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基于基因组和转录组分析对 var. 的木质纤维素降解酶系统的深入了解。

Insights into the Lignocellulose-Degrading Enzyme System of var. Based on Genome and Transcriptome Analysis.

机构信息

Laboratory of Genetics and Biotechnology, EMBRAPA Agroenergy, Brasília, Brazil.

Graduate Program of Microbial Biology, Department of Cell Biology, Institute of Biology, University of Brasília, Brasília, Brazil.

出版信息

Microbiol Spectr. 2021 Oct 31;9(2):e0108821. doi: 10.1128/Spectrum.01088-21. Epub 2021 Sep 15.

Abstract

Humicola grisea var. is a thermophilic ascomycete and important enzyme producer that has an efficient enzymatic system with a broad spectrum of thermostable carbohydrate-active (CAZy) enzymes. These enzymes can be employed in lignocellulose biomass deconstruction and other industrial applications. In this work, the genome of H. grisea var. was sequenced. The acquired sequence reads were assembled into a total length of 28.75 Mbp. Genome features correlate with what was expected for thermophilic Sordariomycetes. The transcriptomic data showed that sugarcane bagasse significantly upregulated genes related to primary metabolism and polysaccharide deconstruction, especially hydrolases, at both pH 5 and pH 8. However, a number of exclusive and shared genes between the pH values were found, especially at pH 8. expresses an average of 211 CAZy enzymes (CAZymes), which are capable of acting in different substrates. The top upregulated genes at both pH values represent CAZyme-encoding genes from different classes, including acetylxylan esterase, endo-1,4-β-mannosidase, exoglucanase, and endoglucanase genes. For the first time, the arsenal that the thermophilic fungus var. possesses to degrade the lignocellulosic biomass is shown. Carbon source and pH are of pivotal importance in regulating gene expression in this organism, and alkaline pH is a key regulatory factor for sugarcane bagasse hydrolysis. This work paves the way for the genetic manipulation and robust biotechnological applications of this fungus. Most studies regarding the use of fungi as enzyme producers for biomass deconstruction have focused on mesophile species, whereas the potential of thermophiles has been evaluated less. This study revealed, through genome and transcriptome analyses, the genetic repertoire of the biotechnological relevant thermophile fungus . Comparative genomics helped us to further understand the biology and biotechnological potential of . The results demonstrate that this fungus possesses an arsenal of carbohydrate-active (CAZy) enzymes to degrade the lignocellulosic biomass. Indeed, it expresses more than 200 genes encoding CAZy enzymes when cultivated in sugarcane bagasse. Carbon source and pH are key factors for regulating the gene expression in this organism. This work shows, for the first time, the great potential of as an enzyme producer and a gene donor for biotechnological applications and provides the base for the genetic manipulation and robust biotechnological applications of this fungus.

摘要

灰色长蠕孢菌变种是一种嗜热子囊菌,也是一种重要的酶类生产菌,其具有高效的、能广泛作用于热稳定碳水化合物活性(CAZy)酶的酶系。这些酶可用于木质纤维素生物质的解构和其他工业应用。在这项工作中,我们对 H. grisea var. 的基因组进行了测序。获得的序列读段组装成长度为 28.75 Mbp 的基因组。基因组特征与嗜热子囊菌的预期特征相符。转录组数据表明,在 pH 值为 5 和 8 时,甘蔗渣均能显著上调与初级代谢和多糖解构相关的基因,尤其是水解酶。然而,在这两种 pH 值下都发现了一些特有和共享的基因,尤其是在 pH 值为 8 时。该菌平均表达 211 种 CAZy 酶(CAZymes),这些酶能够作用于不同的底物。在这两种 pH 值下,上调幅度最大的基因均代表不同类别 CAZyme 编码基因,包括乙酰木聚糖酯酶、内切 1,4-β-甘露聚糖酶、外切葡聚糖酶和内切葡聚糖酶基因。这是首次展示嗜热真菌 var. 降解木质纤维素生物质的酶库。碳源和 pH 值对该生物体内基因表达的调控至关重要,碱性 pH 值是甘蔗渣水解的关键调控因素。这项工作为该真菌的遗传操作和强大的生物技术应用铺平了道路。大多数关于利用真菌作为酶类生产菌来进行生物质解构的研究都集中在中温菌物种上,而对嗜热菌的潜力的评估则较少。通过基因组和转录组分析,本研究揭示了生物技术相关嗜热真菌 的遗传谱。比较基因组学帮助我们进一步了解 的生物学和生物技术潜力。结果表明,该真菌拥有一套碳水化合物活性(CAZy)酶,可用于降解木质纤维素生物质。事实上,当用甘蔗渣培养时,该菌表达了超过 200 种编码 CAZy 酶的基因。碳源和 pH 值是调控该生物体内基因表达的关键因素。这项工作首次表明, 作为酶类生产菌和生物技术应用的基因供体具有巨大的潜力,并为该真菌的遗传操作和强大的生物技术应用提供了基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c8dd/8557918/262e43b12772/spectrum.01088-21-f001.jpg

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