Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul, 143-701, Republic of Korea.
Biotechnol Lett. 2013 Sep;35(9):1487-93. doi: 10.1007/s10529-013-1231-5. Epub 2013 May 21.
Recombinant Escherichia coli, expressing the oleate hydratase gene of Stenotrophomonas maltophilia, was permeabilized by sequential treatments with 0.125 M NaCl and 2 mM EDTA. The optimal conditions for the production of 10-hydroxy-12,15(Z,Z)-octadecadienoic acid from α-linolenic acid by permeabilized cells were 35 °C and pH 7.0 with 0.1 % (v/v) Tween 40, 50 g permeabilized cells l(-1), and 17.5 g α-linolenic acid l(-1). Under these conditions, permeabilized cells produced 14.3 g 10-hydroxy-12,15(Z,Z)-octadecadienoic acid l(-1) after 18 h, with a conversion yield of 82 % (g/g) and a volumetric productivity of 0.79 g l(-1) h(-1). These values were 17 and 168 % higher than those obtained by nonpermeabilized cells, respectively. The concentration, yield, and productivity of 10-hydroxy-12,15(Z,Z)-octadecadienoic acid obtained by permeabilized cells are the highest reported thus far.
表达嗜麦芽寡养单胞菌油酸水解酶基因的重组大肠杆菌经 0.125 M NaCl 和 2 mM EDTA 顺序处理后进行渗透化处理。在 35℃和 pH 7.0 下，用 0.1％（v/v）吐温 40、50 g 渗透化细胞 l（-1）和 17.5 g α-亚麻酸 l（-1）的条件下，从α-亚麻酸生产 10-羟基-12,15（Z，Z）-十八碳二烯酸的产率最佳。在此条件下，渗透化细胞在 18 h 后产生了 14.3 g 10-羟基-12,15（Z，Z）-十八碳二烯酸，转化率为 82％（g/g），比体积生产率为 0.79 g l（-1）h（-1）。这些值分别比非渗透化细胞高 17 和 168％。与非渗透化细胞相比，渗透化细胞获得的 10-羟基-12,15（Z，Z）-十八碳二烯酸的浓度、产率和生产率最高。
