Polyol Dehydrogenases of Gluconobacter oxydans (original) (raw)
The oxidation of glycols by acetic acid bacteria
Biochimica et Biophysica Acta, 1963
SU3IMAI/Y I. Resting cells of 14 different strains of acetic acid bacteria oxidized 1,2-ethanediol, DL-i,2-propanediol, DL-I,3-butanediol, meso-2,3-butanediol and 1,4-butanediol. 2. The oxidation of 22 different glycols was studied with resting cells of Ghtconobacter oxydans (suboxydans). 3. The end products of the oxidation of the following glycols with resting cells of either Gluconobacter oxydans (suboxydans) or Acetobacter aceti (liquefaciens) have been isolated and chemically identified: r,2-ethanediol to glycollic acid, 1,3-propanediol to fl-hydroxypropionic acid, 1,4-butanediol to succinie acid, 1,5-pczt.?-odiol to glutaric acid, 1,6-hexanediol to adipic acid, 1,7-heptanediol to pimelic acid and DL-I,3-butanediol to DL-fl-hydroxybutyric acid. The oxidation of 1,4-butanediol and 1,5-pentanediol occurred in two steps. 4. Acetobacter aceti (liquefaciens) w~.~ unable to grow in a medium with DL-1,3-butanediol as sole carbon source. This compound inhibited growth in culture media containing either ethanol or glycerol. 5. All glycols which were oxidized by resting cells were also oxidized by the particulate fraction. D(-)-and L(+)-I ,2-propanediol, D(-)-and L(+)-2,3-butanediol were oxidized to acetol, D(-)-and L(+)-acetylmethylcarbinol, respectively. 6. A soluble NAD-linked primary alcohol dehydrogenase oxidized monohydric primary alcohols and oJ-diols. DL-I,3-Butanedi6i Was oxidized slowly at C-I. 7. A soluble NAD-linked secondary alcohol dehydrogenase oxidized monohydric secondary alcohols and the secondary alcohol function of the following glycols: meso-2,3-butanediol, DL-2,3-butanediol, DL-i,z-propanedio!, L(+)-i,2-propanediol, meso-3,4-hexanediol and (-)-3,4-hexanediol. Meso-2,3-butanediol and meso-3,4-hexanediol were oxidized to L(+)-acetylmethylcarbinol and (+)ethylpropionylcarbinol. 8. Both soluble dehydrogenases were purified and separated by chromatography. INTRODUCTION Acetic acid bacteria, which appear to be uniquely endowed with the capacity for oxidizing a great variety of carbohydrates and derivatives, also oxidize several glycols. BROWN 1 showed that Acetobacter aceti oxidized 1,2-ethanediol to glycollic acid. This was later confirmed with A. pasteuriamts and A. kiitzingianus by SEIFERT 2 and with A. xylinum, A. aceti, Gluconobacter suboxydans and G. melanogenus by VlSSER 'T HOOFT 3. BANNING 4 reported that many strains were able to form oxalic acid from 1,2-ethanediol. Biochim. Biophys..4cta, 71 (i963) 3 i i 331 ~I2 K. KERSTERS, J. DE LEY With unshaken cultures of a variety of strains KLING 5 showed that the D(-) form of DL-i,2-propanediol was oxidized to acetol. This 5o % conversion was probably a coincidence since VISSER 'T HOOFT a gave evidence that both isomers were oxidized by A. a:vlimm~ and G. suboxydans. BUTLIN AND WINCE 6 convincingly showed that thoroughly aerated cultures of G. suboxydans oxidized both isomers nearly quantitatively to acetol. COPET, FIERENs-SNoECK AND VAN RISSEGHEM 7 and VAN RlSSE-GHEM s, again using poorly aerated unshaken cultures, reported that A. xylinum, A. aceli and G. subo:Lvdans oxidized D(-)-I,2-propanediol, another strain of G. sub. oxydans oxidized the L(+) form. From KLING'S results 9 with unshaken cultures of A. xylinum and A. aceti it can be deduced that the o(-) form of DL-2,3-butanediol was oxidized preferentially to D(-)-acetoin. His results were confirmed by GRIVSKY l° using the same species. UNDERKOFLER, FULMER, BANTZ AND KOOI n confirmed that the n(-) form was oxidized quantitatively by G. suboxydans, but the L(+) form was not attacked. Meso-2,3-butanediol was oxidized by G. suboxydans ~2 with the formation of L(+)acetoin. From the results of VISSER 'T HOOFT 3 with A. xylinum and G. suboxydans the same conclusion can be drawn (although this author thought that he was using the DL-form). According to GRIVSKY 1° only D(-)-acetoin would be oxidized further to diacetyl by A. aceti. VAN P, ISSEGHEM x3 found that A. acegi and A. xylinum oxidized meso-3,4-hexanediol to L(+)-ethylpropionylcarbinol. D(+)-3,4-Hexanediol was oxidized to D(-)ethylpropionylcarbinol. The L(-) isomer was not attacked. Both isomers of ethylpropionylcarbinol were oxidized to dipropionyl. Growing cultures of A. acai, A. xylinum and G. suboxydans did not oxidize 1,2-butanedioF ,°, L2-pentanedioF and 1,2-hexanedioF. COMMI~S ~" reported on the oxidation of the following glycols by resting cells or ce!l-free extracts: nL-I,2-propanediol, 2-methyl-2-nitro-I,3-propanediol, 2-butyne-1,4-diol, 1,2,4-butanetriol, 2-butene-x,4-diol, 1,3-pentanediol, 1,5-pentanediol, hexyleneglycol, 1,2,6-hexanetriol, 2,5-hexanediol, cyclohexane-I,4-diol. The following compounds were not oxidized: 2,5-dimethyl-hexyne-3-diol-2,5, pentaerythritol, dipropylenegiycol, diethyleneglycol, thiodiethyleneglycol, styrenegiycol and cyclohexane-I,4-diol. GOLDSCHMIDT AND KRAMPITZ 15 have briefly reported on an NAD-linked 2,3butanediol dehydrogenase from G. suboxydans which lacks specificity. Many of the previous experiments have been carried out with grovAng cultures, often poorly aerated and extending over periods up to 3-6 months. Knowledge on the enzymology of these glycols is negligible. In the present paper we want to report on the oxidation of several straight-chain glycols by acetic acid bacteria and on the nature of the enzymes which effect the primary catabolic step. It is an extension of the previous work of this laboratory on the biochenfisti T-and enzy~nolog~-of these bacteria (for reviews, see DE LEYlS,I~). METHODS AND MATERIALS Bacteria used We used the same strains of acetic acid bacteria which have previously been studied by DE J..EY 17. The same cultural conditions were adopted, as well as the
Microbiology, 2010
The growth of Gluconobacter oxydans DSM 7145 on meso-erythritol is characterized by two stages: in the first stage, meso-erythritol is oxidized almost stoichiometrically to l-erythrulose according to the Bertrand–Hudson rule. The second phase is distinguished from the first phase by a global metabolic change from membrane-bound meso-erythritol oxidation to l-erythrulose assimilation with concomitant accumulation of acetic acid. The membrane-associated erythritol-oxidizing enzyme was found to be encoded by a gene homologous to sldA known from other species of acetic acid bacteria. Disruption of this gene in the genome of G. oxydans DSM 7145 revealed that the membrane-bound polyol dehydrogenase not only oxidizes meso-erythritol but also has a broader substrate spectrum which includes C3–C6 polyols and d-gluconate and supports growth on these substrates. Cultivation of G. oxydans DSM 7145 on different substrates indicated that expression of the polyol dehydrogenase was not regulated, i...
Oxidative Fermentation of Acetic Acid Bacteria and Its Products
Frontiers in Microbiology
Acetic acid bacteria (AAB) are a group of Gram-negative, strictly aerobic bacteria, including 19 reported genera until 2021, which are widely found on the surface of flowers and fruits, or in traditionally fermented products. Many AAB strains have the great abilities to incompletely oxidize a large variety of carbohydrates, alcohols and related compounds to the corresponding products mainly including acetic acid, gluconic acid, gulonic acid, galactonic acid, sorbose, dihydroxyacetone and miglitol via the membrane-binding dehydrogenases, which is termed as AAB oxidative fermentation (AOF). Up to now, at least 86 AOF products have been reported in the literatures, but no any monograph or review of them has been published. In this review, at first, we briefly introduce the classification progress of AAB due to the rapid changes of AAB classification in recent years, then systematically describe the enzymes involved in AOF and classify the AOF products. Finally, we summarize the applica...
Acetic Acid Bacteria: Physiology and Carbon Sources Oxidation
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Applied Microbiology and Biotechnology, 2004
A 5-ketogluconate (5-KGA)-forming membrane quinoprotein, gluconate dehydrogenase, was isolated from Gluconobacter suboxydans strain IFO 12528 and partially sequenced. Partial sequences of five internal tryptic peptides were elucidated by mass spectrometry and used to isolate the two adjacent genes encoding the enzyme (EBI accession no. AJ577472). These genes share close homology with sorbitol dehydrogenase from another strain of G. suboxydans (IFO 3255). Substrate specificity of gluconate 5-dehydrogenase (GA 5-DH) turned out to be quite broad, covering many polyols, amino derivatives of carbohydrates, and simple secondary alcohols. There is a broad correlation between the substrate specificity of GA 5-DH and the empirical Bertrand-Hudson rule that predicts the specificity of oxidation of polyols by acetic acid bacteria. Escherichia coli transformed with the genes encoding gluconate dehydrogenase were able to convert gluconic acid into 5-KGA at 75% yield. Furthermore, it was found that 5-KGA can be converted into tartaric acid semialdehyde by a transketolase. These results provide a basis for designing a direct fermentation-based process for conversion of glucose into tartaric acid.
European journal of biochemistry / FEBS, 1972
A reinvestigation of the catabolic pathway(s) used by Pseudomonas putida NCIB 10015 (Dagley's strain) for the degradation of phenol and the cresols has proved the existence of a metabolic divergence after meta cleavage of the eatechols formed by hydroxylation of the primary substrates. The ring-fission products of catechol and 4-methylcatechol are shown to be simultaneously catabolized by two different enzymic activities, an NADf-dependent dehydrogenase and a cofactor-independent hydrolase. The metabolizing activitics of bot,h ring-fission products in extracts of cells grown on phenol and the cresols (0-, m-and p-cresol) were found to be nonspecific ; thermal inactivation of extracts of phenol-grown cells has shown that this nonspecificity is attributable to only one enzyme expressing each activity and that the two activities are locatcd on separate proteins.
Applied Microbiology and Biotechnology, 2014
Acetic acid bacteria such as Gluconobacter oxydans are used in several biotechnological processes due to their ability to perform rapid incomplete regio-and stereoselective oxidations of a great variety of carbohydrates, alcohols, and related compounds by their membrane-bound dehydrogenases. In order to understand the growth physiology of industrial strains such as G. oxydans ATCC 621H that has high substrate oxidation rates but poor growth yields, we compared its genome sequence to the genome sequence of strain DSM 3504 that reaches an almost three times higher optical density. Although the genome sequences are very similar, DSM 3504 has additional copies of genes that are absent from ATCC 621H. Most importantly, strain DSM 3504 contains an additional type II NADH dehydrogenase (ndh) gene and an additional triosephosphate isomerase (tpi) gene. We deleted these additional paralogs from DSM 3504, overexpressed NADH dehydrogenase in ATCC 621H, and monitored biomass and the concentration of the representative cell components as well as O 2 and CO 2 transfer rates in growth experiments on mannitol. The data revealed a clear competition of membrane-bound dehydrogenases and NADH dehydrogenase for channeling electrons in the electron transport chain of Gluconobacter and an important role of the additional NADH dehydrogenase for increased growth yields. The less active the NADH dehydrogenase is, the more active is the membrane-bound polyol dehydrogenase. These results were confirmed by introducing additional ndh genes via plasmid pAJ78 in strain ATCC 621H, which leads to a marked increase of the growth rate. Keywords Gluconobacter. Acetic acid bacteria. Growth. NADH dehydrogenase. Membrane-bound dehydrogenases. Electron transport chain. RAMOS Electronic supplementary material The online version of this article
Evidence of a plasmid-encoded oxidative xylose-catabolic pathway in Arthrobacter nicotinovorans pAO1
Research in Microbiology, 2013
Due to its high abundance, the D-xylose fraction of lignocellulose provides a promising resource for production of various chemicals. Examples of efficient utilization of D-xylose are nevertheless rare, mainly due to the lack of enzymes with suitable properties for biotechnological applications. The genus Arthrobacter, which occupies an ecological niche rich in lignocellulosic materials and containing species with high resistance and tolerance to environmental factors, is a very suitable candidate for finding D-xylose-degrading enzymes with new properties. In this work, the presence of the pAO1 megaplasmid in cells of Arthrobacter nicotinovorans was directly linked to the ability of this microorganism to ferment D-xylose and to sustain longer log growth. Three pAO1 genes (orf32, orf39, orf40) putatively involved in degradation of xylose were identified and cloned, and the corresponding proteins purified and characterized. ORF40 was shown to be a homotetrameric NADP þ /NAD þ sugar dehydrogenase with a strong preference for D-xylose; ORF39 is a monomeric aldehyde dehydrogenase with wide substrate specificity and ORF32 is a constitutive expressed transcription factor putatively involved in control of the entire catabolic pathway. Based on analogies with other pentose degradation pathways, a putative xylose oxidative pathway similar to the Weimberg pathway is postulated.