枯草芽孢杆菌WD-23 β-甘露聚糖酶的纯化及其酶学性质

doi:10.3969/j.issn.1000-2006.2014.03.017

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

南京林业大学学报（自然科学版） ›› 2014, Vol. 38 ›› Issue (03): 88-92.doi: 10.3969/j.issn.1000-2006.2014.03.017

枯草芽孢杆菌WD-23 β-甘露聚糖酶的纯化及其酶学性质

张冰,向冰冰,崔岱宗,赵敏^*

东北林业大学生命科学学院,黑龙江哈尔滨 150040

出版日期:2014-05-15 发布日期:2014-05-15
基金资助:
收稿日期:2013-10-26 修回日期:2014-03-06
基金项目:哈尔滨市科技局攻关项目(2008AA6B6BN025,GJ2011GG002112)
第一作者:张冰,硕士生; 向冰冰,硕士生。*通信作者:赵敏,教授。E-mail:82191513@163.com。
引文格式:张冰,向冰冰,崔岱宗,等. 枯草芽孢杆菌WD-23 β-甘露聚糖酶的纯化及其酶学性质[J]. 南京林业大学学报:自然科学版,2014,38(3):88-92.

Inrestigation on the purification methods of β-mannase from Bacillus subtilis WD-23 and studies on enzymatic properties

ZHANG Bing, XIANG Bingbing, CUI Daizong, ZHAO Min^*

College of Life Science, Northeast Forestry University, Harbin 150040, China

Online:2014-05-15 Published:2014-05-15

摘要/Abstract

摘要： 利用硫酸铵盐析缓冲液透析、聚乙二醇浓缩、DEAE-Sepharose FF窝子交换层析等方法,纯化了枯草芽孢杆菌WD-23 β-甘露聚糖酶,并研究了其酶学性质。结果表明:纯化的枯草芽孢杆菌WD-23 β-甘露聚糖酶的分子质量约为40 ku,酶的纯化倍数为14.1倍; 最适反应pH和pH稳定范围分别为5.6和5.0～7.0; 最适反应温度和温度稳定范围分别为55 ℃和40～70 ℃; Ca²⁺对该酶的促进作用最为明显,Li⁺的抑制作用最明显。

Abstract: The objective of this study was to purify β-mannase from Bacillus subtilis WD-23 and examine its enzymology properties. This study used the combining methods of ammonium sulfate, semi-permeable membrane dialysis, polyethylene glycol concentration, DEAE-Sepharose FF ion exchange chromatography, Sephadex G-75 gel filtration and SDS-PAGE gel electrophoresis to purify β-mannase. The result showed that the purified ratio of β-mannase of B. subtilis WD-23 was 14.1 by using the above methods, and the molecular mass was about 40 ku, the optimal pH and the stability range of pH were 5.6 and 5.0-7.0 respectively, and optimal temperature and the stability range of temperature were 55 ℃ and 40-70 ℃ separately, Ca^{2 +} was the most obvious effect on the activation of the enzyme, on the contrary, the inhibitory effect of Li⁺ was the most obvious.

中图分类号:

Q936

张冰,向冰冰,崔岱宗,等. 枯草芽孢杆菌WD-23 β-甘露聚糖酶的纯化及其酶学性质[J]. 南京林业大学学报（自然科学版）, 2014, 38(03): 88-92.

ZHANG Bing, XIANG Bingbing, CUI Daizong, ZHAO Min. Inrestigation on the purification methods of β-mannase from Bacillus subtilis WD-23 and studies on enzymatic properties[J].Journal of Nanjing Forestry University (Natural Science Edition), 2014, 38(03): 88-92.DOI: 10.3969/j.issn.1000-2006.2014.03.017.

参考文献

[1] 王静. β-甘露聚糖酶的制备与分离纯化的研究[D]. 南京:南京林业大学,2012.Wang J. The study on preparation, separation and purification of β-mannase[D]. Nanjing:Nanjing Forestry University, 2012.
[2] 武玉永,于敏,武芝. 产甘露聚糖酶菌株的筛选·鉴定及酶活性的初步研究[J]. 安徽农业科学, 2012, 40(8): 4496-4498.Wu Y Y, Yu M, Wu Z. Preliminary study on selection and identification of a mannanase-producing strain and its enzymatic properties[J]. Anhui Agricultural Sciences, 2012, 40(8): 4496-4498.
[3] Dhawan S, Kaur J. Microbial mannanases: an over-view of production and applications[J]. Critical Reviews in Biotechnology, 2008, 27(4): 197-216.
[4] 胡文波. β-甘露聚糖酶基因的克隆、表达及酶学性质研究[D]. 新乡:河南师范大学,2012.Hu W B. The cloning and expression of β-mannase gene and its recombined enzymatic properties[D]. Xinxiang: Henan Normal University, 2012.
[5] Lu H Q, Zhang H T, Shi P J. A family 5 β-mannanase from the thermophilic fungus Thielavia arenaria XZ7 with typical thermophilic enzyme features[J]. Biotechnologically Relevant Enzymes And Proteins, 2013,97(10): 8121-8128.
[6] 杨阳,张付云,苍桂璐,等. 地衣芽胞杆菌生物活性物质应用研究进展[J]. 生物技术进展, 2013, 3(1): 22-26.Yang Y, Zhang F Y, Cang G L,et al. Research advances in the application of bioactive substances produced by Bacillus licheniformis [J]. Curr Biotech, 2013, 3(1): 22-26.
[7] 高海有,刘正初,段盛文,等. β-甘露聚糖酶和木聚糖酶基因在大肠杆菌中共表达[J]. 微生物学通报, 2012, 39(3):344-352. Gao H Y, Liu Z C, Duan S W,et al. Coexpression of β-mannanase and xylanase genes in Escherichia coli[J]. Journal of Microbiology, 2012, 39(3): 344-352.
[8] Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding[J]. Analytical Biochemistry, 1976, 72(5): 248-254.
[9] Civas A, Eberhard R, Le Dizet P, et al. Glycosidases induced in Aspergillus tamarii secreted alpha-D-galactosidase and beta-D-mannanase[J]. Biochem J, 1984, 219: 857-863.
[10] 陈一平,龙健儿,廖连华,等. 芽孢杆菌M50产生的β-甘露聚糖酶的条件研究[J]. 微生物学报, 2000, 40(1): 62-68.Chen Y P, Long J E, Liao L H,et al. Conditions of β-mannanase produced by Bacillus M50[J]. Acta Microbiologica Sinica, 2000, 40(1): 62-68.
[11] Rihei T, Isal K, Hideyuki K, et al. Purification and some properties of mannanase from Streptomycessr sp.[J]. Agricultural Biochemistry, 1984, 48(9): 2189-2195.
[12] 杨文博,伶树敏,沈庆. 地衣芽抱杆菌β-甘露聚糖酶的纯化及酶学性质[J]. 微生物学通报, 1995, 22(6): 338-342.Yang W B, Ling S M, Shen Q. Purification and properties of β-mannanase from Bacillus licheniformis[J]. Journal of Microbiology, 1995, 22(6): 338-342.
[13] 余红英,孙远明,王炜军,等. 枯草芽抱杆菌SA-22 β-甘露聚糖酶的纯化及其特性[J]. 生物工程学报, 2003, 19(3):327-331.Yu H Y, Sun Y M, Wang W J,et al. Purification and properties of Bacillus subtilis SA-22 Endo-1,4-β-D-mannanase [J]. Chinese Journal of Biotechnology, 2003, 19(3): 327-331.
[14] 吴襟,何秉旺. 诺卡氏菌形放线菌β-甘露聚糖酶的纯化和性质[J]. 微生物学报, 2000, 16(2): 227-230.Wu J, He B W. Purification and properties of nocardia shaped actinomycetes β-mannanase[J]. Acta Microbiologica Sinica, 2000, 16(2): 227-230.
[15] 李小明. 甘露聚糖酶产生菌的筛选及酶学性质研究[D]. 大连:大连理工大学,2012.Li X M. Screening and Enzymatic property of mannanase producing strains[D]. Dalian: Dalian University of Technology,2012.
[16] 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.Liu X J, Xu M J, Yang T W,et al. Cloning and expression of β-mannanase gene in Bacillus subtilis[J]. Chinese Journal of Applied & Environmental Biology, 2012, 18(4): 672-677.

枯草芽孢杆菌WD-23 β-甘露聚糖酶的纯化及其酶学性质

Inrestigation on the purification methods of β-mannase from Bacillus subtilis WD-23 and studies on enzymatic properties

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价

作者

读者

审稿

[1]	樊奔,陈晟,李昱龙. 细菌Hfq蛋白的结构、功能及作用机制[J]. 南京林业大学学报（自然科学版）, 2016, 40(05): 155-162.
[2]	李飞,解静聪,李琦,张雪松,赵林果. 木聚糖酶XynB分子Ser/Thr 平面导入精氨酸对酶热稳定性的影响[J]. 南京林业大学学报（自然科学版）, 2014, 38(04): 107-112.