JOURNAL OF TEXTILE RESEARCH ›› 2012, Vol. 33 ›› Issue (8): 82-86.

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Cloning and expression of β-1,4-endoglucanas gene from Symbiobacterium thermophilum

 JIANG  Hai-Qin, BI  Yun-Feng, HUA  Xin-Chun, CHEN  Li-Li, XU  Xue-Fei, SHEN  Ming-Hao   

  1. College of Food Science and Engineering, Jilin Agricultural University
  • Received:2011-09-14 Revised:2012-01-04 Online:2012-08-15 Published:2012-08-08
  • Contact: SHEN Ming-Hao E-mail:shenmh2003@yahoo.com.cn

Abstract: Abstract: [Objective]To screen endoglucanas gene with thermostability,and construct themostable engineering bacteria with high enzyme activity.[Methods]A gene encoding endoglucanase from S.thermophilum IAM 14863 was cloned and recombinanted into expression vector pET32a(+),then the recombinant vector pET32a-Glu was transformed and expressed in E.coli BL21(DE3)PLYS.[Results]The whole length of is 1101bp,which encodes 366 amino acids.The result of SDS-PAGE showed that the gene was effectively expressed in E.coli BL21(DE3)PLYS,and the enzyme showed that a molecular weight of 60kDa approximately.The enzyme activity was 103 U/mL after purification.Characterization of the expression product showed that the optimum pH value was about 5,and the optimum reaction temperature was at 55℃,and expression product displayed better thermal stability(about 75℃)and pH stability. [Conclusion]The research first recombinanted β-1,4-endoglucanas gene from S.thermophilum into expression vector pET32a(+) and expressed highly in E. coli. It will lay the foundation for food,textile,chemical and other fields by widely using the enzyme.

Key words: cellulase ,  , &beta, -1,4-endoglucanas ,  , Symbiobacterium thermophilum , cloning , prokaryotic expression

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