共表达伴侣蛋白对重组毕赤酵母产米根霉(Rhizopus oryzae)脂肪酶发酵过程的影响

摘要

[Objective] The aim of this study is to improve the yield of Rhizopus oryzae lipase in Pichia pastoris by co-expression of chaperones. [Methods] By high-density fermentation in 7 L bioreactor, the expression of lipase in strain BH128 co-expressed with two chaperones Erolp and PDI was compared with that of H238 co-expressed without any chaperone. [Results] The results showed that the highest lipase activity, the specific growth rate, the specific production rate and the specific consumption rate in BH128 reached 2 388.7 U/mL, 0.02 h-1, 944.5 U/(gDCW·h) and 0.15 gmethanoi/(gDCW·h), which is 1.7, 0.5, 4.1 and 1.3 fold higher than those in H238, respectively. Moreover, the fermentation period in BH128 was 20 hours shorter than in H238. [Conclusion] The high-level expression of Rhizopus oryzae lipase could be achieved by co-expression of chaperones Erolp and PDI, which provided the potential application of Rhizopus oryzae lipase in industry.%[目的]通过共表达伴侣蛋白Erolp和PDI获得米根霉(Rhizopus oryzae)脂肪酶在毕赤酵母中的高效表达.[方法]利用毕赤酵母基因重组菌高密度发酵的方法,在7L发酵罐水平上分析共表达伴侣蛋白菌株(BH128)与非共表达伴侣蛋白菌株(H238)对脂肪酶表达的差异.[结果]在相同条件下,BH128发酵产酶能力高于H238,最高酶活可达到2 338.7U/mL,最大比生长速率达到0.02 h-1,最大产物比形成速率达到944.5 U/(gDCW·h),最大底物比消耗速率也能达到0.15 gmethanol/(gDCW·h),分别是H238的1.7、0.5、4.1和1.3倍,且发酵周期缩短了20h.[结论]毕赤酵母基因重组菌BH128通过共表达伴侣蛋白Ero1p和PDI,提高了米根霉脂肪酶的产量,而且缩短了发酵周期,为工业化生产奠定了基础.