High production of valencene in Saccharomyces cerevisiae through metabolic engineering
Abstract Background The biological synthesis of high value compounds in industry through metabolically engineered microorganism factories has received increasing attention in recent years. Valencene is a high value ingredient in the flavor and fragrance industry, but the low concentration in nature...
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12934-019-1246-2 |
| _version_ | 1818115195421065216 |
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| author | Hefeng Chen Chaoyi Zhu Muzi Zhu Jinghui Xiong Hao Ma Min Zhuo Shuang Li |
| author_facet | Hefeng Chen Chaoyi Zhu Muzi Zhu Jinghui Xiong Hao Ma Min Zhuo Shuang Li |
| author_sort | Hefeng Chen |
| collection | DOAJ |
| description | Abstract Background The biological synthesis of high value compounds in industry through metabolically engineered microorganism factories has received increasing attention in recent years. Valencene is a high value ingredient in the flavor and fragrance industry, but the low concentration in nature and high cost of extraction limits its application. Saccharomyces cerevisiae, generally recognized as safe, is one of the most commonly used gene expression hosts. Construction of S. cerevisiae cell factory to achieve high production of valencene will be attractive. Results Valencene was successfully biosynthesized after introducing valencene synthase into S. cerevisiae BJ5464. A significant increase in valencene yield was observed after down-regulation or knock-out of squalene synthesis and other inhibiting factors (such as erg9, rox1) in mevalonate (MVA) pathway using a recyclable CRISPR/Cas9 system constructed in this study through the introduction of Cre/loxP. To increase the supplement of the precursor farnesyl pyrophosphate (FPP), all the genes of FPP upstream in MVA pathway were overexpressed in yeast genome. Furthermore, valencene expression cassettes containing different promoters and terminators were compared, and PHXT7-VS-TTPI1 was found to have excellent performance in valencene production. Finally, after fed-batch fermentation in 3 L bioreactor, valencene production titer reached 539.3 mg/L with about 160-fold improvement compared to the initial titer, which is the highest reported valencene yield. Conclusions This study achieved high production of valencene in S. cerevisiae through metabolic engineering and optimization of expression cassette, providing good example of microbial overproduction of valuable chemical products. The construction of recyclable plasmid was useful for multiple gene editing as well. |
| first_indexed | 2024-12-11T04:02:46Z |
| format | Article |
| id | doaj.art-e34463a252e44fe19a549f589abbb036 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-11T04:02:46Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-e34463a252e44fe19a549f589abbb0362022-12-22T01:21:36ZengBMCMicrobial Cell Factories1475-28592019-11-0118111410.1186/s12934-019-1246-2High production of valencene in Saccharomyces cerevisiae through metabolic engineeringHefeng Chen0Chaoyi Zhu1Muzi Zhu2Jinghui Xiong3Hao Ma4Min Zhuo5Shuang Li6School of Biology and Biological Engineering, South China University of Technology, Higher Education Mega CenterSchool of Biology and Biological Engineering, South China University of Technology, Higher Education Mega CenterState Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Institute of MicrobiologySchool of Biology and Biological Engineering, South China University of Technology, Higher Education Mega CenterSchool of Biology and Biological Engineering, South China University of Technology, Higher Education Mega CenterSchool of Biology and Biological Engineering, South China University of Technology, Higher Education Mega CenterSchool of Biology and Biological Engineering, South China University of Technology, Higher Education Mega CenterAbstract Background The biological synthesis of high value compounds in industry through metabolically engineered microorganism factories has received increasing attention in recent years. Valencene is a high value ingredient in the flavor and fragrance industry, but the low concentration in nature and high cost of extraction limits its application. Saccharomyces cerevisiae, generally recognized as safe, is one of the most commonly used gene expression hosts. Construction of S. cerevisiae cell factory to achieve high production of valencene will be attractive. Results Valencene was successfully biosynthesized after introducing valencene synthase into S. cerevisiae BJ5464. A significant increase in valencene yield was observed after down-regulation or knock-out of squalene synthesis and other inhibiting factors (such as erg9, rox1) in mevalonate (MVA) pathway using a recyclable CRISPR/Cas9 system constructed in this study through the introduction of Cre/loxP. To increase the supplement of the precursor farnesyl pyrophosphate (FPP), all the genes of FPP upstream in MVA pathway were overexpressed in yeast genome. Furthermore, valencene expression cassettes containing different promoters and terminators were compared, and PHXT7-VS-TTPI1 was found to have excellent performance in valencene production. Finally, after fed-batch fermentation in 3 L bioreactor, valencene production titer reached 539.3 mg/L with about 160-fold improvement compared to the initial titer, which is the highest reported valencene yield. Conclusions This study achieved high production of valencene in S. cerevisiae through metabolic engineering and optimization of expression cassette, providing good example of microbial overproduction of valuable chemical products. The construction of recyclable plasmid was useful for multiple gene editing as well.http://link.springer.com/article/10.1186/s12934-019-1246-2ValenceneSynthetic biologyMetabolic engineeringSaccharomyces cerevisiaeRecyclable plasmidExpression cassette |
| spellingShingle | Hefeng Chen Chaoyi Zhu Muzi Zhu Jinghui Xiong Hao Ma Min Zhuo Shuang Li High production of valencene in Saccharomyces cerevisiae through metabolic engineering Microbial Cell Factories Valencene Synthetic biology Metabolic engineering Saccharomyces cerevisiae Recyclable plasmid Expression cassette |
| title | High production of valencene in Saccharomyces cerevisiae through metabolic engineering |
| title_full | High production of valencene in Saccharomyces cerevisiae through metabolic engineering |
| title_fullStr | High production of valencene in Saccharomyces cerevisiae through metabolic engineering |
| title_full_unstemmed | High production of valencene in Saccharomyces cerevisiae through metabolic engineering |
| title_short | High production of valencene in Saccharomyces cerevisiae through metabolic engineering |
| title_sort | high production of valencene in saccharomyces cerevisiae through metabolic engineering |
| topic | Valencene Synthetic biology Metabolic engineering Saccharomyces cerevisiae Recyclable plasmid Expression cassette |
| url | http://link.springer.com/article/10.1186/s12934-019-1246-2 |
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