Improvement of Functions of Novel Enzymes in the Desulfurization Metaoblism of Petroleum by Protein Engineering and Molecular Genetics

蛋白质工程和分子遗传学改进石油脱硫代谢新型酶的功能

基本信息

批准号：
15580063
负责人：
IZUMI Yoshikazu
金额：
$ 2.43万
依托单位：
National University Corporation Tottori University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580063/
关键词：
desulfurization dibenzothiophene Rhodococcus erythropolis desulfinase DszB DszC Bds Bfl BdsA desuofurization

项目摘要

It is difficult to remove polycyclic sulfur compounds in fossil fuels. Dibenzothiophene(DBT) is considered as a model polycyclic sulfur compound contained in fossil fuels. Some bacteria such as Rhodococcus erythropolis D-1 utilize DBT as a sole source of sulfur without breaking its carbon-carbon backbone through the sulfur-specific pathway. In this pathway, DBT is oxidized to DBT sulfone via DBT sulfoxide by DszC,DBT sulfone is converted to 2-hydroxybiphenyl 2-sulfinic acid (HBPSi) by DszA, and HBPSi is desulfurized to 2-hydroxybiphenyl by DszB. Flavin reductase is necessary for monooxygenase reactions by DszC and DszA.The research results of this study are summarized as follows.1.DszB of R.erythropolis KA2-5-1 and its C27S mutant complexed with biphenyl-2-sulfinic acid were crystallized and preliminary X-ray crystallographic analyses were conducted.2.The gene of thermostable falvin reductase (Bfl) from Bacillus sp.DSM411, which coupled with DszC, was overproduced in Escherichia coli, and the recombinant enzymes were purified and characterized.3.Following the above 1, the mutant Q65H of DszB showed higher thermostability and optimal temperature than the parent DszB enzyme.4.Three enzymes (BdsC,A,B) involved in the DBT desulfurization were purified from the moderately thermophilic bacterium, Bacillus subtilis WU-S2B and characterized.

很难去除化石燃料中的多环硫化合物。 Dibenzothiophene（DBT）被认为是化石燃料中包含的多环硫化合物。一些细菌（例如红粒红球菌D-1）利用DBT作为硫的唯一来源，而无需通过硫的特异性途径破坏其碳碳主链。在这一途径中，DBT通过DSZC通过DSZC氧化为DBT磺酮，DBT磺酮被DSZA转化为2-羟基甲苯基2-硫代磺酸（HBPSI），而HBPSI将HBPSI置于DSZB中，将HBPSI溶于2-羟基二甲基二甲基。黄素还原酶是DSZC和DSZA对单加氧酶反应所必需的。这项研究的研究结果总结如下。1.dszbr。r。thropolis ka2-5-1及其C27S及其C27S突变体与双苯基 - 2-硫酸 - 2-硫酸 - 硫酸 - 硫酸 - 二硫酸酸的相关性，并进行了X型X型 - X射线晶型晶体学分析。（BFL）与DSZC相结合的芽孢杆菌SP.DSM411在大肠杆菌中产生过量，并纯化并表征重组酶。3。遵循上述1，DSZB的突变Q65H显示出比dszbemem的高温和最佳温度。从中等嗜热细菌，枯草芽孢杆菌WU-S2B中纯化DBT脱硫化。