Acetobacter Strains Research Articles

Characterization of the microflora of high acid submerged vinegar fermenters by distinct plasmid profiles

Since isolation of the Acetobacter strains responsible for ethanol oxidation in high acid, submerged vinegar fermentations is still extremely difficult, characterization of the vinegar microflora was approached by isolation and gel electrophoresis of plasmids extractable from the harvested microorganisms. Distinct plasmid profiles have been detected in all of 19 investigated fermenters from 5 different locations. Plasmid sizes varied between 1.3 and 133 Megadalton. Between 3 and 11 different plasmids were recognized in particular profiles. Comparison of these profiles from different sources proves that the plasmid profile of a vinegar fermenter is a unique property of the inherent microflora which allows definite conclusions regarding its stability, origin, identity and composition.

Occurrence of Extracellular (1->2)- -D-Glucans and (1->2)- -D-Gluco-oligosaccharides in Acetobacter

Summary: Of 11 strains of Acetobacter tested, three strains of A. xylinum (IFO 3288, IFO 13693 and IFO 13772), two strains of A. aceti (IFO 3281 and IFO 3283), one strain of A. pasteurianus (IFO 3223) and one strain of A. rancens (IFO 3297) were found to produce 2–25 mg per 100 ml culture filtrate of a mixture of (1→2)-β-d-glucans and (1→2)-β-d-gluco-oligosaccharides as low-molecular-weight products. The glucans and oligosaccharides were linear without branching structures and their degrees of polymerization (DPs) ranged from 6 to about 42. The compounds with lower DPs did not seem to be produced by hydrolysis of those with higher DPs. A. xylinum IFO 3288 and A. xylinum IFO 13693 also produced extracellular acidic polysaccharides containing l-rhamnose, d-glucose, d-mannose and uronic acid.

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 Electrophoregrams of a Wide Variety of Acetobacter strains. Proposal for the Improvement of the Taxonomy of the Genus Acetobacter Beijerinck 1898, 215

Ninety-eight strains, representing all Acetobacter species and subspecies from the Approved Lists of Bacterial Names (Skerman et al., 1980), were examined in a numerical analysis of 177 phenotypic features and compared to ninety-eight Gluconobacter and seven Frateuria strains. Four phenons could be delineated, corresponding to Frateuria (phenon 1), A. aceti subsp. liquefaciens (phenon 2), Gluconobacter (phenon 3) and Acetobacter minus A. aceti subsp. liquefaciens (phenon 4). Acetobacter, Frateuria and Gluconobacter are well- could be distinguished. Comparison of the protein electrophoregrams of Acetobacter strains revealed a fairly high internal homogeneity within phenon 2, subphenons C and D. Strains of the subphenon E gave very divergent protein patterns. The following classificatory changes are proposed within the genus Acetobacter: (1) Acetobacter liquefaciens sp. nov. is proposed for the homogeneous phenon 2, containing all 12 A. aceti subsp. liquefaciens strains (% G + C range of 62.3 to 64.6; IAM 1834 as type strain); (2) for the homogeneous subphenon D containing 8 A. aceti subsp. aceti strains, the name Acetobacter aceti emend, should be retained (% G + C range of 55.9 to 59.5; NCIB 8621 as type strain); (3) for subphenon E, a heterogeneous group, containing a variety of Acetobacter subspecies (all with their type strain) the species name Acetobacter pasteurianus emend, is preserved with LMD 22.1 as type strain; this species has the broad % G + C range of 52.8 to 62.5; (4) for subphenon C, a new species, Acetobacter hansenii sp. nov. is proposed (% G + C range of 58.1 to 62.6, NCIB 8746 as type strain). Minimal descriptions and differentiating keys are provided.

Lysogenie und lysogene Konversion bei methylotrophen Bakterien I. Nachweis des lysogenen Zustandes des fakultativ methanolassimilierenden Stammes Acetobacter MB 58/1 und Charakterisierung seines temperenten Phagen MO 1

AbstractLysogeny could until now not be shown for methylotrophic bacteria or for Acetobacter. In this paper we demonstrate the lysogenic state in a methylotrophic strain of the genus Acetobacter. The responsible temperate phage MO 1 belongs to group A of BRADLEY's classification system and contains double stranded DNA. Phage MO 1 was characterized by the following properties: Host range, sensitivity against environmental factors, serological behaviour, burst size, and latent period in one‐step growth experiments, and especially by its response to inducing factors.

Lysogenie und lysogene Konversion bei methylotrophen Bakterien II. Lysogene Konversion bei fakultativ methanolassimilierenden Acetobacter‐Stämmen

AbstractThe lysogenic state of the methylotrophic strain Acetobacter MB 58/1 is completely demonstrated by curing and lysogenization experiments. During these investigations we found that some phenotypic characteristics are modified by the presence or loss of the prophage MO 1. It could be shown that changes of the serological behaviour, the adsorption of the phages and the sensitivity against oxytetracycline are caused by lysogenic conversion. The phenotypic alterations of the bacterial cells induced by the phage genome are the result of modifications of the lipopolysaccharide structures on the cell surface. In the case of oxytetracycline resistance, interactions between the modified lipopolysaccharide structures and specific transport proteins of the cell membranes must be assumed.

Lysogenie und lysogene Konversion bei methylotrophen Bakterien II. Lysogene Konversion bei fakultativ methanolassimilierendenAcetobacter-Stämmen

The lysogenic state of the methylotrophic strain Acetobacter MB 58/1 is completely demonstrated by curing and lysogenization experiments. During these investigations we found that some phenotypic characteristics are modified by the presence or loss of the prophage MO 1. It could be shown that changes of the serological behaviour, the adsorption of the phages and the sensitivity against oxytetracycline are caused by lysogenic conversion. The phenotypic alterations of the bacterial cells induced by the phage genome are the result of modifications of the lipopolysaccharide structures on the cell surface. In the case of oxytetracycline resistance, interactions between the modified lipopolysaccharide structures and specific transport proteins of the cell membranes must be assumed.

Biosynthesis of monobactam compounds: origin of the carbon atoms in the beta-lactam ring.

The biosynthesis of monobactams by strains of Chromobacterium violaceum, Acetobacter sp., and Agrobacterium radiobacter was studied. Monobactams were produced during logarithmic growth by C. violaceum and Acetobacter sp. and during late log growth on glycerol and in stationary phase by A. radiobacter. The addition of various amino acids failed to significantly stimulate monobactam production in any of the producing organisms. Several 14C-amino acids and pyruvate were incorporated in vivo into monobactams. Serine, glycine, and cysteine were better incorporated than alanine or aspartate, whereas an excess of nonradioactive serine depressed the incorporation of labelled cysteine, glycine, and pyruvate. A comparison of [1-14C] glycine and [2-14C] glycine incorporation data suggests that glycine was first converted to serine. With a mixture of [U-14C[serine and [3-3H]serine, C. violaceum synthesized a monobactam with a complete retention of tritium, whereas with a [U-14C] cystine and [3-3H] cystine mixture, there was an extensive loss of C-3 tritium. Acetobacter sp. and A. radiobacter also utilized the double-labeled serine without the loss of tritium in their respective monobactams. It appears, therefore that in the three organisms, the carbon atoms of the beta-lactam ring of the monobactam are derived directly from serine without the loss of the C-3 hydrogen atoms, probably by an SN2 ring closure mechanism. With [methyl-14C] methionine, most of the radioactivity in the monobactam from Acetobacter sp. was in the methyl moiety of the beta-lactam ring methoxyl group.

Loss of acetic acid resistance and ethanol oxidizing ability in an Acetobacter strain.

A thermophilic strain, No. 1023, of Acetobacter aceti was isolated, by which it became possible to carry out submerged vinegar production at 35°C. The thermophilic property of the strain correlated closely with its acetic acid resistance. Strain No. 1023 lost acetic acid resistance at high frequency (more than 55% in isolated strains) and lost ethanol oxidizing ability at lower frequency (about 40%) compared to the above in the stationary growth phase in a liquid medium containing ethanol. The loss of these properties during the stationary phase was confirmed using a genetically marked strain (pro−) of No. 1023. Such frequent loss of acetic acid resistance and ethanol oxidizing ability suggested a relationship between these properties and a plasmid. A plasmid, pTA 5001, was found in strain No. 1023, and its molecular weight was determined to be 17 × 106 daltons by electron microscopy. The plasmid, however, seemed to have no direct connection with acetic acid resistance or ethanol oxidizing ability, becaus...

Acetic acid bacteria as causal agents of browning and rot of apples and pears

Summary Acetic acid bacteria are associated with apples and pears and have been isolated frequently from rotting fruits. The present work reports a) on the involvement of acetic acid bacteria in the rotting process, b) on the artificial infection of apple disks and entire apples by 159 acetic acid bacteria and c) on the susceptibility of 15 apple and 3 pear varieties. Nearly all Gluconobacter (101) and Acetobacter (58) strains tested were found to cause a type cf rot and browing in apples and pears not accompanied by a softening of the tissue. The rot inducing capacity is not restricted to acetic acid bacteria or to any of its taxonomic subdivisions. All the apple and pear varieties were susceptible, Golden delicious being the most resistant apple variety. It is suggested that the role of acetic acid bacteria in the post-harvest rotting and browning processes of apples and pears has been underestimated up to now.

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.

Isolation and identification of acetic acid bacteria for submerged acetic acid fermentation at high temperature.

Industrial vinegar production by submerged acetic acid fermentation has been carried out using Acetobacter strains at about 30°C. To obtain strains suitable for acetic acid fermentation at higher temperature, about 1,100 strains of acetic acid bacteria were isolated from vinegar mash, soils in vinegar factories and fruits, and their activities to oxidize ethanol at high temperature were examined. One of these strains, No. 1023, identified as Acetobacter aceti, retained full activity to produce acetic acid in continuous submerged culture at 35°C and produced 45% of activity at 38°C, while the usual strain of A. aceti completely lost its activity at 35°C. Thus the use of this strain may reduce the cooling costs of industrial vinegar production.

Acetobacter bacteriophage A-1

A bacteriophage ofAcetobacter suboxydans was isolated and found to correspond to type A phage according to Bradley's classification. The phage contains double stranded DNA. The length of the latency period and burst size could not be precisely determined because of apparent non-synchronous release of phage from single infective cycles. The host range was determined using 24 strains ofAcetobacter andGluconobacter species. Evidence for a probable occurence of host determined restriction and modification was obtained withAcetobacter suboxydans strain ATCC 621. The phage is designated A-1 and it is the first one to be reported forAcetobacter.

CHARACTERIZATION OF ACETOBACTER XYLINUM BY UBIQUINONE SYSTEM

All the Acetobacter xylinum strains examined, including the cellulose-forming and celluloseless ones, were found to have the ubiquinone system comprising Q-10 and were discriminated in this respect from any other members of the genus Acetobacter, especially of the mesoxydans group. The significance of difference in the quinone system is discussed from the taxonomic point of view. It is probable that these Acetobacter strains having such a Q-10 system should be re-accepted as a separatespecies A. xylinum from A. aceti, whether they produce the cellulosic leathery pellicles or not.

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.

Characterization of Acetobacter xylinum by Ubiquinone System

All the Acetobacter xylinum strains examined, including the cellulose-forming and celluloseless ones, were found to have the ubiquinone system comprising Q-10 and were discriminated in this respect from any other members of the genus Acetobacter, especially of the mesoxydans group. The significance of difference in the quinone system is discussed from the taxonomic point of view. It is probable that these Acetobacter strains having such a Q-10 system should be re-accepted as a separatespecies A. xylinum from A. aceti, whether they produce the cellulosic leathery pellicles or not.

Comparative carbohydrate catabolism in corynebacteria.

SUMMARY The catabolic pathways for the utilization of glucose and gluconate by one strain of each of five species of the genus Corynebacterium were examined. The relative participation of concurrent glucose pathways in each of these five strains was estimated. The findings indicated that Corynebacterium hoagii ATCC 7005 and C. tritici ATCC 11408 displayed catabolic behaviour similar to that of Arthrobacter globiformis ATCC 8010 in that the glycolytic pathway played a major role in glucose catabolism. In contrast, in C. equi ATCC 7698 and C. sepedonicum ATCC 9850, the glycolytic pathway and the pentose cycle pathway played almost equally important roles in glucose utilization. The catabolic pathway functioning in C. xerosis ATCC 7084 was distinct from these other strains of Corynebacteria and resemble that of some Acetobacter strains in that the pentose cycle pathway played a predominant role in the utilization of glucose and gluconate.

LETHAL ACTION BY AN ACETOBACTER ON YEASTS.

IT has been found that a recently isolated strain of Acetobacter caused yeasts to die when both organisms were present in a nutrient medium in full screw-capped bottles. In a typical test full McCartney bottles of beer were inoculated with 5 × 105 cells per ml. of Saccharomyces cerevisiae and amounts of a suspension of a washed culture of Acetobacter were added to different bottles to give counts of from 0 to 50 × 105 cells per ml. The numbers of viable yeasts were determined daily by methylene blue staining during incubation at 25°, with the results shown in Fig. 1.