Although, several microorganisms were employed in bioethanol production process, but direct bioconversion of cellulosic materials for example by
is not attractive because of low ethanol productivity and high energy requirements compared to ethanol production by
is a yeast, which is believed to beamong the most effective lignocellulosic biomass degrading microorganisms for hexose sugars suchas glucose, mannose and galactose, it has a capability of rapid rates of glycolysis and ethanol production under optimal conditions, producing over 50 mmol of ethanol per h per g of cellulose
.However, this high rate is maintained for only a brief period during batch fermentation and declines progressively as ethanol accumulates in the surrounding broth
.To date numerous research studies regarding the fermentative activities of
in stirredtank reactors utilizing cellulosic biomass have been reported, alas! There are limited citations onkinetic studies of the process; which we believed will help in understanding the overall process for maximum optimization.
2. Materials and Methods2.1 Fermentation media and Culture
The substrate used for this study is carboxymethylcellulose (CMC) from laboratory stock to serve asthe cellulose substrate, S. cerevisiae that was used was also obtained from laboratory stock. Whilechemicals and reagents such as sodium alginate, calcium chloride, yeast extract, ammonium sulfate, potassium phosphate, hydrated magnesium sulfate, hydrated calcium chloride, DNS reagents,Rochelle salt solution reagents were all obtained from Merck and are of pure quality.2.2 Microbial culture and ImmobilizationThe strain was subcultured on basic media containing yeast extract 3gl
O 0.7 gl
O 0.1 gl
, which was incubated at 30
C for 24 hours.5ml hypodermic syringe was used to pour 100ml of (1% v/v) sodium alginate containing thecultured yeast into a beaker containing 200ml of 0.75% w/v calcium chloride, in a drop wise fashion.The solution in the beaker was then discarded and replaced with 0.11% w/v calcium chloride andwas incubated at 4
C for an hour. And the beads were filtered for fermentation process.
2.3 Standard Glucose Assay
From the standard glucose stock solution, Different samples concentrations (0.1 gl
), were made; following standard DNS glucose assay protocols by Miller
, thespectrophotometric absorbance (OD
)of each sample was recorded and plotted against eachconcentration, which formed the glucose standard curve.
2.4 Enzymatic Hydrolysis