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Combined use of GAP and AOX1 promoters and optimization of culture conditions to enhance expression of Rhizomucor miehei lipase
He, Dong1,2; Luo, Wen1; Wang, Zhiyuan1; Lv, Pengmei1; Yuan, Zhenhong1
2015-08-01
发表期刊JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
卷号42期号:8页码:1175-1182
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.
文章类型Article
关键词Rml Pichia Pastoris P-gap P-aox1 Optimization
WOS标题词Science & Technology ; Life Sciences & Biomedicine
DOI10.1007/s10295-015-1633-6
研究领域[WOS]Biotechnology & Applied Microbiology
关键词[WOS]PICHIA-PASTORIS ; RECOMBINANT PROTEINS ; GENE-EXPRESSION ; COPY NUMBER ; IDENTIFICATION ; CULTIVATION ; STRATEGIES
收录类别SCI
语种英语
WOS类目Biotechnology & Applied Microbiology
WOS记录号WOS:000358291900009
引用统计
文献类型期刊论文
条目标识符http://ir.giec.ac.cn/handle/344007/10537
专题中国科学院广州能源研究所
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
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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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