Center for Molecular Biology and Genetic Engineering (CBMEG), University of Campinas (UNICAMP), Cidade Universitária Zeferino Vaz, Avenida Cândido Rondon 400, Campinas, SP, 13083-875, Brazil; Graduate Program in Genetics and Molecular Biology, Institute of Biology, UNICAMP, Campinas, SP, 13083-970, Brazil.
Center for Molecular Biology and Genetic Engineering (CBMEG), University of Campinas (UNICAMP), Cidade Universitária Zeferino Vaz, Avenida Cândido Rondon 400, Campinas, SP, 13083-875, Brazil; Graduate Program in Genetics and Molecular Biology, Institute of Biology, UNICAMP, Campinas, SP, 13083-970, Brazil; Holzforschung München, TUM School of Life Sciences Weihenstephan, Technical University of Munich, Freising, 85354, Germany.
J Biotechnol. 2021 Jun 20;334:1-10. doi: 10.1016/j.jbiotec.2021.05.001. Epub 2021 May 14.
Bioprospecting genes and proteins related to plant biomass degradation is an attractive approach for the identification of target genes for biotechnological purposes, especially those with potential applications in the biorefinery industry that can enhance second-generation ethanol production technology. Trichoderma harzianum is a potential candidate for cellulolytic enzyme prospection and production. Herein, the enzymatic activities, transcriptome, exoproteome, and coexpression networks of the T. harzianum strain CBMAI-0179 were examined under biomass degradation conditions. We identified differentially expressed genes (DEGs) and carbohydrate-active enzyme (CAZyme) genes related to plant biomass degradation and compared them with those of strains from congeneric species (T. harzianum IOC-3844 and T. atroviride CBMAI-0020). T. harzianum CBMAI-0179 harbors strain- and treatment-specific CAZyme genes and transcription factors. We detected important proteins related to biomass degradation, including β-glucosidases, endoglucanases, cellobiohydrolases, lytic polysaccharide monooxygenases, endo-1,4-β-xylanases and β-mannanases. Based on coexpression networks, an enriched cluster with degradative enzymes was described, and the subnetwork of CAZymes revealed strong correlations among important secreted proteins and differentially expressed CAZyme genes. Our results provide valuable information for future studies on the genetic regulation of plant cell wall-degrading enzymes. This knowledge can be exploited for the improvement of enzymatic reactions in biomass degradation for bioethanol production.
生物勘探与植物生物质降解相关的基因和蛋白质是鉴定生物技术应用目标基因的一种有吸引力的方法，特别是那些在生物炼制工业中有潜在应用的基因，这些基因可以提高第二代乙醇生产技术。哈茨木霉是纤维素酶勘探和生产的潜在候选者。本文研究了哈茨木霉 CBMAI-0179 菌株在生物质降解条件下的酶活性、转录组、外分泌组和共表达网络。我们鉴定了与植物生物质降解相关的差异表达基因（DEGs）和碳水化合物活性酶（CAZyme）基因，并将其与同属种（哈茨木霉 IOC-3844 和哈茨木霉 CBMAI-0020）的菌株进行了比较。哈茨木霉 CBMAI-0179 具有菌株特异性和处理特异性的 CAZyme 基因和转录因子。我们检测到与生物质降解相关的重要蛋白质，包括β-葡萄糖苷酶、内切葡聚糖酶、纤维二糖水解酶、溶菌多糖单加氧酶、内切-1,4-β-木聚糖酶和β-甘露聚糖酶。基于共表达网络，描述了一个具有降解酶的富集簇，CAZymes 的子网络揭示了重要分泌蛋白和差异表达 CAZyme 基因之间的强烈相关性。我们的研究结果为植物细胞壁降解酶的遗传调控提供了有价值的信息。这些知识可以用于提高生物乙醇生产中生物质降解的酶反应。
