Gluconobacter Strains Research Articles

Deoxyribonucleic Acid Homologies Among Organisms in the Genus Gluconobacter

The taxonomy of the genus Gluconobacter has undergone many changes during the past 40 years. Based on phenotypic properties, this genus has changed from numerous species to one species containing five subspecies and then to a single species, Gluconobacter oxydans, with no subspecies. The present study was designed to test the validity of this latter view. Nucleotide sequence similarites were determined for 54 strains of Gluconobacter by using an S1 nuclease procedure. Three distinct deoxyribonucleic acid homology groups were obtained. The average level of relatedness among these groups was 16%. Homology group 1 contained 32 strains and included the type strain of G. oxydans and the type strains of all previously recognized subspecies. Homology group II contained 12 strains that had an intragroup homology level of 44 to 87% (average, 65%) with reference strain IFO 3264. Homology group III contained six strains with an average intragroup homology level of 86% with reference strain IFO 3276a. Reference strains IFO 3264 and IFO 3276 were previously recognized as G. oxydans. The remaining four strains of Gluconobacter had from 0 to 23% homology with reference strains used to delineate the three homology groups. Although these data show that the genus Gluconobacter is composed of at least three species, they also support elimination of the previously designated subspecies. Three isolates implicated in pink disease of pineapples were shown either not to be gluconobacters or to be mixed with gluconobacters. The occurrence of colony variants within many of the Gluconobacter strains is described, and the significance of this observation is discussed.

Distribution and characterization of plasmids in acetic acid bacteria.

Eighty-seven strains of acetic acid bacteria were surveyed for plasmids by CsCl-ethidium bromide equilibrium centrifugation of cell lysates. Twenty-seven of the 33 strains of Acetobacter were found to harbor plasmid DNA and most strains contained multiple species of low-molecular-weight plasmids. On the other hand, plasmid DNAs were detected in 23 of the 36 strains of Gluconobacter and most of them had molecular weights of more than 5 megadaltons. Of the 18 strains newly isolated from a vinegar factory, some of which were examined taxonomically, 17 contained plasmid DNA. The molecular weights of plasmids detected in this study were in the range of about one to over 17 megadaltons. The plasmids with molecular weights of less than 5 megadaltons were characterized with restriction endonucleases. The physical maps of two plasmids designated as pMV1O1 and pMV102, which were found in the isolated strains, were constructed.

Numerical Analysis of Phenotypic Features and Protein Gel Electropherograms of Gluconobacter Asai 1935 emend. mut. char. Asai, Iizuka, and Komagata 1964

Ninety-eight Gluconobacter strains of various origins were examined by numerical analysis of 177 phenotypic features and by gel electrophoresis of soluble proteins. Gluconobacter was phenotypically quite different from Acetobacter and Frateuria. An extensive phenotypic description and a minimal description of the genus Gluconobacter are given. The genus Gluconobacter contained two groups, A and B, by both techniques. Phenons A and B could be differentiated only by the requirement for nicotinic acid and by their electrophoretic patterns. Protein electrophoresis showed clearly that Gluconobacter strains are genetically stable over several decades. The strains of all five subspecies of Gluconobacter oxydans cited in the Approved Lists of Bacterial Names (Skerman et al., Int. J. Syst. Bacteriol. 30:225–420, 1980) were distributed randomly over the phenotypic and electrophoretic clusters and subclusters, and the type strains of the subspecies all fell in phenon B. We conclude that the single species Gluconobacter oxydans should not be further divided into subspecies.

Distribution of beta-lactam and beta-lactone producing bacteria in nature.

Over one million bacteria were isolated from a large variety of soil, plant and water samples collected from different environments and examined in an extremely sensitive and highly specific screen for beta-lactam production. A group of seven related monocyclic beta-lactams (monobactams) were isolated from strains representing four genera-Agrobacterium, Chromobacterium, Gluconobacter and Pseudomonas. Monobactam-producing strains of Agrobacterium and Pseudomonas were isolated only rarely. Producing strains of Chromobacterium were isolated from a relatively limited number of habitats while the Gluconobacter strains appeared to be widespread in nature. In addition, three closely related beta-lactone-containing molecules were isolated from strains representing three genera-Arthrobacter, Bacillus and Pseudomonas. The Bacillus and Pseudomonas strains were isolated infrequently but from a variety of samples. The producing strain of Arthrobacter was isolated only once.

Host Range of a Bacteriophage of Acetic Acid Bacteria

We determined the host range of “Acetobacter bacteriophage A-1.” Only Gluconobacter strains were susceptible to this phage; no Acetobacter strain was susceptible. Thus, the correct name of the phage should be Gluconobacter phage A-1. Susceptibility to this phage appears to be a general characteristic of the members of the genus Gluconobacter. The single species of the genus Gluconobacter oxydans contains four subspecies, and susceptibility to the phage was not confined to strains of any particular subspecies.

Gluconobacters from honey bees.

Fifty-six Gluconobacter strains and one Acetobacter strain were isolated from honey bees and their environment in three different regions in Belgium and identified phenotypically. Polyacrylamide gel electrophoresis of the soluble cell proteins showed that two different types exist within the Gluconobacter isolates: strains from type A were found in samples of the three regions, whereas strains from type B were only isolated in two of the three regions. Both types could occur in bees from the same region, from several hives of one bee keeper and from one hive. Strains from type A were almost identical with collection strain G. oxydans subsp. suboxydans NCIB 9018, whereas strains from type B constituted a new protein electrophoretic type within the genus Gluconobacter. Although Gluconobacter is apparently associated with honey bees, it is not known whether it is important or required for the bees or any hive product.

Growth factor requirements of Gluconobacter

Summary The growth factor requirements of ninety-five Gluconobacter strains are examined: 58% of them require pantothenic acid only, 28% require pantothenic acid and nicotinic acid and 6% require pantothenic acid, nicotinic acid and thiamin. Growth factor requirements are strain-specific and are not correlated with the present subdivisions within the genus.

A rapid, simple and simultaneous detection of 2-keto-, 5-keto-and 2,5-diketogluconic acids by thin-layer chromatography in culture media of acetic acid bacteria

A rapid and simple test for the simultaneous detection of 2-keto-, 5-keto- and 2.5-diketo-gluconic acids by thin-layer chromatography and their revelation with o-phenylene-diamine. HCl is described. 32 Acetobacter and 83 Gluconobacter strains were scanned with this method. The formation of keto-gluconic acids appears to be a stable and reliable feature for the classification of acetic accid bacteria.

Distribution and solubilization of particulate gluconate dehydrogenase and particulate 2-ketogluconate dehydrogenase in acetic acid bacteria.

The distribution of two particulate enzymes, gluconate dehydrogenase (GDH) and 2-ketogluconate dehydrogenase (2KGDH), was investigated with cell free extract through 26 strains of genus Acetobacter and genus Gluconobacter. GDH activity was found in the cell free extracts from all strains of genus Gluconobacter and two species of genus Acetobacter, A. aceti and A. aurantium. High activity of 2KGDH was also found in the pigment-producing strains of genus Gluconobacter.Best solubilization of particulate enzymes was attained with the highest recovery when 10 mg of Triton X–100 and 30 mg of protein of particulate fractions in 1 ml of 0.01 m phosphate buffer, pH 6.0, are incubated for 9 hr at 5°C with continuous stirring.By comparison of the total enzyme activity of particulate enzymes with that of NAD(P)-linked enzymes in the cell free extract, it was obvious that the formation of ketogluconates by particulate enzymes was much more predominant, roughly over 100 times higher, as that of NAD(P)-linked enzymes.

Cytochrome Difference Spectra of Acetic Acid Bacteria

The cytochrome difference spectra of 15 strains of the genus Acetobacter were found to differ from those of 7 strains of the genus Gluconobacter. The Acetobacter strains contained cytochrome a 1, which was lacking in the Gluconobacter strains. Within the genus Acetobacter, the Peroxydans group of Frateur could be separated through its cytochrome d content from the Oxydans and Mesoxydans groups.

Localisation and distribution of alcohol-cytochrome 553 reductase in acetic acid bacteria

Alcohol-cytochrome 553 reductase was detected in several strains of the mesoxydans and oxydans group ofAcetobacter. A similar enzyme, but with a higher optimum pH, was detected inAcetobacter aceti (liquefaciens) and in two strains ofGluconobacter. Intermittent ultrasonic disruption ofAcetobacter aceti cells, strainsrancens T-5 andmobilis 6428, showed that the alcohol-cytochrome 553 reductase was mainly localized on the cell hull. The NADP-linked aldehyde dehydrogenase appeared to be present as a cytoplasmic component. The oxidation rate of ethanol and acetaldehyde by intact resting cells which have been grown with either glucose or ethanol as a carbon source under either neutral or acidic conditions was nearly identical. The ethanol oxidizing enzyme system thus behaved as constitutive enzymes, as would be expected if they were bound to the cell hull. The results support the hypothesis that the alcohol-cytochrome 553 reductase is one of the important components of the enzyme system responsible for the physiological production of acetic acid from ethanol by acetic acid bacteria.

DEOXYRIBONUCLEIC ACID BASE COMPOSITION OF ACETIC ACID BACTERIA.

SUMMARY: The base composition of purified DNA from 28 strains of acetic acid bacteria was determined. Most strains of the genus Gluconobacter clustered closely together at 60.6–63.4 % (guanine + cytosine) of total base. All strains of the Acetobacter aceti biotype lay within the range 55.4–64.0 % (guanine + cytosine). The close relationship and possible common phylo-genetic origin of the genera Gluconobacter and Acetobacter is again stressed by these results. The base composition of DNA from acetic acid bacteria and from species of Pseudomonas was very similar, confirming the suspected close relationship between these groups. There is a noticeable agreement between the sequences of Acetobacter strains, arranged according to increasing % (guanine + cytosine) and arranged according to increasing enzymic equipment: strains with greater biochemical activity have on the whole also a higher % (guanine + cytosine) in DNA. The range of the compositional distribution of DNA molecules is on the whole broader in Acetobacter than in Gluconobacter. The results corroborate previous conclusions that both biotypes contain clusters of strains without species differentiation. A comparison of the paper chromatographic analysis with the method of thermal denaturation (‘melting point’) for estimating base composition of DNA showed that the latter method was to be preferred in routine analysis because of its ease, rapidity and reproducibility.

Lactate and pyruvate catabolism in acetic acid bacteria.

SUMMARY: The enzymic mechanism of D-and L-lactate catabolism was the same in several strains of Acetobacter and Gluconobacter. This was taken to indicate a close relationship between these groups. Both isomers were broken down by way of pyruvate and acetaldehyde to acetate. Identical reactions were carried out by two sets of enzymes of a different nature and localization, one particulate, the other soluble. All strains contained a constitutive, soluble pyruvate decarboxylase which required thiamine pyrophosphate and Mg++. When grown on lactate, a pyruvate decarboxylase was also present in the particulate fraction. No evidence was found for a pyruvate oxidase system. Acetaldehyde was oxidized by a particulate oxidase system and by NADP-and NAD-linked dehydrogenases. This last enzyme was activated by coenzyme A and glutathione. The particulate pyruvate decarboxylase was not very tightly bound to the particles, as it could easily be detached by washing with buffer. The oxidase systems could not be removed under these conditions.

Studies on the Formation of Glucosamine Acid byAcetobacter melanogenumBeijerinck andPseudomonas fluorescens liquefaciens

d-Glucosaminic acid has recently been found to be an oxidized product of d-glucosamine formed by Ps. fluorescens. It has been revealed that many strains of oxidative bacteria can oxidize glucosamine. The formation of glucosamine acid has been recognized among a large number of strains of Pseudomonas, Acetobacter and Gluconobacter, by means of paper chromatography. Furthermore, one of these strains, A. melanogenum Beijerinck, oxidized glucosamine to glucosaminic acid with the theoretical consumption of oxygen as Ps. fluorescens liquefaciens. Glucosaminic acid was proved by isolation and identification by means of using resting cells.The experiment of growth shows that Ps. fluorescens liq. could not secure any energy by means of the oxidation of glucosamine.

Studies on the Formation of Glucosaminic Acid by Acetobacter melanogenum Beigerinck and Pseudomonas fluorescens liquefaciens

D-Glucosaminic acid has recently been found to be an oxidized product of D-glucosamine formed by Ps. fluorescens. It has been revealed that many strains of oxidative bacteria can oxidize glucosamine. The formation of glucosaminic acid has been recognized among a large number of strains of Pseudomonas, Acetobacter and Gluconobacter, by means of paper chromatography. Furthermore, one of these strains, A. melanogenum Beijerinck, oxidized glucosamine to glucosaminic acid with the theoretical consumption of oxygen as Ps. fluorescens liquefaciens. Glucosaminic acid was proved by isolation and identification by means of using resting cells. The experiment of growth shows that Ps. fluorescens liq. could not secure any energy by means of the oxidation of glucosamine.