Genetic engineering and raising temperature enhance recombinant protein production with the cdna1 promoter in Trichoderma reesei

Jiang, Shanshan; Wang, Yue; Liu, Qin; Zhao, Qinqin; Gao, Liwei; Song, Xin; Li, Xuezhi; Qu, Yinbo; Liu, Guodong

doi:10.1186/s40643-022-00607-2

Cited by 4 publications

(1 citation statement)

References 30 publications

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“…To further investigate whether interference of Trrab7 affects a step downstream of cellulase gene transcription, and to evidence that the reduced cellulase production of P tcu-rab7 KD is not due to the compromised secretory capability, the major endoglucanase, EG1, was expressed from the constitutive promoter cdna1 at the pyr4 locus of the P tcu - rab7 KD strain and QM9414, respectively [ 40 ]. Both extracellular CMCase activity and SDS–PAGE analyses showed that EG1 produced in P tcu - rab7 KD was comparable with that in QM9414 & P cdna - eg1 when cultured on glucose (Fig.…”

Section: Resultsmentioning

confidence: 99%

Small GTPase Rab7 is involved in stress adaptation to carbon starvation to ensure the induced cellulase biosynthesis in Trichoderma reesei

Liu,

Wang,

Fang

et al. 2024

Biotechnol Biofuels

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Background The saprophytic filamentous fungus Trichoderma reesei represents one of the most prolific cellulase producers. The bulk production of lignocellulolytic enzymes by T. reesei not only relies on the efficient transcription of cellulase genes but also their efficient secretion after being translated. However, little attention has been paid to the functional roles of the involved secretory pathway in the high-level production of cellulases in T. reesei. Rab GTPases are key regulators in coordinating various vesicle trafficking associated with the eukaryotic secretory pathway. Specifically, Rab7 is a representative GTPase regulating the transition of the early endosome to the late endosome followed by its fusion to the vacuole as well as homotypic vacuole fusion. Although crosstalk between the endosomal/vacuolar pathway and the secretion pathway has been reported, the functional role of Rab7 in cellulase production in T. reesei remains unknown. Results A TrRab7 was identified and characterized in T. reesei. TrRab7 was shown to play important roles in T. reesei vegetative growth and vacuole morphology. Whereas knock-down of Trrab7 significantly compromised the induced production of T. reesei cellulases, overexpression of the key transcriptional activator, Xyr1, restored the production of cellulases in the Trrab7 knock-down strain (Ptcu-rab7KD) on glucose, indicating that the observed defective cellulase biosynthesis results from the compromised cellulase gene transcription. Down-regulation of Trrab7 was also found to make T. reesei more sensitive to various stresses including carbon starvation. Interestingly, overexpression of Snf1, a serine/threonine protein kinase known as an energetic sensor, partially restored the cellulase production of Ptcu-rab7KD on Avicel, implicating that TrRab7 is involved in an energetic adaptation to carbon starvation which contributes to the successful cellulase gene expression when T. reesei is transferred from glucose to cellulose. Conclusions TrRab7 was shown to play important roles in T. reesei development and a stress response to carbon starvation resulting from nutrient shift. This adaptation may allow T. reesei to successfully initiate the inducing process leading to efficient cellulase production. The present study provides useful insights into the functional involvement of the endosomal/vacuolar pathway in T. reesei development and hydrolytic enzyme production.

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Section: Resultsmentioning

confidence: 99%