|本期目录/Table of Contents|

[1]黄敏,徐辉,曹瑜,等.大肠杆菌植酸酶的原核表达及酶学性质[J].应用与环境生物学报,2009,15(03):371-375.[doi:10.3724/SP.J.1145.2009.00371]
 HUANG Min,XU Hui,CAO Yu,et al.Characteristics of Phytase from Escherichia coli[J].Chinese Journal of Applied & Environmental Biology,2009,15(03):371-375.[doi:10.3724/SP.J.1145.2009.00371]
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大肠杆菌植酸酶的原核表达及酶学性质()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
15卷
期数:
2009年03期
页码:
371-375
栏目:
研究论文
出版日期:
2009-05-15

文章信息/Info

Title:
Characteristics of Phytase from Escherichia coli
作者:
黄敏徐辉曹瑜朱俊张飞伟尤芳芳乔代蓉曹毅
1四川大学生命科学学院 成都 610065
2四川大学微生物与代谢工程四川省重点实验室 成都 610065
3四川阳光博睿生物技术有限公司 成都 610016
Author(s):
HUANG MinXU HuiCAO YuZHU JunZHANG FeiweiYOU FangfangQIAO DairongCAO Yi
1College of Life Sciences, Sichuan University, Chengdu 610065, China
2Sichuan Public Exponential Platform of Bioinformatics and Metabolic Engineering, Sichuan University, Chengdu 610065, China
3Sichuan Bioshine Life Technology Co. Ltd., Chengdu 610016, China
关键词:
appA植酸酶原核表达亲和纯化酶学性质
Keywords:
appA-encoded phytase prokaryotic expressionnickel-chelating sepharose chromatography enzymatic characterization
分类号:
Q556.9
DOI:
10.3724/SP.J.1145.2009.00371
文献标志码:
A
摘要:
摘 要 将大肠杆菌植酸酶appA基因重组于表达载体pET32a中,导入大肠杆菌Origami (DE3)构建工程菌 Origami (DE3)-pET32a-appA. 对表达的appA重组植酸酶经Ni柱亲和纯化、SDS-PAGE分析表明,纯化后条带单一,酶的纯度较高. 对其酶学性质的研究表明,该酶反应的比活力为3.36×106 U/mg;最适pH为4.5;最适温度为60 ℃;在80 ℃和85 ℃的耐干热能力超过耐湿热能力;在37 ℃下,Mg2+﹑Ca2+﹑Mn2+ 对酶活性有促进作用,Co2+﹑Cu2+﹑K+对酶活性有不同程度的抑制作用,而Fe3+ 和Zn2+对酶活性有强烈的抑制作用;此外,该酶还具有非常好的抗胃蛋白酶水解的能力和部分抗胰蛋白酶水解的能力. 图8 表1 参21
Abstract:
Abstract Gene appA was cloned into prokaryotic expression vector pET32a to construct pET32a-appA and gene appA was successfully expressed in Escherichia coli Origami (DE3). Recombinant phytase was purified and its enzymatic characterization was studied. The results of SDS-PAGE showed higher purity of the enzyme after purified by nickel-chelating sepharose chromatography. Its specific activity was 3.36×106 U/mg, and optimal pH and temperature for its activity were 4.5 and 60 ℃, respectively. The dry thermal stability of phytase was stronger than wet thermal stability at 80 ℃ and 85 ℃. The enzyme activity was activated by Mg2+, Ca2+ and Mn2+ , and inhibited by Co2+, Cu2+ and K+. However, it almost lost activity when influenced by Fe3+ and Zn2+. In addition, it had strong ability to resist hydrolyzation by pepsin and partial ability to resist hydrolyzation by trypsin. Fig 8, Tab 1, Ref 21

参考文献/References:

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备注/Memo

备注/Memo:
*四川省重点项目(No. 07GG111-03)资助 Supported by the Key Project of the Department of Science and Technology of Sichuan, China (No. 07GG111-03)
**通讯作者 Corresponding author (E-mail: caoyi_01@163.com)
更新日期/Last Update: 2009-07-03