Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701, South Korea.
Biotechnol Lett. 2012 Mar;34(3):483-9. doi: 10.1007/s10529-011-0786-2. Epub 2011 Nov 4.
Pyrococcus furiosus β-glucosidase converted rutin to quercetin and rutinose disaccharide with a ratio of 1:1, with no glucose, L-rhamnose, and isoquercitrin, indicating that the enzyme is a β-rutinosidase. The specific activity for flavonoid glycosides followed the order of isoquercitrin > quercitrin > rutin. The conversion of rutin to quercetin was optimal at pH 5.0 and 95°C in the presence of 0.5% dimethyl sulfoxide with a half-life of 101 h, a k(cat) of 1.6 min(-1), and a K(m) of 0.3 mM. Under the improved conditions, the enzyme produced 6.5 mM quercetin from 10 mM rutin after 150 min, with a molar yield of 65% and a productivity of 2.6 mM/h. This productivity is the highest reported thus far among enzymatic transformations.
Pyrococcus furiosus β-葡萄糖苷酶将芦丁转化为槲皮素和芦丁二糖,比例为 1:1,没有葡萄糖、L-鼠李糖和异槲皮苷,表明该酶是一种β-芦丁糖苷酶。黄酮糖苷的比活性顺序为异槲皮苷>槲皮苷>芦丁。在 pH 5.0 和 95°C 下,在 0.5%二甲亚砜存在下,芦丁转化为槲皮素的反应最佳,半衰期为 101 h,kcat 为 1.6 min-1,Km 为 0.3 mM。在优化条件下,酶在 150 分钟内从 10 mM 芦丁产生 6.5 mM 槲皮素,摩尔收率为 65%,生产能力为 2.6 mM/h。这一生产能力是迄今为止报道的酶转化中最高的。