Isolation of a cellulase-producing bacterium,cloning and expression of its novel β-glucosidase gene
会议名称：《重庆微生物学会第九届会员代表大会暨学术年会》
会议日期：2009年
学科分类：081703[工学-生物化工] 08[工学] 0817[工学-化学工程与技术] 0836[工学-生物工程] 082203[工学-发酵工程] 0822[工学-轻工技术与工程]
关 键 词：Cellulase Novelβ-Glucosidase Screen Identification Activity
摘 要：[Objective]In order to get a cellulase-high-yielding bacterium,and try to obtain some novel cellulase genes.[Methods]CMCNa plate screening method was used to screen cellulose-producing bacteria strains,and one obtained strain was identified according to its 16S rDNA sequence,physiological and biochemical ***,a gene encoding a novel cellulolytic enzyme was cloned from this strain, and its sequence characters were ***,the heterologous expression of this gene was tested in a prokaryotic expression system.[Results]A facultative anaerobic strain which produced cellulase was obtained by the CMCNa plate *** strain was identified as Klebsiella sp..By a shotgun cloning method and with Escherichia coli as the host cell,we isolated a novelβ-glucosidase gene,nglu02,from the *** nglu02 was then successfully expressed in transformant *** strain BL21,and the recombinant enzyme was partially characterized.β-glucosidase activity could be detected in the supernatant of the transformant ***’s worth noting that the recombinant enzyme had certain alkali resistance and could keep its activity with the exist of Cu,Fe,EDTA and urea.[Conclusion]We successfully cloned a novelβ-glucosidase gene,nglu02 from the cellulase-producing *** also expressed the gene in prokaryotic expression system and testified that the produced protein was *** study offered some useful information for the endeavor to understand the molecular structure and mechanism of cellulolytic enzymes.