Tataruch M, Heider J, Bryjak J, Nowak P, Knack D, Czerniak A, Liesiene J, Szaleniec M
Jerzy Haber Institute of Catalysis and Surface Chemistry, PAS, Niezapominajek 8, 30-239 Kraków, Poland.
Laboratory for Microbial Biochemistry and Synmikro Center for Synthetic Microbiology, Philipps University of Marburg, 35043 Marburg, Germany.
J Biotechnol. 2014 Dec 20;192 Pt B:400-9. doi: 10.1016/j.jbiotec.2014.06.021. Epub 2014 Jul 3.
The molybdenum/iron-sulfur/heme protein ethylbenzene dehydrogenase (EbDH) was successfully applied to catalyze enantiospecific hydroxylation of alkylaromatic and alkylheterocyclic compounds. The optimization of the synthetic procedure involves use of the enzyme in a crude purification state that saves significant preparation effort and is more stable than purified EbDH without exhibiting unwanted side reactions. Moreover, immobilization of the enzyme on a crystalline cellulose support and changes in reaction conditions were introduced in order to increase the amounts of product formed (anaerobic atmosphere, electrochemical electron acceptor recycling or utilization of ferricyanide as alternative electron acceptor in high concentrations). We report here on an extension of effective enzyme activity from 4h to more than 10 days and final product yields of up to 0.4-0.5g/l, which represent a decent starting point for further optimization. Therefore, we expect that the hydrocarbon-hydroxylation capabilities of EbDH may be developed into a new process of industrial production of chiral alcohols.
钼/铁硫/血红素蛋白乙苯脱氢酶（EbDH）已成功应用于催化烷基芳烃和烷基杂环化合物的对映体特异性羟基化反应。合成方法的优化包括使用粗纯化状态的酶，这节省了大量的制备工作，并且比纯化的EbDH更稳定，且不会出现不必要的副反应。此外，将酶固定在结晶纤维素载体上并改变反应条件，以增加产物的生成量（厌氧气氛、电化学电子受体循环或使用高浓度铁氰化物作为替代电子受体）。我们在此报告，有效酶活性从4小时延长至超过10天，最终产物产量高达0.4 - 0.5g/l，这为进一步优化提供了一个不错的起点。因此，我们期望EbDH的烃羟基化能力能够发展成为手性醇工业生产的新工艺。
