Cloning, Expression and Characterization of β-glucosidase from Stachybotrys chartarum

Abstract

Abstract: To express β-glucosidase from Stachybotrys chartarum using Aspergillus niger G1 as host. Through genome sequencing and analysis, the β-glucosidase gene(g10158)was isolated from the genome of Stachybotrys chartarum through PCR. The coding region of the gene was inserted to the vector pGm. The expression plasmid was transformed into A. niger G1 strain. An β-glucosidase producing strain(G1-pGm-g10158-13)were selected after screening several transformants using amdS selection plates and confirmed by PCR validation. Results showed that the molecular mass of the enzyme was 87.9 kD by SDS-PAGE gel analysis. Our results indicated that the recombinant enzyme exhibited optimum activity at pH4.8 and 50℃. The β-glucosidase activity was 1 856.48 U/mL under the optimized conditions. The β-glucosidase from Stachybotrys chartarum was expressed in A. niger G1 strain and it was proved to have a high activity.

Key words: β-Glucosidase, Stachybotrys chartarum, Aspergillus niger, G1 Prokaryotic expression

Yu Hailing, Li Shuwei, Wang Huaming. Cloning, Expression and Characterization of β-glucosidase from Stachybotrys chartarum[J]. Biotechnology Bulletin, 2013, 0(6): 128-132.

References

[1] 陈卫红, 李多川.嗜热子囊菌光孢变种β-葡萄糖苷酶的基因克隆、表达及酶学性质分析[J].应用与环境生物学报, 2009, 15(4):549-553.
[2] 陈水莲.深海细菌Exiguobacterium oxidotolerans A011葡萄糖苷酶基因的克隆、表达与酶学分析[D].武汉:华中农业大学, 2011.
[3] 李远华. β-葡萄糖苷酶的研究进展[J].安徽农业大学学报, 2002, 29(4):421-425.
[4] 韩笑, 陈介南, 王义强, 等. β-葡萄糖苷酶基因的克隆与表达研究进展[J].生物技术通报, 2008(3):8-13.
[5] Bhatia Y, Mishra S, Bisaria VS. Microbial beta-glucosidases:cloning, properties, and applications [J]. Crit Rev Biotechnol, 2002, 22(4):375-407.
[6] 宛晓春.水果风味及风味酶的研究[D].无锡:无锡轻工大学, 1992.
[7] Mandels M. 发酵法生产纤维素酶的某些问题和解决办法[J].应用微生物, 1976(6):53-62.
[8] 顾方媛, 陈朝银, 石家骥.纤维素酶的研究进展与发展趋势[J].微生物学杂志, 2008, 28(1):83-87.
[9] Zhou X, Smith JA, Oi FM. Correlation of cellulase gene expression and celluLolytic activity throughout the gut of the termite Reticuliter-mes flavipes [J]. Gene, 2007, 395(1-2):29-39.
[10] 陈阿娜, 汤斌.纤维素酶高产菌株选育研究进展[J].安徽农学通报, 2006, 12(10):64.
[11] Auer N, Hedger J, Evans C. Degradation of nitrocelluLose by fungi [J]. Biodegradation, 2005, 16(3):229-236.
[12] Rabinovich ML, Melnik MS, Bolobova AV. Microbial celluLases [J]. Applied Biochemistry and Microbiology, 2002, 38(4):305.321.
[13] Piontek M, Hagedom J, Hollenberg CP, et al. Two novel gene expr-ession systems based on the yeasts Schwanniomyces occidentalis and Pichai stipitis [J]. Applied Microbiol Biotechnol, 1998, 50(3):331-338.
[14] 孟宪文, 宋小红, 陈历俊. β-葡萄糖苷酶的研究进展[J].中国乳业, 2009(10):42-44.
[15] Amouri B, Gargouri A. Characterization of a novel β-glucosidase fr-om a Stachybotrys strain [J]. Biochem Eng J, 2006, 32:191-197.
[16] Saibi W, Amouri B, Gargouri A. Purifcation and biochemical chara-cterization of a transglucosilating β-glucosidase of Stachybotrys strain [J]. Appl Microbiol Biotechnol, 2007, 77(2):293-300.
[17] 赵颖, 巴再华, 张春艳.双向选择标记amdS在黑曲霉中自发突变及amdS-检测[J].济宁医学院学报, 2008, 31(4):283-285.