Mar 9, 2020

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

bioRxiv
Lijuan HanXu Fang

Abstract

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 _Cre1S387V, Tr _Cre1S388V, Tr _Cre1T389V, and Tr _Cre1T390V, respectively. Transcription of cel7a in Tr _Cre1S388V 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 ( p NPCase) activities, as well as soluble proteins from Tr _Cre1S388V 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...Continue Reading

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Mentioned in this Paper

Transcription Factor
Tissue Engineering
Mental State (Observable Entity)
Down-Regulation
Carbon
Protein Dephosphorylation
Species
Carbon Catabolite Repression
Fungi
ZNF2 gene

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