白腐真菌组合培养提高漆酶酶活的作用机制研究  

Enhancement of Laccase Activity by Combining White Rot Fungal Strains

作　　者：何荣玉[1,2] 刘晓风[1] 闫志英[1] 袁月祥[1] 廖银章[1] 李旭东[1] 

机构地区：[1]中国科学院成都生物研究所,成都610041 [2]中国科学院研究生院,北京100049

出　　处：《环境科学》2010年第2期465-471,共7页Environmental Science

基　　金：国家高技术研究发展计划(863)项目(2007AA1007002);中国科学院知识创新工程重大项目(KSC2-YW-G-008);国家科技支撑技术项目(2006BAD07A02)

摘　　要：采用白腐真菌组合培养的方式提高漆酶酶活,并对组合后菌株相互作用机制进行了研究.菌株55(Trametes trogii)和菌株m-6(Trametes versicolor)组合后漆酶酶活较菌株55和m-6分别提高了24.13倍和4.07倍;组合后菌株间不存在抑制作用;平板培养时,两菌株菌丝生长止于菌丝交界处,该处漆酶酶活最高并分泌褐色色素;液体培养时,菌株m-6对组合后漆酶酶活的提高起着更为重要的作用:向菌株55的培养物中添加菌块m-6,其酶活比向菌株m-6培养物中添加菌块55时的酶活高7.03倍,并且菌株m-6胞外物对菌株55的漆酶分泌也有明显的刺激作用,其中加入20 mL过滤灭菌胞外物可使菌株55漆酶酶活提高6.79倍,而且胞外物高温灭菌后仍能刺激菌株55的漆酶分泌,加入20mL后酶活比对照高4.60倍;Native-PAGE活性染色结果表明组合后同工酶种类未发生变化,但有3种同工酶的浓度升高.The method of combining white rot fungal strains was used to enhance laccase activity, and the interaction mechanism between strains was also studied. The laccase activity of combined fungi of strain 55 （ Trametes trogii） and strain m-6 （ Trametes versicolor） were 24. 13 and 4.07-fold higher than that of strain 55 and strain m-6, respectively. No inhibitory effect was observed when the two strains were co-cultivated. On plate cultivation, there was hyphal interference in the contact area, where laccase activity was the highest followed by brown pigmentation. In liquid cultivation, strain m-6 played much more important role on enhancement of laccase activity, and the laccase activity of strain 55 by adding strain m-6 was 7.03-fold higher than that of strain m-6 by adding strain 55, furthermore, filter sterilized- and high temperature autoclaved-extracellular substances of strain m-6 could also stimulate strain 55 to excrete more laccase, which led to 6. 79-fold and 4. 60-fold increase in laccase activity by adding 20mL, respectively. The native staining results of Native-PAGE showed that the types of laccase isozymes were not changed when strains were co-cultured, but the concentration of three types increased.

关 键 词：漆酶 组合白腐真菌 胞外物 同工酶 

分 类 号：X172[环境科学与工程—环境科学]