Midorikawa Glaucia Emy Okida, Correa Camila Louly, Noronha Eliane Ferreira, Filho Edivaldo Ximenes Ferreira, Togawa Roberto Coiti, Costa Marcos Mota do Carmo, Silva-Junior Orzenil Bonfim, Grynberg Priscila, Miller Robert Neil Gerard
Departamento de Biologia Celular, Universidade de Brasília, Brasília, Brazil.
Embrapa Recursos Genéticos e Biotecnologia, Parque Estação Biológica, Brasília, Brazil.
Front Bioeng Biotechnol. 2018 Sep 18;6:123. doi: 10.3389/fbioe.2018.00123. eCollection 2018.
The production of bioethanol from non-food agricultural residues represents an alternative energy source to fossil fuels for incorporation into the world's economy. Within the context of bioconversion of plant biomass into renewable energy using improved enzymatic cocktails, Illumina RNA-seq transcriptome profiling was conducted on a strain of , efficient in biomass polysaccharide degradation, in order to identify genes encoding proteins involved in plant biomass saccharification. Enzyme production and gene expression was compared following growth in liquid and semi-solid culture with steam-exploded sugarcane bagasse (SB) (1% ) and glucose (1% ) employed as contrasting sole carbon sources. Enzyme production following growth in liquid minimum medium supplemented with SB resulted in 0.626 and 0.711 UI.mL xylanases after 24 and 48 h incubation, respectively. Transcriptome profiling revealed expression of over 7120 genes, with groups of genes modulated according to solid or semi-solid culture, as well as according to carbon source. Gene ontology analysis of genes expressed following SB hydrolysis revealed enrichment in xyloglucan metabolic process and xylan, pectin and glucan catabolic process, indicating up-regulation of genes involved in xylanase secretion. According to carbohydrate-active enzyme (CAZy) classification, 209 CAZyme-encoding genes were identified with significant differential expression on liquid or semi-solid SB, in comparison to equivalent growth on glucose as carbon source. Up-regulated CAZyme-encoding genes related to cellulases (CelA, CelB, CelC, CelD) and hemicellulases (XynG1, XynG2, XynF1, XylA, AxeA, arabinofuranosidase) showed up to a 10-fold log2FoldChange in expression levels. Five genes from the AA9 (GH61) family, related to lytic polysaccharide monooxygenase (LPMO), were also identified with significant expression up-regulation. The transcription factor gene XlnR, involved in induction of hemicellulases, showed up-regulation on liquid and semi-solid SB culture. Similarly, the gene ClrA, responsible for regulation of cellulases, showed increased expression on liquid SB culture. Over 150 potential transporter genes were also identified with increased expression on liquid and semi-solid SB culture. This first comprehensive analysis of the transcriptome of contributes to our understanding of genes and regulatory systems involved in cellulose and hemicellulose degradation in this fungus, offering potential for application in improved enzymatic cocktail development for plant biomass degradation in biorefinery applications.
利用非粮农业残余物生产生物乙醇是一种可替代化石燃料的能源,有望融入世界经济。在利用改良酶混合物将植物生物质转化为可再生能源的背景下,对一株高效降解生物质多糖的菌株进行了Illumina RNA测序转录组分析,以鉴定编码参与植物生物质糖化的蛋白质的基因。在以蒸汽爆破甘蔗渣(SB)(1%)和葡萄糖(1%)作为对比单一碳源的液体和半固体培养基中培养后,比较了酶的产生和基因表达。在补充了SB的液体基本培养基中培养后,分别在培养24小时和48小时后产生的木聚糖酶活性为0.626和0.711 UI/mL。转录组分析揭示了超过7120个基因的表达,基因组根据固体或半固体培养以及碳源进行调节。对SB水解后表达的基因进行基因本体分析,发现木葡聚糖代谢过程以及木聚糖、果胶和葡聚糖分解代谢过程中基因富集,表明参与木聚糖酶分泌的基因上调。根据碳水化合物活性酶(CAZy)分类,与以葡萄糖作为碳源的同等生长相比,在液体或半固体SB上鉴定出209个具有显著差异表达的CAZyme编码基因。与纤维素酶(CelA、CelB、CelC、CelD)和半纤维素酶(XynG1、XynG2、XynF1、XylA、AxeA、阿拉伯呋喃糖苷酶)相关的上调CAZyme编码基因在表达水平上显示出高达10倍的log2倍变化。还鉴定出五个来自与裂解多糖单加氧酶(LPMO)相关的AA9(GH61)家族的基因,其表达显著上调。参与半纤维素酶诱导的转录因子基因XlnR在液体和半固体SB培养中上调。同样,负责纤维素酶调节的基因ClrA在液体SB培养中表达增加。还鉴定出超过150个潜在转运蛋白基因在液体和半固体SB培养中表达增加。对该菌株转录组的首次全面分析有助于我们了解该真菌中参与纤维素和半纤维素降解的基因和调控系统,为生物炼制应用中改进用于植物生物质降解的酶混合物开发提供了潜力。