Engineering of the metabolism of Saccharomyces cerevisiae for anaerobic production of mannitol
Artikel i vetenskaplig tidskrift, 2003

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.

delta gpd1 delta gpd2 double-null mutant

gpd1

glycerol-3-phosphate dehydrogenase mutants

cultures

escherichia-coli

glycerol

yeast

nadph production

growth

redox balance

gene

osmotic-stress

mannitol-1-phosphate dehydrogenase

glycerol formation

nadh-coupled reduction

Författare

R. Costenoble

Lennart Adler

Göteborgs universitet

Claes Niklasson

Chalmers, Centrum för kunskapsbildning och kommunikation (CKK)

G. Liden

FEMS Yeast Research

1567-1356 (ISSN) 1567-1364 (eISSN)

Vol. 3 1 17-25

Ämneskategorier

Biologiska vetenskaper

DOI

10.1111/j.1567-1364.2003.tb00134.x

PubMed

12702242