Quantitative Site-Specific Phosphoproteomics of <i>Trichoderma reesei</i> Signaling Pathways upon Induction of Hydrolytic Enzyme Production

Nguyen, Elizabeth V.; Imanishi, Susumu Y.; Haapaniemi, Pekka; Yadav, Avinash; Saloheimo, Markku; Corthals, Garry L.; Pakula, Tiina

doi:10.1021/acs.jproteome.5b00796

Cited by 33 publications

(42 citation statements)

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“…Recently, a phosphoproteomic investigation of T. reesei exposed to sophorose and distiller’s spent grain showed that Tmk3 is phosphorylated under these cellulase-inducing conditions [64]. What’s particularly interesting is that Tmk3 was also the only MAPK that is found to be phosphorylated when exposed to sophorose or distiller’s spent grain.…”

Section: Discussionmentioning

confidence: 99%

Role of Trichoderma reesei mitogen-activated protein kinases (MAPKs) in cellulase formation

Wang

Zhang

et al. 2017

Biotechnol Biofuels

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BackgroundDespite being the most important cellulase producer, the cellulase-regulating carbon source signal transduction processes in Trichoderma reesei are largely unknown. Elucidating these processes is the key for unveiling how external carbon sources regulate cellulase formation, and ultimately for the improvement of cellulase production and biofuel production from lignocellulose.ResultsIn this work, the role of the mitogen-activated protein kinase (MAPK) signal transduction pathways on cellulase formation was investigated. The deletion of yeast FUS3-like tmk1 in T. reesei leads to improved growth and significantly improved cellulase formation. However, tmk1 deletion has no effect on the transcription of cellulase-coding genes. The involvement of the cell wall integrity maintenance governing yeast Slt2-like Tmk2 in cellulase formation was investigated by overexpressing tmk3 in T. reesei Δtmk2 to restore cell wall integrity. Transcriptional analysis found little changes in cellulase-coding genes between T. reesei parent, Δtmk2, and Δtmk2::OEtmk3 strains. Cell wall integrity decreased in T. reesei Δtmk2 over the parent strain and restored in Δtmk2::OEtmk3. Meanwhile, cellulase formation is increased in T. reesei Δtmk2 and then decreased in T. reesei Δtmk2::OEtmk3. ConclusionsThese investigations elucidate the role of Tmk1 and Tmk2 on cellulase formation: they repress cellulase formation, respectively, by repressing growth and maintaining cell wall integrity, while neither MAPK regulates the transcription of cellulase-coding genes. This work, together with the previous investigations, suggests that all MAPKs are involved in cellulase formation, while Tmk3 is the only MAPK involved in signal transduction for the regulation of cellulase expression on the transcriptional level.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-017-0789-x) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Role of Trichoderma reesei mitogen-activated protein kinases (MAPKs) in cellulase formation

Wang

Zhang

et al. 2017

Biotechnol Biofuels

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“…Production of galactaric acid was much higher in these 4 mL batch cultures than has previously been reported for flask cultures with filamentous fungi (around 4 g L −1 ; [8, 10]). The medium in the 24-well plates contained spent grain extract, which was expected to induce the CBH1 promoter [16], but not necessarily other promoters. Spent grain extract also provides organic nitrogen, resulting in a richer medium, which may contribute to the good conversion of d -galacturonic acid into galactaric acid.…”

Section: Discussionmentioning

confidence: 99%

Scaling up and scaling down the production of galactaric acid from pectin using Trichoderma reesei

2017

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BackgroundBioconversion of d-galacturonic acid to galactaric (mucic) acid has previously been carried out in small scale (50–1000 mL) cultures, which produce tens of grams of galactaric acid. To obtain larger amounts of biologically produced galactaric acid, the process needed to be scaled up using a readily available technical substrate. Food grade pectin was selected as a readily available source of d-galacturonic acid for conversion to galactaric acid.ResultsWe demonstrated that the process using Trichoderma reesei QM6a Δgar1 udh can be scaled up from 1 L to 10 and 250 L, replacing pure d-galacturonic acid with commercially available pectin. T. reesei produced 18 g L−1 galactaric acid from food-grade pectin (yield 1.00 g [g d-galacturonate consumed]−1) when grown at 1 L scale, 21 g L−1 galactaric acid (yield 1.11 g [g d-galacturonate consumed]−1) when grown at 10 L scale and 14 g L−1 galactaric acid (yield 0.77 g [g d-galacturonate consumed]−1) when grown at 250 L scale. Initial production rates were similar to those observed in 500 mL cultures with pure d-galacturonate as substrate. Approximately 2.8 kg galactaric acid was precipitated from the 250 L culture, representing a recovery of 77% of the galactaric acid in the supernatant. In addition to scaling up, we also demonstrated that the process could be scaled down to 4 mL for screening of production strains in 24-well plate format. Production of galactaric acid from pectin was assessed for three strains expressing uronate dehydrogenase under alternative promoters and up to 11 g L−1 galactaric acid were produced in the batch process.ConclusionsThe process of producing galactaric acid by bioconversion with T. reesei was demonstrated to be equally efficient using pectin as it was with d-galacturonic acid. The 24-well plate batch process will be useful screening new constructs, but cannot replace process optimisation in bioreactors. Scaling up to 250 L demonstrated good reproducibility with the smaller scale but there was a loss in yield at 250 L which indicated that total biomass extraction and more efficient DSP would both be needed for a large scale process.

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“…Most of these conserved regions contain up to three continuous S/T phosphorylation sites. Moreover, phosphorylation of the peptide sequence extending from amino acids 387 to 402 (SSTTGSLAGGDLMDRM) at the C-terminus of Cre1 from T. reesei (Tr_Cre1) was identified with LC-MS/MS by Nguyen et al (2016).…”

Section: Phosphorylation Of Cre1mentioning

confidence: 99%

“…Meanwhile, it was assumed that phosphorylation of Cre1 in T. reesei affects its repressing activity on cellulase genes under CCR (Cziferszky et al, 2002;Nguyen et al, 2016).…”

Section: Figmentioning

confidence: 99%

“…Horta et al (2019) revealed that there is large-scale phosphorylation and dephosphorylation of substrate-specific proteins, including Cre1, in Neurospora crassa, according to MS/MS-based peptide analysis. Nguyen et al (2016) suggested that phosphorylation of Cre1 plays a role in the onset of CCR induction and sensing of the carbon source. Cziferszky et al (2002) reported that phosphorylation of Cre1 at S241 in T. reesei has a significant effect on the repression of cellulolytic enzyme genes at the transcriptional level under CCR.…”

Section: Introductionmentioning

confidence: 99%

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Precision engineering of the transcription factor Cre1 inHypocrea jecorina (Trichoderma reesei)for efficient cellulase production in the presence of glucose

Tan

Niu

et al. 2020

Preprint

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In Trichoderma reesei, carbon catabolite repression (CCR) significantly downregulates the transcription of cellulolytic enzymes, which is usually mediated by the zinc finger protein Cre1. It was found that there is a conserved region at the C-terminus of Cre1/CreA in several cellulase-producing fungi that contains up to three continuous S/T phosphorylation sites. Here, S387, S388, T389, and T390 at the C-terminus of Cre1 in T.reesei were mutated to valine for mimicking an unphosphorylated state, thereby generating the transformants Tr_Cre1 S387V , Tr_Cre1 S388V , Tr_Cre1 T389V , and Tr_Cre1 T390V , respectively. Transcription of cel7a in Tr_ Cre1 S388V was markedly higher than that of the parent strain when grown in glucose-containing media. Under these conditions, both filter paperase (FPase) and p-nitrophenyl-β-D-cellobioside (pNPCase) activities, as well as soluble proteins from Tr_Cre1 S388V were significantly increased by up to 2-to 3-fold compared with that of other transformants and the parent strain. To our knowledge, this is the first report demonstrating an improvement of cellulase production in fungal species under CCR by mimicking dephosphorylation at the C-terminus of Cre1. Taken together, we developed a precision engineering strategy based on the modification of phosphorylation sites of Cre1 transcription factor to enhance the production of cellulase in fungal species under CCR. AbbreviationsCCR: carbon catabolite repression; RT-qPCR: reverse transcription quantitative polymerase chain reaction; PKA: cyclic adenosine monophosphate (cAMP)-dependent protein kinase A; FP: filter paper

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Quantitative Site-Specific Phosphoproteomics of Trichoderma reesei Signaling Pathways upon Induction of Hydrolytic Enzyme Production

Cited by 33 publications

References 42 publications

Role of Trichoderma reesei mitogen-activated protein kinases (MAPKs) in cellulase formation

Role of Trichoderma reesei mitogen-activated protein kinases (MAPKs) in cellulase formation

Scaling up and scaling down the production of galactaric acid from pectin using Trichoderma reesei

Precision engineering of the transcription factor Cre1 inHypocrea jecorina (Trichoderma reesei)for efficient cellulase production in the presence of glucose

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