Division of Biotechnology, The Catholic University of Korea, Bucheon 14662, Republic of Korea.
C1Chem Co, Ltd., 405 Sogang Business Center, Seoul 04107, Republic of Korea.
J Microbiol Biotechnol. 2022 May 28;32(5):672-679. doi: 10.4014/jmb.2112.12055.
Microbial lipases are used widely in the synthesis of various compounds due to their substrate specificity and position specificity. 4-Ethyl malate (4-EM) made from diethyl malate (DEM) is an important starting material used to make argon fluoride (ArF) photoresist. We tested several microbial lipases and found that M37 lipase position-specifically hydrolyzed DEM to produce 4-EM. We purified the reaction product through silica gel chromatography and confirmed that it was 4-EM through nuclear magnetic resonance analysis. To mass-produce 4-EM, DEM hydrolysis reaction was performed using an enzyme reactor system that could automatically control the temperature and pH. Effects of temperature and pH on the reaction process were investigated. As a result, 50°C and pH 4.0 were confirmed as optimal reaction conditions, meaning that M37 was specifically an acid lipase. When the substrate concentration was increased to 6% corresponding to 0.32 M, the reaction yield reached almost 100%. When the substrate concentration was further increased to 12%, the reaction yield was 81%. This enzyme reactor system and position-specific M37 lipase can be used to mass-produce 4-EM, which is required to synthesize ArF photoresist.
微生物脂肪酶由于其底物特异性和位置特异性，被广泛应用于各种化合物的合成中。由二乙酯马来酸（DEM）合成的 4-乙基马来酸（4-EM）是一种重要的起始原料，用于合成氩氟（ArF）光刻胶。我们测试了几种微生物脂肪酶，发现 M37 脂肪酶能特异性地水解 DEM 生成 4-EM。我们通过硅胶色谱法对反应产物进行了纯化，并通过核磁共振分析确认其为 4-EM。为了大规模生产 4-EM，我们使用能够自动控制温度和 pH 值的酶反应器系统进行 DEM 水解反应。考察了温度和 pH 值对反应过程的影响。结果表明，50°C 和 pH 值 4.0 为最佳反应条件，这意味着 M37 是一种特异性的酸性脂肪酶。当底物浓度增加至对应 0.32M 的 6%时，反应收率几乎达到 100%。当底物浓度进一步增加至 12%时，反应收率为 81%。这种酶反应器系统和位置特异性的 M37 脂肪酶可用于大规模生产 4-EM，这是合成 ArF 光刻胶所必需的。
