菌物研究2012，Vol.10Issue(4)：234-239,6.
黑木耳漆酶纯化及部分漆酶特性的研究
Purification and Characterization of Laccase from Auricularia auricula
摘要
Abstract
Laccase of Auricularia auricula were studied for further separation and purification, gene ex- pression and mass production applications. Native SDS-PAGE for laccase extract by (NH4)2SO4 step clas- sified showed that three kinds of isoenzymes existed in "Hei 29". The molecular weight of the three isoenzymes was determined, LacA (60 kD), LacB (34 kD), LacC (19 kD). The LacC purifications were purified by ammonium sulphate fractionation, DEAE-Sephacel chromatography. Purification of about 7.60 fold was achieved with an overall yield of 4.28 %. The effects of pH, temperature and metal ion on "Hei-29" strain of Auricularia auricular laccase activity and stability were studied. Meanwhile, the sub- strate concentration effect of laccase was also studied and Km measured. The results showed that: The LacC catalyzed the value of Km of oxidation of ABTS was 1.18 ×10-6 mol/L. The optimum pH for LacC activity were 3.8, respectively in catalytic reaction of oxidizing ABTS. LacC showed a good stability when the pH of the buffer varied from 3.0 to 4.6. The optimum temperature for LacC activity was 55℃, and the laccase could work continuously under 50℃. Various metal ions showed different effects on the LacC ac- tivity. The activity was enhanced by Ag+ , and was strongly inhibited by Fe3+ , Mn2+ and Co2+ .关键词
黑木耳/漆酶/同工酶/酶学性质Key words
Auricularia auricula/laccase/isoenzyme/enzymatic property分类
农业科技引用本文复制引用
韩增华,刘佳宁,党阿丽,张丕奇,戴肖东,张介驰..黑木耳漆酶纯化及部分漆酶特性的研究[J].菌物研究,2012,10(4):234-239,6.基金项目
黑龙江省科学院基金项目 （）
