Solubility Analysis, Cloning and Functional Overexpression of the Lipase from Aneurinibacillus thermoaerophilus strain HZ, the First Member of True Lipases Subfamily I.9


如何引用文章

全文:

开放存取 开放存取
受限制的访问 ##reader.subscriptionAccessGranted##
受限制的访问 订阅存取

详细

Lipase from Aneurinibacillus thermoaerophilus strain HZ (HZ lipase) represents the first member of subfamily I.9 true lipases. The resultant of unique characteristics and structural features of HZ lipase has affirmed that subfamily I.9 is located between mesophilic and thermostable lipases. In advance to clone and express the HZ lipase gene, protein solubility of fusion HZ lipase was predicted and analyzed using different software. Then, to overexpress the target gene, high-level expression was performed in a prokaryotic system using different strains and vectors, and production conditions were optimized. HZ lipase was expressed under the control of strong and chemically inducible T7 promoter for high-level expression. It was fused to Trx-, His- and S-tags to solubilize the protein and also to specify and accelerate the purification procedure. The high amount of the HZ lipase protein was obtained as the soluble form (72.5 U/mL) using IPTG final concentration of 0.025 mM after 8 h induction at 30ºC. The expression was analyzed by SDS-PAGE and presence of His-tag was confirmed by Western blotting of protein. As the HZ lipase is the only member of subfamily I.9 that yet cloned and overexpressed, this procedure could be applied to the other close members.

作者简介

M. Masomian

Enzyme and Microbial Technology Research Center; Faculty of Biotechnology and Biomolecular Science

Email: rnzaliha@upm.edu.my
马来西亚, Serdang, Selangor; Serdang, Selangor

R. Rahman

Enzyme and Microbial Technology Research Center; Faculty of Biotechnology and Biomolecular Science

编辑信件的主要联系方式.
Email: rnzaliha@upm.edu.my
马来西亚, Serdang, Selangor; Serdang, Selangor

A. Salleh

Enzyme and Microbial Technology Research Center; Faculty of Biotechnology and Biomolecular Science

Email: rnzaliha@upm.edu.my
马来西亚, Serdang, Selangor; Serdang, Selangor

M. Basri

Enzyme and Microbial Technology Research Center; Faculty of Science

Email: rnzaliha@upm.edu.my
马来西亚, Serdang, Selangor; Serdang, Selangor

补充文件

附件文件
动作
1. JATS XML

版权所有 © Pleiades Publishing, Inc., 2018