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.

从光合非硝化杆菌中分离出的二甲基亚硫氧化物还原酶（DMSO-R），Rhodobacter Sphaeroides F.Sp. Denitrificans不仅将硫氧化物的变种还原为相应的硫化物，而且还将N-氧化物减少到其还原形式。在减少亚硫氧化物的情况下，该酶识别不对称亚硫氧化物的立体化学，即，即硫化物和100％E.E. E.E.（r）E.E.（r）的异构体的甲基苯基硫氧化物（MPSO）。合成MPSO和其他烷基亚硫亚硫氧化亚磺氧化物（RARSO），并分析DMSO-R的立体选择性以及动力学参数，VMAX和KM。使用胶束电动色谱模式通过毛细管电泳对反应速率进行很好的监测。为合成的底物清楚地溶解了酶的立体特异性反应模式。 （2）通过将底物添加到事先诱导DMSO-R的sphaeroides培养物中的生物转化，以及（3）使用固定的DMSO-R.MTSO和甲基甲基磺基硫氧化物（MTSO）使用固定的甲基甲基硫氧化物（MTSO），对生物反应剂的降低是适用于纯正的纯度纯度的，该方法是有机化的，是有机化的 - 以革兰氏级的产量获得。

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

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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。