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Transcriptome Profiling-Based Analysis of Carbohydrate-Active Enzymes in Aspergillus terreus Involved in Plant Biomass Degradation
Frontiers in Bioengineering and Biotechnology ( IF 5.7 ) Pub Date : 2020-10-06 , DOI: 10.3389/fbioe.2020.564527
Camila L. Corrêa , Glaucia E. O. Midorikawa , Edivaldo Ximenes Ferreira Filho , Eliane Ferreira Noronha , Gabriel S. C. Alves , Roberto Coiti Togawa , Orzenil Bonfim Silva-Junior , Marcos Mota do Carmo Costa , Priscila Grynberg , Robert N. G. Miller

Given the global abundance of plant biomass residues, potential exists in biorefinery-based applications with lignocellulolytic fungi. Frequently isolated from agricultural cellulosic materials, Aspergillus terreus is a fungus efficient in secretion of commercial enzymes such as cellulases, xylanases and phytases. In the context of biomass saccharification, lignocellulolytic enzyme secretion was analyzed in a strain of A. terreus following liquid culture with sugarcane bagasse (SB) (1% w/v) and soybean hulls (SH) (1% w/v) as sole carbon source, in comparison to glucose (G) (1% w/v). Analysis of the fungal secretome revealed a maximum of 1.017 UI.mL–1 xylanases after growth in minimal medium with SB, and 1.019 UI.mL–1 after incubation with SH as carbon source. The fungal transcriptome was characterized on SB and SH, with gene expression examined in comparison to equivalent growth on G as carbon source. Over 8000 genes were identified, including numerous encoding enzymes and transcription factors involved in the degradation of the plant cell wall, with significant expression modulation according to carbon source. Eighty-nine carbohydrate-active enzyme (CAZyme)-encoding genes were identified following growth on SB, of which 77 were differentially expressed. These comprised 78% glycoside hydrolases, 8% carbohydrate esterases, 2.5% polysaccharide lyases, and 11.5% auxiliary activities. Analysis of the glycoside hydrolase family revealed significant up-regulation for genes encoding 25 different GH family proteins, with predominance for families GH3, 5, 7, 10, and 43. For SH, from a total of 91 CAZyme-encoding genes, 83 were also significantly up-regulated in comparison to G. These comprised 80% glycoside hydrolases, 7% carbohydrate esterases, 5% polysaccharide lyases, 7% auxiliary activities (AA), and 1% glycosyltransferases. Similarly, within the glycoside hydrolases, significant up-regulation was observed for genes encoding 26 different GH family proteins, with predominance again for families GH3, 5, 10, 31, and 43. A. terreus is a promising species for production of enzymes involved in the degradation of plant biomass. Given that this fungus is also able to produce thermophilic enzymes, this first global analysis of the transcriptome following cultivation on lignocellulosic carbon sources offers considerable potential for the application of candidate genes in biorefinery applications.

中文翻译:

基于转录组分析的土曲霉中碳水化合物活性酶参与植物生物质降解

鉴于全球丰富的植物生物质残留物,木质纤维素分解真菌在基于生物精炼的应用中存在潜力。土曲霉经常从农业纤维素材料中分离出来,是一种能有效分泌商业酶如纤维素酶、木聚糖酶和植酸酶的真菌。在生物质糖化的背景下,在以甘蔗渣 (SB) (1% w/v) 和大豆壳 (SH) (1% w/v) 为唯一液体培养的土星菌菌株中分析木质纤维素分解酶的分泌碳源,与葡萄糖 (G) (1% w/v) 相比。真菌分泌组分析显示,在含有 SB 的基本培养基中生长后最多产生 1.017 UI.mL-1 木聚糖酶,在以 SH 作为碳源孵育后最多产生 1.019 UI.mL-1。真菌转录组的特征在于 SB 和 SH,与作为碳源的 G 的等效生长相比,检查了基因表达。鉴定了超过 8000 个基因,包括许多参与植物细胞壁降解的编码酶和转录因子,根据碳源具有显着的表达调节。在 SB 上生长后鉴定了 89 个碳水化合物活性酶 (CAZyme) 编码基因,其中 77 个差异表达。这些包括 78% 的糖苷水解酶、8% 的碳水化合物酯酶、2.5% 的多糖裂解酶和 11.5% 的辅助活性。糖苷水解酶家族的分析揭示了编码 25 种不同 GH 家族蛋白的基因的显着上调,其中家族 GH3、5、7、10 和 43 占优势。对于 SH,来自总共 91 个 CAZyme 编码基因,83 与 G 相比也显着上调。这些包括 80% 糖苷水解酶、7% 碳水化合物酯酶、5% 多糖裂解酶、7% 辅助活性 (AA) 和 1% 糖基转移酶。类似地,在糖苷水解酶中,观察到编码 26 种不同 GH 家族蛋白的基因显着上调,家族 GH3、5、10、31 和 43 再次占优势。A. terreus 是一种有前途的生产相关酶的物种在植物生物量的降解中。鉴于这种真菌也能够产生嗜热酶,在木质纤维素碳源上培养后对转录组的首次全球分析为候选基因在生物精炼应用中的应用提供了相当大的潜力。7% 碳水化合物酯酶、5% 多糖裂解酶、7% 辅助活性 (AA) 和 1% 糖基转移酶。类似地,在糖苷水解酶中,观察到编码 26 种不同 GH 家族蛋白的基因显着上调,家族 GH3、5、10、31 和 43 再次占优势。A. terreus 是一种有前途的生产相关酶的物种在植物生物量的降解中。鉴于这种真菌也能够产生嗜热酶,在木质纤维素碳源上培养后对转录组的首次全球分析为候选基因在生物精炼应用中的应用提供了相当大的潜力。7% 碳水化合物酯酶、5% 多糖裂解酶、7% 辅助活性 (AA) 和 1% 糖基转移酶。类似地,在糖苷水解酶中,观察到编码 26 种不同 GH 家族蛋白的基因显着上调,家族 GH3、5、10、31 和 43 再次占优势。A. terreus 是一种有前途的生产相关酶的物种在植物生物量的降解中。鉴于这种真菌也能够产生嗜热酶,在木质纤维素碳源上培养后对转录组的首次全球分析为候选基因在生物精炼应用中的应用提供了相当大的潜力。观察到编码 26 种不同 GH 家族蛋白的基因显着上调,GH3、5、10、31 和 43 家族再次占优势。A. terreus 是生产参与植物生物量降解的酶的有前途的物种。鉴于这种真菌也能够产生嗜热酶,在木质纤维素碳源上培养后对转录组的首次全球分析为候选基因在生物精炼应用中的应用提供了相当大的潜力。观察到编码 26 种不同 GH 家族蛋白的基因显着上调,GH3、5、10、31 和 43 家族再次占优势。A. terreus 是生产参与植物生物量降解的酶的有前途的物种。鉴于这种真菌也能够产生嗜热酶,在木质纤维素碳源上培养后对转录组的首次全球分析为候选基因在生物精炼应用中的应用提供了相当大的潜力。
更新日期:2020-10-06
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