{"title":"具有高ATP再生系统的工程大肠杆菌高效全细胞合成s -腺苷- l-蛋氨酸。","authors":"Zhang Pengfei, Siqi Zuo, Peijie Shen, Zhengjun Si, Peilian Wei, Zhinan Xu","doi":"10.1080/10826068.2025.2509892","DOIUrl":null,"url":null,"abstract":"<p><p>S-adenosyl-L-methionine (SAM) is an important intermediate metabolite and widely used in the treatment of liver disease, arthritis, and depression. In this work, a whole-cell catalysis strategy was employed to enhance SAM production by combining the SAM biosynthetic pathway with an adenosine triphosphate (ATP) regeneration system in <i>Escherichia coli.</i> Specifically, the <i>ado1</i>, <i>ack</i>, and <i>adk</i> genes were previously introduced into the genome of the host strain. We then confirmed the availability of the ATP regeneration system under the condition of adding adenosine monophosphate (AMP) and acetyl phosphate (ACP) as supplements. To improve the SAM production, the <i>sam2</i> gene derived from <i>Saccharomyces cerevisiae</i> was overexpressed using the plasmid pGEX-2TK in the strain and the conditions of biocatalytic process were optimized. Under the optimal biocatalytic conditions, the recombinant strain RS01 (pGEX-2TK-SAM2) achieved a SAM titer of 11.4 g/L after 10 h cultivation. This work not only provides a new platform for the efficient production of SAM but also offers insights into the biosynthesis of other ATP-dependent products.</p>","PeriodicalId":20401,"journal":{"name":"Preparative Biochemistry & Biotechnology","volume":" ","pages":"1-8"},"PeriodicalIF":2.0000,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Efficient whole-cell biosynthesis of S-adenosyl-L-methionine by the engineered <i>Escherichia coli</i> with high ATP regenerating system.\",\"authors\":\"Zhang Pengfei, Siqi Zuo, Peijie Shen, Zhengjun Si, Peilian Wei, Zhinan Xu\",\"doi\":\"10.1080/10826068.2025.2509892\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>S-adenosyl-L-methionine (SAM) is an important intermediate metabolite and widely used in the treatment of liver disease, arthritis, and depression. In this work, a whole-cell catalysis strategy was employed to enhance SAM production by combining the SAM biosynthetic pathway with an adenosine triphosphate (ATP) regeneration system in <i>Escherichia coli.</i> Specifically, the <i>ado1</i>, <i>ack</i>, and <i>adk</i> genes were previously introduced into the genome of the host strain. We then confirmed the availability of the ATP regeneration system under the condition of adding adenosine monophosphate (AMP) and acetyl phosphate (ACP) as supplements. To improve the SAM production, the <i>sam2</i> gene derived from <i>Saccharomyces cerevisiae</i> was overexpressed using the plasmid pGEX-2TK in the strain and the conditions of biocatalytic process were optimized. Under the optimal biocatalytic conditions, the recombinant strain RS01 (pGEX-2TK-SAM2) achieved a SAM titer of 11.4 g/L after 10 h cultivation. This work not only provides a new platform for the efficient production of SAM but also offers insights into the biosynthesis of other ATP-dependent products.</p>\",\"PeriodicalId\":20401,\"journal\":{\"name\":\"Preparative Biochemistry & Biotechnology\",\"volume\":\" \",\"pages\":\"1-8\"},\"PeriodicalIF\":2.0000,\"publicationDate\":\"2025-05-23\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Preparative Biochemistry & Biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1080/10826068.2025.2509892\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMICAL RESEARCH METHODS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Preparative Biochemistry & Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1080/10826068.2025.2509892","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
摘要
s -腺苷- l-蛋氨酸(SAM)是一种重要的中间代谢物,广泛应用于肝病、关节炎和抑郁症的治疗。在这项工作中,采用全细胞催化策略,通过将SAM生物合成途径与大肠杆菌的三磷酸腺苷(ATP)再生系统相结合,来提高SAM的产生。具体来说,ado1, ack和adk基因先前被引入宿主菌株的基因组中。然后,在添加单磷酸腺苷(AMP)和乙酰磷酸(ACP)作为补充物的条件下,我们证实了ATP再生系统的有效性。为了提高SAM的产量,利用质粒pGEX-2TK在菌株中过表达酿酒酵母的sam2基因,并优化了生物催化过程的条件。在最佳生物催化条件下,重组菌株RS01 (pGEX-2TK-SAM2)培养10 h后,SAM滴度达到11.4 g/L。这项工作不仅为SAM的高效生产提供了一个新的平台,而且为其他atp依赖性产物的生物合成提供了新的见解。
Efficient whole-cell biosynthesis of S-adenosyl-L-methionine by the engineered Escherichia coli with high ATP regenerating system.
S-adenosyl-L-methionine (SAM) is an important intermediate metabolite and widely used in the treatment of liver disease, arthritis, and depression. In this work, a whole-cell catalysis strategy was employed to enhance SAM production by combining the SAM biosynthetic pathway with an adenosine triphosphate (ATP) regeneration system in Escherichia coli. Specifically, the ado1, ack, and adk genes were previously introduced into the genome of the host strain. We then confirmed the availability of the ATP regeneration system under the condition of adding adenosine monophosphate (AMP) and acetyl phosphate (ACP) as supplements. To improve the SAM production, the sam2 gene derived from Saccharomyces cerevisiae was overexpressed using the plasmid pGEX-2TK in the strain and the conditions of biocatalytic process were optimized. Under the optimal biocatalytic conditions, the recombinant strain RS01 (pGEX-2TK-SAM2) achieved a SAM titer of 11.4 g/L after 10 h cultivation. This work not only provides a new platform for the efficient production of SAM but also offers insights into the biosynthesis of other ATP-dependent products.
期刊介绍:
Preparative Biochemistry & Biotechnology is an international forum for rapid dissemination of high quality research results dealing with all aspects of preparative techniques in biochemistry, biotechnology and other life science disciplines.