Optimization of Expression Conditions of Zea mays Transglutaminase Gene in Pichia pastoris
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(College of Food Science, Northeast Agricultural University, Harbin 150030)

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    Abstract:

    In order to determine the optimal inducible expression conditions of zea mays transglutaminase gene in Pichia pastoris, pPIC9K-TGZ was introduced into the component P. pastoris GS115 strains by the electroporation method, screening positive clones with antibiotics, PCR amplification verification, IPTG induced expression, optimize the best expression conditions. In this study, the optimal expression conditions of engineering bacteria were determined by orthogonal experiment. The expression medium was BMMY and the induced expression time was 96 h, initial expression of pH 6.0, the initial induction OD600nm of the bacteria was 4.0; the induction temperature was 24 ℃; the methanol content was 0.6% (volume fraction); methanol flow was added once/ 12 h; glycerol of 1 g/L was added every 24 hours after 48 h induction. The maximum enzyme activity of medium supernatant under optimized conditions was 7.51 U/mL.

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  • Received:April 18,2021
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  • Online: May 26,2022
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