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, Dong1,2; Luo, Wen1; Wang, Zhiyuan1; Lv, Pengmei1; Yuan, Zhenhong1
2015-08-01
Source PublicationJOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
Volume42Issue:8Pages:1175-1182
AbstractRhizo 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.
SubtypeArticle
KeywordRml Pichia Pastoris P-gap P-aox1 Optimization
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1007/s10295-015-1633-6
WOS Subject ExtendedBiotechnology & Applied Microbiology
WOS KeywordPICHIA-PASTORIS ; RECOMBINANT PROTEINS ; GENE-EXPRESSION ; COPY NUMBER ; IDENTIFICATION ; CULTIVATION ; STRATEGIES
Indexed BySCI
Language英语
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000358291900009
Citation statistics
Document Type期刊论文
Identifierhttp://ir.giec.ac.cn/handle/344007/10537
Collection中国科学院广州能源研究所
Affiliation1.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
First Author AffilicationGuangZhou Institute of Energy Conversion,Chinese Academy of Sciences
Recommended Citation
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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