中国革孔菌属Coriolopsis种质资源收集及漆酶活性研究

Laccase has been paid much attention because of the widely used in many fields, such as biology and chemical industry. Large scale laccase production has become important from an economic point of view although there are some technical difficulties. Studies on fungal laccase have been focused mostly on a few model species, and there is almost no study on utilization of new resources. China has a vast territory and abundant fungal resources that provide favorable conditions for new fungal resources. Coriolopsis is a widely distributed fungal genus and important laccase producer in China, which has great potential applications. However, species diversity and laccase of Coriolopsis are not well known. In this project, extensive resource investigation, isolation and identification of Coriolopsis species will be carried out. All the studied stains will be identified by molecular data. Higher laccase-producing strains from species of Coriolopsis will be screened. The laccase activities from different species and different strains of the same species will be analyzed. Meanwhile, the fermentation conditions of the strains with higher laccase-producing will be optimized. The laccase will be purified and the enzymatic properties will be analyzed. The results will provide a reliable theoretical basis for laccase applications.

漆酶在生物、化工等各领域均具有极为广泛的应用，一直备受关注，但是漆酶的商业生产还存在一定的技术困难，而漆酶的规模化生产十分重要。目前对漆酶的研究大部分都集中在少数模式种上，对新资源的开发利用比较缺乏。我国幅员辽阔，菌种资源众多，为新资源的开发提供了有利条件。革孔菌属真菌在我国分布广泛且是重要的漆酶生产者，极具应用潜力。但目前对革孔菌属真菌资源及其产漆酶的研究不够，因此可研究空间较大。本项目拟对我国革孔菌属真菌进行广泛的资源调查和菌种的分离并鉴定，筛选革孔菌属真菌种类的高产漆酶菌株以分析同属不同种、同种不同菌株及不同生境下菌株漆酶的活性差异并确定这些菌株的分类地位。同时，优化高产漆酶菌株的发酵条件，对漆酶进行分离纯化和酶学性质的分析，研究结果将为漆酶广泛使用提供理论依据。

漆酶，被誉为“绿色催化剂”，其在分子氧的协助下能够将酚类、芳胺类化合物等氧化为水及终产物，因其在众多领域，如食品工业、能源开发和环境保护，均具有极为广泛的应用潜力而一直备受关注。但漆酶活性差和生产成本高等缺点限制了其的广泛应用。此外，目前对漆酶的研究大部分都集中在少数白腐真菌的种类，缺乏对新资源的开发利用，而革孔菌属真菌在我国分布广泛且是重要的漆酶生产者，极具开发和应用的潜力。本项目对我国革孔菌属真菌进行了标本采集、菌种分离和保藏，采用形态学和分子生物学相结合的方法，按照现代分类观点，对采集的革孔菌属真菌标本和菌种进行了鉴定。共采集大型真菌标本750余份，其中革孔菌属及其近缘属等真菌标本295份，分离、纯化及保存革孔菌属及其近缘属等真菌菌种共118株，隶属于31种，为我国革孔菌属真菌的多样性分析奠定了一定的基础。利用愈创木酚显色法初筛和发酵复筛的方式，获得高产漆酶的菌株——硬毛革孔菌，为工业生产漆酶提供了可供选择的菌株；同时，发现革孔菌属真菌及其近缘属（Earliella、Neofomitella和Trametes）具普遍的产漆酶特性。同属不同种以及同种不同菌株的产漆酶能力存在差异，即便是采自同一地域的、同样寄主的同种不同菌株也是如此。此外，发现革孔菌属的不同种或同种不同菌株在分泌漆酶时对木质纤维素材料的偏好性存在差异。发现新种2个，丰富了我国乃至世界大型真菌的物种多样性；发现混合金属离子诱导和混合白腐真菌这两种新的能够提升漆酶活性的方法，为漆酶工厂化快速生产低成本漆酶提供了新思路。