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NAMA : SUJASMIN

NPM : G1J216022
INTRODUCTION
These pectinase enzymes have importance in industries
like fruit industry for clarification, textile industry for
bioscouring, in waste water treatment and therefore have
promising future aspects
MATERIALS AND METHODS
Materials
1. Carrot Waste
2. Nutrient Broth (NB)
3. Nutrien Agar (NA)
4. Potato Dextrose Broth (PDB)
5. Potato Dextrose Agar (PDA)

Methods
A. Culture Isolation
5g of carrot waste collected from local vegetable market was
suspended in 50ml D/W.
1ml was dispensed in 100ml Nutrient Broth (NB) & Potato Dextrose
Broth (PDB) respectively
The flasks wereincubated at shaker conditions of 600 rpm for
48 hr at 26oC
one loopful of suspended broth was isolated onto Nutrient
Agar (NA) and Potato Dextrose Agar (PDA) and incubated at 26oC for
48hrs
B. Screening of Isolates
Colonies that were grown on NA and PDA
media were picked up and spotted on plates
containing Hankinss agar medium (Hi-Media).
These plates were incubated at 26oC for 48 hrs.
1 % cetrimide was overlaid on these plates and kept for
half an hour
Cells from these colonies were then picked up and sub-
cultured on NA slants and maintained for further studies
C. Mediaum and Culture condition

The Hankins broth medium was prepared in


500ml Erlenmeyer flasks and the flask was
inoculated with loopful of cells grown for 48hr on NA
slants.
The flasks were kept on shaker conditions of 600 rpm
for 48hr at 26oC with media pH of 7.0
2. Production Of Pectinase Using Carrot waste as substrat

In 500ml Erlenmeyer flasks, 300g of crushed


carrot waste was added and was dispensed
with 100ml of D/W
To this flask, 10ml of bacterial suspension of O.D. 0.05
from Inoculum flask was added
Similar set up was followed for studying the effect of
temperature (15oC to 60oC), pH (3 to 9) and
concentration of vegetable waste (0.4kg to 2.0kg) on
enzyme production
The flask was incubated on ashaker at 600rpm for
48hr
Result and Discussion
Fifty isolates showing polygalacturonase activity were
obtained based on zones of translucent clearance seen
around them. The nine isolates showing good Cx ratio
on agar plate were selected from 50 isolates, of which
three isolates showing maximum Cx ratio were than
selected and were named Isolate 1, Isolate 2, and
Isolate 3 (Table 1)
Effect of temperature on enzyme activity
Effect of pH on enzyme production
Effect of substrate on enzyme
production
conclusion
So the present investigation reports a new,
effective and easy method for obtaining
pectinase producing organisms from vegetable wastes.
Thus in future, similar cheaper sources such as carrot
waste can be employed for the
extraction of enzyme pectinase which holds
numerous industrial applications