食品科学
• 生物工程 • 上一篇 下一篇
侯进慧,张 翔,乔高翔
出版日期:
发布日期:
HOU Jinhui, ZHANG Xiang, QIAO Gaoxiang
Online:
Published:
摘要: 利用分子克隆的方法,对一种源于菠萝泛菌的内切葡聚糖酶(即β-1,4-内切葡聚糖酶)基因进行克隆、原核表达,利用Ni-NTA吸附柱对表达的重组内切葡聚糖酶进行纯化,分析重组酶的活性。研究结果显示,此重组内切葡聚糖酶含1 个1 002 bp的开放阅读框,编码334 个氨基酸序列,重组酶在大肠杆菌细胞中的表达量占可溶性蛋白的50%以上,经过纯化获得了纯度高于95%的内切葡聚糖酶蛋白,酶活力可以达到2 245 U/mL。
关键词: 内切葡聚糖酶, 原核表达, 活性分析
Abstract: The endo-1,4-β-D-glucanase gene from Pantoea ananatis was cloned and expressed in Escherichia coli. The recombinant enzyme was purified with Ni-NTA affinity chromatography and its activity was analyzed. The results showed that the recombinant glucanase gene contained a 1 002 bp-length open reading frame encoding two putative peptides of 334 amino acids. The expression of the recombinant enzyme in E. coli reached up to 50% of the total soluble protein. After purification, the purity was higher than 95% and glucanase activity was as high as 2 245 U/mL.
Key words: endo-1,4-β-D-glucanase, prokaryotic expression, activity analysis
中图分类号:
Q939.9
侯进慧,张 翔,乔高翔. 菠萝泛菌β-1,4-内切葡聚糖酶基因克隆、表达与酶活性分析[J]. 食品科学, doi: 10.7506/spkx1002-6630-201623035.
HOU Jinhui, ZHANG Xiang, QIAO Gaoxiang. Cloning, Expression and Activity of an Endo-1,4-β-D-Glucanase from Pantoea ananatis[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201623035.
0 / / 推荐
导出引用管理器 EndNote|Ris|BibTeX
链接本文: https://www.spkx.net.cn/CN/10.7506/spkx1002-6630-201623035
https://www.spkx.net.cn/CN/Y2016/V37/I23/211