Dataset Information

Low-Cost Cellulase-Hemicellulase Mixture Secreted by Trichoderma harzianum EM0925 with Complete Saccharification Efficacy of Lignocellulose.

ABSTRACT: Fermentable sugars are important intermediate products in the conversion of lignocellulosic biomass to biofuels and other value-added bio-products. The main bottlenecks limiting the production of fermentable sugars from lignocellulosic biomass are the high cost and the low saccharification efficiency of degradation enzymes. Herein, we report the secretome of Trichoderma harzianum EM0925 under induction of lignocellulose. Numerously and quantitatively balanced cellulases and hemicellulases, especially high levels of glycosidases, could be secreted by T. harzianum EM0925. Compared with the commercial enzyme preparations, the T. harzianum EM0925 enzyme cocktail presented significantly higher lignocellulolytic enzyme activities and hydrolysis efficiency against lignocellulosic biomass. Moreover, 100% yields of glucose and xylose were obtained simultaneously from ultrafine grinding and alkali pretreated corn stover. These findings demonstrate a natural cellulases and hemicellulases mixture for complete conversion of biomass polysaccharide, suggesting T. harzianum EM0925 enzymes have great potential for industrial applications.

SUBMITTER: Zhang Y

PROVIDER: S-EPMC7014229 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Low-Cost Cellulase-Hemicellulase Mixture Secreted by <i>Trichoderma harzianum</i> EM0925 with Complete Saccharification Efficacy of Lignocellulose.

Zhang Yu Y Yang Jinshui J Luo Lijin L Wang Entao E Wang Ruonan R Liu Liang L Liu Jiawen J Yuan Hongli H

International journal of molecular sciences 20200107 2

Fermentable sugars are important intermediate products in the conversion of lignocellulosic biomass to biofuels and other value-added bio-products. The main bottlenecks limiting the production of fermentable sugars from lignocellulosic biomass are the high cost and the low saccharification efficiency of degradation enzymes. Herein, we report the secretome of <i>Trichoderma harzianum</i> EM0925 under induction of lignocellulose. Numerously and quantitatively balanced cellulases and hemicellulases ...[more]

PMID: 31936000

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Project description:BackgroundThe soft rot ascomycetal fungus Trichoderma reesei is utilized for industrial production of secreted enzymes, especially lignocellulose degrading enzymes. T. reesei uses several different enzymes for the degradation of plant cell wall-derived material, including 9 characterized cellulases, 15 characterized hemicellulases and at least 42 genes predicted to encode cellulolytic or hemicellulolytic activities. Production of cellulases and hemicellulases is modulated by environmental and physiological conditions. Several regulators affecting the expression of cellulase and hemicellulase genes have been identified but more factors still unknown are believed to be present in the genome of T. reesei.ResultsWe have used transcriptional profiling data from T. reesei cultures in which cellulase/hemicellulase production was induced by the addition of different lignocellulose-derived materials to identify putative novel regulators for cellulase and hemicellulase genes. Based on this induction data, supplemented with other published genome-wide data on different protein production conditions, 28 candidate regulatory genes were selected for further studies and they were overexpressed in T. reesei. Overexpression of seven genes led to at least 1.5-fold increased production of cellulase and/or xylanase activity in the modified strains as compared to the parental strain. Deletion of gene 77513, here designated as ace3, was found to be detrimental for cellulase production and for the expression of several cellulase genes studied. This deletion also significantly reduced xylanase activity and expression of xylan-degrading enzyme genes. Furthermore, our data revealed the presence of co-regulated chromosomal regions containing carbohydrate-active enzyme genes and candidate regulatory genes.ConclusionsTranscriptional profiling results from glycoside hydrolase induction experiments combined with a previous study of specific protein production conditions was shown to be an effective method for finding novel candidate regulatory genes affecting the production of cellulases and hemicellulases. Recombinant strains with improved cellulase and/or xylanase production properties were constructed, and a gene essential for cellulase gene expression was found. In addition, more evidence was gained on the chromatin level regional regulation of carbohydrate-active enzyme gene expression.

Project description:Filamentous fungi are widely used for producing cellulolytic enzymes to degrade lignocellulosic biomass. Microbial resources from Tibet have received great attention due to the unique geographic and climatic conditions in the Qinghai-Tibet Plateau. However, studies on cellulase producing fungal strains originated from Tibet remain very limited, and so far no studies have been focused on regulation of cellulase production of the specific strains thereof. Here, filamentous fungal strains were isolated from soil, plant, and other environments in Tibet, and cellulase-producing strains were further investigated. A total of 88 filamentous fungal strains were identified, and screening of cellulase-producing fungi revealed that 16 strains affiliated with the genera Penicillium, Trichoderma, Aspergillus, and Talaromyces exhibited varying cellulolytic activities. Among these strains, T. harzianum isolate LZ117 is the most potent producer. Comparative transcriptome analysis using T. harzianum LZ117 and the control strain T. harzianum K223452 cultured on cellulose indicated an intensive modulation of gene transcription related to protein synthesis and quality control. Furthermore, transcription of xyr1 which encodes the global transcriptional activator for cellulase expression was significantly up-regulated. Transcription of cre1 and other predicted repressors controlling cellulase gene expression was decreased in T. harzianum LZ117, which may contribute to enhancing formation of primary cellulases. To our knowledge, this is the first report that the transcription landscape at the early enzyme production stage of T. harzianum was comprehensively described, and detailed analysis on modulation of transporters, regulatory proteins as well as protein synthesis and processing was presented. Our study contributes to increasing the catalog of publicly available transcriptome data from T. harzianum, and provides useful clues for unraveling the biotechnological potential of this species for lignocellulosic biorefinery.

Project description:BACKGROUND:A complete saccharification of plant polymers is the critical step in the efficient production of bio-alcohols. Beta-glucosidases acting in the degradation of intermediate gluco-oligosaccharides produced by cellulases limit the yield of the final product. RESULTS:In the present work, we have identified and then successfully cloned, expressed, purified and characterised 4 highly active beta-glucosidases from fibre-adherent microbial community from the cow rumen. The enzymes were most active at temperatures 45-55°C and pH 4.0-7.0 and exhibited high affinity and activity towards synthetic substrates such as p-nitrophenyl-beta-D-glucopyranoside (pNPbetaG) and pNP-beta-cellobiose, as well as to natural cello-oligosaccharides ranging from cellobiose to cellopentaose. The apparent capability of the most active beta-glucosidase, herein named LAB25g2, was tested for its ability to improve, at low dosage (31.25 units g-1 dry biomass, using pNPbetaG as substrate), the hydrolysis of pre-treated corn stover (dry matter content of 20%; 350 g glucan kg-1 dry biomass) in combination with a beta-glucosidase-deficient commercial Trichoderma reseei cellulase cocktail (5 units g-1 dry biomass in the basis of pNPbetaG). LAB25g2 increased the final hydrolysis yield by a factor of 20% (44.5 ± 1.7% vs. 34.5 ± 1.5% in control conditions) after 96-120 h as compared to control reactions in its absence or in the presence of other commercial beta-glucosidase preparations. The high stability (half-life higher than 5 days at 50°C and pH 5.2) and 2-38000 fold higher (as compared with reported beta-glucosidases) activity towards cello-oligosaccharides may account for its performance in supplementation assays. CONCLUSIONS:The results suggest that beta-glucosidases from yet uncultured bacteria from animal digestomes may be of a potential interest for biotechnological processes related to the effective bio-ethanol production in combination with low dosage of commercial cellulases.

Dataset Information

Low-Cost Cellulase-Hemicellulase Mixture Secreted by Trichoderma harzianum EM0925 with Complete Saccharification Efficacy of Lignocellulose.

Publications

Low-Cost Cellulase-Hemicellulase Mixture Secreted by <i>Trichoderma harzianum</i> EM0925 with Complete Saccharification Efficacy of Lignocellulose.

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