乙酰化修饰在里氏木霉纤维素酶合成调控中的功能研究

项目名称： 乙酰化修饰在里氏木霉纤维素酶合成调控中的功能研究

项目编号： No.31470201

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 周志华

作者单位： 中国科学院上海生命科学研究院

项目金额： 85万元

中文摘要： 里氏木霉是应用最广泛的纤维素酶工业生产菌种，它具有强大的纤维素酶诱导分泌能力。近年来鉴定了多个纤维素酶合成相关的转录调控因子及非转录因子功能蛋白。但对其纤维素酶合成诱导机制仍缺乏深入的了解。最近的研究表明，蛋白质的可逆乙酰化修饰，在丝状真菌的生命活动中也扮演着重要角色，这类修饰也可能参与了纤维素酶合成诱导调控。本项目中，我们将从里氏木霉基因组中注释为乙酰转移酶或去乙酰化酶的编码基因中确定候选基因，敲除或过表达这些基因，观测突变菌株在不同产酶条件下生长与产酶的表型，确定产酶表型差异显著的突变株，通过表达谱分析确定表达水平变化显著的已知及未知功能蛋白，并确认它们表达水平的差异与乙酰化修饰水平的相关性，找到参与纤维素酶合成诱导的新功能蛋白。这些研究将为全面认识里氏木霉中纤维素酶的合成诱导调控机制及解析乙酰化修饰与蛋白合成分泌的相关性积累基础数据，并为里氏木霉菌种的设计与进一步改造提供理论根据。

中文关键词： 丝状真菌；功能基因；基因表达调控；纤维素酶；乙酰化修饰

英文摘要： Trichoderma reesei is widely used in cellulase industry with its high production of secreted proteins. Although many regulators including transcriptional and non-transcriptional factors involved in synthesis of cellulase were elucidated in the past years, the substantial mechanism to induce the cellulase production is still unknown. The reversible acetylation of protein plays an important role in fungal life cycle, and evidences implying that acetylation may take parts in the regulations of cellulase synthesis and secretion in T. reesei have accumulated recently. In this study, we will select the candidates from all the annotated genes of acetyltransferases and deacetylases in T. reesei genome. Mutated strains would be built by deleting or overexpressing the candidate genes in order to characterize their functions in the regulations of cellulase synthesis and secretion in T. reesei. The mutants with significantly different phenotypes (especially for the cellulase production) would be chosen for the digital gene expression profiling analyses to identify the known regulators or function-unknown proteins with significant expression difference. And the correlation between the transcriptional level of the target proteins and the acetylation status would be further testified. Novel regulators or other functional proteins involved in cellulase synthesis and secretion are expected to be obtained based on the above investigations. These results will help us to know the regulating network of cellulase synthesis and secretion in T. reesei comprehensively. The revelation of the novel mechanisms to regulating cellulase synthesis and secretion in T. reesei will provide clues and foundation for further strain engineering for industry applications.

英文关键词： filamentous fungus;functional genes;transcription regulator;cellulase;acetylation