Effect Of Miciroclustered Water On RhaR Production By Microorganism,and Enzymatic Reaction Conditions Of RhaR

In this study,the Absidia sp.R9g was cultured in water clusters.Based on the activity of rutin-glycosidase,the efffedcts of water clusters on the growth of microorganism were assayed.The results showed that the RhaR produced by Absidia sp.R9g cultured in water clusters was much more stableale to the environment,PH,and temperature.In addition,rutin dissolved better due to higher PH,which permitted lutin to be catalyzed into soquercitrin at a higher solubility.First of all,we founded that the solublility of rutin in different pH rised as the resulta of a higer PH.Moreover,the Absidia sp.R9g cultured in cluster water for 5-6d in 30℃ induced by 2.5%Flos Sophorae showed a higer enzyme activitySecondly,the reaction condition of the rutin-a-rhamnosidase hydrolyzing rutin to isoquercitrin and rhamnose was optimized:the optimal temperature was 50℃;the optimal substrate concentration was 1.5%;the optimal pH was 10;the optimal reaction time was 14 hours.Finally,the enzyme was purified using DEAE-cellulose column.The optimum conditions of the enzyme were as follows:pH 6.0 stable at 3-10;temperature was 60℃,;stable below 60 ℃.The pure enzyme purity and molecular weight of the rutin-a-rhamnosidase was determined by SDS-PAGE,and the molecular mass was 67.5 kDa.