Elucidation of functions of novel enzymes catalyzing the formation and the cleavage of cabond and their application

新型酶催化碳键形成和裂解的功能阐明及其应用

基本信息

批准号：
09650877
负责人：
IZUMI Yoshikazu
金额：
$ 2.05万
依托单位：
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：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09650877/
关键词：
biotin BioB bioB dibenzothiophen (DBT)DszC,A 脱硫菌 DszA DszC

项目摘要

The research results of this study are summarized as follows.(1) Studies on biotin synthase(1-1) Biotin sythase (BioB) was purified with a high yield and with a high purity from cell-free extract transformant of Bacillus sphaericus which could overexpress BioB.Using the purified BioB enzyme biotin analogs, thiohene valeric acid (TVA) and acidomycin (ACM), were found to completely in synthase reaction at a concentration of 20 mM and 10 mM, respectively.(1-2) Stimulatory factors essential for biotin synthase reactions were found. Through ammonium sulfate fi* and column chromatographies of cell-free extract of a wild type strain of Bacillus aphaericus, a low weight fraction (LF), which was heat-stable and passed through ultramembrane of cut-off MW=10,000, molecular weight fraction (HF), which was heat-lable and did not pass through the membrane, wen* Neither of LFand HF alone showed stimulatory activity toward biotin synthase reaction, but activity w* both of them were added together in the reaction mixutre of biotin synthase. Since mon* stimulatory proteins may exist in the HF.further purification of HF was considered to be necessary(2) Studies on the purification of enzymes of dibenzothiophene (DRI) desulfurization metabolism(2-1) DBT monoxygenase (DszC) and DBT sulfone monooxygenase (DszA) were purified from a DBT ck Rhodxoccus eryrhmpolis D-1 and their reactions were found to essentially rerpuire reductase. I DszC and DszA were successfully ciystallized.(2-2) Sulfinase (DszB) which catalyzes the last step of DBT desulfurization metabolism, was also purl degree from another DBT desulfurizing Rhod2coccus eythmpolis KA2-5-l by various chromatoraphies.

（1）对生物素合酶（1-1）生物素Sythase（Biob）的研究总结为以下。纯化的生物素Sythase（Biob）的产量很高，并且具有较高的细胞提取物转化为芽孢杆菌的无细胞提取物，可以过度表达生物蛋白的生物蛋白（thio Biobin ot Actir）（thio thio thio anticion，actio）酸（thio thio anticion）（TTVA）酸（TVA）酸性（TVEA）酸性（TVA），这是纯化的。浓度分别为20 mm和10 mm的合酶反应。（1-2）发现生物素合酶反应必不可少的刺激因子。通过硫酸铵FI*和无细胞提取物的无细胞提取物的野生型植菌菌株的无细胞提取物，低体重分数（LF），它是热稳定的，并通过截止MW = 10,000的超脑，分子量分数（HF）都不是热量的，并且均无法通过刺激性刺激，并且均无法通过本节均匀，wen**反应，但它们的活性W*在生物素合酶的反应混合物中加在一起。由于HF中可能存在Mon*刺激性蛋白，因此认为HF的纯化被认为是必要的（2）研究二苯噻吩（DRI）脱硫代谢（2-1）DBT dbt一氧化酶（DSZC）和DBT Sulfone sulfone sulfone sulfied a a a a a a a a a a a a a a d d d d d d d d d d d d d d d d d ccys purs pur pur pur pur pur purifience（2）研究了二苯噻吩（DRI）脱硫代谢（2-1）dbt solfied a ddciSai depcys a d coska） Eryrhmpolis D-1及其反应基本上是旋转还原酶。 I DSZC和DSZA已成功地构成。（2-2）硫酸盐酶（DSZB）催化DBT DEBT去硫化代谢的最后一步，也从另一个DBT Desulfulizing Rhodfulizing Rhod2coccus eythmpolis eythmpolis ka2-5-l中获得了Purl学位。