Expression of linoleate isomerase gene from Lactobacillus reuteri PYR8 in Pichia pastoris
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 de...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2006-09-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/1008-9209.2006.05.0505 |
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| Summary: | 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·mg<sup>-1</sup>. |
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| ISSN: | 1008-9209 2097-5155 |