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HERO ID
1228911
Reference Type
Journal Article
Title
Expression of linoleate isomerase gene from Lactobacillus reuteri PYR8 in Pichia pastoris
Author(s)
Zhang, YH; Zhang, LW; Hu, S; Bu, ZG
Year
2006
Is Peer Reviewed?
0
Journal
Zhejiang Daxue Xuebao (Nongye yu Shengming Kexue Ban)
ISSN:
1008-9209
Volume
32
Issue
5 (Sep 2006)
Page Numbers
505-510
Abstract
Linoleate isomerase gene was amplified by PCR from chromosome of Lactobacillus reuteri PYR8, then the gene was cloned into Pichia pastoris expression vector pPIC9K. The recombinant plasmid pPIC9K-LI was transformed into P. pastoris GS115 by electroporation and was induced with 1% methanol, it was demonstrated by SDS-PAGE that a 67 kD protein which was equall to LI in molecular weight was expressed in supernatant culture. The recombined LI protein with purity up to 95% was finally obtained after purification through two-step chromatography: SP Sepharose Fast Flow and Phenyl Sepharose Fast Flow. The analysis of gas chromatography showed that the recombinant LI protein exhibited the specific activity of conveting linoleic acid to conjugated linoleic acid, which was 6.8 U super(.) mg super(-1).
Keywords
Linoleic acid; Gas chromatography; Chromosomes; Linoleate isomerase; Electroporation; Plasmids; protein purification; Methanol; Lactobacillus reuteri; Pichia pastoris; Polymerase chain reaction; Substance P; Expression vectors; Molecular weight
Tags
IRIS
•
Methanol (Non-Cancer)
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