{"title":"工程微生物细胞工厂利用 L-色氨酸生物合成褪黑素","authors":"Lijuan Wang, Yongdong Deng, Jianjie Gao, Bo Wang, Hongjuan Han, Zhenjun Li, Wenhui Zhang, Yu Wang, Xiaoyan Fu, Rihe Peng, Quanhong Yao, Yongsheng Tian, Jing Xu","doi":"10.1186/s13068-024-02476-7","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>The demand for melatonin is increasing due to its health-promoting bioactivities such as antioxidant and sleep benefits. Although melatonin is present in various organisms, its low content and high extraction cost make it unsustainable. Biosynthesis is a promising alternative method for melatonin production. However, the ectopic production of melatonin in microorganisms is very difficult due to the low or insoluble expression of melatonin synthesis genes. Hence, we aim to explore the biosynthesis of melatonin using <i>Escherichia coli</i> as a cell factory and ways to simultaneously coordinated express genes from different melatonin synthesis pathways.</p><h3>Results</h3><p>In this study, the <i>mXcP4H</i> gene from <i>Xanthomonas campestris</i>, as well as the <i>HsAADC</i>, <i>HsAANAT</i> and <i>HIOMT</i> genes from human melatonin synthesis pathway were optimized and introduced into <i>E. coli</i> via a multi-monocistronic vector. The obtained strain BL7992 successfully synthesized 1.13 mg/L melatonin by utilizing L-tryptophan (<span>l</span>-Trp) as a substrate in a shake flask. It was determined that the rate-limiting enzyme for melatonin synthesis is the arylalkylamine N-acetyltransferase, which is encoded by the <i>HsAANAT</i> gene. Targeted metabolomics analysis of <span>l</span>-Trp revealed that the majority of <span>l</span>-Trp flowed to the indole pathway in BL7992, and knockout of the <i>tnaA</i> gene may be beneficial for increasing melatonin production.</p><h3>Conclusions</h3><p>A metabolic engineering approach was adopted and melatonin was successfully synthesized from low-cost <span>l</span>-Trp in <i>E. coli</i>. This study provides a rapid and economical strategy for the synthesis of melatonin.</p></div>","PeriodicalId":494,"journal":{"name":"Biotechnology for Biofuels","volume":null,"pages":null},"PeriodicalIF":6.1000,"publicationDate":"2024-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://biotechnologyforbiofuels.biomedcentral.com/counter/pdf/10.1186/s13068-024-02476-7","citationCount":"0","resultStr":"{\"title\":\"Biosynthesis of melatonin from l-tryptophan by an engineered microbial cell factory\",\"authors\":\"Lijuan Wang, Yongdong Deng, Jianjie Gao, Bo Wang, Hongjuan Han, Zhenjun Li, Wenhui Zhang, Yu Wang, Xiaoyan Fu, Rihe Peng, Quanhong Yao, Yongsheng Tian, Jing Xu\",\"doi\":\"10.1186/s13068-024-02476-7\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>The demand for melatonin is increasing due to its health-promoting bioactivities such as antioxidant and sleep benefits. Although melatonin is present in various organisms, its low content and high extraction cost make it unsustainable. Biosynthesis is a promising alternative method for melatonin production. However, the ectopic production of melatonin in microorganisms is very difficult due to the low or insoluble expression of melatonin synthesis genes. Hence, we aim to explore the biosynthesis of melatonin using <i>Escherichia coli</i> as a cell factory and ways to simultaneously coordinated express genes from different melatonin synthesis pathways.</p><h3>Results</h3><p>In this study, the <i>mXcP4H</i> gene from <i>Xanthomonas campestris</i>, as well as the <i>HsAADC</i>, <i>HsAANAT</i> and <i>HIOMT</i> genes from human melatonin synthesis pathway were optimized and introduced into <i>E. coli</i> via a multi-monocistronic vector. The obtained strain BL7992 successfully synthesized 1.13 mg/L melatonin by utilizing L-tryptophan (<span>l</span>-Trp) as a substrate in a shake flask. It was determined that the rate-limiting enzyme for melatonin synthesis is the arylalkylamine N-acetyltransferase, which is encoded by the <i>HsAANAT</i> gene. Targeted metabolomics analysis of <span>l</span>-Trp revealed that the majority of <span>l</span>-Trp flowed to the indole pathway in BL7992, and knockout of the <i>tnaA</i> gene may be beneficial for increasing melatonin production.</p><h3>Conclusions</h3><p>A metabolic engineering approach was adopted and melatonin was successfully synthesized from low-cost <span>l</span>-Trp in <i>E. coli</i>. This study provides a rapid and economical strategy for the synthesis of melatonin.</p></div>\",\"PeriodicalId\":494,\"journal\":{\"name\":\"Biotechnology for Biofuels\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":6.1000,\"publicationDate\":\"2024-02-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://biotechnologyforbiofuels.biomedcentral.com/counter/pdf/10.1186/s13068-024-02476-7\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology for Biofuels\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://link.springer.com/article/10.1186/s13068-024-02476-7\",\"RegionNum\":1,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology for Biofuels","FirstCategoryId":"5","ListUrlMain":"https://link.springer.com/article/10.1186/s13068-024-02476-7","RegionNum":1,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
Biosynthesis of melatonin from l-tryptophan by an engineered microbial cell factory
Background
The demand for melatonin is increasing due to its health-promoting bioactivities such as antioxidant and sleep benefits. Although melatonin is present in various organisms, its low content and high extraction cost make it unsustainable. Biosynthesis is a promising alternative method for melatonin production. However, the ectopic production of melatonin in microorganisms is very difficult due to the low or insoluble expression of melatonin synthesis genes. Hence, we aim to explore the biosynthesis of melatonin using Escherichia coli as a cell factory and ways to simultaneously coordinated express genes from different melatonin synthesis pathways.
Results
In this study, the mXcP4H gene from Xanthomonas campestris, as well as the HsAADC, HsAANAT and HIOMT genes from human melatonin synthesis pathway were optimized and introduced into E. coli via a multi-monocistronic vector. The obtained strain BL7992 successfully synthesized 1.13 mg/L melatonin by utilizing L-tryptophan (l-Trp) as a substrate in a shake flask. It was determined that the rate-limiting enzyme for melatonin synthesis is the arylalkylamine N-acetyltransferase, which is encoded by the HsAANAT gene. Targeted metabolomics analysis of l-Trp revealed that the majority of l-Trp flowed to the indole pathway in BL7992, and knockout of the tnaA gene may be beneficial for increasing melatonin production.
Conclusions
A metabolic engineering approach was adopted and melatonin was successfully synthesized from low-cost l-Trp in E. coli. This study provides a rapid and economical strategy for the synthesis of melatonin.
期刊介绍:
Biotechnology for Biofuels is an open access peer-reviewed journal featuring high-quality studies describing technological and operational advances in the production of biofuels, chemicals and other bioproducts. The journal emphasizes understanding and advancing the application of biotechnology and synergistic operations to improve plants and biological conversion systems for the biological production of these products from biomass, intermediates derived from biomass, or CO2, as well as upstream or downstream operations that are integral to biological conversion of biomass.
Biotechnology for Biofuels focuses on the following areas:
• Development of terrestrial plant feedstocks
• Development of algal feedstocks
• Biomass pretreatment, fractionation and extraction for biological conversion
• Enzyme engineering, production and analysis
• Bacterial genetics, physiology and metabolic engineering
• Fungal/yeast genetics, physiology and metabolic engineering
• Fermentation, biocatalytic conversion and reaction dynamics
• Biological production of chemicals and bioproducts from biomass
• Anaerobic digestion, biohydrogen and bioelectricity
• Bioprocess integration, techno-economic analysis, modelling and policy
• Life cycle assessment and environmental impact analysis