双酶法生物合成L-苯基乙酰基甲醇的研究

双酶偶联体系越来越受到人们重视,广泛应用于医药、食品等领域,是实现高效生物合成的重要方法。本文利用乳酸氧化酶(Lactate Oxidase,LOD)和丙酮酸脱羧酶(Pyruvatedecarboxylase,PDC)偶联构建双酶体系,催化乳酸钠和苯甲醛合成L-苯基乙酰基甲醇(L-Phenylacetylcarbinol,L-PAC),后者是合成L-麻黄素的重要中间体。转化反应的最佳温度为34℃,pH 6.8,转化时间6 h,苯甲醛用量160 mmol/L,乳酸钠用量330 mmol/L,葡萄糖用量3 g/100 mL,L-PAC产量达到8.79 g/L。在L-PAC的制备过程中,以廉价的乳酸钠代替丙酮酸大大降低了生产成本,缩短了反应时间,简化了操作步骤,有利于工业生产。双酶偶联体系为生物合成L-PAC提供了一个新的思路和新方法。

Study on Biotransformation of L-Phenylacetylcarbinol Using Dual-Enzyme Coupled Reaction System

With the development of biotechnology,Dual-enzyme Coupled Reaction System has been put on increaseing emphasis in the study.As an important method for biotransformation,it has been widely used in the respect of medicine and foodstuff.In this paper,biotransformation of L-phenylacetylcarbinol(L-PAC),a key intermediate for L-ephedrine synthesis,has been evaluated by coupled Lactate Oxidase and Pyruvatedecarboxylase system,using benzaldehyde and sodium lactate as substrate.The results suggested that the yield reached the highest for six hour’s biotransformation at 34 ℃ and pH 6.8,with L-PAC yield of 8.79 g/L by adding 160 mmol/L benzaldehyde,330 mmol/L sodium lactate and 3 g/100 mL glucose in batches.In the preparation of L-PAC,compared with the consumption of pyruvic acid,sodium lactate as the substitute can largely reduce the production cost,cut down biotransformation time and simplify the operational processes in L-PAC industry.Dual-enzyme Coupled System provides a novel method and a new direction of biotransformation of L-PAC.