de Eugenio Laura I, Méndez-Líter Juan A, Nieto-Domínguez Manuel, Alonso Lola, Gil-Muñoz Jesús, Barriuso Jorge, Prieto Alicia, Martínez María Jesús
Department of Environmental Biology, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid, Spain.
Genetic and Molecular Epidemiology Group, Human Cancer Genetics Programme, Spanish National Cancer Research Centre, CNIO, Melchor Fernández Almagro 3, 28029 Madrid, Spain.
Biotechnol Biofuels. 2017 Jun 23;10:161. doi: 10.1186/s13068-017-0844-7. eCollection 2017.
Genomic and proteomic analysis are potent tools for metabolic characterization of microorganisms. Although cellulose usually triggers cellulase production in cellulolytic fungi, the secretion of the different enzymes involved in polymer conversion is subjected to different factors, depending on growth conditions. These enzymes are key factors in biomass exploitation for second generation bioethanol production. Although highly effective commercial cocktails are available, they are usually deficient for β-glucosidase activity, and genera like and are being explored for its production.
This article presents the description of as a cellulase-producer fungus that secretes high levels of β-glucosidase. β-1,4-endoglucanase, exoglucanase, and β-glucosidase activities were quantified in the presence of different carbon sources. Although the two first activities were only induced with cellulosic substrates, β-glucosidase levels were similar in all carbon sources tested. Sequencing and analysis of the genome of this fungus revealed multiple genes encoding β-glucosidases. Extracellular proteome analysis showed different induction patterns. In all conditions assayed, glycosyl hydrolases were the most abundant proteins in the supernatants, albeit the ratio of the diverse enzymes from this family depended on the carbon source. At least two different β-glucosidases have been identified in this work: one is induced by cellulose and the other one is carbon source-independent. The crudes induced by Avicel and glucose were independently used as supplements for saccharification of slurry from acid-catalyzed steam-exploded wheat straw, obtaining the highest yields of fermentable glucose using crudes induced by cellulose.
The genome of contains several genes encoding β-glucosidases and the fungus secretes high levels of this activity, regardless of the carbon source availability, although its production is repressed by glucose. Two main different β-glucosidases have been identified from proteomic shotgun analysis. One of them is produced under different carbon sources, while the other is induced in cellulosic substrates and is a good supplement to Celluclast in saccharification of pretreated wheat straw.
基因组和蛋白质组分析是用于微生物代谢特征表征的有力工具。虽然纤维素通常会触发纤维素分解真菌产生纤维素酶,但参与聚合物转化的不同酶的分泌受不同因素影响,这取决于生长条件。这些酶是第二代生物乙醇生产中生物质利用的关键因素。尽管有高效的商业酶混合物可用,但它们通常缺乏β - 葡萄糖苷酶活性,因此正在探索如[具体属名缺失]等属来生产该酶。
本文描述了[具体菌种名缺失]作为一种能分泌高水平β - 葡萄糖苷酶的纤维素酶产生真菌。在不同碳源存在的情况下对β - 1,4 - 内切葡聚糖酶、外切葡聚糖酶和β - 葡萄糖苷酶活性进行了定量分析。虽然前两种活性仅在纤维素底物诱导下产生,但在所测试的所有碳源中β - 葡萄糖苷酶水平相似。对该真菌基因组的测序和分析揭示了多个编码β - 葡萄糖苷酶的基因。细胞外蛋白质组分析显示了不同的诱导模式。在所有测定条件下,糖基水解酶是上清液中最丰富的蛋白质,尽管该家族中不同酶的比例取决于碳源。在这项研究中至少鉴定出两种不同的β - 葡萄糖苷酶:一种由纤维素诱导产生,另一种与碳源无关。由微晶纤维素(Avicel)和葡萄糖诱导产生的粗酶分别用作酸催化蒸汽爆破麦草浆糖化的补充剂,使用纤维素诱导产生的粗酶可获得最高产量的可发酵葡萄糖。
[具体菌种名缺失]的基因组包含多个编码β - 葡萄糖苷酶的基因,该真菌能分泌高水平的这种活性,无论碳源是否可用,尽管其产生受葡萄糖抑制。通过蛋白质组鸟枪法分析鉴定出两种主要不同的β - 葡萄糖苷酶。其中一种在不同碳源下产生,而另一种在纤维素底物中诱导产生,并且是预处理麦草糖化中对纤维素酶(Celluclast)的良好补充。