Cellulose is the most abundant biomass on earth. The major players in cellulose degradation in nature are cellulases produced by microorganisms.
Aerobic filamentous fungi are the main sources of commercial cellulase. Trichoderma reesei has been explored extensively for cellulase production; however, its major limitations are its low beta-glucosidase activity and inefficiency in biomass degradation.
The aim of this work was to isolate new fungal strains from subtropical and tropical forests in China, which produce high levels of cellulase in order to facilitate development of improved commercial cellulases.
Results:
We isolated 305 fungal strains from 330 samples collected from subtropical and tropical virgin forests in China. Of these, 31 strains were found to have Avicelase activity of more than 0.2 U/ml in liquid batch cultivation.
Molecular analyses of the 31 strains based on internal transcribed spacer sequences revealed that 18 were Trichoderma and 13 were Penicillium species. The best-performing isolate was Trichoderma koningiopsis FCD3-1, which had similar Avicelase activity to T.
reesei Rut-C30. Most interestingly, strain FCD3-1 exhibited extracellular beta-glucosidase activity of 1.18 U/ml, which was approximately 17 times higher than that of Rut-C30.
One beta-glucosidase secreted by FCD3-1 was purified, and its gene was cloned and identified. The beta-glucosidase belonged to glycosyl hydrolase (GH) family 3, sharing the highest identity of 94% with a GH family 3 protein from Trichoderma atroviride IMI 206040, and was designated TkBgl3A.
The optimal pH and temperature of TkBgl3A were 4.5 and 65[degree sign]C, respectively. The enzyme retained over 90% activity for 360 hours at pH 4.0 and 30[degree sign]C, which are the usual conditions used for simultaneous saccharification and fermentation (SSF) of cellulose to ethanol.
The enzyme showed significantly higher specific activity toward natural substrate cellobiose (141.4 U/mg) than toward artificial substrate p-nitrophenyl-beta-D-glucopyranoside (108.0 U/mg).
Conclusions:
Strains of Trichoderma and Penicillium were the predominant cellulolytic fungi in subtropical and tropical forests in China. T.
koningiopsis FCD3-1 was the most efficient producer of cellulase, and also produced a high level of beta-glucosidase. The high specific activity toward cellobiose and stability under SSF conditions of the purified beta-glucosidase from FCD3-1 indicates its potential application in SSF of cellulose to bioethanol.
Author: Zheng ZhangJun-Liang LiuJian-Yi LanCheng-Jie DuanQing-Sheng MaJia-Xun Feng
Credits/Source: Biotechnology for Biofuels 2014, 7:107
Published on: 2014-07-17
Tweet
News Provider: 7thSpace Interactive / EUPB Press Office
Social Bookmarking
RETWEET This! | Digg this! | Post to del.icio.us | Post to Furl | Add to Netscape | Add to Yahoo! | Rojo
There are no comments available. Be the first to write a comment.