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Overexpression and Identification of Bioactivity of L.reurei linoleite isomerase in Escherichia coli |
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Abstract The recombinant expression of linoleite isomerase using Escherichia coli BL21as host cell was studied. A pair of primers were designed and synthesized according to the linoleic isomerase(gLI) gene sequence published by Genbank. Linoleic isomerase gene was amplified by PCR from genome of L.reuteriPYR8. Recombinant pET-gLI was constructed after the gLI gene was sequenced. BL21(DE3) was induced with 1.0mmol/L IPTG, SDS-PAGE showed recombinant protein was about 67 KD. After inducted and purified, the gLI can converts linolenic acid to CLA in phosphate buffer(pH7.3) 30℃ activity enzyme 1377U, which gave a foundation for biological activity and machnism。
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Received: 10 April 2006
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