Understanding the Role of Trichoderma reesei Vib1 in Gene Expression during Cellulose Degradation

Chen, Xiuzhen; Song, Bingran; Liu, Minglu; Qin, Lu‐Ping; Dong, Zhiyang

doi:10.3390/jof7080613

Cited by 6 publications

(8 citation statements)

References 48 publications

(62 reference statements)

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“…3f, g and Table 2), which are vital positive TFs that regulate cellulase genes in T. reesei [9,12]. Furthermore, vib1 was prominently enhanced (Table 2), whose deletion caused the decreased expression of almost all cellulase and hemicellulase genes, crucial sugar transporter genes, and main transcriptional activators xyr1 and ace3 in T. reesei [52]. In Neurospora crassa, Vib1 modulated cellulase synthesis by regulating the expression of the essential cellulase regulator CLR2 [53].…”

Section: Discussionmentioning

confidence: 99%

Mechanism of Zn2+ regulating cellulase production in Trichoderma reesei Rut-C30

Chen

et al. 2023

Preprint

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Background Trichoderma reesei Rut-C30 is a hypercellulolytic mutant strain that degrades abundant sources of lignocellulosic plant biomass into renewable biofuels. Zn2+ is an activator of enzymes in almost all organisms; however, its effect on cellulase activity has not been reported in T. reesei. Results Although high concentrations of Zn2+ severely hampered the extension of T. reesei mycelia, adding 1–4 mM Zn2+ improved cellulase production in T. reesei high-yielding cellulase-producing strain Rut-C30. The expression levels of the major cellulase genes and two critical transcription activators (xyr1 and ace3) increased significantly on Zn2+ addition. Transcriptome analysis revealed that the mRNA level of plc-e encoding phospholipase C, which is involved in the calcium signaling pathway, was enhanced by Zn2+ addition. The disruption of plc-e abolished the cellulase-positive influence of Zn2+ in the early phase, indicating that plc-e is involved in Zn2+-induced cellulase production; furthermore, supplementation with LaCl3 (a plasma membrane Ca2+ channel blocker) and deletion of crz1 (calcineurin-responsive zinc finger transcription factor 1) demonstrated that calcium signaling is partly involved in this process. Moreover, we identified the zinc-responsive transcription factor zafA; the transcriptional level of zafA decreased significantly with Zn2+ stress, and its deletion indicated that zafA mainly mediates Zn2+-induced cellulase production. Conclusions For the first time, we have demonstrated that Zn2+ was toxic to T. reesei and it remarkably promoted cellulase production. This positive influence of Zn2+ was facilitated by the gene plc-e and the transcription factor zafA. These findings provide insights into the role of Zn2+ in T. reesei and a mechanistic study of signal transduction in cellulase synthesis.

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Section: Discussionmentioning

confidence: 99%

Mechanism of Zn2+ regulating cellulase production in Trichoderma reesei Rut-C30

Chen

et al. 2023

Preprint

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“…2). Previously, the deletion of these factors was reported to cause loss or reduction of cellulase expression [38][39][40]45 . These factors were also affected by enhanced expression of XYR1 and ACE3 (for example, crt1 is regulated by ACE3 38,47 ).…”

Section: Discussionmentioning

confidence: 99%

“…In T. reesei , VIB1 is also essential for cellulase production and acts as an important regulator of cellulase induction 36 , 44 . A comparison of the Δ vib1 and Δ xyr1 transcriptomes using cellulose as a carbon source showed a high frequency of overlap between cellulose-induced genes regulated by VIB1 and XYR1 targets 45 . This suggests that VIB1 partially regulates cellulase gene expression via XYR1 45 .…”

Section: Introductionmentioning

confidence: 99%

Inducer-free cellulase production system based on the constitutive expression of mutated XYR1 and ACE3 in the industrial fungus Trichoderma reesei

Arai

Ichinose

Shibata

et al. 2022

Sci Rep

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Trichodermareesei is a widely used host for producing cellulase and hemicellulase cocktails for lignocellulosic biomass degradation. Here, we report a genetic modification strategy for industrial T.reesei that enables enzyme production using simple glucose without inducers, such as cellulose, lactose and sophorose. Previously, the mutated XYR1V821F or XYR1A824V was known to induce xylanase and cellulase using only glucose as a carbon source, but its enzyme composition was biased toward xylanases, and its performance was insufficient to degrade lignocellulose efficiently. Therefore, we examined combinations of mutated XYR1V821F and constitutively expressed CRT1, BGLR, VIB1, ACE2, or ACE3, known as cellulase regulators and essential factors for cellulase expression to the T.reesei E1AB1 strain that has been highly mutagenized for improving enzyme productivity and expressing a ß-glucosidase for high enzyme performance. The results showed that expression of ACE3 to the mutated XYR1V821F expressing strain promoted cellulase expression. Furthermore, co-expression of these two transcription factors also resulted in increased productivity, with enzyme productivity 1.5-fold higher than with the conventional single expression of mutated XYR1V821F. Additionally, that productivity was 5.5-fold higher compared to productivity with an enhanced single expression of ACE3. Moreover, although the DNA-binding domain of ACE3 had been considered essential for inducer-free cellulase production, we found that ACE3 with a partially truncated DNA-binding domain was more effective in cellulase production when co-expressed with a mutated XYR1V821F. This study demonstrates that co-expression of the two transcription factors, the mutated XYR1V821F or XYR1A824V and ACE3, resulted in optimized enzyme composition and increased productivity.

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“…Fungal transformation was based on infection using the GV3101 strain of Agrobacterium tumefaciens [ 51 ]. T. reesei Rut-C30 (ATCC 56765) [ 52 ] was used as the host strain for genetic transformation. Luria broth (LB) was used to culture E. coli and A. tumefaciens , and MA medium [ 12 ] containing 2% (w/v) glucose and MM [ 46 ] containing 1% (w/v) Avicel or 2% (w/v) glucose were used for general fungal culture.…”

Section: Methodsmentioning

confidence: 99%

Mechanism of Zn2+ regulation of cellulase production in Trichoderma reesei Rut-C30

Chen

et al. 2023

Biotechnol Biofuels

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Background Trichoderma reesei Rut-C30 is a hypercellulolytic mutant strain that degrades abundant sources of lignocellulosic plant biomass, yielding renewable biofuels. Although Zn2+ is an activator of enzymes in almost all organisms, its effects on cellulase activity in T. reesei have yet to be reported. Results Although high concentrations of Zn2+ severely suppressed the extension of T. reesei mycelia, the application of 1–4 mM Zn2+ enhanced cellulase and xylanase production in the high-yielding cellulase-producing Rut-C30 strain of T. reesei. Expression of the major cellulase, xylanase, and two essential transcription activator genes (xyr1 and ace3) increased in response to Zn2+ stimulation. Transcriptome analysis revealed that the mRNA levels of plc-e encoding phospholipase C, which is involved in the calcium signaling pathway, were enhanced by Zn2+ application. The disruption of plc-e abolished the cellulase-positive influence of Zn2+ in the early phase of induction, indicating that plc-e is involved in Zn2+-induced cellulase production. Furthermore, treatment with LaCl3 (a plasma membrane Ca2+ channel blocker) and deletion of crz1 (calcineurin-responsive zinc finger transcription factor 1) indicated that calcium signaling is partially involved in this process. Moreover, we identified the zinc-responsive transcription factor zafA, the transcriptional levels of which declined in response to Zn2+ stress. Deletion of zafA indicates that this factor plays a prominent role in mediating the Zn2+-induced excessive production of cellulase. Conclusions For the first time, we have demonstrated that Zn2+ is toxic to T. reesei, although promotes a marked increase in cellulase production. This positive influence of Zn2+ is facilitated by the plc-e gene and zafA transcription factor. These findings provide insights into the role of Zn2+ in T. reesei and the mechanisms underlying signal transduction in cellulase synthesis.

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Understanding the Role of Trichoderma reesei Vib1 in Gene Expression during Cellulose Degradation

Cited by 6 publications

References 48 publications

Mechanism of Zn2+ regulating cellulase production in Trichoderma reesei Rut-C30

Mechanism of Zn2+ regulating cellulase production in Trichoderma reesei Rut-C30

Inducer-free cellulase production system based on the constitutive expression of mutated XYR1 and ACE3 in the industrial fungus Trichoderma reesei

Mechanism of Zn2+ regulation of cellulase production in Trichoderma reesei Rut-C30

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