产琼胶酶菌株NBRC102603发酵条件优化及酶的分离纯化  

Conditions for Enzyme Production and Purification of Agarase Produced by Agarivorans albus NBRC102603

作　　者：朱慧文[1] 孙延娜[1] 周晓龙[1] 郝梅[1] 郭振博[1] 施超[1] 朱启忠[1] 

机构地区：[1]山东大学威海分校海洋学院,山东威海264209

出　　处：《食品与发酵工业》2011年第4期120-124,共5页Food and Fermentation Industries

基　　金：国家大学创新性基金项目(101042239);山东大学(威海)科技立项重点项目专项基金(A10010)

摘　　要：通过正交试验对产琼胶酶海洋菌株NBRC102603发酵条件进行了优化,优化得到的发酵产酶条件为:蛋白胨浓度5.0 g/L、酵母膏浓度1.25 g/L、琼胶浓度4.0 g/L、装瓶量100 mL、接种量1%、转速150 r/min,28℃下发酵48 h后酶活力达到58.94 U/mL,比未优化前提高了2.08倍。经纯化得到琼胶酶A和琼胶酶B,琼胶酶A纯化倍数为17.29倍,酶比活力为870.51 U/mg;琼胶酶B纯化倍数为16.65倍,酶比活力为838.39 U/mg。纯化后琼胶酶经SDS-PAGE检测,显示为单一条带,其相对分子质量酶A约为83.6 ku、酶B约为36.8 ku。The fermentation conditions of an agar-degrading bacterium NBRC102603 isolated from the coastal water were optimized by using orthogonal experiment. The optimum condition for agarase production consisted of peptone 5.0g/L, yeast extract t. 25 g/L, agar 4.0g/L, 1% inoculating quantity,rate of shaking flask with 100 mL liquid in a 500 mL flask was 150 r/rain. After 48h fermentation at 28~C ,the enzyme activity was 58.94U/mL ,which was 2.08 times higher than before optimization. After a serious of purification processes, two types of purified agarases of NBRC102603 were obtained,identified as agarase A and agarase B. Agarase A was purified 17.29-fold, with a specific activity of 870.51U/mg; while B was purified 16.65-fold ,with a specific activity of 838.39U/mg . The purified enzyme A and B appeared to be homogeneous under the inspection of SDS-PAGE, and they had molecular masses about 83.6 ku and 36.8 ku.

关 键 词：海洋细菌 琼胶酶 正交试验 分离纯化 

分 类 号：TQ925[轻工技术与工程—发酵工程]