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Cloning and Expression of an Exo-β-1，3 glucanaseGene from Coprinopsis cinerea(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2016年03期
Page:
96-
Research Field:
·生命科学·
Publishing date:

Info

Title:
Cloning and Expression of an Exo-β-1，3 glucanaseGene from Coprinopsis cinerea
Author(s):
Wang Jun; Kang Liqin; Liu Zhonghua; Yuan Sheng
School of Life Sciences，Nanjing Normal University，Jiangsu Key Laboratory for Microbes and Microbial Functional Genomics，Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources，Nanjing 210023，China
Keywords:
exo-β-1; 3 glucanase; gene cloning; heterologous expression; biochemical characterization; Coprinopsis cinerea
PACS:
Q786
DOI:
10.3969/j.issn.1001-4616.2016.03.016
Abstract:
To acquire enough β-1，3 glucanase for investigating its role in morphological changes of fungus，an exo-β-1，3 glucanase gene（exg）was cloned from cDNA of Coprinopsis cinerea okayama 7#130 and inserted into plasmid pET28A（+）to generate the recombinant expression plasmid pET28A（+）-exg. pET28A（+）-exg was transformed into Escherichia coli Rosetta strain for heterologous expression. The result showed that the length of exg gene is 2 415 bp，which encodes 786 amino acids. SDS-PAGE analysis showed that the exo-β-1，3 glucanase was effectively expressed in E. coli Rosetta with an apparent molecular masses of 85 kDa. The specific enzyme activity was 45 U/mg after purification. Characterization of the recombinant enzyme showed that it had an exo-hydrolases activity and the optimum pH value was 6.0，and the optimum reaction temperature was at 60 ℃ when laminarin as substrate. Besides，the thermostability and pH stability give it some prospect in the future.

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Last Update: 2016-09-30