离子液体存在下D-氨基酸氧化酶的性能

Performance of D-amino acid oxidase in presence of ionic liquids.

作者信息

Lutz-Wahl S, Trost E-M, Wagner B, Manns A, Fischer L

机构信息

Department of Biotechnology, Institute of Food Technology, University of Hohenheim, Stuttgart, Germany.

出版信息

J Biotechnol. 2006 Jun 25;124(1):163-71. doi: 10.1016/j.jbiotec.2006.01.023. Epub 2006 Mar 3.

DOI:10.1016/j.jbiotec.2006.01.023

PMID:16516324

Abstract

The activity and stability of free and immobilized D-amino acid oxidase (DAAO, EC 1.4.3.3) from Trigonopsis variabilis CBS 4095 in different water-soluble and water-insoluble ionic liquids (ILs) as well as in organic solvents were studied for comparison. The most promising ILs ([BMIM][BF(4)] and [MMIM][MMPO(4)]) were investigated in detail. The kinetic parameters (v(max) = 187 nkat/g dry weight, K(M) = 1.38 mM) with D-phenylalanine as substrate were calculated in 40% [BMIM][BF(4)]. Bioconversions of D/L-phenylalanine in 40% [BMIM][BF(4)] and 20% [MMIM][MMPO(4)] on a 3 ml scale using immobilized DAAO were performed by addition of free catalase from Micrococcus lysodeikticus. After total conversion of substrate in presence of 20% [MMIM][MMPO(4)] the residual activity of the immobilized DAAO was 79% and 100% of the free catalase.

摘要

为作比较，研究了来自可变三角酵母CBS 4095的游离和固定化D-氨基酸氧化酶（DAAO，EC 1.4.3.3）在不同水溶性和水不溶性离子液体（ILs）以及有机溶剂中的活性和稳定性。对最具前景的离子液体（[BMIM][BF(4)]和[MMIM][MMPO(4)]）进行了详细研究。在40%的[BMIM][BF(4)]中，以D-苯丙氨酸为底物计算动力学参数（v(max)=187 nkat/g干重，K(M)=1.38 mM）。使用固定化DAAO在3 ml规模上于40%的[BMIM][BF(4)]和20%的[MMIM][MMPO(4)]中进行D/L-苯丙氨酸的生物转化，通过添加来自溶壁微球菌的游离过氧化氢酶来实现。在20%的[MMIM][MMPO(4)]存在下底物完全转化后，固定化DAAO的残余活性分别为游离过氧化氢酶的79%和100%。