Characterization of benzaldehyde lyase from Pseudomonas fluorescens:: A versatile enzyme for asymmetric C-C bond formation
Characterization of benzaldehyde lyase from Pseudomonas fluorescens:: A versatile enzyme for asymmetric C-C bond formation
复制标题
DOI：
10.1016/j.bioorg.2006.09.002
复制
发表时间：
2006-12-01
影响因子：
5.1
通讯作者：
Pohl, Martina
中科院分区：
文献类型：
--
作者：
Janzen, Elena;Mueller, Michael;Pohl, Martina
二磷酸硫胺素依赖的苯甲醛裂解酶是化学酶法合成(R)-羟基酮类化合物的重要催化剂。我们研究了重组酶及其变异体对pH、温度、缓冲盐、辅因子和有机共溶剂的稳定性。在化学酶法合成中，BAL的稳定性需要在缓冲液中加入辅因子。反应温度不应超过37℃，酶在pH 6~8范围内稳定，pH 8是裂解酶和连接酶反应的最适pH。结果表明，磷酸二氢钾和Tris是最佳的反应缓冲剂，加入20%DMSO有利于提高芳香族底物和产物的溶解度和BAL的稳定性。BAL催化的反应最初的广泛产物范围已经扩大到包括高取代度的羟基丁苯酮和脂肪族酰内酯。(C)2006 Elsevier Inc.保留所有权利。
The thiamin-diphosphate-dependent enzyme benzaldehyde lyase is a very import catalyst for chemoenzymatic synthesis catalyzing the formation and cleavage of (R)-hydroxy ketones. We have studied the stability of the recombinant enzyme and some enzyme variants with respect to pH, temperature, buffer salt, cofactors and organic cosolvents. Stability of BAL in chemoenzymatic synthesis requires the addition of cofactors to the buffer. Reaction temperature should not exceed 37 degrees C. The enzyme is stable between pH 6 and 8, with pH 8 being the pH-optimum of both the lyase and the ligase reaction. Potassium phosphate and Tris were identified as optimal reaction buffers and the addition of 20 vol% DMSO is useful to enhance both the solubility of aromatic substrates and products and the stability of BAL. The initial broad product range of BAL-catalyzed reactions has been enlarged to include highly substituted hydroxybutyrophenones and aliphatic acyloins. (c) 2006 Elsevier Inc. All rights reserved.