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Choline stimulates synthesis of extracellular proteins in Trichoderma reesei QM 9414

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Abstract

A correlation between intracellular phospholipid levels and the rate of exoprotein synthesis was investigated in the filamentous fungus Trichoderma reesei QM 9414 during growth on cellulose. When the incubation temperature was varied between 20 and 37°C, the exoprotein synthesis rate correlated with the total cellular amount of phospholipids, but not with an individual phospholipid component. In contrast, when phospholipid bases were added exogenuously, a significant stimulation of exoprotein synthesis was observed with choline. The addition of the surfactant Tween 80—which also stimulates exoprotein secretion in T. reesei QM 9414—prevented choline stimulation. Optimal stimulation occurred around 20 mM choline. Choline stimulated exoprotein synthesis in general as shown by increased activities of several extracellular enzymes. Mycelia required preincubation for at least 20 h before stimulation of choline could be seen. Mycelia pregrown in the absence or presence of choline were equally effective in formation of β-glucosidase upon induction with methyl-β-d-glucoside, and the addition of choline to the induction medium had no effect. Choline did not alter the osmotic stability of protoplasts of T. reesei. Electron microscopic examinations and analysis of chemical constituents as well as marker enzymes from choline grown and non-choline grown mycelia revealed higher contents of mitochondria and endoplasmic reticula in choline grown mycelia. The possibility is discussed that choline may stimulate exoprotein synthesis by increasing the cellular content of endoplasmic reticula.

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References

  • Biely P, Vrsanaska M, Kratky Z (1980) Xylan degrading enzymes of the yeast Cryptococcus albidus. Identification and cellular localization. Eur J Biochem 108:313–321

    Google Scholar 

  • Broekhuyse RM (1968) Phospholipids in tissues of the eye. I. Isolation, characterization and quantitative analysis by two-dimensional thin-layer chromatography of diacly- and vinyl-ether lipids. Biochim Biophys Acta 152:307–315

    Google Scholar 

  • Davis BD, Tai PC (1980) The mechanism of protein secretion across membranes. Nature (Lond) 283:433–438

    Google Scholar 

  • De Duve C (1967) Criteria of homogeneity and purity of mitochondria. In: Estabrook RW, Pullman ME (eds) Methods in enzymology 10. Academic Press, New York London, pp 7–18

    Google Scholar 

  • Fishman Y, Rottem S, Citri N (1980) Preferential suppression of normal exoenzyme formation by membrane-modifying agents. J Bacteriol 141:1435–1438

    Google Scholar 

  • Folch J, Lees M, Sloane-Stanley GH (1957) A simple method for the isolation and purification of total lipids from animal cultures. J Biol Chem 226:497–509

    Google Scholar 

  • Ghosh A, Al-Rabiai S, Ghosh BK, Trimino-Vasquez H, Eveleigh DE, Montenecourt BS (1982) Increased endoplasmic reticulum content of a mutant of Trichoderma reesei (RUT C-30) in relation to cellulase synthesis. Enzyme Microb Technol 4:110–113

    Google Scholar 

  • Ghosh A, Ghosh BK, Trimino-Vasquez H, Eveleigh DE, Montenecourt BS (1984) Cellulase secretion from a hypercellulolytic mutant of Trichoderma reesei Rut-C 30. Arch Microbiol 140:126–133

    Google Scholar 

  • Gong CS, Tsao GT (1979) Cellulase and biosynthesis regulation. Annu Rep Ferment Proc 3:111–140

    Google Scholar 

  • Hase A (1982) Changes in phospholipid composition during the development of Dictyostelium discoideum. Arch Biochem Biophys 219:21–29

    Google Scholar 

  • Hossack JA, Sharpe VJ, Rose AH (1977) Stability of the plasma membrane in Saccharomyces cerevisiae enriched with phosphatidylcholine or phosphatidylethanolamine. J Bacteriol 129:1144–1147

    Google Scholar 

  • Jacques NA (1983) Membrane perturbation by cerulenin modulates glucosyl-transferase secretion and acetate uptake by Streptococcus salivarius. J Gen Microbiol 129:3293–3302

    Google Scholar 

  • Jacques NA, Jacques VL, Wolf AC, Wittenberger CL (1985) Does an increase in membrane unsaturated fatty acids account for Tween 80 stimulation of glucosyltransferase secretion by Streptococcus salivarius? J Gen Microbiol 131:67–72

    Google Scholar 

  • Kammel WP, Kubicek CP (1985) Absence of postsecretional modification of extracellular proteins in Trichoderma reesei during growth on cellulose. J Appl Biochem 7:138–144

    Google Scholar 

  • Kikkoman CO (1968) Brit Patent 1 101 440 Cited. In: Eveleigh DE, Montenecourt BS (eds) (1979) Increasing yields of extracellular enzymes. Adv Appl Microbiol 25:57–74

  • Kolar H, Mischak H, Kammel WP, Kubicek CP (1985) Carboxymethyl-cellulase and β-glucosidase secretion by protoplasts from Trichoderma reese. J Gen Microbiol 131:1339–1347

    Google Scholar 

  • Koliander B, Hampel W, Röhr M (1984) Indirekt determination of biomass by rapid ribonucleic acid determination. Appl Microbiol Biotechnol 19:272–276

    Google Scholar 

  • Kubicek CP (1981) Release of carboxymethyl-cellulase and β-glucosidase from cell walls of Trichoderma reesei. Eur J Appl Microbiol Biotechnol 13:226–231

    Google Scholar 

  • Kubicek CP (1982) β-glucosidase excretion by Trichoderma pseudokoningii: correlation with cell-wall bound β-1,3-glucanase activities. Arch Microbiol 132:349–354

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Mandels M (1982) Cellulases. Annu Rep Ferm Proc 5:35–78

    Google Scholar 

  • Meixner-Monori B, Kubicek CP, Habison A, Kubicek-Pranz E, Röhr M (1985) Presence and regulation of the α-ketoglutarate dehydrogenase multienzyme complex in the filamentous fungus Aspergillus niger. J Bacteriol 161:265–271

    Google Scholar 

  • Meyer DI (1982) The signal hypothesis—a working model. Trends Biochem Sci 7:320–321

    Google Scholar 

  • Montenecourt BS, Nhlapo SD, Trimino-Vasquez H, Cuskey S, Schamhart DHJ, Eveleigh DE (1981) Regulatory controls in relation to overproduktion of fungal cellulases. In: Hollaender A, Rabson R, Rogers PV and Wolfe S (eds) Trends in the biology of fermentations for fuels and chemicals. Plenum Press, New York, pp. 33–53

    Google Scholar 

  • Morrisey JH (1981) Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310

    Google Scholar 

  • Novikoff AB, Heus M (1963) A microsomal nucleoside diphosphatase. J Biol Chem 238:710–719

    Google Scholar 

  • Palade GE (1975) Intracellular aspects of the process of protein synthesis. Science 189:347–358

    Google Scholar 

  • Petit-Glatron MF, Chambert R (1981) Levansucrase of Bacillus subtilis: conclusive evidence that its production and export are unrelated to fatty acid synthesis but modulated by membrane modifying agents. Eur J Biochem 119:603–611

    Google Scholar 

  • Reese ET, Maguire A (1971) Increase in cellulase yields by addition of surfactants to cellobiose cultures of Trichoderma viride. Devel Ind Microbiol 12:212–224

    Google Scholar 

  • Riezman H, Hay R, Gasser S, Daum G, Schneider G, Witte C, Schatz G (1983) The outer membrane of yeast mitochondria: isolation of outside-out sealed vesicles. EMBO Journal 2:1105–1111

    Google Scholar 

  • Santos TM, Sanchez M, Villanueva JR, Nombela C (1979) Derepression of β-1,3-glucanases of Penicillium italicum: localization of the various enzymes and correlation with cell wall glucan mobilization and autolysis. J Bacteriol 137:6–12

    Google Scholar 

  • Schekman R (1982) The secretory pathway in yeast. Trends Biochem Sci 7:243–246

    Google Scholar 

  • Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct 26:31–43

    Google Scholar 

  • Wittenberger CL, Beaman AJ, Lee LN (1978) Tween 80 effect on glucosyltransferase synthesis by Streptococcus salivarius. J Bacteriol 133:231–239

    Google Scholar 

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Schreiber, G., Kolar, H., Foisner, R. et al. Choline stimulates synthesis of extracellular proteins in Trichoderma reesei QM 9414. Arch. Microbiol. 144, 41–47 (1986). https://doi.org/10.1007/BF00454954

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  • DOI: https://doi.org/10.1007/BF00454954

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