生物技术通报 ›› 2020, Vol. 36 ›› Issue (1): 45-59.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0974

• 研究报告 • 上一篇    下一篇

白腐真菌落叶松锈迷孔菌产漆酶液体培养基的优化及其对染料的脱色作用

吴怡, 马鸿飞, 曹永佳, 司静, 崔宝凯   

  1. 北京林业大学生态与自然保护学院,北京 100083
  • 收稿日期:2019-10-14 出版日期:2020-01-26 发布日期:2020-01-08
  • 作者简介:吴怡,女,硕士研究生,研究方向:大型真菌漆酶的开发利用;E-mail:1792354604@qq.com
  • 基金资助:
    国家自然科学基金项目(31700016),中央高校基本科研业务费专项资金资助项目(2019ZY37,2016ZCQ04)

Medium Optimization for the Laccase Production by White Rot Fungus Porodaedalea laricis and Its Dye Decolorizing Capacity

WU Yi, MA Hong-fei, CAO Yong-jia, SI Jing, CUI Bao-kai   

  1. School of Ecology and Nature Conservation,Beijing Forestry University,Beijing 100083
  • Received:2019-10-14 Published:2020-01-26 Online:2020-01-08

摘要: 以白腐真菌落叶松锈迷孔菌(Porodaedalea laricis)胞外漆酶为响应值,通过将Plackett-Burman设计、最陡爬坡设计和Box-Behnken设计相结合,获得了P. laricis产胞外漆酶的最适培养基为:去皮马铃薯365.61 g/L、蛋白胨5.0 g/L、葡萄糖20.0 g/L、KH2PO4 1.0 g/L、MgSO4·7H2O 0.5 g/L、MnSO4·H2O 0.15 g/L、CaCl2·2H2O 0.03 g/L、酒石酸铵6.68 g/L、琥珀酸钠1.5 g/L、吐温80 0.48 mL/L、玉米芯46.43 g/L、维生素B1 0.01 g/L。在该条件下,P. laricis漆酶活性为3.29 U/mL,相比于优化前提高了2.81倍,与理论值3.32 U/mL相近,说明该模型准确可靠。此外,将漆酶应用于降解多种合成染料包括活性亮蓝X-BR、雷马素亮蓝R、酸性黑172、刚果红、亚甲基蓝、中性红、靛蓝、萘酚绿B和结晶紫,反应168 h后脱色率分别可达到95.64%、97.21%、36.11%、91.63%、61.42%、74.65%、48.60%、25.13%和68.80%。

关键词: 落叶松锈迷孔菌, 漆酶, 液体培养, 响应面设计, 染料脱色能力

Abstract: In this study,Plackett-Burman design,path of the steepest ascent,and Box-Behnken design were used to optimize the liquid medium composition for laccase production of white rot fungus Porodaedalea laricis with the laccase activity set as a response. The optimal compositions for production of P. laricis laccase was obtained as follows:peeled potato 365.61 g/L,peptone 5.0 g/L,glucose 20.0 g/L,KH2PO4 1.0 g/L,MgSO4·7H2O 0.5 g/L,MnSO4·H2O 0.15 g/L,CaCl2·2H2O 0.03 g/L,ammonium tartrate 6.68 g/L,sodium succinate 1.5 g/L,Tween 80 0.48 mL/L,corncob 46.43 g/L,and vitamin B1 0.01 g/L. Under the optimized conditions,the laccase activity was 3.29 U/mL,which was 2.81-fold of that given in the basal medium,and this yield was very in good agreement with the theoretical result of 3.32 U/mL,indicating the model was accurate and reliable. Moreover,the laccase exerted notably decolorizing capacity for many synthetic dyes including Reactive Brilliant Blue X-BR,Remazol Brilliant Blue R,Acid Black 172,Congo Red,Methylene Blue,Neutral Red,Indigo Blue,Naphthol Green B,and Crystal Violet,with the respective decolorization rate of 95.64%,97.21%,36.11%,91.63%,61.42%,74.65%,48.60%,25.13%,and 68.80% after 168 h.

Key words: Porodaedalea laricis, laccase, liquid cultivation, response surface methodology, dye decolorizing capacity