Purification and identification of bacterial cellulase activity of Bacillus subtilis W48 for biofuel production

Abstract

Lignocellulosic biomass is a renewable, inexpensive, and abundant resource with high potential for biofuel production to implement the sustainable energy worldwide. An

important key of biofuel

production

is the hydrolysis of

lignocellulosic biomass to

fermentable sugars. Screening for a novel cellulase, as a biocatalyst, is challenge for

development of

biofuel production to

be the

economic

and environmental friendly

process. Natural environment is an

important

source to

find an efficient cellulase

producing bacteria. Previously, Bacillus subtilis strain W48 was screened and isolated

from

rainforest

park at Assumption

University,

Bangkok,

Thailand. In this study, we

focused on the

purification

and characterization

of cellulase enzyme

produced

by B.

subtilis strain W48. The crude cellulase enzyme was collected by culture grown at 45°C in carboxy-methyl-cellulose (CMC) media. Then, it is concentrated by ammonium sulfate precipitation and fractionated by using size exclusion chromatography using sephacryl S-

100

HR column. In this study, the fractions that has highest CMCase activity (0.017

mg/ml), as endoglucanase, and FPase activity (0.013 mg/ml), exoglucanase, are fraction no. 13 and 11, respectively.