香菇漆酶高产菌株筛选及漆酶基因的表达研究

发布时间：2018-10-31 13:49

【摘要】：漆酶是一种含铜的多酚氧化酶,其作用底物范围广,在食品、能源和环保等领域具有重要的应用价值,因此筛选高产漆酶菌株对于其工业应用具有重要意义。本实验以78株香菇菌株为实验材料,利用愈创木酚平板法初筛,液体发酵复筛的方法筛选高产漆酶香菇菌株,并通过实时定量PCR分析香菇漆酶同工酶在不同菌株之间转录表达的差异性和特异性。结果表明,不同香菇菌株的生长速度、变色圈大小存在差异,以菌丝圈和变色圈大小及其比例为依据,从78株香菇菌株中筛选出19株潜在的漆酶高产菌株;液体发酵从这19株菌株中复筛出3株高产漆酶的菌株:香240、15和冕宁浅灰,其中香240在培养第3天酶活达到最高(3.583U/m L),香15和冕宁浅灰则在培养第7天酶活达到最高(分别为3.842U/m L和2.806U/m L);实时荧光定量PCR结果显示,10个漆酶同工酶基因在两株香菇菌株中的表达存在差异性和特异性,其中Lelac5和Lelac9只在菌株香240中检测到转录本;Lelac1 4在菌株15中表达量较高,而Lelac8、Lelac10和Lelac11在香240中的表达量较高。研究结果将为香菇漆酶的进一步研究提供基础。
[Abstract]:Laccase is a copper-containing polyphenol oxidase, which has a wide range of substrates and has important application value in food, energy and environmental protection. Therefore, it is of great significance to screen high-yield laccase strains for its industrial application. In this experiment, 78 strains of lentinus edodes were used to screen laccase strains with guaiacol plate method and liquid fermentation method. The transcriptional expression of laccase isozyme in Lentinus edodes was analyzed by real-time quantitative PCR. The results showed that there were differences in growth speed and chromotropic circle size among different strains of Lentinus edodes. According to the size and proportion of hyphal and chromotropic circles, 19 potential laccase producing strains were screened from 78 strains of Lentinus edodes. Three laccase producing strains were screened from the 19 strains by liquid fermentation: Xiang240P15 and Mianning light ash. The enzyme activity of Xiang240 reached the highest level on the 3rd day of culture (3.583U/m L),). On the 7th day of culture, the enzyme activity of Xiang15 and Mianning reached the highest level (3.842U/m L and 2.806U/m L);, respectively). The results of real-time fluorescence quantitative PCR showed that the expression of 10 laccase isozyme genes in two strains of Lentinus edodes was different and specific, among which Lelac5 and Lelac9 only detected transcripts in strain Xiang240. The expression of Lelac1 4 was higher in strain 15, while the expression of Lelac8,Lelac10 and Lelac11 in Xiang240 was higher than that in strain 15. The results will provide a basis for further study of laccase in Lentinus edodes.
【作者单位】：四川农业大学资源学院微生物系;成都市农林科学院园艺研究所;
【基金】：四川省教育厅重点项目(15ZA0013) 四川省科技支撑计划项目四川农业大学科研兴趣培养计划(20150316)~~
【分类号】：S646.12

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