Incapability of <i>Gluconobacter oxydans</i> to produce tartaric acid

Klasen, Ralf; Bringer-Meyer, Staphanie; Sahm, Hermann

doi:10.1002/bit.260400126

Cited by 29 publications

(11 citation statements)

References 16 publications

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“…Protein concentrations were determined by the method of Bradford (8). Gluconic acid and 5-ketogluconic acid were analyzed by high-pressure liquid chromatography (HPLC) with a Nucleosil 10-NH 2 column (4.6 by 250 mm; CS, Langerwehe, Germany), with 25 mM potassium phosphate buffer, pH 2.9, and UV detection at 215 nm (21).…”

Section: Methodsmentioning

confidence: 99%

Biochemical characterization and sequence analysis of the gluconate:NADP 5-oxidoreductase gene from Gluconobacter oxydans

1995

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Gluconate:NADP 5-oxidoreductase (GNO) from the acetic acid bacterium Gluconobacter oxydans subsp. oxydans DSM3503 was purified to homogeneity. This enzyme is involved in the nonphosphorylative, ketogenic oxidation of glucose and oxidizes gluconate to 5-ketogluconate. GNO was localized in the cytoplasm, had an isoelectric point of 4.3, and showed an apparent molecular weight of 75,000. In sodium dodecyl sulfate gel electrophoresis, a single band appeared corresponding to a molecular weight of 33,000, which indicated that the enzyme was composed of two identical subunits. The pH optimum of gluconate oxidation was pH 10, and apparent K m values were 20.6 mM for the substrate gluconate and 73 M for the cosubstrate NADP. The enzyme was almost inactive with NAD as a cofactor and was very specific for the substrates gluconate and 5-ketogluconate. D-Glucose, D-sorbitol, and D-mannitol were not oxidized, and 2-ketogluconate and L-sorbose were not reduced. Only D-fructose was accepted, with a rate that was 10% of the rate of 5-ketogluconate reduction. The gno gene encoding GNO was identified by hybridization with a gene probe complementary to the DNA sequence encoding the first 20 N-terminal amino acids of the enzyme. The gno gene was cloned on a 3.4-kb DNA fragment and expressed in Escherichia coli. Sequencing of the gene revealed an open reading frame of 771 bp, encoding a protein of 257 amino acids with a predicted relative molecular mass of 27.3 kDa. Plasmidencoded gno was functionally expressed, with 6.04 U/mg of cell-free protein in E. coli and with 6.80 U/mg of cell-free protein in G. oxydans, which corresponded to 85-fold overexpression of the G. oxydans wild-type GNO activity. Multiple sequence alignments showed that GNO was affiliated with the group II alcohol dehydrogenases, or short-chain dehydrogenases, which display a typical pattern of six strictly conserved amino acid residues.Gluconobacter oxydans, a gram-negative, strictly aerobic, rod-shaped bacterium, belongs to the family of acetic acid bacteria (11) and oxidizes glucose via two alternative pathways (28): one requires an initial phosphorylation followed by oxidation via the hexose monophosphate pathway (28); the second involves the nonphosphorylative formation of gluconic acid and of ketogluconic acids (12,22). The enzyme catalyzing the formation of gluconic acid, glucose dehydrogenase, is located in the cytoplasmic membrane of G. oxydans and is coupled to the respiratory chain (3). Further oxidation of gluconic acid by the membrane-bound and respiratory chain-coupled enzymes gluconate dehydrogenase and 2-ketogluconate dehydrogenase leads to the formation of 2-ketogluconic and 2,5-diketogluconic acids (39, 40). The enzymatic activity converting gluconic acid to 5-ketogluconic acid, however, is located in the cytoplasm of G. oxydans and depends on the presence of the cofactor NADP (12, 27). The enzymes catalyzing this reaction have been purified from G. suboxydans and Gluconobacter liquefaciens (1, 2). However, these enzymes were termed 5-ketogluconate...

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Section: Methodsmentioning

confidence: 99%

Biochemical characterization and sequence analysis of the gluconate:NADP 5-oxidoreductase gene from Gluconobacter oxydans

1995

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“…QGA5DH is also described to be involved in D -sorbitol, glycerol, and D -arabitol oxidation in G. oxydans Salusjärvi et al, 2004;Sugisawa and Hoshino, 2002] and leads to the formation of 5-KGA when D -gluconate is used as a substrate. 5-KGA is the industrially important precursor of L -(+)-tartaric acid [Klasen et al, 1992;Matzerath et al, 1995]. Beside the membrane-bound enzyme system, G. oxydans is also equipped with a cytosolic-oxidizing enzyme system made up of NAD(P)-dependent dehydrogenases.…”

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confidence: 99%

Glucose Oxidation and PQQ-Dependent Dehydrogenases in <i>Gluconobacter oxydans</i>

et al. 2008

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Gluconobacter oxydans is famous for its rapid and incomplete oxidation of a wide range of sugars and sugar alcohols. The organism is known for its efficient oxidation of D-glucose to D-gluconate, which can be further oxidized to two different keto-D-gluconates, 2-keto-D-gluconate and 5-keto-D-gluconate, as well as 2,5-di-keto-D-gluconate. For this oxidation chain and for further oxidation reactions, G. oxydans possesses a high number of membrane-bound dehydrogenases. In this review, we focus on the dehydrogenases involved in D-glucose oxidation and the products formed during this process. As some of the involved dehydrogenases contain pyrroloquinoline quinone (PQQ) as a cofactor, also PQQ synthesis is reviewed. Finally, we will give an overview of further PQQ-dependent dehydrogenases and discuss their functions in G. oxydans ATCC 621H (DSM 2343).

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“…In the early time, the use of NH 4 VO 3 as a catalyst has been researched and some patent applications have resulted [12][13][14]. The associated catalytic mechanism is quite clear [19] although, the production of L-TA and the molar conversion ratio obtained from this compound were relatively low at 24.66 g/L and 38.…”

Section: The Effect Of the Different Metal Catalystsmentioning

confidence: 99%

“…The conversion of 5-KGA to L-TA using vanadate in an alkaline buffer has been reported [13,14] and a mechanism has also been proposed [19]. However, the mechanism of the oxidation reaction by metal catalyst, especially by CuSO 4 ·5H 2 O,…”

Section: Qualitative Detection Of 5-kga During Oxidation Reactionmentioning

confidence: 99%

“…Soon after, the direct biological conversion of glucose to L-TA by Gluconobacter suboxydans in the presence of a trace metal catalyst, in particular ammonium vanadate, was patented [12]. However, Klasen later determined that the oxidation of 5-KGA to L-TA did not actually require the addition of G. suboxydans, but rather was promoted by the ammonium vanadate [13]. Moreover, Matzerath reported the production of L-TA from 5-KGA using ammonium vanadate as a catalyst in phosphate or carbonate buffer solutions over eight days, obtaining a conversion rate and selectivity of 80% and 82%, respectively [14].…”

Section: Introductionmentioning

confidence: 99%

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Selective oxidation of 5-keto-d-gluconate to l-(+)-tartaric acid on transition metal chelate catalyst

Yuan

Lin

et al. 2016

Inorganica Chimica Acta

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Incapability of Gluconobacter oxydans to produce tartaric acid

Cited by 29 publications

References 16 publications

Biochemical characterization and sequence analysis of the gluconate:NADP 5-oxidoreductase gene from Gluconobacter oxydans

Biochemical characterization and sequence analysis of the gluconate:NADP 5-oxidoreductase gene from Gluconobacter oxydans

Glucose Oxidation and PQQ-Dependent Dehydrogenases in <i>Gluconobacter oxydans</i>

Selective oxidation of 5-keto-d-gluconate to l-(+)-tartaric acid on transition metal chelate catalyst

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