Journal Article

PreJuSER-16090

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Expression of glf(Z.m.) increases D-mannitol formation in whole cell biotransformation with resting cells of Corynebacterium glutamicum

Bäumchen, C.FZJ* ; Bringer-Meyer, S.FZJ*

2007
Springer Berlin

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Please use a persistent id in citations: doi:10.1007/s00253-007-0987-8

Abstract: A recombinant oxidation/reduction cycle for the conversion of D-fructose to D-mannitol was established in resting cells of Corynebacterium glutamicum. Whole cells were used as biocatalysts, supplied with 250 mM sodium formate and 500 mM D-fructose at pH 6.5. The mannitol dehydrogenase gene (mdh) from Leuconostoc pseudomesenteroides was overexpressed in strain C. glutamicum ATCC 13032. To ensure sufficient cofactor [nicotinamide adenine dinucleotide (reduced form, NADH)] supply, the fdh gene encoding formate dehydrogenase from Mycobacterium vaccae N10 was coexpressed. The recombinant C. glutamicum cells produced D-mannitol at a constant production rate of 0.22 g (g cdw)(-1) h(-1). Expression of the glucose/fructose facilitator gene glf from Zymomonas mobilis in C. glutamicum led to a 5.5-fold increased productivity of 1.25 g (g cdw)(-1) h(-1), yielding 87 g l(-1) D-mannitol from 93.7 g l(-1) D-fructose. Determination of intracellular NAD(H) concentration during biotransformation showed a constant NAD(H) pool size and a NADH/NAD(+) ratio of approximately 1. In repetitive fed-batch biotransformation, 285 g l(-1) D-mannitol over a time period of 96 h with an average productivity of 1.0 g (g cdw)(-1) h(-1) was formed. These results show that C. glutamicum is a favorable biocatalyst for long-term biotransformation with resting cells.

Keyword(s): Bacterial Proteins: genetics (MeSH) ; Bacterial Proteins: metabolism (MeSH) ; Base Sequence (MeSH) ; Biotechnology (MeSH) ; Biotransformation (MeSH) ; Corynebacterium glutamicum: genetics (MeSH) ; Corynebacterium glutamicum: metabolism (MeSH) ; DNA Primers: genetics (MeSH) ; DNA, Bacterial: genetics (MeSH) ; Formate Dehydrogenases: genetics (MeSH) ; Formate Dehydrogenases: metabolism (MeSH) ; Fructose: metabolism (MeSH) ; Gene Expression (MeSH) ; Genes, Bacterial (MeSH) ; Leuconostoc: enzymology (MeSH) ; Leuconostoc: genetics (MeSH) ; Mannitol: metabolism (MeSH) ; Mannitol Dehydrogenases: genetics (MeSH) ; Mannitol Dehydrogenases: metabolism (MeSH) ; Mutation (MeSH) ; Mycobacterium: genetics (MeSH) ; Mycobacterium: metabolism (MeSH) ; NAD: metabolism (MeSH) ; Oxidation-Reduction (MeSH) ; Recombinant Proteins: genetics (MeSH) ; Recombinant Proteins: metabolism (MeSH) ; Zymomonas: genetics (MeSH) ; Zymomonas: metabolism (MeSH) ; Bacterial Proteins ; DNA Primers ; DNA, Bacterial ; Recombinant Proteins ; glucose facilitator protein, Zymomonas ; Fructose ; NAD ; Mannitol ; Mannitol Dehydrogenases ; Formate Dehydrogenases ; J ; whole cell biotransformation (auto) ; NADH regeneration (auto) ; Corynebacterium glutamicum ATCC13032 (auto) ; D-mannitol (auto)

Note: Record converted from VDB: 12.11.2012

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Contributing Institute(s):

1. Biotechnologie 1 (IBT-1)

Research Program(s):

1. Biotechnologie (PBT)

Appears in the scientific report 2007

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Database coverage:
; JCR ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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