Browsing by Subject "Purification"
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ItemIsolation and purification of cellulase from alkaline-tolerant Bacillus subtilisLignocellulosic biomass is one of the most abundant renewable resource in the world. It has high potential to be used as material for biorefinery process because it yields fermentable sugar that can be converted to various bioproducts via microorganism functions. To make the biorefinery process success, the efficient hydrolysis process is needed to be improved to release the maximum amounts of sugars from lignocellulosic biomass. Natural environment is an important source to find an efficient cellulase producing bacteria. Previously, an alkaline-tolerant cellulase-producing bacterium, Bacillus subtilis strain MSB9, was screened and isolated from Botanic garden in Mahasarakham province, Thailand. In this study, we focused on the purification and characterization of cellulase enzyme produced by B. subtilis strain MSB9. The crude cellulase enzyme was partially purified and concentrated by ammonium sulfate precipitation and fractionated by using size exclusion chromatography using sephacryl S-100 HR column. Two of purified cellulase have relative molecular mass of 35 and 45 kDa, as determined by SDS-PAGE combined with CMC-zymogram.
ItemPurification and identification of bacterial cellulase activity of Bacillus subtilis W48 for biofuel production(School of Biotechnology, Assumption University of Thailand, 2014-07) Patchanee YasurinLignocellulosic 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.