期刊论文

中文

中国农业科技导报

1008-0864

2010

12

4

114-120

10.3969/j.issn.1008-0864.2010.04.21

2009ZX08003-020B:转基因生物新品种培育科技重大专项 nycytx-42-G2-05:国家肉鸡产业技术体系项目

本校为第一机构

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 ...