Department of Food Science and Technology, Chung-Ang University, Anseong, Gyeonggi 17546, Republic of Korea.
Department of Food Science and Engineering, Ewha Womans University, Seoul 03760, Republic of Korea.
J Microbiol Biotechnol. 2020 Aug 28;30(8):1244-1251. doi: 10.4014/jmb.2001.01052.
Phospholipase A (PLA) from is a lipolytic enzyme used in a wide range of industrial applications including production of lysolecithins and enzymatic degumming of edible oils. We have therefore investigated expression and secretion of PLA in two workhorse microbes, and . The PLA was produced to an activity of 0.517 ± 0.012 U/ml in the culture broth of the recombinant . On the other hand, recombinant BL21 star (DE3), overexpressing the authentic PLA (P-PLA2), showed activity of 17.0 ± 1.3 U/ml in the intracellular fraction and 21.7 ± 0.7 U/ml in the culture broth. The extracellular PLA activity obtained with the recombinant system was 3.2-fold higher than the corresponding value reached in a previous study, which employed recombinant BL21 (DE3) overexpressing codon-optimized PLA2. Finally, we observed that the extracellular PLA from the recombinant P-PLA culture was able to hydrolyze 31.1 g/l of crude soybean lecithin, an industrial substrate, to a conversion yield of approximately 95%. The newly developed -based PLA expression system led to extracellular production of PLA to a productivity of 678 U/l·h, corresponding to 157-fold higher than that obtained with the -based system. This study will contribute to the extracellular production of a catalytically active PLA.
来自 的磷脂酶 A（PLA）是一种脂解酶，广泛应用于各种工业应用，包括溶血磷脂和食用油脂的酶法脱胶的生产。因此，我们研究了 PLA 在两种工程菌 和 中的表达和分泌。在重组 的培养液中，PLA 的活性达到了 0.517±0.012 U/ml。另一方面，过表达天然 PLA（P-PLA2）的重组 BL21 星（DE3）在胞内部分的活性为 17.0±1.3 U/ml，在培养液中的活性为 21.7±0.7 U/ml。与之前使用过表达密码子优化的 PLA2 的重组 BL21（DE3）的研究相比，用重组 系统获得的细胞外 PLA 活性提高了 3 倍。最后，我们观察到重组 P-PLA 培养物中的细胞外 PLA 能够水解 31.1 g/l 的粗大豆卵磷脂，转化率约为 95%。新开发的基于 的 PLA 表达系统实现了 PLA 的细胞外生产，生产率为 678 U/l·h，比基于 的系统提高了 157 倍。本研究将有助于细胞外生产具有催化活性的 PLA。
