欢迎访问林业科学,今天是

林业科学 ›› 2017, Vol. 53 ›› Issue (1): 94-100.doi: 10.11707/j.1001-7488.20170112

• 论文与研究报告 • 上一篇    下一篇

Paraconiothyium variable GHJ-4木质素降解酶的酶学性质

王凤娟, 李伟庆, 牟志美, 王彦文, 刘庆信, 高绘菊   

  1. 山东农业大学林学院 泰安 271018
  • 收稿日期:2015-08-21 修回日期:2015-11-02 出版日期:2017-01-25 发布日期:2017-03-03
  • 通讯作者: 高绘菊
  • 基金资助:
    国家自然科学基金项目(31200450);中国博士后科学基金项目(2013M541944);山东省现代农业产业技术体系蚕桑产业创新团队建设资助项目(SDAIT-18-05)。

Enzymological Characteristics of Ligninolytic Enzyme from Paraconiothyrium variabile GHJ-4

Wang Fengjuan, Li Weiqing, Mu Zhimei, Wang Yanwen, Liu Qingxin, Gao Huiju   

  1. College of Forestry, Shandong Agricultural University Tai'an 271018
  • Received:2015-08-21 Revised:2015-11-02 Online:2017-01-25 Published:2017-03-03

摘要: [目的] 研究木质素降解子囊菌Paraconiothyrium variabile GHJ-4分泌的漆酶(laccase)、锰过氧化物酶(MnP)和木质素过氧化物酶(LiP)3种木质素降解酶的酶活性变化规律及其酶学性质,为利用该菌株进行木质素降解酶的工业化生产和应用提供依据。[方法] 分别以愈创木酚、2,6-二甲基苯酚和黎芦醇为底物测定laccase,MnP和LiP的酶活性,研究3种酶的最适反应温度和pH、温度和pH稳定性及金属离子对其活性的影响。[结果] GHJ-4菌株发酵18,15和21天后,分别获得laccase,MnP和LiP最高酶活为1 390.3,30.3和52.5 U·mL-1。laccase的最适反应温度为55℃左右,最适反应pH为5.5左右,在55℃以下、pH 4.0~7.0的范围内较为稳定;MnP的最适反应温度为60℃左右,最适反应pH为5.0左右,在55℃以下、pH 4.0~9.0的范围内较为稳定;LiP的最适反应温度为40℃左右,最适反应pH为3.0左右,在40℃以下、pH 2.0~4.0的范围内较为稳定。Mg2+,Zn2+,Cu2+,K+对laccase起促进作用,Na+和Zn2+对LiP起促进作用,Mn2+对MnP起激活作用,而Fe3+,Ca2+,Pb2+,Co2+,Al3+对3种酶均起抑制作用,其中尤以5 mmol·L-1Fe3+对3种酶的抑制作用最强。[结论] 温度、pH及金属离子对Paraconiothyrium variabile GHJ-4 3种木质素降解酶的酶学特性均有着不同程度的影响。

关键词: Paraconiothyrium variabile GHJ-4, 漆酶, 锰过氧化物酶, 木质素过氧化物酶, 酶学性质

Abstract: [Objective] The production and enzymological characteristics of ligninolytic enzymes, including laccase, manganese peroxidase (MnP) and lignin peroxidase (LiP) produced by Paraconiothyrium variabile GHJ-4 were investigated. This study could provide theoretical data for commercial process and application of Paraconiothyrium variabile GHJ-4 ligninolytic enzymes.[Method] Laccase, MnP and LiP activity were measured with guaiacol, 2,6-Dimethylphenol and veratryl alcohol as substrate respectively. Effects of temperature, pH value and metal irons on three ligninolytic enzymes were assayed. [Result] Laccase, MnP and LiP activity reached the peak value of 1 390.3 U·mL-1, 30.3 U·mL-1 and 52.5 U·mL-1 respectively on the 18th, 15th and 21st day during the fermentation. The optimum temperature and pH of laccase were 55℃ and 5.5 respectively, and the enzyme activity was stable under 55℃ and pH 4.0-7.0. The optimum temperature and pH of MnP were 60℃ and 5.0 respectively, and the enzyme activity was stable under 55℃ and pH 4.0-9.0. The optimum temperature and pH of LiP were 40℃ and 3.0 respectively, and the enzyme activity was stable under 40℃ and pH 2.0-4.0. Laccase activity was enhanced by the metal ions Mg2+,Zn2+,Cu2+,K+, LiP activity was enhanced by Na+ and Zn2+, and MnP activity was enhanced by Mn2+. Whereas three ligninolytic enzymes were inhibited by Fe3+, Ca2+, Pb2+, Co2+ and Al3+, especially the effect of Fe3+ was the strongest.[Conclusion] The effects of temperature, pH and metal ions on activities of ligninolytic enzymes from Paraconiothyrium variabile GHJ-4 were different.

Key words: Paraconiothyrium variabile GHJ-4, laccase, manganese peroxidase(MnP), lignin peroxidase(LiP), enzyme characterization

中图分类号: