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  

Gene Cloning and Characterization of a Protease-resistant β-mannanase Rooted in Streptomyces fradiae var.k11

作　　者：邱振华[1,2] 石鹏君[2] 刘素纯[1] 姚斌[2] 

机构地区：[1]湖南农业大学食品科技学院,湖南省食品科学与生物技术重点实验室,长沙410128 [2]中国农业科学院饲料研究所,农业部饲料生物技术重点开放实验室,北京100081

出　　处：《中国农业科技导报》2010年第4期114-120,共7页Journal of Agricultural Science and Technology

基　　金：国家转基因生物新品种培育重大专项(2009ZX08003-020B);国家肉鸡产业技术体系(nycytx-42-G2-05)资助

摘　　要：通过设计简并引物和TAIL-PCR的方法从Streptomyces fradiaevar.k11中克隆得到一个β-l,4-甘露聚糖酶编码基因manS221,其全长1 359 bp,编码452个氨基酸,前端28个氨基酸为预测信号肽。构建表达载体pET-28a-manS221并转化大肠杆菌BL2l(DE3),特异性表达manS221基因,重组的甘露聚糖酶通过Ni-NTA亲和层析纯化。酶学性质分析表明,重组甘露聚糖酶最适温度为52℃,最适pH值为6.0,在pH5.0～9.0的范围内有良好的稳定性,并且对中性蛋白酶具有高抗性。这些优良的性质使得ManS221在食品、饲料和纺织工业生产中具有良好的应用前景。The gene manS221 encoding a β-l,4-mananase was cloned from Streptomyces fradiae var.k11 by degenerate primers and TAIL-PCR technology.manS221 gene contains 1 359 bp,encoding a 452 amino acid peptide consisting of a putative N-terminal signal peptide of 28 residues.The expression vector pET-28a-manS221 was constructed and transformed into Eschetichia coli BL21.The expressed ManS221 was purified by Ni-NTA chromatography.The optimum temperature and pH of the recombinant mannose were 52℃ and 6.0,respectively.The enzyme showed stability over a pH range of 5.0～9.0,and resistance to neutral protease.These favorable properties would make ManS221 as a good candidate in various industrial applications.

关 键 词：链霉菌Streptomyces fradiae var.k11 甘露聚糖酶 manS221基因 蛋白酶抗性 

分 类 号：Q786[生物学—分子生物学]