caa a22 4500 606212752 CHVBK 20210128101044.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s10295-014-1549-6 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1549-6 Molecular cloning and expression of thermostable glucose-tolerant β-glucosidase of Penicillium funiculosum NCL1 in Pichia pastoris and its characterization [Elektronische Daten] [Gurusamy Ramani, Balasubramanian Meera, Chinnathambi Vanitha, Jeyaprakash Rajendhran, Paramasamy Gunasekaran] A partial peptide sequence of β-glucosidase isoform (Bgl4) of Penicillium funiculosum NCL1 was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The cDNA (bgl4) encoding Bgl4protein was cloned from P. funiculosum NCL1 RNA by consensus RT-PCR. The bgl4 gene encoded 857 amino acids that contained catalytic domains specific for glycoside hydrolase family 3. The cDNA was over-expressed in Pichia pastoris KM71H and the recombinant protein (rBgl4) was purified with the specific activity of 1,354.3U/mg. The rBgl4 was a glycoprotein with the molecular weight of ~130kDa and showed optimal activity at pH 5.0 and 60°C. The enzyme was thermo-tolerant up to 60°C for 60min. The rBgl4 was highly active on aryl substrates with β-glucosidic, β-xylosidic linkages and moderately active on cellobiose and salicin. It showed remarkably high substrate conversion rate of 3,332 and 2,083μmol/min/mg with the substrates p-nitrophenyl β-glucoside and cellobiose respectively. In addition, the rBgl4 showed tolerance to glucose concentration up to 400mM. It exhibited twofold increase in glucose yield when supplemented with crude cellulase of Trichoderma reesei Rut-C30in cellulose hydrolysis. These results suggested that rBgl4 is a thermo- and glucose-tolerant β-glucosidase and is a potential supplement for commercial cellulase in cellulose hydrolysis and thereby assures profitability in bioethanol production. Society for Industrial Microbiology and Biotechnology, 2015 Penicillium funiculosum NCL1 nationallicence β-Glucosidase nationallicence Pichia pastoris nationallicence Thermo-tolerant nationallicence Glucose-tolerant nationallicence Ramani Gurusamy Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut Meera Balasubramanian Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut Vanitha Chinnathambi Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut Rajendhran Jeyaprakash Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut Gunasekaran Paramasamy Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/4(2015-04-01), 553-565 1367-5435 42:4<553 2015 42 10295 https://doi.org/10.1007/s10295-014-1549-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-014-1549-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ramani Gurusamy Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut NATIONALLICENCE 700 1- Meera Balasubramanian Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut NATIONALLICENCE 700 1- Vanitha Chinnathambi Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut NATIONALLICENCE 700 1- Rajendhran Jeyaprakash Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut NATIONALLICENCE 700 1- Gunasekaran Paramasamy Department of Genetics, Centre for Excellence in Genomic Sciences, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/4(2015-04-01), 553-565 1367-5435 42:4<553 2015 42 10295