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Characterization of a new Cu/Zn-superoxide dismutase from Beauveria bassiana and two site-directed mutations crucial to its antioxidation activity without chaperon

Paper ID Volume ID Publish Year Pages File Format Full-Text
17632 42686 2010 6 PDF Available
Title
Characterization of a new Cu/Zn-superoxide dismutase from Beauveria bassiana and two site-directed mutations crucial to its antioxidation activity without chaperon
Abstract

A Cu/Zn-superoxide dismutase (SOD) was characterized for the first time from Beauveria bassiana by gene cloning, heterogeneous expression and function analysis. This 154-aa SOD (BbSod1) was deduced from a 465-bp gene cloned, showing 49–96% sequence identity to Cu/Zn-SODs from other 57 fungi. BbSod1 and its form engineered with two site-directed mutations P143S and P145L (BbSod1-Mut) or a fused copper chaperon Lys7 (BbSod1-Lys7) were expressed well in Escherichia coli. Crude extracts and purified BbSod1-Mut from cell cultures exhibited much higher antioxidation activities than the counterparts of BbSod1-Lys7 whereas BbSod1 showed no substantial activity. The engineered enzymes were best induced by overnight incubation at 20 °C in Luria-Bertani medium including 2.5 mM Cu2+, 0.5 mM Zn2+ and 0.5 mM isopropyl-d-thiogalactopyranoside after 5-h growth to log-phase at 37 °C. Our results highlight alternative means to producing highly active fungal Cu/Zn-SOD in E. coli by making use of the two site-directed mutations without chaperon.

Keywords
Superoxide dismutase; Beauveria bassiana; Site-directed mutation; Antioxidation activity enhancement; Copper chaperon
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Characterization of a new Cu/Zn-superoxide dismutase from Beauveria bassiana and two site-directed mutations crucial to its antioxidation activity without chaperon
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 46, Issues 3–4, 5 March 2010, Pages 217–222
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us