毕赤酵母X33/pGAPZαA-MLcc1产漆酶的高密度发酵
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Study of High Cell-Density Fermentation of Laccase Produced by Pichia pastoris X33/pGAPZαA-MLcc1
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    摘要:

    在5 L的发酵罐中进行了重组毕赤酵母X33/pGAPZα A-MLcc1表达重组漆酶的发酵试验,漆酶活性的变化趋势与菌体干质量变化趋势相一致,当菌体增长进入稳定期46 h左右时,产酶达至高峰47.2 U/L。进一步研究了工程菌在5 L容积发酵罐中的高密度发酵工艺,并获得了高水平表达漆酶的优化策略:在补料阶段采用溶氧反馈控制方式,并且保持溶氧水平在15~30 g/dL,发酵过程中通过在线补加氨水和盐酸来调整pH恒定在5。通过高密度发酵可获得最高酶活力达216.3 U/L,是分批发酵获得最高酶活力的4.6倍。

    Abstract:

    The expression of laccase gene by Pichia pastoris X33/pGAPZα A-MLcc1 was investigated in a 5 L fermenter. The change trend of laccase activity was consistent with the change trend dry cell weight. The extracellular laccase activity reached the highest value of 47.2 U/L when the yeast X33 entered the stationary phage at 46 h. High-density fermentation in a 5 L fermenter was further investigated and the optimized method for high-level expression was obtained as follows:by adopting dissolved oxygen(DO) feed-back feeding strategy and maintaining DO at 15% to 30% at feeding stage,as well as adding ammonia solution and hydrochloride to maintain pH value at 5 in the fermentation stage,the laccase activity reached 216.3 U/L,which was 4.6 times higher than that of fed batch fermentation.

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黄亮,刘逸寒,刘晓光,李玉,路福平.毕赤酵母X33/pGAPZαA-MLcc1产漆酶的高密度发酵[J].食品与生物技术学报,2016,35(3):324-329.

HUANG Liang, LIU Yihan, LIU Xiaoguang, LI Yu, LU Fuping. Study of High Cell-Density Fermentation of Laccase Produced by Pichia pastoris X33/pGAPZαA-MLcc1[J]. Journal of Food Science and Biotechnology,2016,35(3):324-329.

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  • 在线发布日期: 2016-11-01
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