Cloning and Expression of Exo-β-1,4-Cellobiohydrolase Gene from Bacillus pumilus

Abstract:

The exo-β-1,4-cellobiohydrolase gene(cex) of Bacillus pumilus AC-6 was cloned. The cex gene from AC-6 is 2100bp, and encodes 700 amino acids. The cex gene has 94% similarities to the exo-β-1,4-cellobiohydrolase gene sequence from other Bacillus strains. The cex gene was linked with the vector pGEX-4T-1, and transferred into the competent E. coli BL21 for expression. The result of protein electrophoresis showed that the protein has expressed, whose molecular weight was about 104kDa. Measured the enzyme activity of the expression protein of the recombinant strains was 5.40U/mL, which is 1.20 times as the original strains.

• Periodical: Advanced Materials Research (Volume 647)
• Pages: 150-154
• DOI: https://doi.org/10.4028/www.scientific.net/AMR.647.150
• Online since: January 2013
• Authors: Meng Jiao Yan, Song Wei, Ya Li Dai, Lin Yuan
• Keywords: Bacillus pumilus, Exo-β-1,4-Cellobiohydrolase, Expression, Gene Cloning

References

[1] Ding Xinli, Wang Tianhong, Study of the expressions of cellulase from Trichoderma reesei in Saccharomyces cerevisiae, Liguor-making Science & Technology. 9 (2005)28-35.

[2] GaoFengju, Chen Hui, Wu Qi, Selection and identification of cellulase-producing Bacillus C-36, Journal of Sichuan Agricultural University. 24 (2006)175-177.

[3] Jing Yinghui, Liu Xiangmei, Chen Shicheng, Screening of an alkali-tolerant β-glycanases producing strain and its optimization of their enzyme production, Chin. J. Appl. Environ. Biol. 5 (1999) 404-410.

[4] WU Ruiqing, DU Junqi, LI Ming, Deinking technology for mixed wastepaper with highly efficient deinking agent, Modern Chemical Industry. 30 (2010) 56-60.

[5] JI Yachao, YANGYaru, WANG Zhonglin, LEI Dengying, WEI Yongqin, ZHANG Xiangming, Pilot test on application of enzymatic deinking agent in ONP deinking line, Paper Chemicals. 22 (2010) 28-30.

[6] Wang Zhonglin, Yang Yaru, Zhang Xiangming, Xie Bifeng, Li Jiang, Lei Yichao, Studies on the Bio-enzymatic deinking of old newsprint, Paper Chemicals. 41 (2010) 58-60.

[7] Wu Qiong, Yuan Lin, Lu Fuping, Shen Minghua, Liu Chunyan, Bai Yuchen, Screening and identification of an alkaline cellulase-producing strain, Biotechnology Bulletin. 9 (2010) 205-209.

[8] Chenyan Zhou, Jianyu Bai, Shanshan Den, Cloning of a xylanase gene from Aspergillu susamii and its expression in Escherichia coli, Bioresource Technology. 99 (2008) 831–838.

DOI: 10.1016/j.biortech.2007.01.035

[9] J Sambrook, DW Russell. Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, New York, (2001).

[10] CSIC-QB, QB2583-2003, China standard press, (2003).