Sharma Ghimire Prakriti, Ouyang Haomiao, Wang Qian, Luo Yuanming, Shi Bo, Yang Jinghua, Lü Yang, Jin Cheng
State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences , Beijing 100101, China.
University of Chinese Academy of Sciences , Beijing 100101, China.
J Proteome Res. 2016 Dec 2;15(12):4387-4402. doi: 10.1021/acs.jproteome.6b00465. Epub 2016 Oct 10.
Lignocelluloses contained in animal forage cannot be digested by pigs or poultry with 100% efficiency. On contrary, Aspergillus fumigatus, a saprophytic filamentous fungus, is known to harbor 263 glycoside hydrolase encoding genes, suggesting that A. fumigatus is an efficient lignocellulose degrader. Hence the present study uses corn, wheat, or soybean as a sole carbon source to culture A. fumigatus under animal physiological condition to understand how cellulolytic enzymes work together to achieve an efficient degradation of lignocellulose. Our results showed that A. fumigatus produced different sets of enzymes to degrade lignocelluloses derived from corn, wheat, or soybean cell wall. In addition, the cellulolytic enzymes produced by A. fumigatus were stable under acidic condition or at higher temperatures. Using isobaric tags for a relative and absolute quantification (iTRAQ) approach, a total of ∼600 extracellular proteins were identified and quantified, in which ∼50 proteins were involved in lignocellulolysis, including cellulases, hemicellulases, lignin-degrading enzymes, and some hypothetical proteins. Data are available via ProteomeXchange with identifier PXD004670. On the basis of quantitative iTRAQ results, 14 genes were selected for further confirmation by RT-PCR. Taken together, our results indicated that the expression and regulation of lignocellulolytic proteins in the secretome of A. fumigatus were dependent on both nature and complexity of cellulose, thus suggesting that a different enzyme system is required for degradation of different lignocelluloses derived from plant cells. Although A. fumigatus is a pathogenic fungus and cannot be directly used as an enzyme source, as an efficient lignocellulose degrader its strategy to synergistically degrade various lignocelluloses with different enzymes can be used to design enzyme combination for optimal digestion and absorption of corn, wheat, or soybean that are used as forage of pig and poultry.
动物饲料中含有的木质纤维素不能被猪或家禽完全有效地消化。相反,烟曲霉是一种腐生丝状真菌,已知其含有263个编码糖苷水解酶的基因,这表明烟曲霉是一种高效的木质纤维素降解菌。因此,本研究以玉米、小麦或大豆作为唯一碳源,在动物生理条件下培养烟曲霉,以了解纤维素分解酶如何协同作用以实现对木质纤维素的高效降解。我们的结果表明,烟曲霉产生不同的酶系来降解源自玉米、小麦或大豆细胞壁的木质纤维素。此外,烟曲霉产生的纤维素分解酶在酸性条件或较高温度下稳定。使用相对和绝对定量的等压标签(iTRAQ)方法,共鉴定和定量了约600种细胞外蛋白质,其中约50种蛋白质参与木质纤维素分解,包括纤维素酶、半纤维素酶、木质素降解酶和一些假定蛋白质。数据可通过ProteomeXchange获得,标识符为PXD004670。基于定量iTRAQ结果,选择14个基因通过RT-PCR进行进一步验证。综上所述,我们的结果表明,烟曲霉分泌组中木质纤维素分解蛋白的表达和调控取决于纤维素的性质和复杂性,因此表明降解源自植物细胞的不同木质纤维素需要不同的酶系统。虽然烟曲霉是一种致病真菌,不能直接用作酶源,但作为一种高效的木质纤维素降解菌,其利用不同酶协同降解各种木质纤维素的策略可用于设计酶组合,以优化猪和家禽用作饲料的玉米、小麦或大豆的消化和吸收。