Increased endoplasmic reticulum content of a mutant of Trichoderma reesei (RUT-C30) in relation to cellulase synthesis

Ghosh, Arati; Al-Rabiai, S.; Ghosh, B. K.; Trimiño-Vazquez, H.; Eveleigh, D. E.; Montenecourt, Bland S.

doi:10.1016/0141-0229(82)90093-x

Cited by 61 publications

(24 citation statements)

References 11 publications

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“…Braell, 1988). This explanation is strongly supported by the findings that the hypersecretory T. reesei mutant RUTC-30 -which contains an increased cellular amount of endoplasmic reticulum (Ghosh et al, 1982(Ghosh et al, , 1984 -exhibited almost no stimulation of exoprotein secretion by choline or Tween 80. It is also supported by the fact that this mutant already contains elevated activities of Dol-P-Man synthase.…”

Section: Discussionsupporting

confidence: 73%

Stimulation of exoprotein secretion by choline and Tween 80 in Trichoderma reesei QM 9414 correlates with increased activities of dolichol phosphate mannose synthase

Kruszewska

Palamarczyk

Kubicek

1990

Journal of General Microbiology

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~Addition of choline (20 mM) or Tween 80 (0.06 %) to the culture medium of Trichoderma reesei QM 9414 increased (a) the secretion of protein under both carbon-catabolite-repressed and -derepressed conditions, and (b) cellulase secretion under carbon-catabolite-derepressed conditions. In contrast, no stimulation by choline or Tween 80 was observed with the hypersecretory strain T. reesei RUT C-30. In view of the obligatory role of 0-glycosylation in protein secretion by this fungus, an investigation was made into the effects on this process of choline and Tween 80.A membrane preparation was isolated from both strains of T. reesei and used to assay enzymes involved in 0-glycosylation. Significant differences were observed with respect to the activity of dolichol phosphate mannose (Dol-P-Man) synthase only. Strain QM 9414, grown on media supplemented with choline or Tween 80 exhibited a two-to threefold higher activity of Dol-P-Man synthase compared to a control lacking these supplements. This stimulatory effect was observed during growth under both carbon-catabolite-repressed and -derepressed conditions. In contrast, strain RUT C-30 exhibited decreased activities of Dol-P-Man synthase when grown in media supplemented with choline. Choline had no effect on Dol-P-Man synthase in vifro, whereas Tween 80 decreased the activity. Thus the effect of Tween 80 or choline on protein secretion by T. reesei may be due to a stimulation of formation and/or activity of Dol-P-Man synthase, thereby elevating the level of 0-glycosylation and protein secretion.

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Section: Discussionsupporting

confidence: 73%

Stimulation of exoprotein secretion by choline and Tween 80 in Trichoderma reesei QM 9414 correlates with increased activities of dolichol phosphate mannose synthase

Kruszewska

Palamarczyk

Kubicek

1990

Journal of General Microbiology

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“…Early electron microscopic analysis of RUT-C30 grown on Avicel cellulose exposed a six-to sevenfold higher endoplasmic reticulum (ER) content in RUT-C30 during secretion than in QM6a. Furthermore, the phospholipid content of mycelium and the amount of the ER marker enzyme nucleoside diphosphatase was approximately double that of QM6a on the fourth day of growth (Ghosh et al, 1982). Mycelial protein content of RUT-C30 was four-five times higher than that of QM6a, the endoglucanase activity was 30 times higher and b-glucosidase activity three times higher in RUT-C30 cells.…”

Section: Physiological Changes In Rut-c30mentioning

confidence: 85%

Trichoderma reesei RUT-C30 – thirty years of strain improvement

2012

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The hypersecreting mutant Trichoderma reesei RUT-C30 (ATCC 56765) is one of the most widely used strains of filamentous fungi for the production of cellulolytic enzymes and recombinant proteins, and for academic research. The strain was obtained after three rounds of random mutagenesis of the wild-type QM6a in a screening program focused on high cellulase production and catabolite derepression. Whereas RUT-C30 achieves outstanding levels of protein secretion and high cellulolytic activity in comparison to the wild-type QM6a, recombinant protein production has been less successful. Here, we bring together and discuss the results from biochemical-, microscopic-, genomic-, transcriptomic-, glycomic-and proteomic-based research on the RUT-C30 strain published over the last 30 years.

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“…For example, defined cell wall components can be used as antigens for preparation of vaccines or immunodiagnostic reagents [91], or as nonspecific immunostimulants [ 921. Indeed, it has been observed that the hypersecretory mutant of Trichoderma reesei Rut-C 30 producing cellulase, contains increased content of ER elements in its cytoplasm [97]. Furthermore, the knowledge on the cell wall structure is necessary for prediction of methods for covalent immobilization of fungal cells on various supports [93] or for their precipitation with natural or artificial flocculants [94].…”

Section: Biotechnological Aspectsmentioning

confidence: 99%