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Combined use of gap and aox1 promoters and optimization of culture conditions to enhance expression of rhizomucor miehei lipase

文献类型:期刊论文

作者He, Dong1,2; Luo, Wen1; Wang, Zhiyuan1; Lv, Pengmei1; Yuan, Zhenhong1
刊名Journal of industrial microbiology & biotechnology
出版日期2015-08-01
卷号42期号:8页码:1175-1182
关键词Rml Pichia pastoris P-gap P-aox1 Optimization
ISSN号1367-5435
DOI10.1007/s10295-015-1633-6
通讯作者Lv, pengmei(lvpm@ms.giec.ac.cn)
英文摘要Rhizo mucor miehei lipase (rml) is an industrially important enzyme, but its application is limited due to its high cost. in this study, a series of measures such as codon optimization, propeptide addition, combined use of gap and aox1 promoters, and optimization of culture conditions were employed to increase the expression of rml. three transformants of the constitutive-inducible combined pichia pastoris strains were generated by transforming the pgapz alpha a-rml vector into the ppic9k-rml/gs115 strain, which resulted in high-expression yields of rml. using the shake flask method, highest enzyme activity corresponding to 140 u/ml was observed in the strain 3-17, which was about sixfold higher than that of ppic9k-rml/gs115 or pgapz alpha a-rml/gs115. after optimization of culture conditions by response surface methodology, the lipolytic activity of strain 3-17 reached 175 u/ml in shake flasks. an increase in the copy number simultaneously with the synergistic effect provided by two promoters led to enhanced degree of protein expression.
WOS关键词PICHIA-PASTORIS ; RECOMBINANT PROTEINS ; GENE-EXPRESSION ; COPY NUMBER ; IDENTIFICATION ; CULTIVATION ; STRATEGIES
WOS研究方向Biotechnology & Applied Microbiology
WOS类目Biotechnology & Applied Microbiology
语种英语
WOS记录号WOS:000358291900009
出版者SPRINGER HEIDELBERG
URI标识http://www.irgrid.ac.cn/handle/1471x/2374060
专题物理研究所
通讯作者Lv, Pengmei
作者单位1.Chinese Acad Sci, Guangzhou Inst Energy Convers, Key Lab Renewable Energy, Guangzhou 510640, Guangdong, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714
He, Dong,Luo, Wen,Wang, Zhiyuan,et al. Combined use of gap and aox1 promoters and optimization of culture conditions to enhance expression of rhizomucor miehei lipase[J]. Journal of industrial microbiology & biotechnology,2015,42(8):1175-1182.
APA He, Dong,Luo, Wen,Wang, Zhiyuan,Lv, Pengmei,&Yuan, Zhenhong.(2015).Combined use of gap and aox1 promoters and optimization of culture conditions to enhance expression of rhizomucor miehei lipase.Journal of industrial microbiology & biotechnology,42(8),1175-1182.
MLA He, Dong,et al."Combined use of gap and aox1 promoters and optimization of culture conditions to enhance expression of rhizomucor miehei lipase".Journal of industrial microbiology & biotechnology 42.8(2015):1175-1182.

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来源:物理研究所

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