抗生物降解药渣中高效水解木质素的菌株筛选  

Screening of strains for highly efficient hydrolyzing lignin in anti-biodegradable drug dregs

作　　者：程敏 王海洁 黄劲 赵亮 王迪 廖万清 康颖倩 CHENG Min;WANG Haijie;HUANG Jin;ZHAO Liang;WANG Di;LIAO Wanqing;KANG Yingqian(Department of Nosocomial Infection,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Microbiology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;Research Talent Base for the Relationship between Microbiology and Human Health in Guizhou Province&Key Laboratory of Pathogen Biology in Universities in Guizhou Province,Guiyang 550025,Guizhou,China;Department of Biochemistry and Molecular Biology,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou,China;Department of Dermatology,Changzheng Hospital,Naval Military Medical University,Shanghai 200003,Shanghai,China;Shanghai Key Laboratory of Molecular Medical Mycology,Shanghai 200003,China)

机构地区：[1]贵州医科大学附属医院医院感染管理科,贵州贵阳550004 [2]贵州医科大学基础医学院微生物学教研室,贵州贵阳550025 [3]贵州省微生物与人类健康关系研究人才基地&贵州省普通高校病原生物学特色重点实验室,贵州贵阳550025 [4]贵州医科大学基础医学院生物化学与分子生物学教研室,贵州贵阳550025 [5]海军军医大学长征医院皮肤科,上海200003 [6]上海市医学真菌分子生物学重点实验室,上海200003

出　　处：《贵州医科大学学报》2025年第1期24-30,共7页Journal of Guizhou Medical University

基　　金：国家自然科学基金(32060034);国家科技部及教育部“111计划”(D20009);贵州省第六批人才基地项目(RCJD2018-22);贵州省科技厅社会发展攻关支撑项目(黔科合支撑[2019]2873);贵州省科学技术厅国际科技合作基地(黔科合平台人才[2020]4101);贵州省科技计划项目(黔科合支撑[2020]4Y220)。

摘　　要：目的从抗生物降解药渣中筛选对木质素水解能力强的真菌菌株。方法通过分离鉴定、纯培养及以木质素为唯一碳源液体发酵后测定木质素过氧化物酶(lignin peroxidase,LiP)与漆酶(laccase,Lac)酶活筛选堆放药渣中高效水解木质素的真菌。结果从药渣中分离出42株真菌,经过木质素碳源平板筛选进一步发现了8株水解效果较好的丝状真菌,其中ZYJHYZ246、ZYJHYZ257、ZYJHYZ280在平板上脱色效果最明显;将以上8株真菌进行分子鉴定及LiP及Lac酶活检测发现多数菌株在发酵第5~7天时达峰值,其中阳性对照NDM3-2在第7天时产LiP酶最高,为223.59 U/L,其次是ZYJHYZ268,生长第3天时达峰值(176.85 U/L);Lac酶活结果表明,ZYJHYZ268是产酶最高的一株菌,在第5天时达到250.83 U/L,是阳性对照最高产酶时期的1.8倍。结论筛选出的ZYJHYZ268经鉴定为Piloderma sp.,其能产较高水平LiP与Lac,具有良好的水解木质素的能力,为降解木质素菌株提供了更多选择。Objective To screen fungal strains with strong hydrolyzing lignin ability from anti-biodegradable drug dregs.Methods The enzymatic activities of lignin peroxidase(LiP)and laccase(Lac)were determined to screen for fungi that efficiently hydrolyze lignin in stacked drug dregs via isolation,identification,pure cultivation and liquid fermentation using lignin as a sole carbon source.Results Forty-two strains of fungi were isolated from drug dregs,and 8 filamentous fungi with good hydrolysis effects were further found through lignin carbon source plate screening.Among them,ZYJHYZ246,ZYJHYZ257,and ZYJHYZ280 showed the most obvious decolorization effect on the plates.After molecular identification,LiP and Lac enzyme activity tests of the above 8 strains,most strains were found to reach the peak values on the 5^(th)-7^(th)days of fermentation,among which,positive control NDM3-2 had the highest LiP activity(223.59 U/L)on the 7^(th)day,followed by ZYJHYZ268,which reached the peak value(176.85 U/L)on the 3^(rd)day.Lac activity showed that ZYJHYZ268 had the highest activity,reaching 250.83 U/L on the 5^(th)day,which was 1.8 times higher than the peak enzymatic activity of the positive control.Conclusion Screened ZYJHYZ268 is identified as Piloderma sp.,which can produce high levels of LiP and Lac and has good ability to hydrolyze lignin,providing more choices for degrading lignin.

关 键 词：木质素 中草药渣 丝状真菌 水解 酶活 

分 类 号：R917[医药卫生—药物分析学]