甾醇酯酶菌株Chryseobacterium+sp的筛选和酶学性质研究.pdfVIP

甾醇酯酶菌株Chryseobacterium sp 的筛选及酶学性质研究 曾诚 范远鑫 丁少军*1 （南京林业大学，化学工程学院，江苏南京 210037） 摘要：设计了一种以植物甾醇酯为底物，罗丹明 B 为荧光指示剂的平板筛选方法,从土壤中筛选得到 24 株具有 甾醇酯酶活性的菌株，其中编号为 jxpLT 的菌株具有最高产酶活性，经 16S rDNA 鉴定表明菌株 jxpLT 属于金 黄杆菌属，命名为 Chryseobacterium sp.jxpLT。在优化的产酶条件下，菌株 jxpLT 培养 18 h 产酶活性最高可 达到 0.194 IU/ml。初步纯化了 Chryseobacterium sp.jxpLT 的甾醇酯酶并研究其部分酶学性质，该酶的酶促 o o 反应最适温度为 40 C，最适 pH 为 7.5，其在 45 C 以下，pH5.0-9.0 具有较好的稳定性。该酶对不同碳链长度 的脂肪酸胆固醇酯和对硝基苯酚酯具有广泛的底物特异性，显示该甾醇酯酶具有解决机械制浆造纸工艺中树脂 障碍问题的潜力。 关键词：甾醇酯酶;Chryseobacterium sp;酶学性质 Screening of Sterol Esterase-producing Strain Chryseobacterium sp and Its Emzymatic Properties ZENG Cheng, FAN Yuan-xin, DING Shao-jun (Faculty of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, Jangsu, China) Abstract ：Twenty four strains producing sterol esterase were screened from soil by using a plate assay in a medium containing sterol ester and the fluorescent dye rhodamine B. Among these strains, jxpLT, which was found to produce the highest level of sterol esterase,belongs to gennus Chryseobacterium based on 16S rDNA sequencing analysis and then was designated as Chryseobacterium sp.jxpLT. The enzyme activity of this strain reached 0.194 IU/ml after 18 hours induction under the optimized culture conditions. The sterol esterase was partially purified by ion exchange chromatography on DEAE Sepharose Fast Flow column. This enzyme showed an optimum activity at pH of 7.5 and temperature of 40 oC. The enzyme had good thermostability below 45 oC and pH stability at 5.0-9.0. Chryseobacterium sp.jxpLT sterol esterase can hydrolyze cholesterol esters and pNP esters of different fatty acids. It may have the potential to solve the pitch problems caused by lipophi