5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae
Abstract Background 5′-Aminolevulinic acid (ALA) is widely used in the pharmaceutical industry, healthcare, and food production, and is a substrate for the biosynthesis of heme, which is required for respiration and photosynthesis. Enhancement of ALA biosynthesis has never been developed in Saccharo...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-11-01
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| Series: | Microbial Cell Factories |
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| Online Access: | http://link.springer.com/article/10.1186/s12934-019-1242-6 |
| _version_ | 1818523346949636096 |
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| author | Kiyotaka Y. Hara Masaru Saito Hiroko Kato Kana Morikawa Hiroshi Kikukawa Hironari Nomura Takanori Fujimoto Yoko Hirono-Hara Shigeyuki Watanabe Kengo Kanamaru Akihiko Kondo |
| author_facet | Kiyotaka Y. Hara Masaru Saito Hiroko Kato Kana Morikawa Hiroshi Kikukawa Hironari Nomura Takanori Fujimoto Yoko Hirono-Hara Shigeyuki Watanabe Kengo Kanamaru Akihiko Kondo |
| author_sort | Kiyotaka Y. Hara |
| collection | DOAJ |
| description | Abstract Background 5′-Aminolevulinic acid (ALA) is widely used in the pharmaceutical industry, healthcare, and food production, and is a substrate for the biosynthesis of heme, which is required for respiration and photosynthesis. Enhancement of ALA biosynthesis has never been developed in Saccharomyces cerevisiae, which is a well-known model microorganism used for bioproduction of many value-added compounds. Results We demonstrated that metabolic engineering significantly improved ALA production in S. cerevisiae. First, we found that overexpression of HEM1, which encodes ALA synthetase, increased ALA production. Furthermore, addition of an optimal amount of glycine, a substrate for ALA biosynthesis, or levulinic acid, an inhibitor of ALA dehydrogenase, effectively increased ALA production. Next, we developed an assay for multiple metabolites including ALA and found that aconitase, encoded by ACO1 and ACO2, is the rate-limiting enzyme of ALA biosynthesis when sufficient glycine is supplied. Overexpression of ACO2 further enhanced ALA production in S. cerevisiae overexpressing HEM1. Conclusions In this study, ALA production in S. cerevisiae was enhanced by metabolic engineering. This study also shows a strategy to identify the rate-limiting step of a target synthetic pathway by assay for multiple metabolites alongside the target product. This strategy can be applied to improve production of other valuable products in the well-studied and well-industrialized microorganism S. cerevisiae. |
| first_indexed | 2024-12-11T05:43:51Z |
| format | Article |
| id | doaj.art-0853b3c32a5f422aa840ddf320c66736 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-11T05:43:51Z |
| publishDate | 2019-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-0853b3c32a5f422aa840ddf320c667362022-12-22T01:19:02ZengBMCMicrobial Cell Factories1475-28592019-11-011811810.1186/s12934-019-1242-65-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiaeKiyotaka Y. Hara0Masaru Saito1Hiroko Kato2Kana Morikawa3Hiroshi Kikukawa4Hironari Nomura5Takanori Fujimoto6Yoko Hirono-Hara7Shigeyuki Watanabe8Kengo Kanamaru9Akihiko Kondo10Department of Environmental and Life Sciences, School of Food and Nutritional Sciences, University of ShizuokaCosmo Oil Co., Ltd.Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe UniversityOrganization of Advanced Science and Technology, Kobe UniversityDepartment of Environmental and Life Sciences, School of Food and Nutritional Sciences, University of ShizuokaOrganization of Advanced Science and Technology, Kobe UniversityCosmo Oil Co., Ltd.Department of Environmental and Life Sciences, School of Food and Nutritional Sciences, University of ShizuokaCosmo Oil Co., Ltd.Department of Applied Chemistry in Bioscience, Graduate School of Agricultural Science, Kobe UniversityDepartment of Chemical Science and Engineering, Graduate School of Engineering, Kobe UniversityAbstract Background 5′-Aminolevulinic acid (ALA) is widely used in the pharmaceutical industry, healthcare, and food production, and is a substrate for the biosynthesis of heme, which is required for respiration and photosynthesis. Enhancement of ALA biosynthesis has never been developed in Saccharomyces cerevisiae, which is a well-known model microorganism used for bioproduction of many value-added compounds. Results We demonstrated that metabolic engineering significantly improved ALA production in S. cerevisiae. First, we found that overexpression of HEM1, which encodes ALA synthetase, increased ALA production. Furthermore, addition of an optimal amount of glycine, a substrate for ALA biosynthesis, or levulinic acid, an inhibitor of ALA dehydrogenase, effectively increased ALA production. Next, we developed an assay for multiple metabolites including ALA and found that aconitase, encoded by ACO1 and ACO2, is the rate-limiting enzyme of ALA biosynthesis when sufficient glycine is supplied. Overexpression of ACO2 further enhanced ALA production in S. cerevisiae overexpressing HEM1. Conclusions In this study, ALA production in S. cerevisiae was enhanced by metabolic engineering. This study also shows a strategy to identify the rate-limiting step of a target synthetic pathway by assay for multiple metabolites alongside the target product. This strategy can be applied to improve production of other valuable products in the well-studied and well-industrialized microorganism S. cerevisiae.http://link.springer.com/article/10.1186/s12934-019-1242-65′-Aminolevulinic acidYeastSaccharomyces cerevisiaeMetabolic engineeringCell factory |
| spellingShingle | Kiyotaka Y. Hara Masaru Saito Hiroko Kato Kana Morikawa Hiroshi Kikukawa Hironari Nomura Takanori Fujimoto Yoko Hirono-Hara Shigeyuki Watanabe Kengo Kanamaru Akihiko Kondo 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae Microbial Cell Factories 5′-Aminolevulinic acid Yeast Saccharomyces cerevisiae Metabolic engineering Cell factory |
| title | 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae |
| title_full | 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae |
| title_fullStr | 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae |
| title_full_unstemmed | 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae |
| title_short | 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae |
| title_sort | 5 aminolevulinic acid fermentation using engineered saccharomyces cerevisiae |
| topic | 5′-Aminolevulinic acid Yeast Saccharomyces cerevisiae Metabolic engineering Cell factory |
| url | http://link.springer.com/article/10.1186/s12934-019-1242-6 |
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