Replacement of the carbon catabolite regulator (cre1) and fed-batch cultivation as strategies to enhance cellulase production in Trichoderma harzianum

Delabona, Priscila da Silva; Lima, Deise Juliana da Silva; Codima, Carla Aloia; Ramoni, Jonas; Gelain, Lucas; Melo, Vandierly Sampaio de; Farinas, Cristiane S.; Pradella, José Geraldo da Cruz; Schink, Bernhard

doi:10.1016/j.biteb.2021.100634

Cited by 4 publications

(9 citation statements)

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“…Although the importance of XYR1 and CRE1 in the expression of CAZyme-encoding genes and other proteins required for lignocellulose degradation is evident, the transcriptional regulation mediated by both proteins in T. harzianum strains remains poorly explored ( Delabona et al, 2017 ; Delabona et al, 2021 ). A previous study demonstrated that some genes encoding regulatory proteins, such as xyr1 , evolved by vertical gene transfer in Trichoderma spp.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, the effect of cre1 deletion on the gene expression profile of its targets varies among species. In T. reesei RUT-C30, the deletion of a full cre1 can lead to pleiotropic effects and strong growth impairment ( Portnoy et al, 2011 ; Mello-de-Sousa et al, 2014 ), whereas in T. harzianum P49P11, it can increase the expression levels of CAZyme genes and xyr1 ( Delabona et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

“…Although T. harzianum strains have shown high cellulolytic activity ( Horta et al, 2018 ; Li J. X. et al, 2020 ), most studies investigating this species have focused on biological control ( Maruyama et al, 2020 ; Yan and Khan 2021 ). Thus, the regulatory mechanisms underlying hydrolytic enzyme production by these fungi are still poorly explored at the transcriptional level ( Delabona et al, 2017 ; Delabona et al, 2020 ; Delabona et al, 2021 ). In addition, given the high degree of genetic variation observed within the genus Trichoderma ( Kubicek et al, 2019 ) along with the complex speciation observed in the T. harzianum species ( Druzhinina et al, 2010 ), it is necessary to explore how the main regulators XYR1 and CRE1 behave across T. harzianum strains.…”

Section: Introductionmentioning

confidence: 99%

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Network Analysis Reveals Different Cellulose Degradation Strategies Across Trichoderma harzianum Strains Associated With XYR1 and CRE1

et al. 2022

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Trichoderma harzianum, whose gene expression is tightly controlled by the transcription factors (TFs) XYR1 and CRE1, is a potential candidate for hydrolytic enzyme production. Here, we performed a network analysis of T. harzianum IOC-3844 and T. harzianum CBMAI-0179 to explore how the regulation of these TFs varies between these strains. In addition, we explored the evolutionary relationships of XYR1 and CRE1 protein sequences among Trichoderma spp. The results of the T. harzianum strains were compared with those of Trichoderma atroviride CBMAI-0020, a mycoparasitic species. Although transcripts encoding carbohydrate-active enzymes (CAZymes), TFs, transporters, and proteins with unknown functions were coexpressed with cre1 or xyr1, other proteins indirectly related to cellulose degradation were identified. The enriched GO terms describing the transcripts of these groups differed across all strains, and several metabolic pathways with high similarity between both regulators but strain-specific differences were identified. In addition, the CRE1 and XYR1 subnetworks presented different topology profiles in each strain, likely indicating differences in the influences of these regulators according to the fungi. The hubs of the cre1 and xyr1 groups included transcripts not yet characterized or described as being related to cellulose degradation. The first-neighbor analyses confirmed the results of the profile of the coexpressed transcripts in cre1 and xyr1. The analyses of the shortest paths revealed that CAZymes upregulated under cellulose degradation conditions are most closely related to both regulators, and new targets between such signaling pathways were discovered. Although the evaluated T. harzianum strains are phylogenetically close and their amino acid sequences related to XYR1 and CRE1 are very similar, the set of transcripts related to xyr1 and cre1 differed, suggesting that each T. harzianum strain used a specific regulation strategy for cellulose degradation. More interestingly, our findings may suggest that XYR1 and CRE1 indirectly regulate genes encoding proteins related to cellulose degradation in the evaluated T. harzianum strains. An improved understanding of the basic biology of fungi during the cellulose degradation process can contribute to the use of their enzymes in several biotechnological applications and pave the way for further studies on the differences across strains of the same species.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Network Analysis Reveals Different Cellulose Degradation Strategies Across Trichoderma harzianum Strains Associated With XYR1 and CRE1

et al. 2022

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“…Although T. harzianum strains have shown high cellulolytic activity (Horta et al, 2018;Li et al, 2020b), most studies investigating this species have focused on biological control (Maruyama et al, 2020;Yan and Khan, 2021). Thus, the regulatory mechanisms underlying hydrolytic enzyme production by these fungi are still poorly explored at the transcriptional level (Delabona et al, 2017(Delabona et al, , 2020(Delabona et al, , 2021. In addition, given the high degree of genetic variation observed within the genus Trichoderma (Kubicek et al, 2019) along with the complex speciation observed in the T. harzianum species (Druzhinina et al, 2010), it is necessary to explore how the main regulators XYR1 and CRE1 behave across T. harzianum strains.…”

Section: Xyr1 and Cre1 Response To Cellulose Degradationmentioning

confidence: 99%

“…Interestingly, the effect of cre1 deletion on the gene expression profile of its targets varies among species. In T. reesei RUT-C30, the deletion of a full cre1 can lead to pleiotropic effects and strong growth impairment (Mello-de-Sousa, et al 2014; Portnoy, et al 2011), whereas in T. harzianum P49P11, it can increase the expression levels of CAZyme genes and xyr1 (Delabona, et al 2021).…”

Section: Introductionmentioning

confidence: 99%

Network Analysis Reveals Different Cellulose Degradation Strategies acrossTrichoderma harzianumStrains Associated with XYR1 and CRE1

Rosolen¹,

Aono²,

Almeida³

et al. 2020

Preprint

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Background: Trichoderma species have attracted attention as alternative plant polysaccharide-degrading enzyme sources, among other reasons. Most Trichoderma spp., such as Trichoderma atroviride and Trichoderma harzianum, are described as mycoparasitic fungi and are widely used in agriculture as biocontrol agents. T. harzianum is also a potential candidate for hydrolytic enzyme production in which gene expression is tightly controlled. Thus, a better understanding of transcriptional regulation related to the main activator (XYR1) and repressor (CRE1) of cellulases and hemicellulases is necessary. Herein, we explore the genetic mechanisms of the key regulators of genes encoding plant cell wall-degrading enzymes by inferring a gene coexpression network for T. harzianum (IOC-3844 and CBMAI-0179) and, for comparative purposes, for T. atroviride CBMAI-0020.Results: Phylogenetic analysis indicated that both regulatory proteins are widely distributed among ascomycete fungi and suggested how T. atroviride is differentiated from T. harzianum. The coexpression network analyses separated the transcripts into several groups according to the expression profile during growth in cellulose or glucose.Groups with xyr1 or cre1 were identified, and within them, transcripts encoding carbohydrate-active enzymes (CAZymes), TFs, sugar and ion transporters, and proteins with unknown function were coexpressed. However, qualitative and quantitative differences among groups and strains were observed. Core transcripts from these groups (hubs) were identified, and they included transcripts not yet characterized or described as related to cellulose degradation. Several metabolic pathways triggered were recognized with high similarity between both regulators during cellulose degradation, but they differed according to the strains. An enrichment related to mycoparasitism in T. atroviride, especially that associated with CRE1, was observed. It was also noticed that the transcripts encoding CAZymes related to polysaccharide degradation in T.harzianum are not necessarily coexpressed with the TFs studied herein. Conclusion: Our results suggest that different strategies related to XYR1 and CRE1 are used by Trichoderma spp. during cellulose degradation and provide new insights into transcripts coexpressed with both TFs in T. harzianum. This knowledge can be exploited to improve the understanding of the genetic mechanisms involved in hydrolytic enzyme regulation, expanding the potential of T. harzianum use in several industrial applications. 3 Due to the expanding population and industrialization, the demand for energy is increasing throughout the world. Currently, the primary energy source is fossil fuels, i.e., nonrenewable sources such as natural gas, oil, and coal [1]. To reduce the dependency on these finite resources, the development of new sustainable alternatives has become critical. In this context, increasing attention has been given to the development of biofuels from biomass [2, 3]. The United States and Brazil are the world's largest bioet...

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Cellulases production optimization and inductors evaluation in pretreated plantain rachis by Penicillium oxalicum

Jiménez Fallas,

Charpentier Alfaro,

Wischral

et al. 2024

Env Prog and Sustain Energy

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Pretreated lignocellulosic residues are suitable substrates for cellulases production by filamentous fungi. In the current work, plantain rachis was pretreated with sequential acid and alkali and then used as the main carbon source for cellulases production. First, a full 23 factorial design and response surface methodology (RSM), based on central composite rotatable design (CCRD), were carried out to cellulases production media optimization from plantain rachis by Penicillium oxalicum. The cellulases production was evaluated in flasks and bioreactor scale; in parallel, the addition of possible cellulases inductors was evaluated in flasks: molasses, beer bran, oat bran, and wheat bran. Results from statistical analyses with a level of confidence of 95% demonstrated that the concentration of ammonium sulfate must be kept at 1.625 g/L. The optimum urea and yeast extract concentrations were 0.560 g/L and 0.250 g/L, respectively. Cellulases volumetric productivities were higher in instrumented bioreactor than in flasks: 78.03% for exoglucanase, 10.87% for endoglucanase, 1.58% for β‐glucosidase, and 44.36% for FPU. Therefore, P. oxalicum was able to produce cellulases from plantain rachis in flasks and bioreactor, and molasses was the additional inductor that presented an increment in cellulases activities: endoglucanase 15%, exoglucanase 81%, and β‐glucosidase 55%.

show abstract

Replacement of the carbon catabolite regulator (cre1) and fed-batch cultivation as strategies to enhance cellulase production in Trichoderma harzianum

Cited by 4 publications

References 42 publications

Network Analysis Reveals Different Cellulose Degradation Strategies Across Trichoderma harzianum Strains Associated With XYR1 and CRE1

Network Analysis Reveals Different Cellulose Degradation Strategies Across Trichoderma harzianum Strains Associated With XYR1 and CRE1

Network Analysis Reveals Different Cellulose Degradation Strategies acrossTrichoderma harzianumStrains Associated with XYR1 and CRE1

Cellulases production optimization and inductors evaluation in pretreated plantain rachis by Penicillium oxalicum

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