Yongchuang Liu , Feilong Ma , Hongfei Zhang , Cuiwei Chu , Xiaohui Wang , Yuehui Tang , Jian He , Jiguo Qiu , Siqiong Xu , Lili Li
{"title":"Cloning and expression of the dimethylsulfoniopropionate lyase gene dddY and the identification of the key amino acids necessary for its activity","authors":"Yongchuang Liu , Feilong Ma , Hongfei Zhang , Cuiwei Chu , Xiaohui Wang , Yuehui Tang , Jian He , Jiguo Qiu , Siqiong Xu , Lili Li","doi":"10.1016/j.crmicr.2025.100444","DOIUrl":null,"url":null,"abstract":"<div><div>The abundant solute dimethylsulfoniopropionate (DMSP) is crucial in marine ecosystems. In this study, a bacterium, <em>Acinetobacter</em> sp. ZS25, capable of completely mineralizing DMSP and producing DMS and acrylate, was isolated. The possible DMSP degradation pathway was primarily identified. The role of DMSP lyases DddY was identified through a combination of genomic comparison, gene knockout and heterologous expression. The <em>K</em><sub>m</sub> and <em>k</em><sub>cat</sub> of AsDddY for DMSP were 2.6 mM and 12.7 × 10<sup>3</sup> s<sup>−1</sup>, respectively. Site-directed mutagenesis was employed to examine the influence of specific amino acid residues (Thr131, Asp181, Tyr225, Gly230, Gly250, His263, His265, Glu269, Tyr271, Leu274, Tyr331, and His338) within AsDddY, elucidating their critical roles in the protein's functionality. Bioinformatics analysis revealed 19 distinct <em>acu-dddY</em> cluster order types, while the number of strains with a complete <em>dddY</em>-<em>acu</em> cluster is limited. Our findings offer novel insights into the mechanisms underlying DMSP biodegradation and enhance our understanding of the diversity of <em>acu</em>-<em>dddY</em> clusters present in natural bacterial populations.</div></div>","PeriodicalId":34305,"journal":{"name":"Current Research in Microbial Sciences","volume":"9 ","pages":"Article 100444"},"PeriodicalIF":5.8000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Research in Microbial Sciences","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666517425001051","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
The abundant solute dimethylsulfoniopropionate (DMSP) is crucial in marine ecosystems. In this study, a bacterium, Acinetobacter sp. ZS25, capable of completely mineralizing DMSP and producing DMS and acrylate, was isolated. The possible DMSP degradation pathway was primarily identified. The role of DMSP lyases DddY was identified through a combination of genomic comparison, gene knockout and heterologous expression. The Km and kcat of AsDddY for DMSP were 2.6 mM and 12.7 × 103 s−1, respectively. Site-directed mutagenesis was employed to examine the influence of specific amino acid residues (Thr131, Asp181, Tyr225, Gly230, Gly250, His263, His265, Glu269, Tyr271, Leu274, Tyr331, and His338) within AsDddY, elucidating their critical roles in the protein's functionality. Bioinformatics analysis revealed 19 distinct acu-dddY cluster order types, while the number of strains with a complete dddY-acu cluster is limited. Our findings offer novel insights into the mechanisms underlying DMSP biodegradation and enhance our understanding of the diversity of acu-dddY clusters present in natural bacterial populations.