The effects of deletion of cellobiohydrolase genes on carbon source-dependent growth and enzymatic lignocellulose hydrolysis in Trichoderma reesei

2020 ◽  
Vol 58 (8) ◽  
pp. 687-695
Author(s):  
Meibin Ren ◽  
Yifan Wang ◽  
Guoxin Liu ◽  
Bin Zuo ◽  
Yuancheng Zhang ◽  
...  
2018 ◽  
Author(s):  
Marcella Fernandes de Souza ◽  
Elba Pinto da Silva Bon ◽  
Ayla Sant’ Ana da Silvab

AbstractThe high cost of commercial cellulases still hampers the economic competitiveness of the production of fuels and chemicals from lignocellulosic biomasses. This cost may be decreased by the on-site production of cellulases with the integrated use of the lignocellulosic biomass as carbon source. This integrated approach was evaluated in the present study whereby steam-pretreated sugarcane bagasse (SPSB) was used as carbon source for the production of cellulases by Trichoderma reesei Rut C30 and the produced enzymes were subsequently used for SPSB hydrolysis. An enzyme preparation with a high cellulase activity, of 1.93 FPU/mL, was obtained, and a significant β-glucosidase activity was achieved in buffered media, indicating the importance of pH control during enzyme production. The hydrolysis of SPSB with the laboratory-made mixture resulted in a glucose yield of 80%, which was equivalent to those observed for control experiments using commercial enzymes. Even though the supplementation of this mixture with external β-glucosidase from Aspergillus awamori was found to increase the initial hydrolysis rates, it had no impact on the final hydrolysis yield. It was shown that SPSB is a promising carbon source for the production of cellulases and β-glucosidases by T. reesei Rut C30 and that the enzyme preparation obtained is effective for the hydrolysis of SPSB, supporting the on-site integrated approach to decrease the cost of the enzymatic hydrolysis of lignocellulosic biomass.


mSphere ◽  
2017 ◽  
Vol 2 (3) ◽  
Author(s):  
Eva Stappler ◽  
Christoph Dattenböck ◽  
Doris Tisch ◽  
Monika Schmoll

ABSTRACT In fungi, most metabolic processes are subject to regulation by light. For Trichoderma reesei, light-dependent regulation of cellulase gene expression is specifically shown. Therefore, we intended to unravel the relationship between regulation of enzymes by the carbon source and regulation of enzymes by light. Our two-dimensional analysis included inducing and repressing carbon sources which we used to compare light-specific regulation to dark-specific regulation and to rule out effects specific for a single carbon source. We found close connections with respect to gene regulation as well as significant differences in dealing with carbon in the environment in light and darkness. Moreover, our analyses showed an intricate regulation mechanism for substrate degradation potentially involving surface sensing and provide a basis for knowledge-based screening for strain improvement. In fungi, most metabolic processes are subject to regulation by light. Trichoderma reesei is adapted to degradation of plant cell walls and regulates production of the required enzymes in a manner dependent on the nutrient source and the light status. Here we investigated the interrelated relevance of two regulation levels of the transcriptome of T. reesei: light regulation and carbon source-dependent control. We show that the carbon source (cellulose, lactose, sophorose, glucose, or glycerol) is the major source of variation, with light having a modulating effect on transcript regulation. A total of 907 genes were regulated under cellulase-inducing conditions in light, and 947 genes were regulated in darkness, with 530 genes overlapping (1,324 in total). Only 218 of the 1,324 induction-specific genes were independent of light and not regulated by the BLR1, BLR2, and ENV1 photoreceptors. Analysis of the genomic distribution of genes regulated by light upon growth on cellulose revealed considerable overlap of light-regulated clusters with induction-specific clusters and carbohydrate-active enzyme (CAZyme) clusters. Further, we found evidence for the operation of a sensing mechanism for solid cellulosic substrates, with regulation of genes such as swo1, cip1, and cip2 or of genes encoding hydrophobins which is related to the cyclic AMP (cAMP)-dependent regulatory output of ENV1. We identified class XIII G-protein-coupled receptors (GPCRs) CSG1 and CSG2 in T. reesei as putative cellulose/glucose-sensing GPCRs. Our data indicate that the cellulase regulation pathway is bipartite, comprising a section corresponding to transcriptional regulation and one corresponding to posttranscriptional regulation, with the two connected by the function of CSG1. IMPORTANCE In fungi, most metabolic processes are subject to regulation by light. For Trichoderma reesei, light-dependent regulation of cellulase gene expression is specifically shown. Therefore, we intended to unravel the relationship between regulation of enzymes by the carbon source and regulation of enzymes by light. Our two-dimensional analysis included inducing and repressing carbon sources which we used to compare light-specific regulation to dark-specific regulation and to rule out effects specific for a single carbon source. We found close connections with respect to gene regulation as well as significant differences in dealing with carbon in the environment in light and darkness. Moreover, our analyses showed an intricate regulation mechanism for substrate degradation potentially involving surface sensing and provide a basis for knowledge-based screening for strain improvement.


Open Biology ◽  
2013 ◽  
Vol 3 (12) ◽  
pp. 130146 ◽  
Author(s):  
Reena Lamichhane-Khadka ◽  
Stéphane L. Benoit ◽  
Susan E. Maier ◽  
Robert J. Maier

Glucarate, an oxidized product of glucose, is a major serum organic acid in humans. Still, its role as a carbon source for a pathogen colonizing hosts has not been studied. We detected high-level expression of a potential glucarate permease encoding gene gudT when Salmonella enterica serovar Typhimurium are exposed to hydrogen gas (H 2 ), a gaseous by-product of gut commensal metabolism. A gudT strain of Salmonella is deficient in glucarate-dependent growth, however, it can still use other monosaccharides, such as glucose or galactose. Complementation of the gudT mutant with a plasmid harbouring gudT restored glucarate-dependent growth to wild-type (WT) levels. The gudT mutant exhibits attenuated virulence: the mean time of death for mice inoculated with WT strain was 2 days earlier than for mice inoculated with the gudT strain. At 4 days postinoculation, liver and spleen homogenates from mice inoculated with a gudT strain contained significantly fewer viable Salmonella than homogenates from animals inoculated with the parent. The parent strain grew well H 2 -dependently in a minimal medium with amino acids and glucarate provided as the sole carbon sources, whereas the gudT strain achieved approximately 30% of the parent strain's yield. Glucarate-mediated growth of a mutant strain unable to produce H 2 was stimulated by H 2 addition, presumably owing to the positive transcriptional response to H 2 . Gut microbiota-produced molecular hydrogen apparently signals Salmonella to catabolize an alternative carbon source available in the host. Our results link a gut microbiome-produced diffusible metabolite to augmenting bacterial pathogenesis.


2017 ◽  
Vol 233 ◽  
pp. 67-73 ◽  
Author(s):  
Takashi Kogo ◽  
Yuki Yoshida ◽  
Keisuke Koganei ◽  
Hitoshi Matsumoto ◽  
Taisuke Watanabe ◽  
...  

2014 ◽  
Vol 14 (2) ◽  
pp. 88-95 ◽  
Author(s):  
Lilian dos Santos Castro ◽  
Amanda Cristina Campos Antoniêto ◽  
Wellington Ramos Pedersoli ◽  
Rafael Silva-Rocha ◽  
Gabriela F. Persinoti ◽  
...  

2008 ◽  
Vol 158 (3) ◽  
pp. 552-560 ◽  
Author(s):  
Pradnya Deshpande ◽  
Sajitha Nair ◽  
Shubhangi Khedkar

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