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Development of Biocatalysts Using Organized Nano-reactors

利用有序纳米反应器开发生物催化剂

基本信息

批准号：
15550093
负责人：
EMA Tadashi
金额：
$ 2.43万
依托单位：
OKAYAMA 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

项目摘要

Kinetic resolution of 1,1-diphenyl-2-propanol was performed at high temperatures using a lipase immobilized on porous ceramics, Toyonite 200M. Reactions proceeded even at 120℃, giving enantiomerically pure ester product. In contrast, the reaction at 100℃ using the same lipase immobilized on Celite resulted in no reaction. The high-temperature biocatalysis in supercritical CO_2 medium at 80℃ gave 42% conversion with an E value of 211. The lipase-catalyzed acylation of a secondary alcohol having tetraphenylporphyrin as a substituent proceeded at 60℃ most efficiently, giving enantiomerically pure ester and alcohol.Lipase mutant was immobilized on the porous ceramics to study how far the enantioselectivity can be controlled on the basis of a reaction mechanism. Substitution of some amino acid residue in the proximity of the active site with another one resulted in a change in enantioselectivity, as expected. Thermodynamic parameters suggested that the change in enantioselectivity was driven by the change in activation enthalpy.A new method for the prediction of absolute configurations of secondary alcohols using immobilized lipase was invented. Using the E values for two kinds of 1-substituted ethanols having different substituents, the E value for another secondary alcohol having the two substituents on both sides was predicted and determined. The experimental results for 19 out of 20 examples were consistent with the prediction.A carbonyl reductase obtained with recombinant E.coli was immobilized on the porous ceramics, which unfortunately showed no remarkable effect. Rather, whole-cell reductions using E.coli coexpressing two genes encoding the carbonyl reductase and a glucose dehydrogenase gave optically active alcohols much more easily and efficiently. Highly enantioselective asymmetric reductions were achieved, and 8 out of 16 ketones were reduced to optically active alcohols with >98% ee.

在高温条件下，用固定化脂肪酶对1，1-二苯基-2-丙醇进行了动力学拆分。反应甚至在120℃下进行，得到了对映体纯净的酯类产品。相反，在100℃的条件下，用相同的固定化脂肪酶固定在Celite上的反应没有发生反应。在超临界CO_2介质中，80℃的高温生物催化转化率为42%，E值为211。脂肪酶催化的以四苯基卟啉为取代基的仲醇的酰化反应在60℃下进行，得到了对映体纯度较高的酯和醇。正如预期的那样，活性中心附近的一些氨基酸残基被另一个氨基酸取代后，对映体选择性发生了变化。热力学参数表明，对映体选择性的变化是由活化热的变化驱动的。发明了一种利用固定化脂肪酶预测仲醇绝对构型的新方法。利用两种具有不同取代基的1-取代乙醇的E值，预测和测定了另一种具有两个取代基的仲醇的E值。20个样品中有19个样品的实验结果与预测一致。用重组大肠杆菌获得的羰基还原酶被固定在多孔陶瓷上，但效果并不显著。相反，使用共表达两个编码羰基还原酶和葡萄糖脱氢酶的基因的大肠杆菌进行全细胞还原，可以更容易、更有效地产生光学活性酒精。实现了高对映选择性的不对称还原，16个酮中有8个被还原为光学活性醇，其ee为98%。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Highly Enantioselective Lipase-Catalyzed Reactions at High Temperatures up to 120℃ in Organic Solvent.

在有机溶剂中在高达 120℃ 的高温下进行高度对映选择性脂肪酶催化反应。

DOI：
发表时间：
2003
期刊：
Tetrahedron : Asymmetry 14
影响因子：
0
作者：
Ema;T.;Kageyama;M.;Korenaga;T.;Sakai;T.
通讯作者：
T.

Asymmetric reduction of a variety of ketones with a recombinant carbonyl reductase: identification of the gene encoding a versatile biocatalyst

DOI：
10.1016/j.tetasy.2005.02.004
发表时间：
2005-03
期刊：
Tetrahedron-asymmetry
影响因子：
0
作者：
T. Ema;Hideo Yagasaki;Nobuyasu Okita;Kumiko Nishikawa;T. Korenaga;T. Sakai
通讯作者：
T. Ema;Hideo Yagasaki;Nobuyasu Okita;Kumiko Nishikawa;T. Korenaga;T. Sakai

Ema, T., Kageyama, M., Korenaga, T., Sakai, T.: "Highly Enantioselective Lipase-Catalyzed Reactions at High Temperatures up to 120℃ in Organic Solvent"Tetrahedron : Asymmetry. 14・24. 3943-3947 (2003)

Ema, T.、Kageyama, M.、Korenaga, T.、Sakai, T.：“有机溶剂中高达 120℃ 的高温下的高度对映选择性脂肪酶催化反应”四面体 14・24。 2003）