Martina Kiel, Israel Barrantes, Dietmar H. Pieper, Karl-Heinrich Engesser
{"title":"(2R,3S)-2,3-二羟基-2,3-二氢苯甲酸酯的生物技术生产、分离与表征","authors":"Martina Kiel, Israel Barrantes, Dietmar H. Pieper, Karl-Heinrich Engesser","doi":"10.1111/1751-7915.70228","DOIUrl":null,"url":null,"abstract":"<p>Bacterial Rieske non-heme iron oxygenases catalyse the transformation of a wide range of aromatic compounds to vicinal <i>cis</i>-dihydrodiols. Such compounds have been successfully applied in chemoenzymatic synthetic routes for, for example, pharmaceuticals, natural products and polymers. In the case of benzoate, only (1<i>S</i>,2<i>R</i>)-<i>cis</i>-1,2-dihydroxy-2-hydrobenzoate is readily accessible via enzymatic transformation, but not the regioisomeric <i>cis</i>-2,3-dihydroxy-2,3-dihydrobenzoate (2,3-DD) or <i>cis</i>-3,4-dihydroxy-3,4-dihydrobenzoate. While trace amounts of putative <i>cis</i>-2,3-DD have been obtained before by using <i>p</i>-cumate 2,3-dioxygenase (PCDO) or a combination of chlorobenzene dioxygenase and nitrilase, none of these approaches enabled its production and isolation at a greater scale for potential use as a chiral building block in organic synthesis. We here provide a protocol for biotransformation of benzoate yielding (2<i>R</i>,3<i>S</i>)-2,3-dihydroxy-2,3-dihydrobenzoate using the PCDO of <i>Pseudomonas citronellolis</i> strain EB200 with negligible formation of side products. An isolation procedure suitable for production of the 2,3-DD sodium salt monohydrate at high purity (> 95%) at a gram scale, and a comprehensive characterisation of this novel metabolite is given.</p>","PeriodicalId":209,"journal":{"name":"Microbial Biotechnology","volume":"18 9","pages":""},"PeriodicalIF":5.2000,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12442788/pdf/","citationCount":"0","resultStr":"{\"title\":\"Biotechnological Production, Isolation and Characterisation of (2R,3S)-2,3-Dihydroxy-2,3-Dihydrobenzoate\",\"authors\":\"Martina Kiel, Israel Barrantes, Dietmar H. Pieper, Karl-Heinrich Engesser\",\"doi\":\"10.1111/1751-7915.70228\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Bacterial Rieske non-heme iron oxygenases catalyse the transformation of a wide range of aromatic compounds to vicinal <i>cis</i>-dihydrodiols. Such compounds have been successfully applied in chemoenzymatic synthetic routes for, for example, pharmaceuticals, natural products and polymers. In the case of benzoate, only (1<i>S</i>,2<i>R</i>)-<i>cis</i>-1,2-dihydroxy-2-hydrobenzoate is readily accessible via enzymatic transformation, but not the regioisomeric <i>cis</i>-2,3-dihydroxy-2,3-dihydrobenzoate (2,3-DD) or <i>cis</i>-3,4-dihydroxy-3,4-dihydrobenzoate. While trace amounts of putative <i>cis</i>-2,3-DD have been obtained before by using <i>p</i>-cumate 2,3-dioxygenase (PCDO) or a combination of chlorobenzene dioxygenase and nitrilase, none of these approaches enabled its production and isolation at a greater scale for potential use as a chiral building block in organic synthesis. We here provide a protocol for biotransformation of benzoate yielding (2<i>R</i>,3<i>S</i>)-2,3-dihydroxy-2,3-dihydrobenzoate using the PCDO of <i>Pseudomonas citronellolis</i> strain EB200 with negligible formation of side products. An isolation procedure suitable for production of the 2,3-DD sodium salt monohydrate at high purity (> 95%) at a gram scale, and a comprehensive characterisation of this novel metabolite is given.</p>\",\"PeriodicalId\":209,\"journal\":{\"name\":\"Microbial Biotechnology\",\"volume\":\"18 9\",\"pages\":\"\"},\"PeriodicalIF\":5.2000,\"publicationDate\":\"2025-09-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12442788/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microbial Biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.70228\",\"RegionNum\":2,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microbial Biotechnology","FirstCategoryId":"5","ListUrlMain":"https://enviromicro-journals.onlinelibrary.wiley.com/doi/10.1111/1751-7915.70228","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Biotechnological Production, Isolation and Characterisation of (2R,3S)-2,3-Dihydroxy-2,3-Dihydrobenzoate
Bacterial Rieske non-heme iron oxygenases catalyse the transformation of a wide range of aromatic compounds to vicinal cis-dihydrodiols. Such compounds have been successfully applied in chemoenzymatic synthetic routes for, for example, pharmaceuticals, natural products and polymers. In the case of benzoate, only (1S,2R)-cis-1,2-dihydroxy-2-hydrobenzoate is readily accessible via enzymatic transformation, but not the regioisomeric cis-2,3-dihydroxy-2,3-dihydrobenzoate (2,3-DD) or cis-3,4-dihydroxy-3,4-dihydrobenzoate. While trace amounts of putative cis-2,3-DD have been obtained before by using p-cumate 2,3-dioxygenase (PCDO) or a combination of chlorobenzene dioxygenase and nitrilase, none of these approaches enabled its production and isolation at a greater scale for potential use as a chiral building block in organic synthesis. We here provide a protocol for biotransformation of benzoate yielding (2R,3S)-2,3-dihydroxy-2,3-dihydrobenzoate using the PCDO of Pseudomonas citronellolis strain EB200 with negligible formation of side products. An isolation procedure suitable for production of the 2,3-DD sodium salt monohydrate at high purity (> 95%) at a gram scale, and a comprehensive characterisation of this novel metabolite is given.
期刊介绍:
Microbial Biotechnology publishes papers of original research reporting significant advances in any aspect of microbial applications, including, but not limited to biotechnologies related to: Green chemistry; Primary metabolites; Food, beverages and supplements; Secondary metabolites and natural products; Pharmaceuticals; Diagnostics; Agriculture; Bioenergy; Biomining, including oil recovery and processing; Bioremediation; Biopolymers, biomaterials; Bionanotechnology; Biosurfactants and bioemulsifiers; Compatible solutes and bioprotectants; Biosensors, monitoring systems, quantitative microbial risk assessment; Technology development; Protein engineering; Functional genomics; Metabolic engineering; Metabolic design; Systems analysis, modelling; Process engineering; Biologically-based analytical methods; Microbially-based strategies in public health; Microbially-based strategies to influence global processes