Construction of High Efficient Expression System of the β-mannanase Gene from Erwinia carotovora CXJZ95-198

doi:10.11924/j.issn.1000-6850.casb14110094

Chinese Agricultural Science Bulletin ›› 2015, Vol. 31 ›› Issue (10): 240-245.doi: 10.11924/j.issn.1000-6850.casb14110094

Special Issue: 生物技术

Previous Articles Next Articles

Construction of High Efficient Expression System of the β-mannanase Gene from Erwinia carotovora CXJZ95-198

Cheng Lifeng, Feng Xiangyuan, Duan Shengwen, Zheng Ke, Liu Zhengchu

(Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205)

Received:2014-11-17 Revised:2015-03-06 Accepted:2015-01-23 Online:2015-05-05 Published:2015-05-05

Abstract

Abstract: High efficient expression system of the β-mannanase gene was built. Recombinant plasmid slp-man-pET28a was extracted from the genetic engineering strain slp-man-pET28a/JM, and then transformed to E. coli BL21 (DE3). β-mannanase of the new genetic engineering strain was tested by comparing hydrolysis circle, determination of β-mannanase activity and degradation effect of the natural hemi-cellulose. New genetic engineering strain slp-man-pET28a/BL produced significant hydrolysis circles in selective medium, and secreted β-mannanase up to 645.5 U/mL, as 1.28 times and 1.97 times as that of genetic engineering strain slp-man-pET28a/JM and original strains E. carotovora CXJZ95-198, respectively. COD value of the fermented liquid in ramie raw material by the new genetic engineering strain increased by 5.13 g/L, as 1.5 times and 1.2 times as that of slp-man-pET28a/JM and E. carotovora CXJZ95-198, respectively. Reducing sugar of the fermented liquid in ramie raw material by new genetic engineering strain was 0.721 mg/mL, higher than that of slp-man-pET28a/JM strain and E. carotovora CXJZ95-198 strain by 26.7% and 10.1%, respectively. β-mannanase activity is increased greatly by the construction of high efficient expression system of the new β-mannanase gene, and it may provide scientific basis for preparation used for bast fiber bio-degumming.

Cheng Lifeng,Feng Xiangyuan,Duan Shengwen,Zheng Ke and Liu Zhengchu. Construction of High Efficient Expression System of the β-mannanase Gene from Erwinia carotovora CXJZ95-198[J]. Chinese Agricultural Science Bulletin, 2015, 31(10): 240-245.

References

[1] Chauhan P S, Puri N, Sharma P, et al. Mannanases: microbial sources, production, properties and potential biotechnological applications[J]. Applied Microbiology and Biotechnology, 2012(93): 1817-1830.
[2] Wang M, You S, Zhang S, et al. Purification, characterization, and production of β-mannanase from Bacillus subtilis TJ-102 and its application in gluco-mannooligosaccharides preparation[J]. European Food Research and Technology, 2013(237): 399-408.
[3] 张敏,江正强,李里特.重组海栖热袍菌极耐热甘露聚糖酶的纯化和性质研究[J].微生物学通报,2008,35(10):1565-1571.
[4] Van Zyl W H, Rose S H, Trollope K, et al. Fungal β-mannanases: mannan hydrolysis, heterologous production and biotechnological applications[J]. Process Biochemistry, 2010(45): 1203-1213.
[5] Liu Z H, Qi W, He Z M. Optimization of β-mannanase production from Bacillus licheniformis TJ-101 using response surface Methodology[J]. Chemical and Biochemical Engineering Quarterly, 2008, 22(3):355-362.
[6] 何艳喜,宋影,曹红,等.Armillariella tabescens EJLY2098 β-甘露聚糖酶atMAN47的纯化及酶学性质分析[J].微生物学通报,2009,36(10):1455-1459.
[7] Bhoria P, Singh G, Hoondal G S. Optimization of mannanase production from Streptomyces sp. PG-08-03 in submerged fermentation[J]. Bioresources, 2009, 4: 1130-1138.
[8] 吴襟,段子渊,高启禹,等.诺卡氏菌形放线菌β-甘露聚糖酶的水解特性[J].微生物学通报,2006,33(4):31-33.
[9] Yang P L, Li Y N, Wang Y R, et al. A novel β-mannanase with high specific activity from Bacillus circulans CGMCC1554: gene cloning, expression and enzymatic characterization[J]. Applied Biochemistry and Biotechnology, 2009(159): 85-94.
[10] Duruksu G, Ozturk B, Biely P, et al. Cloning, expression and characterization of endo-beta-1, 4-mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris[J]. Biotechnology Progress, 2009(25):271-276.
[11] Bai W Q, Xue Y F, Zhou C, et al. Cloning, expression and characterization of a novel salt-tolerant xylanase from Bacillus sp. SN5[J]. Biotechnology Letters, 2012, 34: 2093-2099.
[12] Wang J, Zeng D S, Liu G, et al. Truncation of a mannanase from Trichoderma harzianum improves its enzymatic properties andexpression efficiency in?Trichoderma reesei[J]. Journal of Industrial Microbiology & Biotechnology, 2014, 41: 125-133.
[13] Zhang Y X, Liu Z C, Chen X B. Cloning and expression of a mannanase gene from Erwinia carotovora CXJZ95-198[J]. Annals of Microbiology, 2007, 57: 623-628.
[14] 李斌.基因串联与β-甘露聚糖酶基因高效表达体系构建[D].北京:中国农业科学院,2009.
[15] Akino T, Nakamura N, Horikoshi K. Production of β-mannosidase and β-mannanase by an alkalophilic Bacillus sp. [J]. Applied?Microbiology and Biotechnology, 1987(26): 323-327.
[16] Laemmli U K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4[J]. Nature, 1970(277): 680-685.
[17] 邱振华,石鹏君,刘素纯,等.来源于链霉菌Streptomyces fradiae var k11的抗蛋白酶甘露聚糖酶的基因克隆与鉴定[J].中国农业科技导报,2010,12(4):114-120.
[18] 刘小丹,杨培龙,刘永超,等.一种来源于Paenibacillus sp. A1的中性β-甘露聚糖酶基因克隆及其酶学性质研究[J].中国农业科技导报,2009,11(5):60-65.
[19] 乔宇,陈小兵,丁宏标,等.甘露聚糖酶基因在毕赤酵母中的表达及酶学性质研究[J].中国生物工程杂志,2006,26(7):52-56.
[20] 刘项羽,徐美娟,杨套伟,等.β-甘露聚糖酶基因在枯草芽孢杆菌中的克隆及表达[J].应用与环境生物学报,2012,18(4):672-677.
[21] 田雪,解鑫,周晓杭,等.地衣芽孢杆菌产β-甘露聚糖酶分批发酵动力学模型的建立[J].中国农学通报,2013,29(18):193-199.
[22] 谢春芳,黎玉凤,刘大岭,等.利用N-糖基化修饰对β-甘露聚糖酶Man47的稳定性改造[J].中国生物工程杂志,2013,33(12):79-85.

Construction of High Efficient Expression System of the β-mannanase Gene from Erwinia carotovora CXJZ95-198

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 0

Recommended Articles

Metrics

Comments

For authors

For reviewers

Reader center

Contact us