Converting Agricultural Biomass Wastes into

Cellulosic Ethanol by Fermentation

H. Leng1, J. Yang1, C.F. Yu2,*
1Hong Kong Baptist University, Hong Kong, China
2United International College, Zhuhai, China

Introduction In the control fermentation, sucrose was the only carbon source for the yeasts. All the
three types of yeasts could efficiently utilize sucrose in the production of ethanol. The
ethanol production rates among the natural grape yeasts and two commercial yeasts
Converting waste agricultural biomass into a reusable material or recovered energy re-
were as follows: grape yeasts > white wine yeast > red wine yeast.
source would not only greatly reduce the extent of air and soil contamination, but also
significantly lower the waste disposal cost and generate considerable revenue from the
In the cellulase enzymatic hydrolysis of the two crop wastes to remove lignin and di-
produced material and renewable energy[1,2]. Crop waste is abundant in China and
gest cellulose into simple sugars, sugar bagasse would produce more sugar per unit
the recovery of the sun energy stored in the waste by converting it into cellulosic
mass for yeast fermentation into ethanol than corn stalks under the same conditions.
ethanol as renewable energy is one of the feasible solutions in integrated solid waste
management[3,4].
All the ethanol samples collected after distillation were first determined by the alcohol
meter and then further analyzed with GC-MS to confirm their chemical components
In this study, the common biomass wastes generated during the cultivation of corn and
(Figure 2).
sugarcane, corn stalks and sugarcane bagasse, were collected from local farms. They
were first pretreated by cellulase enzymatic hydrolysis before conversion into cellulosic
ethanol by fermentation using two commercial wine yeasts and the natural yeast cells
isolated from grapes. The ethanol conversion efficiency from different wastes was eval-
uated and compared among different fermentation conditions. The amount of ethanol
produced was determined with an alcohol meter and gas chromatography-mass spec-
trometry (GC-MS)[5].

Materials and Methods

Corn stalks and sugarcane bagasse were collected as agricultural waste after harvest
from local farms. The natural yeast cells were isolated from the surface of Jufeng grapes
purchased from local fruit market. The commercial red and white wine yeasts were pro-
duced by Angel Yeast Company Limited in Yichang city. The cellulase enzyme was
produced by Donghua Qiangsheng Biotechnology Company in Beijing city. The water
used was pure water generated by Millipore Purification System and all chemicals were
of analytical grade. The alcohol meter (hydrometer) for measuring ethanol concentra-
tion was manufactured by Shanghai Huan Ao Technology Company Limited.
Figure 2. The chemical components of each ethanol sample were evaluated with GC-MS.
The fermentation of each type of crop wastes (1000 g wet weight and cut into pieces of
2-3 cm long) was conducted in a 3-liter fermentation bottle equipped with an airlock.
The activation of commercial yeast cells and cellulose was conducted according to It was observed that the particle size of the crop wastes was critical in the enzymatic
manufacturers’ manuals before use. After boiling the crop wastes in water for 60 min- hydrolysis and fermentation as the smaller particle size would enhance both the hy-
utes, they were immersed into enzyme solution (0.1% by weight and adjusted to pH drolysis and fermentation rates which would increase ethanol production efficiency
4.5 with 1M HCl) for 24 hours at 45°C. They were filtered to remove the enzyme so- eventually. Moreover, the types of yeast used in fermentation would play a very impor-
lution and then filled into the fermentation bottle containing 3 g of activated yeast tant role and it was found that the natural grape yeasts performed better in the conver-
cells. Water was added into each bottle until a total volume of 3 L before closing with sion of the waste biomass into fuel ethanol by fermentation.
an airlock. After a course of 8- day fermentation at 25°C, the ethanol produced in each
bottle was extracted by (1) filtration to remove yeast cells and crop wastes, and (2) dis-
tillation of filtrate to collect the 95% ethanol, which was determined by an alcohol Conclusion
meter and its total volume was recorded. The ethanol samples were further investigated
by gas chromatography-mass spectrometry (Agilent 5975i GC-MS system). Each fer- The stored energy in waste cellulosic biomass could be recovered and converted into
mentation condition was conducted in triplicate. fuel ethanol by two main steps, (1) cellulase enzymatic hydrolysis to remove lignin and
digest cellulose into simple sugars, and (2) fermentation of sugars with yeasts. The two
common crop wastes, under the same pretreatment and fermenting conditions, sugar-
Results and Discussion cane bagasse gave higher alcohol production rate than corn stalks, whereas the decreas-
ing order of ethanol fermentation efficiency by different yeast cells was natural yeasts >
The ethanol production in the fermentation of different crop wastes compared with the white wine yeast > red wine yeast. In order to further reduce the solid wastes, the com-
control fermentation (each using 300 g of sucrose) was shown in Figure 1. posting of the fermenting wastes into fertilizer was under investigation.

Acknowledgements
The authors are grateful to the generous sponsorship from the College Research Grant
(R201008) of Beijing Normal University – Hong Kong Baptist University United In-
ternational College to support this study.

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