, CDRI,
Lucknow and ID, New Delhi 1: 530.
Processed Products
Preserve: On the basis of sensory evaluation, it was observed
that amla preserve scored 7.0, 7.5 and 7.5 for colour, flavour
and body characteristics respectively (Plate 2a). On the basis
of taste, the sample scored 8.0 thus emphasizing its overall
acceptability (Table 4).
Amla pickle: To improve the texture of the fruit and also to
remove astringency, brining is important in pickling. Sensory
evaluation of the amla pickle showed that the pickle scored 7.0
for colour and flavour on the basis of 9 point Hedonic Rating
(Table 4). However, the taste perception of the product scored
7.85 out of 9 (Plate 2b). Thus, on the basis of overall
acceptability, amla pickle prepared by using local cultivars
highly acceptable (7.93) apart from meeting the specifications
laid down in FPO (1955).
Table 4: Sensory evaluation of processed amla products
S.No
1.
2.
3.
4.
5.
Attributes*
Colour
Flavour
Body
Taste
Overall Acceptability
Pickle
Preserve
7.00.2
7.00.15
7.450.08
7.850.10
7.930.10
7.00.10
7.50.18
7.50.11
8.00.12
8.00.15
97
Research note
Department of Postharvest technology of Hort. Crops, Bidhan Chandra Krishi Viswavidyalaya,West Bengal,India
Department of Food Science and Technology, Dr Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal
Pradesh,India
2
Email: v.k.joshipht@rediffmail.com
Paper no: 44
Abstract
Colour of a foodstuff is one of the most important elements determining its acceptance besides
enhancing its delicacy. Storage stability of the dried plum anthocyanin powder with 300B maltodextrin
as a carrier in RTS beverage model solution showed more effect of temperature at 350C than at 00C.
There were less changes in dark light condition than in day and UV light. The change in colour was
rapid in the first 1 month than in the later period of storage. By the use of plum pomace and with the
above optimized conditions, crude anthocyanin pigments can be produced and used in processed
food.
2012 New Delhi Publishers. All rights reserved
100
T6 :350C Dark
T5 :250C Dark
T4 :00C Dark
T3 :350C UV
T2 :350C Day
T1 :00CDay
Times (Months)
Treatments
TSS
14.0 14.1
14.0 14.1
14.0 14.2
14.0 13.9
14.0 13.9
14.0 14.2
0.70
0.70
0.40
0.70
0.70
1.40
pH
3
3.15 3.22
L
%
3 change
%
chang
e
3.30
12.8 0.30
0.30 0.29
12.70 0
3.30
12.8 0.30
0.30 0.29
12.70 0
3.30
12.9 0.50
0.30 0.29
12.70 0
3.15 3.20
3.15 3.20
3.15 3.21
10.00
2.20 78.50 73.9 2.50
12.8 0.30
3.15 3.22
0.30 0.27
12.70 0
0
10.00
2.80 78.50 73.7 2.50
12.8 0.30
0
3.15 3.24
0.30 0.27
12.70 0
6.60
12.9 0.50
0.30 0.28
12.70 0
% Total sugar %
%
Acidity
change 0
3 change
3 change 0
a
7.70 6.90 10.40 19.00 16.4 13.70 3.60 1.30 63.90 3.20 4.03
0
7.70 7.00 9.00 19.00 16.3 14.20 3.60 1.80 50.00 3.20 3.99
0
7.70 7.40 3.90 19.00 15.8 16.80 3.60 1.80 50.00 3.20 3.92
0
7.70 5.70 26.00 19.00 16.6 12.60 3.60 1.00 72.20 3.20 4.10
0
7.70 6.60 14.30 19.00 16.5 13.20 3.60 1.10 69.40 3.20 4.10
0
26.00
24.70
22.50
28.00
28.00
23.40
%
b
% Yellow unit % change
Red unit
%
3
3 change 0
3 change 0
3 change 0
7.70 7.30 7.80 19.00 16.0 15.80 3.60 1.70 52.70 3.20 3.95
0
Table 1: Per cent change in various physico-chemical and colour characteristics of the RTS based model solution after three months of storage
Devi et al.
Evaluation of the stability of plum anthocyanin powder in RTS based model solution
Conclusion
Physico-chemical characteristics
There was a slight increase in the TSS, total sugar and pH
during the different storage conditions and storage interval in
RTS based model solution. However, the titratable acidity
showed a slight decrease during different storage conditions
and storage intervals. Although, statistically no significant
differences were observed in this parameters as the variation
were too narrow that almost a constant TSS, total sugar,
titratable acidity and pH were observed along the experiment.
These findings are in accordance with those of Viguera et.al.
(1998). Attri (2004) also conducted stroage study of microbial
Pigment and reported that there were no significant changes
in these parameters in RTS beverage prepared with microbial
pegments during storage at 20, 30 and 370C.
There was a significant change in the colour measurement of
the RTS based model solutions in different light and
temperature conditions as well as during storage for 3 months
interval, which is also in line with the findings of Attri (2004).
The a value which gives the depth of redness was found to
be decreasing as the storage interval increases. These findings
are in agreement with those of Pilando et. al. (1985) who
reported that anthocyanins, the red pigments in raspberry and
other fruits, degrade and polymerized easily with passage of
time with time. These findings are further in conformity with
Timberlake and Bridle (1976) who studied in model system,
found that mixtures of anthocyanins and catechin during
storage gradually lost colour in the red region of the spectrum
while increasing in the brown region.According to Skrede
(1985) day light storage of syrups lowered half-life of Hunter
a values by 10-30% as compared with dark storage which is
in line with the observations recorded in our study.
It is also clear from the values given in table 1 that changes in
TSS and total sugar during storage was not even 1%.It is
desirable from food processing point of view. However, it is
very much evident that the changes in acidity, pH, L, a, b
values of colour measurement and the red and yellow units of
tintometer colour unit were very drastic. The fading of colour
was very much apperent during 3 month of storage. Similarly,
effect of storage in light was greater than in lower temperature
and in dark conditions.The contrasting effect of storage
revealed the occurrence of faster changes during the first one
Book Review
Bio-processing of Foods
ASIA TECH PUBLISHERS INC, NEW DELHI
This book is based on the proceedings of the conference on New Horizons in Bo-Processing of Foods organized by Department
of Food Engineering and Technology held at Sant Longowal Institute of Engineering and Technology from 25th -26th February
2011. The selected papers that were presented in th conference have been compiled and edited in the form of book. It provides
an in-depth understanding on different topics that are of great relevance to food and fermentation technology. This indispensible
outcome is the result of combined efforts of different academic credentials in the area of food technology and biotechnology. To
make the concept more clear, the contents of this treatise have been divided into following sections:
The each sections comprises of many chapters. The chapters clearly demonstrate the recent developments and advances made
in the field of food Industry and applications of biotechnology especially its latest innovations in food processing. The
prominent topics covered in the book are
Importance and Nutitive value,Sole present status and future strategies in Fruit wines in India
Nowadays bio technology plays an important role in the food industry as integral part of processing. This field for food
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quality of foods. During the last years, much research has focused on the improvement of enzyme behaviour in the conditions
in which they were to be used, and especially on the increase of their thermal and operational stability. This book attempts to
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of enzyme and biotechnological interventions in food technology. In this regard, the use of food enzymes have been covered
in the three chapters of this volume.
The text is supported by a number of clear, informative diagrams for better understanding. The book is highly useful for postgraduate students and researchers of food technology, biotechnology, applied biology, microbiology and biochemical engineering.
It presents the basic and applied aspects from all the possible facets to serve as a text-cum-reference book. The book includes
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the topics biotechnology in food processing which would be an asset to all the readers.
Dr ( Mrs) Neerja Rana
Assistant Professor
Department of Basic Sciences
Dr Y.S. Parmar University of Horticulture and Forestry Nauni Solan(HP)