Optimization of expression and fermentation of β-1,4-galactosyltransferase gene from dairy cow in Pichia pastoris GS115
Reeombinant plasmid pPIC9K-GT was transformed into P. pastoris GS115 by electroporation. In order to achieve a high transformation efficiency, the electroporation conditions were optimized. The positive transformant was obtained after phenotype screening and PCR identification. Inducible expression of the reeombinant P. pastoris GS115-GT was conducted. Target protein detected by SDS-PAGE verified β-1,4-galactosyltransferase gene was expressed solubly in P. pastoris GS115. The specific activity of β-1,4-galactosyltransferase after optimization of fermentation conditions for genetically engineered microorganim was 30.909 U/ml increased by 88.47% compared to that before optimization, which was only 16.40 U/ml.
NETL
NSTL
万方数据
维普
