Knowledge Management System Of Guangzhou Institute of Energy Conversion, CAS
| 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
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| 2015-08-01 | |
| Source Publication | JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
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| Volume | 42Issue:8Pages:1175-1182 |
| Abstract | 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. |
| Subtype | Article |
| Keyword | Rml Pichia Pastoris P-gap P-aox1 Optimization |
| WOS Headings | Science & Technology ; Life Sciences & Biomedicine |
| DOI | 10.1007/s10295-015-1633-6 |
| WOS Subject Extended | Biotechnology & Applied Microbiology |
| WOS Keyword | PICHIA-PASTORIS ; RECOMBINANT PROTEINS ; GENE-EXPRESSION ; COPY NUMBER ; IDENTIFICATION ; CULTIVATION ; STRATEGIES |
| Indexed By | SCI |
| Language | 英语 |
| WOS Subject | Biotechnology & Applied Microbiology |
| WOS ID | WOS:000358291900009 |
| Citation statistics | |
| Document Type | 期刊论文 |
| Identifier | http://ir.giec.ac.cn/handle/344007/10537 |
| Collection | 中国科学院广州能源研究所 |
| Affiliation | 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 |
| First Author Affilication | GuangZhou 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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