Characterization of benzaldehyde lyase from Pseudomonas fluorescens:: A versatile enzyme for asymmetric C-C bond formation

被引：65

作者：

Janzen, Elena

Mueller, Michael

Kolter-Jung, Doris

Kneen, Malea M.

McLeish, Michael J.

Pohl, Martina ^{[1
]}

机构：

[1] Univ Dusseldorf, Inst Enzyme Technol, Res Ctr Juelich, D-52426 Julich, Germany

[2] Univ Freiburg, Inst Pharmaceut Sci, D-79104 Freiburg, Germany

[3] Res Ctr Juelich, Inst Biotechnol, D-52425 Julich, Germany

[4] Univ Michigan, Coll Pharm, Ann Arbor, MI 48109 USA

来源：

BIOORGANIC CHEMISTRY | 2006年 / 34卷 / 06期

基金：

美国国家科学基金会;

关键词：

biocatalysis; thiamin diphosphate; stability investigation; benzoin; 2-hydroxy ketones; benzoylformate decarboxylase;

D O I：

10.1016/j.bioorg.2006.09.002

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：345 / 361

页数：17

共 32 条

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