Application of biocatalysts for development of non-natural organosilicon compounds

生物催化剂在非天然有机硅化合物开发中的应用

• 批准号: 04805102
• 负责人: SONOMOTO Kenji
• 金额: $ 1.22万
• 依托单位: KYUSHU UNIVERSITY (1993)Kyushu Institute of Technology (1992)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04805102/
• 关键词: Organosilicon compounds; Sila-substitution effects; Sila-substituted alcohols; stereoselective esterification; Alcohol dehydrogenase; Lipase; Trimethylsilyl group; beta-effect; 電子的安定化効果; エステル化反応; 光学分割; Candida cylindraceaリパーゼ; Pseudomonas sp.リ

Comparative studies were made of the use as acyl acceptor of organosilicon compounds [Me_3Si(CH_2)_nOH] and the corresponding carbon compounds [Me_3C(CH_2)_nOH] in the stereoselective esterification of 2-(4-chlorophenoxy)propanoic acid by the use of lipase OF 360 of candida cylindracea in water-saturated benzene. The organosilicon compounds were effectively used as the substrates for the esterification of D-acid enantiomer. Of the organosilicon compounds of different chain-length between the silicon atom and the hydroxyl group, trimethylsilylmethanol (n=1) enabled the esterification reaction to be both fast and highly stereoselective, which was difficult with conventional substrates such as its carbon counterpart. On the other hand, no difference was observed between trimethylsilylethanol (n=2) and its carbon analogue (3,3-dimethylbutanol) in the enzymatic activity and the stereoselectivity. These results indicate that the silicon atom behaved as a mimic carbon atom for lipase in the c … More ase of trimethylsilylethanol but was effective in enhancing the reactivity of trimethylsilylmethanol. The difference could be explained on the basis of the properties of silicon atom, such as its low electronegativity and big atomic radius compared with the carbon atom.Among the commercially available alcohol dehydrogenases from different sources tested, equine lliver alcohol dehydrogenase (HLADH) recognized several organosilicon compounds, sila-substituted alcohols, as a substrate and exhibited the oxidative activity. Especially, large differences in catalytic rates were observed between trimethylsilyl alcohols [Me_3Si(CH_2)_nOH] but not between the carbon analogues. Thus, the activity of HLADH was extremely and peculiarly affected by trimethylsilyl group in the organosilicon compounds [trimethylsilylmethanol (n=1), 27% ; trimethylsilylethanol (n=2), 212% ; trimethylsilylpropanol (n=3), 65% ; ethanol, 100%]. These differences might result from beta-effect of trimethylsilyl group (stabilization of a negative charge in the carbon to the silicon and ・ a positive charge on the carbon to the silicon) by considering the possible mechanisms for hydrogen transfer at HLADH active site. As a result of kinetic investigations with HLADH, trimethylsilylethanol was found to show high affinity compared with the others. Furthermore, enhancement of HLADH activity derived from sila-substitution was not easily provided even by the use of the corresponding carbon analogues with electron attractive group (for example, 3-methoxy-3-methylbutanol). Less

比较研究了有机硅化合物[Me_3Si(CH_2)_nOH]和相应的碳化合物[Me_3C(CH_2)_nOH]作为酰基受体，利用白茅假丝酵母360脂肪酶在水饱和苯中立体选择性酯化2-（4-氯苯氧基）丙酸。有机硅化合物被有效地用作d -酸对映体酯化反应的底物。在硅原子和羟基之间的不同链长的有机硅化合物中，三甲基硅基甲醇（n=1）使酯化反应既快速又具有高度的立体选择性，这在传统的底物（如碳对应物）中很难实现。另一方面，三甲基硅基乙醇（n=2）与其碳类似物（3,3-二甲基丁醇）在酶活性和立体选择性方面没有差异。这些结果表明，硅原子在三甲基硅基乙醇的c…More酶中充当脂肪酶的模拟碳原子，并能有效地提高三甲基硅基甲醇的反应活性。与碳原子相比，硅原子的电负性低、原子半径大等特性可以解释这种差异。在市售的不同来源的醇脱氢酶中，马肝脏醇脱氢酶（HLADH）识别几种有机硅化合物，硅取代醇作为底物，并表现出氧化活性。三甲基硅醇[Me_3Si(CH_2)_nOH]之间的催化速率差异较大，而碳类似物之间的催化速率差异不大。因此，有机硅化合物中三甲基硅基对HLADH活性的影响是极其特殊的[三甲基硅基甲醇（n=1）， 27%；三甲基硅乙醇（n=2）， 212%；三甲基硅丙醇（n=3）， 65%；乙醇,100%)。考虑氢在HLADH活性位点转移的可能机制，这些差异可能是由于三甲基硅基的β效应（稳定碳对硅的负电荷和碳对硅的正电荷）造成的。通过HLADH的动力学研究，发现三甲基硅基乙醇与其他溶剂相比具有较高的亲和性。此外，即使使用相应的具有电子吸引基团的碳类似物（例如3-甲氧基-3-甲基丁醇），也不容易通过硅取代获得HLADH活性的增强。少

项目成果

C.-S.Rui: "A flow-injection biosensor system for the amperometric determination of creatinine : simultaneous compensation of endogenous interferents." Anal.Biochem.210. 163-171 (1993)

C.-S.Rui：“用于电流测定肌酐的流动注射生物传感器系统：同时补偿内源性干扰物。”

K.Sonomoto: "Integrated bioreactor system for biologically active peptides from isolated soybean protein." Ann.N.Y.Acad.Sci.in press. (1994)

K.Sonomoto：“用于从分离的大豆蛋白中提取生物活性肽的集成生物反应器系统。”

C.-S.Rui: "A multifunctional flow-injection biosensor for the simultaneous determination of ammonia, creatinine and urea." Ann.N.Y.Acad.Sci.in press. (1994)

C.-S.Rui：“一种用于同时测定氨、肌酐和尿素的多功能流动注射生物传感器。”

T.Yamakido: "Enzymatic hydrolysis of castor oil for valuable fatty acid production." Ann.N.Y.Acad.Sci.in press. (1994)

T.Yamakido：“酶法水解蓖麻油以生产有价值的脂肪酸。”

K.Sonomoto: "Integrated bioreactor system for biologically active peptides from isolated soybean protein." Ann. N. Y. Acad. Sci.(in press). (1994)

K.Sonomoto：“用于从分离的大豆蛋白中提取生物活性肽的集成生物反应器系统。”