Applied Biochemistry and Biotechnology

, Volume 137, Issue 1, pp 711–720

Fed-Batch production of glucose 6-Phosphate dehydrogenase using recombinant saccharomyces cerevisiae

  • Luiz Carlos Martins das Neves
  • Adalberto Pessoa
  • Michele Vitolo
Session 4

DOI: 10.1007/s12010-007-9091-x

Cite this article as:
Martins das Neves, L.C., Pessoa, A. & Vitolo, M. Appl Biochem Biotechnol (2007) 137: 711. doi:10.1007/s12010-007-9091-x

Abstract

The strain Saccharomyces cerevisiae W303-181, having the plasmid YEpPGK-G6P (built by coupling the vector YEPLAC 181 with the promoter phosphoglycerate kinase 1), was cultured by fed-batch process in order to evaluate its capability in the formation of glucose 6-phosphate dehydrogenase (EC.1.1.1.49). Two liters of culture medium (10.0 g/L glucose, 3.7 g/L yeast nitrogen broth (YNB), 0.02 g/L l-tryptophan, 0.02 g/L l-histidine, 0.02 g/L uracil, and 0.02 g/L adenine) were inoculated with 1.5 g dry cell/L and left fermenting in the batch mode at pH 5.7, aeration of 2.2 vvm, 30°C, and agitation of 400 rpm. After glucose concentration in the medium was lower than 1.0 g/L, the cell culture was fed with a solution of glucose (10.0 g/L) or micronutrients (l-tryptophan, l-histidine, uracil, and adenine each one at a concentration of 0.02 g/L) following the constant, linear, or exponential mode. The volume of the culture medium in the fed-batch process was varied from 2 L up to 3 L during 5 h. The highest glucose 6-phosphate dehydrogenase activity (350 U/L; 1 U=1 µmol of NADP/min) occurred when the glucose solution was fed into the fermenter through the decreasing linear mode.

Index Entries

Fed-batchglucose 6-phosphate dehydrogenaseglucoserecombinant strainSaccharomyces cerevisiaefermentation

Copyright information

© Humana Press Inc 2007

Authors and Affiliations

  • Luiz Carlos Martins das Neves
    • 1
  • Adalberto Pessoa
    • 1
  • Michele Vitolo
    • 1
  1. 1.Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical SciencesUniversity of São PauloSão Paulo SPBrazil