Simultaneous overexpression of enzymes of the lower part of glycolysis can enhance the fermentative capacity of Saccharomyces cerevisiae.
Recombinant S. cerevisiae strains, with elevated levels of the enzymes of lower glycolysis (glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate mutase, phosphoglycerate kinase, enolase, pyruvate kinase, pyruvate decarboxylase and alcohol dehydrogenase) were physiologically characterized. During growth on glucose the enzyme levels in the recombinant strains (YHM4 and YHM7) were 1.1-3.4-fold higher than in the host strain (CEN.PK.K45). The recombinant strains were grown in aerobic or anaerobic batch cultures on glucose or a mixture of glucose and galactose. The specific ethanol production rates in the recombinant strains were the same as for the host strain and the physiological behaviour of the recombinant strains and the host strain was similar. When the cellular demand for ATP was increased by means of glucose pulses (final concentrations of 3.9 g/l or 2.0 g/l, respectively) to aerobic chemostat cultures maintained at a dilution rate of 0.08/h, the specific carbon dioxide production rate (qCO(2)) of CEN.PK.K45 accelerated at 6x10(-3) mmol/g/min(2) during the first 15 min, whereas during the same time period the qCO(2) of YHM7 accelerated twice as fast at 12x10(-3) mmol/g/min(2), indicating a higher fermentative capacity in the recombinant strain.
Peter Smits
H
H
Center for Process Biotechnology, Department of Biotechnology, Building 223, Technical University of Denmark, DK-2800 Lyngby, Denmark.
Hauf
J
J
Müller
S
S
Hobley
T J
TJ
Zimmermann
F K
FK
Hahn-Hägerdal
B
B
Nielsen
J
J
Olsson
L
L
D000332:Aerobiosis; D000693:Anaerobiosis; D019149:Bioreactors; D002245:Carbon Dioxide / Q000378:metabolism; D003470:Culture Media; D000431:Ethanol / Q000378:metabolism; D005285:Fermentation; D005690:Galactose / Q000378:metabolism; D015870:Gene Expression; D005947:Glucose / Q000378:metabolism*; D006019:Glycolysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D014170:Transformation, Genetic
D003470:Culture Media; D002245:Carbon Dioxide; D000431:Ethanol; D005947:Glucose; D005690:Galactose