Expression, purification and characterization of pectate lyase A from Aspergillus nidulans in Escherichia coli
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- 作者：Qingxin Zhao ; Sheng Yuan ; Yuling Zhang ; Hong Zhu ; Chuanchao Dai ; Fang Yang and Fengmin Han
- 关键词：Aspergillus nidulans ; Pectate lyase A ; Characterization ; Expression ; Purification
- 刊名：World Journal of Microbiology and Biotechnology
- 出版年：2007
- 出版时间：August, 2007
- 年：2007
- 卷：23
- 期：8
- 页码：1057-1064
- 全文大小：316 KB
- 刊物类别：Chemistry and Materials Science
- 刊物主题：Chemistry
Applied Microbiology
Biotechnology
Biochemistry
Environmental Biotechnology
Microbiology - 出版者：Springer Netherlands
- ISSN：1573-0972
文摘
Pectate lyase A (PelA) of Aspergillus nidulans was successfully expressed in Escherichia coli and effectively purified using a Ni2+-nitrilotriacetate-agarose column. Enzyme activity of the recombinant PelA could reach 360 U ml?1 medium. The expressed PelA exhibited its optimum level of activity over the range of pH 7.5–10 at 50°C. Mn2+, Ca2+, Fe2+, Mg2+ and Fe3+ ions stimulated the pectate lyase activity, but Cu2+ and Zn2+ inhibited it. The recombinant PelA had a V max of 77 μmol min?1 mg?1 and an apparent K m of 0.50 mg ml?1 for polygalacturonic acid. Low-esterified pectin was the optimum substrate for the PelA, whereas higher-esterified pectin was hardly cleaved by it. PelA efficiently macerated mung bean hypocotyls and potato tuber tissues into single cells.