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Construction and physiological characterization of glyceraldehyde-3-phosphate dehydrogenase overproducing transformants of Aspergillus nidulans

  • Applied Genetics and Regulation
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Summary

The construction and characterization of glyceraldehyde-3-phosphate-dehydrogenase (GPD) overproducing transformants of Aspergillus nidulans and their behaviour in acetate-limited continuous cultures and glucose-grown batch cultures are described. The A. nidulans acetamidase deletion strain MH1277 was transormed with the homologous gpdA gene on a vector with the homologous acetamidase-gene (amdS) as a selection marker. Transformant Al contains about nine integrated copies of the gpdA gene, and shows a proportional gene-dosage GPD production of about 22% of the total soluble cell protein. Compared to the wild-type MH1277, Al has higher growth yields and reaches higher specific growth rates on both acetate and glucose, which could be due to the key position of GPD in glycolysis and gluconeogenesis.

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References

  • Aar PC van der, Lopes TS, Klootwijk J, Groeneveld Ph, Van Verseveld HW, Stouthamer AH (1990) Consequences of phosphoglycerate kinase overproduction for the growth and physiology of Saccharomyces cerevisiae. Appl Microbiol Biotechnol 32:577–587.

    Google Scholar 

  • Babel WJ, Muller RH (1985) Correlation between cell composition and carbon conversion efficiency in microbial growth: a theoretical study. Appl Microbiol Biotechnol 22:201–207.

    Google Scholar 

  • Bergmeyer HU (1974) Methods of enzymatic analysis, vol 2, 2nd edn. Academic Press, New York, p 467.

    Google Scholar 

  • Brammar WJ, Clarke PH (1964) Induction and repression of Pseudomonas aeruginosa amidase. J Gen Microbiol 37:307–319.

    Google Scholar 

  • Bull AT (1970) Chemical composition of wild type and mutant Aspergillus nidulans cell walls. The nature of polysaccharide and melanin constituents. J Gen Microbiol 63:75–94.

    Google Scholar 

  • Bulthuis BA (1990) Stoichiometry of growth and product formation in Bacillus licheniformis. Ph.D thesis, Vrije Universiteit, Amsterdam.

    Google Scholar 

  • Bulthuis BA, Frankena J, Koningstein GM, Van Verseveld HW, Stouthamer AH (1988) Instability of protease production in a rel+/rel— pair of Bacillus licheniformis and associated morphological and physiological characteristics. Antonie van Leeuwenhoek 54:95–111.

    Google Scholar 

  • Carter BLA, Bull AT (1971) The effect of oxygen tension in the medium on the morphology and growth kinetics of Aspergillus nidulans. J Gen Microbiol 65:265–273.

    Google Scholar 

  • Carter BLA, Bull AT, Pirt SJ, Rowley BI (1971) Relationship between energy substrate utilization and specific growth rate in Aspergillus nidulans. J Bacteriol 108:309–313.

    Google Scholar 

  • Fincham JRS (1989) Transformation in fungi. Microbiol Rev 53:148–170.

    Google Scholar 

  • Finkelstein DB (1987) Improvement of enzyme production in Aspergillus. Antonie van Leeuwenhoek 53:349–352.

    Google Scholar 

  • Gopal CV, Broad D, Lloyd D (1989) Bioenergetic consequences of protein overexpression in Saccharomyces cerevisiae. Appl Microbiol Biotechnol 30:160–165.

    Google Scholar 

  • Hynes MJ, Corrick CM, King JA (1983) Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol Cell Biol 3:1430–1439.

    CAS  PubMed  Google Scholar 

  • Laemmli UK, Farve M (1973) Maturation of the head of bacteriophage T4. I. DNA packing events. J Mol Biol 80:575–599.

    CAS  PubMed  Google Scholar 

  • Lipmann F, Tuttle LC (1945) A specific micromethod for the determination of acylphosphates. J Biol Chem 159:21–28.

    Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275.

    CAS  PubMed  Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  • McCullough W, Roberts CF, Osmani SA, Scrutton MC (1986) Regulation of carbon metabolism in filamentous fungi. In: Morgan MJ (ed) Carbohydrate metabolism in cultured cells. Plenum Press, New York, pp 287–355.

    Google Scholar 

  • Pitt DE, Bull AT (1982) Influence of culture conditions on the physiology and composition of Trichoderma aureoviride. J Gen Microbiol 128:1517–1527.

    Google Scholar 

  • Postma E, Verduyn C, Scheffers WA, Dijken JP van (1989) Enzymatic analysis of the Crabtree effect in glucose-limited chemostat cultures of Saccharomyces cerevisiae. Appl Environ Microbiol 55:468–477.

    CAS  PubMed  Google Scholar 

  • Punt PJ, Dingemanse MA, Jacobs-Meijsing BJM, Pouwels PH, Van den Hondel CAMJJ (1988) Isolation and characterization of the glyceraldehyde-3-phosphate dehydrogenase gene of Aspergillus nidulans. Gene 69:49–57.

    Google Scholar 

  • Roels JA (1981) The application of macroscopic principles to microbial metabolism. Ann NY Acad Sci 369:113–134.

    Google Scholar 

  • Rowley BI, Pirt SJ (1972) Melanin production by Aspergillus nidulans in batch and chemostat cultures. J Gen Microbiol 72:553–563.

    Google Scholar 

  • Rowley BI, Bull AT (1973) Chemostat for the cultivation of moulds. Lab Prac 22:286–289.

    Google Scholar 

  • Saunders G, Picknett TM, Tuite MF, Ward M (1989) Heterologous gene expression in filamentous fungi. Tibtech 7:283–287.

    Google Scholar 

  • Schellart JA (1975) The utilization of ethanol, lactic acid and acetic acid. Fungal protein from corn waste effluents. Ph.D thesis, Landbouw Universiteit, Wageningen, pp 39–45.

    Google Scholar 

  • Schreferl-Kunar G, Grotz M, Rohr M, Kubicek CP (1989) Increased citric acid production by mutants of Aspergillus niger with increased glycolytic capacity. FEMS Microbiol Lett 59:297–300.

    Google Scholar 

  • Stouthamer AH (1973) A theoretical study on the amount of ATP required for synthesis of microbial cell material. Antoine van Leeuwenhoek 39:545–565.

    Google Scholar 

  • Stouthamer AH (1979) The search for correlation between theoretical and experimental growth yields. Microb Biochem 21:1–47.

    Google Scholar 

  • Vries W de, Stouthamer AH (1968) Fermentation of glucose, lactose, galactose, mannitol, and xylose by bifidobacteria. J Bacteriol 96:472–478.

    Google Scholar 

  • Wernars K, Goosen T, Wennekes LMJ, Visser J, Bos CJ, Van den Broek HWJ, Gorcom RFM, Van den Hondel CAMJJ, Pouwels PH (1985) Gene amplification in Aspergillus nidulans by transformation with vectors containing the amdS gene. Curr Genet 9:361–368.

    Google Scholar 

  • Wernars K, Goosen T, Van den Hondel CAMJJ, Wennekes LMJ, Van den Broek HWJ (1986) The effect of the A. nidulans ans1 sequence on Aspergillus transformation. DNA-mediated transformation of the filamentous fungus Aspergillus nidulans. Ph.D thesis, Landbouw Universiteit, Wageningen, pp 100–116.

    Google Scholar 

  • Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119.

    Article  CAS  PubMed  Google Scholar 

  • Yelton MM, Hamer JE, Timberlake WE (1984) Transformation of Aspergillus nidulans using a trpC plasmid. Proc Natl Acad Sci USA 81:1470–1474.

    Google Scholar 

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Authors and Affiliations

  1. Department of Biology, Free University, Amsterdam, The Netherlands

    P. P. F. Hanegraaf, W. Dekker, W. Yap, H. W. van Verseveld & A. H. Stouthamer

  2. TNO Medical Biological Laboratory, Rijswijk, The Netherlands

    P. J. Punt & C. A. M. J. J. van den Hondel

Authors
  1. P. P. F. Hanegraaf
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  2. P. J. Punt
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  3. C. A. M. J. J. van den Hondel
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  4. W. Dekker
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  5. W. Yap
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  6. H. W. van Verseveld
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  7. A. H. Stouthamer
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Offprint requests to: P. P. F. Hanegraaf

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Hanegraaf, P.P.F., Punt, P.J., van den Hondel, C.A.M.J.J. et al. Construction and physiological characterization of glyceraldehyde-3-phosphate dehydrogenase overproducing transformants of Aspergillus nidulans . Appl Microbiol Biotechnol 34, 765–771 (1991). https://doi.org/10.1007/BF00169347

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  • DOI: https://doi.org/10.1007/BF00169347

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