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  

Effect of co-expression of chaperones on Rhizopus oryzae recombinant lipase production in Pichia pastoris

作　　者：卢鑫[1] 喻晓蔚[1] 沙冲[1] 范文来[1] 徐岩[1] 

机构地区：[1]工业生物技术教育部重点实验室江南大学酿酒微生物与酶技术研究室,江苏无锡214122

出　　处：《微生物学通报》2013年第3期407-413,共7页Microbiology China

基　　金：国家973计划项目(No.2011CB710800);国家863计划项目(No.2012AA022207;2010AA101501);中央高校基本科研业务费专项资金(No.JUSRP11014);国家自然科学基金项目(No.20802027)

摘　　要：【目的】通过共表达伴侣蛋白Ero1p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7 L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2 338.7 U/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20 h。【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。[Objective] The aim of this study is to improve the yield of Rhizopus oryzae lipase http：//journals.im.ac.cn/wswxtbcn 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 Ero1p 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 gmethanol/（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 Ero1p and PDI,which provided the potential application of Rhizopus oryzae lipase in industry.

关 键 词：米根霉脂肪酶 高效表达 毕赤酵母 高密度发酵 共表达伴侣蛋白 

分 类 号：TQ925[轻工技术与工程—发酵工程]