高效木质素降解菌的筛选及产漆酶条件的研究  

Study on screening of efficient lignin degrading bacteria and the conditions for laccase production

作　　者：陈建军[1] 刘梁涛 曹香林 CHEN Jian-jun;LIU Liang-tao;CAO Xiang-lin(College of Life Science,Henan Normal University,Xinxiang 453007,China;College of Aquaculture, Henan Normal University,Xinxiang 453007,China)

机构地区：[1]河南师范大学生命科学学院,河南新乡453007 [2]河南师范大学水产学院,河南新乡453007

出　　处：《甘肃农业大学学报》2018年第4期130-136,共7页Journal of Gansu Agricultural University

基　　金：国家自然科学基金项目(5201049120029);河南省重点科技攻关计划项目(122102210168;152102210081)

摘　　要：【目的】为了获得高产漆酶的菌株用于降解木质素.【方法】从新乡周边腐木堆积处、秸秆堆积处和造纸厂废水这些木质素含量丰富的地方取样,以木质素为唯一碳源,初筛出对木质素具有降解能力的菌株.【结果】采用愈创木酚平板法对产漆酶的菌株进行复筛得到4株产漆酶能力较强的菌株,通过生理生化及18S rDNA序列分析对菌株进行鉴定,通过液态发酵培养以2,2′-连氮基-双-(3-乙基苯并二氢噻唑啉-6-磺酸)(简称ABTS)法对漆酶活性和木质素降解能力进行研究.结果表明,黄孢原毛平革菌在培养6d后,漆酶活力达到最高,为233.33U/L.利用液态发酵研究了碳源、氮源、金属离子、pH、温度对其发酵产漆酶的影响,确定了该菌株的最优碳源为乳糖,最优氮源为酵母粉,最适铜离子浓度为0.6mmol/L,最适温度为30℃,最适pH为4,在此条件下木质素去除率达到了84.7%.【结论】试验筛选出的黄孢原毛平革菌具有良好的降解木质素的能力,为研究漆酶基因的克隆表达和酶学研究奠定了基础.【Objective】In order to obtain high laccase production bacteria strains for degraded lignin.【Method】The lignin-rich samples such as rotting wood,straw and paper mill wastewater were collected in Xinxiang,and the lignin was used as the sole carbon source to screen out the strains with lignin decomposition ability.【Result】Four strains of laccase-producing strains were obtained by using the guaiacol plate method.The strains were identified by physiological and biochemical analysis and 18S rDNA sequence analysis.The laccase activity and lignin decomposition ability were studied by liquid fermentation and 2,2'-azobis(3-ethylben-zothiazole-6-sulfonaic acid)(ABTS)method.The results showed that the laccase activity of Phanerochaete chrysosporium reached the highest(233.33 U/L)after 6 days culture.The effects of carbon source,nitrogen source,metal ions,pH and temperature on laccase production were studied by liquid fermentation.The optimal carbon source for the strain was lactose.The optimum nitrogen source was yeast extract,and the optimum copper concentration was 0.6 mmol/L,the optimum temperature of 30℃,the optimum pH of 4.And under these conditions,the lignin removal rate reached 84.7%.【Conclusion】Phanerochaete chrysosporium performed a good ability for lignin decomposition,and the study provided the foundation for the cloning,expression and enzymology of laccase gene.

关 键 词：漆酶 筛选 黄孢原毛平革菌 酶活 

分 类 号：Q939.9[生物学—微生物学]