Application of DMSO reductase to asymmetric synthesis and its large scale praparation of the enzyme.

DMSO还原酶在不对称合成中的应用及其大规模制备。

• 批准号: 07556027
• 负责人: YAMAZAKI Sunao
• 金额: $ 6.14万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07556027/
• 关键词: DMSO reductase; Rhodobacter sphaeroides; reduction of sukfoxide; reduction of N-oxide; asymmetric synthesis; stereoselective synthesis; 不斉合成; キラルスルホキシド; N-オキシド; 不斉還元; 脱窒光合成細菌; Rhodobacter Sphaeroides; スルフォキシド還元; 不斉還元合成; 微生物還元

Dimethyl sulfoxide reductase (DMSO-R) isolated from the photosynthetic denitrifyingbacterium, Rhodobacter sphaeroides f.sp. denitrificans, reduces not only varieties of sulfoxides to the corresponding sulfides but also N-oxides to their reduced forms. In the case of sulfoxide reduction, the enzyme recognizes the stereochemistry of asymmetric sulfoxides, i.e., (S) -methylphenylsulfoxide (MPSO) of the racemic mixtureis exclusivelyreduced to the sulfide and 100% e.e.(R) isomer is recovered. MPSO and other alkylaryl sulfoxides (RArSO) were synthesized and assayd for the stereoselectivity of DMSO-R as well as for the kinetic parameters, Vmax and Km. Reaction rate was well monitored by capillary electrophoresis using the micellar electrokinetic chromatographic mode. Stereospecific reaction pattern of the enzyme was clearly solved for the synthesized substrates, RArSO.Three preparative methods to obtain optically pure sulfoxides were examined : (1) reduction by in vitro enzyme reaction which requires pure enzymes ; (2) biotransformation by adding substrates to the culture of Rhodobacter sphaeroides in which DMSO-R was beforehand induced and (3) reduction in bioreactor using immobilized DMSO-R.MTSO and methyltolylsulfoxide (MTSO) which are important intermediates for the asymmetric organic synthesis were applied to the method (2) and optically pure (R) isomers were obtained in gram-order yield.

从光合细菌Rhodobacter sphaeroides f. sp. p.中分离的二甲基亚砜还原酶（DMSO-R）不仅能将各种亚砜还原成相应的硫化物，而且能将N-氧化物还原成还原态。在亚砜还原的情况下，酶识别不对称亚砜的立体化学，即，(S)外消旋混合物中的-甲基苯亚砜（MPSO）被完全还原为硫化物，其e.e.为100%。(R)异构体被回收。合成了MPSO和其它烷基芳基亚砜（RArSO），并测定了DMSO-R的立体选择性和动力学参数Vmax和Km。采用毛细管电泳胶束电动色谱模式对反应速率进行了监测。研究了三种制备光学纯亚砜的方法：（1）需要纯酶的体外酶还原法;（2）通过向其中预先诱导DMSO-R的球形红细菌培养物中加入底物进行生物转化，和（3）在生物反应器中使用固定化DMSO-将不对称有机合成的重要中间体MTSO和甲基甲苯基亚砜（MTSO）应用于方法（2），得到了克级产率的光学纯（R）异构体。

项目成果

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I.Yamamoto、N.Wada、T.Ujiiye、M.Tachibana、M.Matsuzaki、H.Kajiwara、Y.Watanabe、H.Hirano、A.Okubo、T.Satoh 和 S.Yamazaki：“”克隆和核苷酸序列

K.Sato, H.Sasaki, A.Okubo, M.Tanokura and S.Yamazaki: "Crystallization and Preliminary Crystallographic Studies of Dimethyl Sulfoxide Reductase from Rhodobacter sphaeroides f.sp.denitrificans." Proc.Japan Acad.73, Ser.B. 30-32 (1997)

K.Sato、H.Sasaki、A.Okubo、M.Tanokura 和 S.Yamazaki：“球形红杆菌反硝化细菌二甲亚砜还原酶的结晶和初步晶体学研究”。

安保 充 他: "ジメチルスルフォキシド還元酵素によるスルフォキシドの速度論的分割" 分析化学. 44. 835-838 (1995)

Mitsuru Anbo 等人：“二甲亚砜还原酶对亚砜的动力学拆分”，《分析化学》44. 835-838 (1995)。

M.Abo, M.Tachibana, A.Okubo and S.Yamazaki: ""Enantioselective Deoxygenation of Alkyl Aryl Sulfoxides by DMSO Reductase from Rhodobacter sphaeroides f.s.denitrificans."" Bioorg.Med.Chem.3. 109-112 (1995)

M.Abo、M.Tachibana、A.Okubo 和 S.Yamazaki：“通过 DMSO 还原酶从球形红细菌 f.s.denitificans 中对烷基芳基亚砜进行对映选择性脱氧。”Bioorg.Med.Chem.3。