Combined use of GAP and AOX1 promoters and optimization of culture conditions to enhance expression of Rhizomucor miehei lipase

doi:10.1007/s10295-015-1633-6

GIEC OpenIR > 中国科学院广州能源研究所

	Combined use of GAP and AOX1 promoters and optimization of culture conditions to enhance expression of Rhizomucor miehei lipase
	He, Dong 1,2; Luo, Wen1 ; Wang, Zhiyuan 1; 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
DOI	10.1007/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	被引频次：18[WOS] [WOS记录] [WOS相关记录]
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
第一作者单位	中国科学院广州能源研究所
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.