粗毛革孔菌及其高产漆酶突变株的比较分析  

Comparison of Coriolopsis gallica and High Laccases Production Mutant

作　　者：韩振亚[1] 徐晓立[1] 张爱萍[1] 谢君[1] 

机构地区：[1]农业部能源植物资源与利用重点实验室,华南农业大学新能源与新材料研究所,广东广州510642

出　　处：《现代食品科技》2014年第7期81-86,229,共7页Modern Food Science and Technology

基　　金：广东省科技计划重点项目(2011B090400437);"十二五"国家科技支撑计划(2011BAD22B01)

摘　　要：漆酶是一种广泛分布的多酚氧化酶,在食品、能源和环保等领域具有重要的应用价值。本实验室所筛的高产漆酶粗毛革孔菌经诱变后,生长形态变化大,突变株漆酶活力可达303 U/mL,是野生菌的3倍多。本文比较分析了原始菌株和突变菌株的生理生化差异,并通过RAPD比较分析了它们的差异基因。用140条随机引物扩增获得20条稳定的特异条带,并对其中的3条进行回收、克隆,测序结果与BLASTX、NCBI数据库的比对表明引物S381扩增出的809 bp的特异条带对应于云芝的假想蛋白,引物S225扩增出的654 bp的特异条带对应于30 S核糖体蛋白S4,引物S193扩增的1027 bp的特异条带对应于糖苷水解酶。该试验为探讨粗毛革孔菌突变株高产漆酶的分子机制,以及通过定向突变构建高产漆酶工程菌提供理论依据。Laccase is a kind of polyphenol oxidase, and it has important application values in food, energy, environment industries and other fields. A mutant strain from wild Coriolopsis gallica was selected, and its morphological characteristics were very different from the wild strain. The production of laccase by the mutant was up to 303 U/mL, which was three times higher than that by the wild strain. In this article, the physiological and biochemical differences between the wild and mutant, and their genetic differences with RAPD were compared and analyzed. Twenty stable specific bands were amplified with 140 random primers, and three of them were recovered and cloned. Then comparison analysis of their sequencing results with BLASTX, NCBI database showed that the 809 bp band from primer S381 was corresponding to a hypothetical protein of Trametes versicolor. The 654 bp band from primer S225 was corresponding to 30 S ribosomal protein S4 of Oribacterium sp, and the 1027 bp band from primer S193 was corresponding to glycoside hydrolase of Dichomitus squalens. This test would provide a theoretical basis for investigating molecular mechanism of the mutant yielding high laccase, and building high yield laccase engineering stain by directed mutation.

关 键 词：粗毛革孔菌 变异体 形态学 酶活 生物信息学分析 

分 类 号：TQ925[轻工技术与工程—发酵工程]