In this paper the diameter of inhibition zone to Staphylococcus aureus was used as an indicator to opti-mize the processing conditions of expression of antibacterial peptide in Pichia pastoris. The compositions of culture medium for strain high expressing antibacterial peptide are obtained by single factor and orthogonal tests, which is 2.0% glycerin, 1.5% ammonium citrate, 2.0% tryptone, 1% yeast extract and 0.25% glutamic acid. The recombinant antibacterial peptides could be expressed and obtained with high level while the pH of the culturing medium was 5.5, the yeast was cultured under 30℃ for 72 hours and 1.5% methanol was supplemented into the medium at in-cubating 24h. The result also showed the antibacterial activity of the expressed antibacterial peptide in Pichia pas-toris to both Gram-positive and negative bacteria.%本实验先以抗菌肽对金色葡萄球菌抑菌圈大小为指标,通过单因素和正交试验,对毕赤酵母基因工程菌发酵生产抗菌肽培养基配方和工艺进行了优化,获得工程菌株在2.0%甘油(v/v),1.5%柠檬酸铵,2.0%蛋白胨,1%酵母膏和0.25%谷氨酸组成的培养基中,30℃发酵培养24h补加1.5%甲醇(v/v),培养时间72h,抗菌肽抗菌效果最好,抗菌活性比优化前提高56.8%.最后用革兰阴性大肠杆菌和革兰阳性金色葡萄球菌验证了毕赤酵母基因工程菌发酵生产抗菌肽优化工艺参数的有效性.研究结果表明通过优化发酵工艺,可以提高工程抗菌肽的抗菌活性,同时也为该基因工程菌进一步工业化生产提供了重要的发酵工艺参数.
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