Analysis of microbial metabolism of biosulfur compounds and adaptive evolution for the degradation of xenobiotics

生物硫化合物的微生物代谢分析和外源物质降解的适应性进化

• 批准号: 14360050
• 负责人: OMORI Toshio
• 金额: $ 8.06万
• 依托单位: Shibaura Institute of Technology (2003)The University of Tokyo (2002)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360050/
• 关键词: Dimethyl Sulfide; Dibenzothiophene; Organic sulfur compounds; Dimethyl Sulfone; sigma54; Sulfate starvation; Dimethyl Slufide; Dimethyl Slufone

Microbial dimethyl sulfide (DMS) conversion is thought to be involved in the global sulfur cycle. We isolated Pseudomonas putida strain DS1 from soil as a bacterium utilizing DMS as a sole sulfur source, and tried to elucidate the DMS conversion mechanism of strain DS1 at biochemical and genetic level. Strain DS1 oxidized DMS to dimethyl sulfone ( DMSO_2) via dimethyl sulfoxide, whereas the oxidation was repressed in the presence of sulfate, suggesting that a sulfate starvation response is involved in DMS utilization by starin DS1. Two of the five DMS-utilization-defective mutants isolated by transposon5 (Th5) mutagenesis had a Tn5 insertion in the ssuEAECBF operon, which has been reported to encode a two-component monooxygenase system (SsuED), an ABC-type transporter (SsuABC), and a small protein (SsuF), and also to play a key role in utilization of sulfonates and sulfate esters in another bacterium, P.pulida strain S-313. Disniption of ssuD and SsuD enzymatic activity demonstrated that methanesulfonate is a metabolic intermediate of DMS and desulfonated by SsuD.Disruption of ssuC cr ssuF also led to a DMS-utilization-defective phenotype. Another two mutants bad a defect in a gene homologous to pa2354 from P.aeruginosa PA01, which encodes a putative transcriptional regulator, while the remaining mutant had a defect in cysM encoding O-acetylserine(thio)-lyase B.

微生物二甲基硫化物（DMS）转化被认为参与了全球硫循环。本研究利用DMS作为唯一硫源，从土壤中分离出恶臭假单胞菌DS1，并试图从生化和遗传水平上阐明菌株DS1转化DMS的机制。菌株DS1通过二甲亚砜将DMS氧化为二甲基砜（DMSO_2），而在硫酸盐存在下氧化被抑制，表明菌株DS1对DMS的利用涉及硫酸盐饥饿反应。通过转座子5 （Th5）诱变分离的5个dms利用缺陷突变体中有2个在ssuEAECBF操纵子中有Tn5插入，据报道该突变体编码双组分单加氧酶系统（ssue）， abc型转运体（SsuABC）和小蛋白（SsuF），并且在另一种细菌P.pulida菌株S-313中对硫酸盐和硫酸盐酯的利用起关键作用。ssuD的分解和ssuD的酶活性表明，甲磺酸盐是DMS的代谢中间体，并被ssuD脱硫。ssuC和ssuF的破坏也导致dms利用缺陷表型。另外两个突变体在P.aeruginosa PA01的pa2354同源基因中存在缺陷，该基因编码一种推定的转录调节因子，而其余突变体在编码o -乙酰丝氨酸（硫）裂解酶B的cysM中存在缺陷。

项目成果

Takayuki Endoh, Hiroshi Habe, Hideaki Nojiri, Toshio Omori, et al.: "A CysB-regulated and sigma(54)-dependent regulator, SfnR, is essential for dimethyl sulfone metabolism of Pseudomonas putida strain DS1"Microbiology. 149. 991-1000 (2003)

Takayuki Endoh、Hiroshi Habe、Hideaki Nojiri、Toshio Omori 等人：“CysB 调节和 sigma(54) 依赖性调节器 SfnR 对于恶臭假单胞菌菌株 DS1 的二甲砜代谢至关重要”微生物学。

Takayuki Endoh, Hiroshi Habe, Hideaki Nojiri, Toshio Omori, et al.: "Characterization and identification of genes essential for dimethy1 sulfide utilization in Pseudomonas putida strain DS1"Applied Microbiology and Biotechnology. 62(1). 83-91 (2003)

Takayuki Endoh、Hiroshi Habe、Hideaki Nojiri、Toshio Omori 等人：“恶臭假单胞菌菌株 DS1 中二甲基硫醚利用所必需的基因的表征和鉴定”应用微生物学和生物技术。