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林业科学 ›› 2016, Vol. 52 ›› Issue (8): 157-166.doi: 10.11707/j.1001-7488.20160819

• 研究简报 • 上一篇    下一篇

不同木质底物诱导下白囊耙齿菌胞外木质纤维素酶活性和胞内蛋白质组的差异

吴柯军, 闫绍鹏, 卢宏, 李海峰, 王秋玉   

  1. 东北林业大学生命科学学院 哈尔滨 150040
  • 收稿日期:2015-06-23 修回日期:2015-11-10 出版日期:2016-08-25 发布日期:2016-09-19
  • 通讯作者: 王秋玉
  • 基金资助:
    中央高校基本科研业务费专项资金资助(2572014CA22)。

Difference in the Activity of Extracellular Lignocellulolytic Enzymes and the Intracellular Proteome of Irpex lacteus Induced by Different Wood Substrates

Wu Kejun, Yan Shaopeng, Lu Hong, Li Haifeng, Wang Qiuyu   

  1. College of Life Science, Northeast Forestry University Harbin 150040
  • Received:2015-06-23 Revised:2015-11-10 Online:2016-08-25 Published:2016-09-19

摘要: [目的]研究白囊耙齿菌5种胞外分泌木质纤维素酶活性变化以及胞内蛋白质组的表达差异,探讨该菌对不同基质的利用机制,为筛选用于生物制浆、林木病腐防治和工业污染物降解的高效菌株提供科学依据。[方法]选用白桦、红皮云杉木屑作为诱导物,不加木屑为对照,研究白囊靶齿菌5种主要胞外分泌木质纤维素酶的活性以及胞内蛋白质组的表达差异。[结果]木屑诱导下的木质纤维素相关胞外酶活性大都高于对照样品,且锰过氧化物酶和漆酶活性在处理组与对照组间的差异达显著和极显著水平;木质素降解酶系中漆酶活性最强,木质素过氧化物酶次之,锰过氧化物酶最弱;纤维素降解酶系中葡聚糖外切酶活性远大于葡聚糖内切酶活性,2种酶活性总体上趋于稳定,表达量较高,受培养时间的影响变化不明显;白桦和云杉木屑处理组中检测到32个胞内明显差异表达蛋白点,2个处理组各有3个特异的差异表达蛋白点。白桦木屑诱导下的特异表达蛋白与膜蛋白、膜运输和能量提供有关,而云杉诱导组特异表达蛋白均与蛋白合成有关。[结论]不同树种木屑底物诱导下白囊耙齿菌胞外木质纤维素酶活性存在显著差异。同时胞内蛋白质的表达也存在明显差异,在分解白桦木屑时分泌蛋白的胞内外运输可能是决定木材降解能力的关键,而在分解云杉木屑时其相关分泌蛋白的大量合成将起决定作用。

关键词: 白囊耙齿菌, 胞外蛋白酶, 胞内蛋白质组, 木质纤维素酶, 差异表达

Abstract: [Objective] As a main white-rot fungus, Irpex lacteus shows very strong wood degradation ability. Here, we analyzed and measured the activities of five extracellular lignocellulolytic enzymes and intracellular proteome of Irpex lacteus induced by birch and spruce wood sawdust to explore the mechanism of substrate utilization by this fungus and provide more information and scientific basis for screening high efficient strains in biological pulping, prevention of forest disease and degradation of industrial pollutants.[Method] We measured the activities of five main extracellular lignocellulolytic enzymes and the changes the of intracellular proteome of I. lacteus subjected to birch(Betula playtyphylla) or spruce(Picea korainsis) sawdust or no sawdust (served as control).[Result] The enzyme activities induced by sawdust were higher than that of the control. There were significant differences in the manganese-peroxidase and laccase activities between the sawdust-treatments and control. The laccase activity was the highest among ligninolytic enzymes, followed by lignin-peroxidase activity, and manganese-peroxidase activity was the lowest. The exoglucanase activity was much higher than that of endoglucanase in the cellulose degrading enzymes. Compared with the control, 32 proteins with significantly differential expression were detected after being induced by sawdust. Three differential expression proteins were found in different substrates. Among these differential expression proteins, the proteins induced by birch sawdust were involved with membrane protein, membrane transportation and energy, while the proteins induced by spruce sawdust were all related with protein synthesis.[Conclusion] The lignocellulolytic enzymes activities and intracellular proteome of I. lacteus induced by different substrates were significantly different. Thus, the membrane transportation of secretory proteins could be a key in determining birch wood-decaying ability of I. lacteus, while the protein synthesis would play an important role in degrading spruce.

Key words: Irpex lacteus, extracellular protease, intracellular proteome, lignocellulose enzyme, differential expression

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