{"title":"在枯草芽孢杆菌中生产 2,3-丁二醇的不同脱氢酶的硅学筛选和验证","authors":"Sailee Sanjay Asolkar, M. Anju, Ravindra Kumar, Apoorva Deshmukh, Anand Ghosalkar, Pramod Kumbhar","doi":"10.1007/s12257-024-00053-1","DOIUrl":null,"url":null,"abstract":"<p><i>Bacillus subtilis</i> is a natural producer of 2,3-butanediol (2,3-BDO) and has acquired “Generally Regarded as Safe\" status. It is reported to produce 2,3-BDO from synthetic sugars as well as complex and economic sugar sources such as molasses. However, the rate-limiting step in the formation of 2,3-BDO is its conversion from acetoin to 2,3-BDO by the enzyme butanediol dehydrogenase (2,3-BDH). Such 2,3-BDHs were screened based on higher affinity (lower <i>K</i><sub>m</sub>) towards acetoin as substrate. The in silico docking studies were conducted for further validation, and they showed a high interaction profile for the PpBDH protein towards acetoin. Heterologous expression of these genes was studied in engineered <i>Bacillus subtilis</i> (BS1A1). In this study, it was seen that 2,3-BDH from <i>Paenibacillus polymyxa ZJ-9</i> was reported to have higher enzyme activity levels, and in the fermentation studies, it was seen that the ratio of 2,3-BDO to acetoin was increased by 80.25%. The insights encourage further bioprocess optimization for increasing the fermentative production of 2,3-BDO. Our results provide a potential strategy to avoid the back conversion of 2,3-BDO to acetoin in an engineered <i>Bacillus</i> system.</p>","PeriodicalId":8936,"journal":{"name":"Biotechnology and Bioprocess Engineering","volume":"103 1","pages":""},"PeriodicalIF":2.5000,"publicationDate":"2024-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"In silico screening and validation of different dehydrogenases to produce 2,3-butanediol in Bacillus subtilis\",\"authors\":\"Sailee Sanjay Asolkar, M. Anju, Ravindra Kumar, Apoorva Deshmukh, Anand Ghosalkar, Pramod Kumbhar\",\"doi\":\"10.1007/s12257-024-00053-1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><i>Bacillus subtilis</i> is a natural producer of 2,3-butanediol (2,3-BDO) and has acquired “Generally Regarded as Safe\\\" status. It is reported to produce 2,3-BDO from synthetic sugars as well as complex and economic sugar sources such as molasses. However, the rate-limiting step in the formation of 2,3-BDO is its conversion from acetoin to 2,3-BDO by the enzyme butanediol dehydrogenase (2,3-BDH). Such 2,3-BDHs were screened based on higher affinity (lower <i>K</i><sub>m</sub>) towards acetoin as substrate. The in silico docking studies were conducted for further validation, and they showed a high interaction profile for the PpBDH protein towards acetoin. Heterologous expression of these genes was studied in engineered <i>Bacillus subtilis</i> (BS1A1). In this study, it was seen that 2,3-BDH from <i>Paenibacillus polymyxa ZJ-9</i> was reported to have higher enzyme activity levels, and in the fermentation studies, it was seen that the ratio of 2,3-BDO to acetoin was increased by 80.25%. The insights encourage further bioprocess optimization for increasing the fermentative production of 2,3-BDO. Our results provide a potential strategy to avoid the back conversion of 2,3-BDO to acetoin in an engineered <i>Bacillus</i> system.</p>\",\"PeriodicalId\":8936,\"journal\":{\"name\":\"Biotechnology and Bioprocess Engineering\",\"volume\":\"103 1\",\"pages\":\"\"},\"PeriodicalIF\":2.5000,\"publicationDate\":\"2024-03-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biotechnology and Bioprocess Engineering\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1007/s12257-024-00053-1\",\"RegionNum\":4,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biotechnology and Bioprocess Engineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1007/s12257-024-00053-1","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
In silico screening and validation of different dehydrogenases to produce 2,3-butanediol in Bacillus subtilis
Bacillus subtilis is a natural producer of 2,3-butanediol (2,3-BDO) and has acquired “Generally Regarded as Safe" status. It is reported to produce 2,3-BDO from synthetic sugars as well as complex and economic sugar sources such as molasses. However, the rate-limiting step in the formation of 2,3-BDO is its conversion from acetoin to 2,3-BDO by the enzyme butanediol dehydrogenase (2,3-BDH). Such 2,3-BDHs were screened based on higher affinity (lower Km) towards acetoin as substrate. The in silico docking studies were conducted for further validation, and they showed a high interaction profile for the PpBDH protein towards acetoin. Heterologous expression of these genes was studied in engineered Bacillus subtilis (BS1A1). In this study, it was seen that 2,3-BDH from Paenibacillus polymyxa ZJ-9 was reported to have higher enzyme activity levels, and in the fermentation studies, it was seen that the ratio of 2,3-BDO to acetoin was increased by 80.25%. The insights encourage further bioprocess optimization for increasing the fermentative production of 2,3-BDO. Our results provide a potential strategy to avoid the back conversion of 2,3-BDO to acetoin in an engineered Bacillus system.
期刊介绍:
Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.