Sunday, July 24, 2011
Grand Ballroom, 5th fl (Sheraton New Orleans)
Cellulose is the most abundant and ubiquitous renewable biopolymer consisting of around 8,000 to 12,000 D- anhydro glucopyranose units linked together by the beta (1-->4) glycosidic bonds. Complete hydrolysis of Cellulose to glucose requires cellulase- a multi-enzyme system consisting of 3 different component enzymes; endoglucanases, exoglucanases and beta-glucosidases. The fungus Penicillium funiculosum is well known for cellulase production.
In this work we started screening for the various cellulase component enzymes present in the culture supernatant of P. funiculosum. SDS-zymogram revealed that there are 3 beta-glucosidases (120, 100 and 50 kDa.) and multiple endoglucanases with molecular weights in the range of 19-70 kDa. We investigated the expression profiles of the 3 cellulase components as a function of time when P.funiculosum was cultivated on different carbon sources (disaccharides, monosaccharides, complex cellulosic and non-cellulosic carbohydrates). Our results showed that supplementation of these carbon sources with wheat bran improved the cellulase production significantly. Since the genomic DNA sequence of P.funiculosum is not available we designed degenerate primers based on the cellulase genes available in the database. PCR amplification with these degenerate primers yielded partial sequence (550 bp) of GH3 family beta-glucosidase and a partial sequence (650 bp) of cellobiohydrolase belonging to the GH7 family.
In this work we started screening for the various cellulase component enzymes present in the culture supernatant of P. funiculosum. SDS-zymogram revealed that there are 3 beta-glucosidases (120, 100 and 50 kDa.) and multiple endoglucanases with molecular weights in the range of 19-70 kDa. We investigated the expression profiles of the 3 cellulase components as a function of time when P.funiculosum was cultivated on different carbon sources (disaccharides, monosaccharides, complex cellulosic and non-cellulosic carbohydrates). Our results showed that supplementation of these carbon sources with wheat bran improved the cellulase production significantly. Since the genomic DNA sequence of P.funiculosum is not available we designed degenerate primers based on the cellulase genes available in the database. PCR amplification with these degenerate primers yielded partial sequence (550 bp) of GH3 family beta-glucosidase and a partial sequence (650 bp) of cellobiohydrolase belonging to the GH7 family.