US EPA

1228911 

Journal Article 

Expression of linoleate isomerase gene from Lactobacillus reuteri PYR8 in Pichia pastoris 

Zhang, YH; Zhang, LW; Hu, S; Bu, ZG 

2006 

0 

Zhejiang Daxue Xuebao (Nongye yu Shengming Kexue Ban)
ISSN: 1008-9209 

32 

5 (Sep 2006) 

505-510 

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). 

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