caa a22 4500 465822584 CHVBK 20180323112139.0 cr unu---uuuuu 170327e19900201xx s 000 0 eng 10.1007/BF00173731 doi (NATIONALLICENCE)springer-10.1007/BF00173731 Consequences of phosphoglycerate kinase overproduction for the growth and physiology of Saccharomyces cerevisiae [Elektronische Daten] [P. van der Aar, T. Lopes, J. Klootwijk, Ph. Groeneveld, H. van Verseveld, A. Stouthamer] Summary: The physiological consequences of overproduction of the homologous glycolytic enzyme 3-phosphoglycerate kinase (PGK), integrated in 80 PGK1 gene copies in the genome of Saccharomyces cerevisiae are described. This multiple integration and the strong PGK overproduction (maximum 47% of the total soluble cell protein) do not affect the maximal specific growth rate, but cause 40% reduction of the molar growth yield, compared with that of the wild-type host. The extra energy that is needed for protein overproduction is mainly provided by extra fermentation (respirofermentative growth), but respiration is also elevated compared with the reference strains. The increase in the specific oxygen uptake rate indicates that the respiratory capacity of the yeasts is higher than that in the wild-type host, in which the limited capacity of respiration is generally supposed to be at its maximal level at the critical dilution rate, and is thus responsible for the switch to respirofermentative growth. In a medium PGK1 gene copy integrant (about 25 copies), overproduction of 10%-12% PGK has a stimulating effect on the growth yield and energy efficiency. In these cells the growth benefits of overproduction of the glycolytic enzyme are higher than the disadvantages of extra protein synthesis. The overproduction of PGK has also consequences for the glucose affinity of the yeasts: In the more overproducing strain the K s is increased, compared to its reference strains. Elimination of strong overproducing cells from a glucose-limited chemostat culture is caused by two factors: (a) the excision of the PGK genes from the genome, which is of minor importance for wash-out, but the induction process for this overall decline of overproduction, and (b) the physiological selection process for less overproducing cells, caused by differences in affinity for glucose, most obvious at µ ≈ 1/2µmax. However in batch culture and in a chemostat at low specific growth rates, all the overproducing strains show high genetic stability and constantly provide high PGK quantities. Springer-Verlag, 1990 van der Aar P. Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut Lopes T. Department of Biochemistry, Vrije Universiteit, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands aut Klootwijk J. Department of Biochemistry, Vrije Universiteit, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands aut Groeneveld Ph Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut van Verseveld H. Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut Stouthamer A. Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut Applied Microbiology and Biotechnology Springer-Verlag 32/5(1990-02-01), 577-587 0175-7598 32:5<577 1990 32 253 https://doi.org/10.1007/BF00173731 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00173731 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- van der Aar P. Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Lopes T. Department of Biochemistry, Vrije Universiteit, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Klootwijk J. Department of Biochemistry, Vrije Universiteit, de Boelelaan 1083, 1081 HV, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Groeneveld Ph Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- van Verseveld H. Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut NATIONALLICENCE 700 1- Stouthamer A. Biological Laboratory, Department of Microbiology, Vrije Universiteit, de Boelelaan 1087, 1081 HV, Amsterdam, The Netherlands aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 32/5(1990-02-01), 577-587 0175-7598 32:5<577 1990 32 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer