Department of Bioscience and Biotechnology, Konkuk University, Seoul, Republic of Korea.
Appl Microbiol Biotechnol. 2012 Aug;95(4):929-37. doi: 10.1007/s00253-011-3805-2. Epub 2011 Dec 22.
A recombinant enzyme from Lysinibacillus fusiformis was expressed, purified, and identified as an oleate hydratase because the hydration activity of the enzyme was the highest for oleic acid (with a k (cat) of 850 min(-1) and a K (m) of 540 μM), followed by palmitoleic acid, γ-linolenic acid, linoleic acid, myristoleic acid, and α-linolenic acid. The optimal reaction conditions for the enzymatic production of 10-hydroxystearic acid were pH 6.5, 35 °C, 4% (v/v) ethanol, 2,500 U ml(-1) (8.3 mg ml(-1)) of enzyme, and 40 g l(-1) oleic acid. Under these conditions, 40 g l(-1) (142 mM) oleic acid was converted into 40 g l(-1) (133 mM) 10-hydroxystearic acid for 150 min, with a molar yield of 94% and a productivity of 16 g l(-1) h(-1), and olive oil hydrolyzate containing 40 g l(-1) oleic acid was converted into 40 g l(-1) 10-hydroxystearic acid for 300 min, with a productivity of 8 g l(-1) h(-1).
从溶杆菌中表达、纯化并鉴定出一种脂肪酸水合酶,因为该酶对油酸的水合活性最高(kcat 为 850 min-1,Km 为 540 μM),其次是棕榈油酸、γ-亚麻酸、亚油酸、豆蔻酸和α-亚麻酸。酶法生产 10-羟基硬脂酸的最佳反应条件为 pH 6.5、35°C、4%(v/v)乙醇、2500 U ml-1(8.3 mg ml-1)的酶和 40 g l-1油酸。在此条件下,40 g l-1(142 mM)油酸转化为 40 g l-1(133 mM)10-羟基硬脂酸,反应 150 min,摩尔收率为 94%,生产强度为 16 g l-1 h-1,橄榄油水解物中含有 40 g l-1油酸,转化为 40 g l-1 10-羟基硬脂酸 300 min,生产强度为 8 g l-1 h-1。