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Engineering of the metabolism of Saccharomyces cerevisiae for anaerobic production of mannitol

R. Costenoble ; Lennart Adler ; Claes Niklasson (Centrum för kunskapsbildning och kommunikation (CKK)) ; G. Liden
Fems Yeast Research (1567-1356). Vol. 3 (2003), 1, p. 17-25.
[Artikel, refereegranskad vetenskaplig]

Under anaerobic conditions, Saccharomyces cerevisiae uses NADH-dependent glycerol-3-phosphate dehydrogenase (Gpd1p and Gpd2p) to re-oxidize excess NADH, yielding substantial amounts of glycerol. In a Deltagpd1 Deltagpd2 double-null mutant, the necessary NAD(+) regeneration through glycerol production is no longer possible, and this mutant does not grow under anaerobic conditions. The excess NADH formed can potentially be used to drive other NADH-dependent reactions or pathways. To investigate this possibility, a double-null mutant was transformed with a heterologous gene (mt1D) from Escherichia coli, coding for NADH-dependent mannitol-I-phosphate dehydrogenase. Expression of this gene in S. cerevisiae should result in NADH oxidation by the NADH-requiring formation of mannitol-1-phosphate from fructose-6-phosphate. The strain was characterized using step-change experiments, in which, during the exponential growth phase, the inlet gas was changed from air to nitrogen. It was found that the mutant produced mannitol only under anaerobic conditions. However, anaerobic growth was not regained, which was probably due to the excessive accumulation of mannitol in the cells. (C) 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Nyckelord: delta gpd1 delta gpd2 double-null mutant, redox balance, nadh-coupled reduction, mannitol-1-phosphate dehydrogenase, glycerol formation, glycerol-3-phosphate dehydrogenase mutants, escherichia-coli, nadph production, osmotic-stress, glycerol, yeast, cultures, growth, gpd1, gene



Denna post skapades 2007-04-19. Senast ändrad 2008-04-23.
CPL Pubid: 40668

 

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Institutioner (Chalmers)

Institutionen för cell- och molekylärbiologi (1994-2011)
Centrum för kunskapsbildning och kommunikation (CKK) (2002-2006)

Ämnesområden

Biologiska vetenskaper

Chalmers infrastruktur