灰盖鬼伞中过表达的碱性真菌漆酶在高盐染料废水中的应用潜力  

An alkaline fungal laccase overexpressed in Coprinopsis cinerea shows application potential in treating high-salt dye wastewater

作　　者：王先华 朱雪玲 林嘉钇 谷贤富 彭齐霞 肖亚中[1,2,3] Ursula Kües 方泽民 WANG Xianhua;ZHU Xueling;LIN Jiayi;GU Xianfu;PENG Qixia;XIAO Yazhong;Ursula Kües;FANG Zemin(School of Life Sciences,Anhui University,Hefei 230601,Anhui,China;Anhui Key Laboratory of Modern Biomanufacturing,Hefei 230601,Anhui,China;Anhui Provincial Engineering Technology Research Center of Microorganisms and Biocatalysis,Hefei 230601,Anhui,China;University of Goettingen,Goettingen 237077,Germany)

机构地区：[1]安徽大学生命科学学院,安徽合肥230601 [2]安徽省现代生物制造重点实验室,安徽合肥230601 [3]安徽省微生物与生物催化工程技术研究中心,安徽合肥230601 [4]University of Goettingen,Goettingen 237077,Germany

出　　处：《微生物学报》2024年第8期3014-3029,共16页Acta Microbiologica Sinica

基　　金：国家自然科学基金(32370133,31870068,U22A20442);国家重点研发计划(2021YFC2103000);安徽省杰出青年学者科学基金(2008085J12)。

摘　　要：【目的】本研究利用双孢菇(Agaricus bisporus)gpdII启动子,实现了碱性真菌漆酶PIE5(CcPIE5)在灰盖鬼伞(Coprinopsis cinerea)FA2222中的同源过表达。【方法】在mKjalke液体培养基中37℃培养7 d,漆酶活力达到(24.2±1.1)U/mL。纯化的CcPIE5在pH 8.0和60℃下表现出最佳催化活性。【结果】与已经研究报道的真菌漆酶不同,CcPIE5能耐受高浓度NaCl,当NaCl浓度从0升至1.5 mol/L时,CcPIE5的k_(cat)和K_(m)均呈下降趋势,表示CcPIE5在纺织印染废水脱色中有潜在应用价值。进一步在染料废水脱色应用中,以丁香酸作为介体,CcPIE5在pH 8.5和60℃条件下可高效降解(92.9±2.3)%的靛蓝胭脂红。通过LC-MS分析确认,Isatin 5-sulfonic acid(ISA)是靛蓝胭脂红降解的主要副产物。【结论】CcPIE5在高温、碱性和含盐条件下高效脱色染料,是处理环境和工业特定应用的理想选择。[Objective]To homologously overexpress an alkaline fungal laccase PIE5(CcPIE5)in Coprinopsis cinerea FA2222 under the control of the Agaricus bisporus gpdII promoter.[Methods]The laccase activity reached(24.2±1.1)U/mL in the supernatant after 7 days of cultivation at 37℃in the mKjalke medium.The purified CcPIE5 showcased the best performance at pH 8.0 and 60℃.[Results]Unlike other characterized fungal laccases,CcPIE5 was tolerant to high concentrations of NaCl.Particularly,both k_(cat)and K_(m)decreased when the concentration of NaCl was increased from 0 to 1.5 mol/L,which indicated that CcPIE5 demonstrated application potential in the dye decoloring of texile finishing.In dye decolorization,CcPIE5 efficiently degraded(92.9±2.3)%indigo carmine at pH 8.5 and 60℃,with syringic acid as the mediator.Isatin 5-sulfonic acid(ISA)was identified by LC-MS as the primary byproduct of indigo carmine degradation.[Conclusion]CcPIE5 is best-suited in decolorizing dyes under high temperatures and alkaline and salty conditions.It serves as a good candidate for specific applications in the environment and industry.

关 键 词：真菌漆酶 过表达 灰盖鬼伞 高盐 靛蓝胭脂红 

分 类 号：X703[环境科学与工程—环境工程]