1株真菌漆酶高产菌株的筛选及其漆酶的纯化和性质研究  

Isolation of a high-level laccase-producing fungus strain,and purification and characterization of laccase

作　　者：李松[1] 刘宇[1] 汤文晶[1] 陈阿娜[1] 汤斌[1] 崔凤杰[2] 赵鹏翔 

机构地区：[1]安徽工程大学生物与化学工程学院,安徽芜湖241000 [2]江苏大学食品与生物工程学院,江苏镇江212013 [3]国网节能服务有限公司北京生物质能源技术中心,北京100053

出　　处：《食品与发酵工业》2015年第9期45-50,共6页Food and Fermentation Industries

基　　金：安徽省自然科学青年基金资助项目(1408085QC61);安徽工程大学引进人才科研启动基金(2013YQQ003);国家电网公司科技项目资助(合同号:SGECS56-2014)

摘　　要：使用愈创木酚筛选平板筛选分离获得1株真菌漆酶高产菌株，通过形态学观察和5．8S rDNA—ITS序列分析，将该菌株鉴定为栓菌属真菌。初步摇瓶发酵研究结果表明该菌株发酵至第10天时，漆酶活力达到20．3U／mL。发酵液经硫酸铵盐析和DEAE FF柱层析方法纯化后，该酶蛋白分子质量约为65kDa。性质研究表明，该酶在60℃以下及pH3．5～5．0较为稳定，其Vmax和Km分别为31．06mmol／（L·min）和152μmol／L。筛选得到的栓菌菌株具有产酶速率快、发酵周期短等特点。This study aimed to select high-level laccase-producing fungus strain from natural samples. The laccase producing ability of the selected strain and the properties of produced laccase were further studied. The target strain was selected and purified by guaiacol-containing selecting medium and the isolated strain was identified as Trametes sp. by morphology observation and 5.8S rDNA-ITS sequence analysis method. In shaking flask fermentation, a high laccase producing level of 20.3 u/mL was observed in the supernatant on 10 d. The produced laccase protein was salted out by ammonium sulfate fraction and further purified by DEAE FF chromatographic column. The molecular mass of the purified laccase was around 65 kDa, and the laccase was stable under 60℃ and pH 3.5 - 5.0. The Vmax and K of the laccase were 31.06 mmol/（L · min） and 152 μmol/L, respectively. The isolated fungus strain Trametes sp. has an ability to produce high-level laccase in a short period, which could make it a good candidate for breeding and optimization of industrial laccase producing strain.

关 键 词：真菌漆酶 筛选 发酵 纯化 酶学性质 

分 类 号：TQ925[轻工技术与工程—发酵工程]