Acetobacter tropicalis Is a Major Symbiont of the Olive Fruit Fly (Bactrocera oleae)

ABSTRACT Following cultivation-dependent and -independent techniques, we investigated the microbiota associated with Bactrocera oleae, one of the major agricultural pests in olive-producing countries. Bacterial 16S rRNA gene libraries and ultrastructural analyses revealed the presence of several bacterial taxa associated with this insect, among which Acetobacter tropicalis was predominant. The recent increased detection of acetic acid bacteria as symbionts of other insect model organisms, such as Anopheles stephensi (G. Favia et al., Proc. Natl. Acad. Sci. USA 104:9047-9051, 2007) or Drosophila melanogaster (C. R. Cox and M. S. Gilmore, Infect. Immun. 75:1565-1576, 2007), prompted us to investigate the association established between A. tropicalis and B. oleae. Using an A. tropicalis-specific PCR assay, the symbiont was detected in all insects tested originating from laboratory stocks or field-collected from different locations in Greece. This acetic acid bacterium was successfully established in cell-free medium, and typing analyses, carried out on a collection of isolates, revealed that different A. tropicalis strains are present in fly populations. The capability to colonize and lodge in the digestive system of both larvae and adults and in Malpighian tubules of adults was demonstrated by using a strain labeled with a green fluorescent protein.

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Saccharibacter floricola gen. nov., sp. nov., a novel osmophilic acetic acid bacterium isolated from pollen

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.02911-0 ◽

2004 ◽

Vol 54 (6) ◽

pp. 2263-2267 ◽

Cited By ~ 84

Author(s):

Yasuko Jojima ◽

Yasuhiro Mihara ◽

Sonoko Suzuki ◽

Kenzo Yokozeki ◽

Shigeru Yamanaka ◽

...

Keyword(s):

Acetic Acid ◽

Phylogenetic Analyses ◽

Acetic Acid Bacteria ◽

Cellular Fatty Acid ◽

Acetic Acid Bacterium ◽

Rrna Gene ◽

Bacterial Strains ◽

Growth Properties ◽

Novel Genus And Species ◽

Straight Chain Acid

Three Gram-negative, aerobic, rod-shaped bacterial strains were isolated, from the pollen of Japanese flowers, as producers of xylitol; these strains were subjected to a polyphasic taxonomic study. Phylogenetic analyses of the 16S rRNA gene sequences demonstrated that these three isolates formed a new cluster within a group of acetic acid bacteria in the α-Proteobacteria. The characteristics of the three isolates were as follows: (i) their predominant quinone was Q-10; (ii) their cellular fatty acid profile contained major amounts of 2-hydroxy acids and an unsaturated straight-chain acid (C18 : 1 ω7c); and (iii) their DNA G+C contents were in the range 51·9–52·3 mol%, which is around the lower limit of the reported range for the genera of acetic acid bacteria. The negligible or very weak productivity of acetic acid from ethanol and the osmophilic growth properties distinguished these strains from other acetic acid bacteria. The unique phylogenetic and phenotypic characteristics suggest that the three isolates should be classified within a novel genus and species with the proposed name Saccharibacter floricola gen. nov., sp. nov. The type strain is strain S-877T (=AJ 13480T=JCM 12116T=DSM 15669T).

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Acetobacter ghanensis sp. nov., a novel acetic acid bacterium isolated from traditional heap fermentations of Ghanaian cocoa beans

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.64840-0 ◽

2007 ◽

Vol 57 (7) ◽

pp. 1647-1652 ◽

Cited By ~ 51

Author(s):

Ilse Cleenwerck ◽

Nicholas Camu ◽

Katrien Engelbeen ◽

Tom De Winter ◽

Katrien Vandemeulebroecke ◽

...

Keyword(s):

Acetic Acid ◽

Carbon Sources ◽

16S Rrna Gene Sequencing ◽

Acetic Acid Bacteria ◽

Acetic Acid Bacterium ◽

Rrna Gene ◽

Cocoa Beans ◽

Phenotypic Data ◽

Type Strains ◽

Ketogluconic Acid

Twenty-three acetic acid bacteria, isolated from traditional heap fermentations of Ghanaian cocoa beans, were subjected to a polyphasic taxonomic study. The isolates were catalase-positive, oxidase-negative, Gram-negative rods. They oxidized ethanol to acetic acid and were unable to produce 2-ketogluconic acid, 5-ketogluconic acid and 2,5-diketogluconic acid from glucose; therefore, they were tentatively identified as Acetobacter species. 16S rRNA gene sequencing and phylogenetic analysis confirmed their position in the genus Acetobacter, with Acetobacter syzygii and Acetobacter lovaniensis as their closest phylogenetic neighbours. (GTG)5-PCR fingerprinting grouped the strains in a cluster that did not contain any type strains of members of the genus Acetobacter. DNA–DNA hybridization with the type strains of all recognized Acetobacter species revealed DNA–DNA relatedness values below the species level. The DNA G+C contents of three selected strains were 56.9–57.3 mol%. The novel strains had phenotypic characteristics that enabled them to be differentiated from phylogenetically related Acetobacter species, i.e. they were motile, did not produce 2-ketogluconic acid or 5-ketogluconic acid from glucose, were catalase-positive and oxidase-negative, grew on yeast extract with 30 % glucose, grew on glycerol (although weakly) but not on maltose or methanol as carbon sources, and did not grow with ammonium as sole nitrogen source and ethanol as carbon source. Based on the genotypic and phenotypic data, the isolates represent a novel species of the genus Acetobacter for which the name Acetobacter ghanensis sp. nov. is proposed. The type strain is R-29337T (=430AT=LMG 23848T=DSM 18895T).

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Asaia krungthepensis sp. nov., an acetic acid bacterium in the α-Proteobacteria

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.02734-0 ◽

2004 ◽

Vol 54 (2) ◽

pp. 313-316 ◽

Cited By ~ 62

Author(s):

Pattaraporn Yukphan ◽

Wanchern Potacharoen ◽

Somboon Tanasupawat ◽

Morakot Tanticharoen ◽

Yuzo Yamada

Keyword(s):

Acetic Acid ◽

Enrichment Culture ◽

Acetic Acid Bacteria ◽

Acetic Acid Bacterium ◽

Rrna Gene ◽

Bacterial Strains ◽

16S Rrna Gene Sequences ◽

Dna Base Composition ◽

Dna Base ◽

Type Strains

Three bacterial strains were isolated from flowers collected in Bangkok, Thailand, by an enrichment-culture approach for acetic acid bacteria. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates were located in the lineage of the genus Asaia but constituted a cluster separate from the type strains of Asaia bogorensis and Asaia siamensis. The DNA base composition of the isolates was 60·2–60·5 mol% G+C, with a range of 0·3 mol%. The isolates constituted a taxon separate from Asaia bogorensis and Asaia siamensis on the basis of DNA–DNA relatedness. The isolates had morphological, physiological, biochemical and chemotaxonomic characteristics similar to those of the type strains of Asaia bogorensis and Asaia siamensis, but the isolates grew on maltose. The major ubiquinone was Q10. On the basis of the results obtained, the name Asaia krungthepensis sp. nov. is proposed for the isolates. The type strain is isolate AA08T (=BCC 12978T=TISTR 1524T=NBRC 100057T=NRIC 0535T), which had a DNA G+C content of 60·3 mol% and was isolated from a heliconia flower (‘paksaasawan’ in Thai; Heliconia sp.) collected in Bangkok, Thailand.

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Granulibacter bethesdensis gen. nov., sp. nov., a distinctive pathogenic acetic acid bacterium in the family Acetobacteraceae

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.64412-0 ◽

2006 ◽

Vol 56 (11) ◽

pp. 2609-2616 ◽

Cited By ~ 73

Author(s):

David E. Greenberg ◽

Stephen F. Porcella ◽

Frida Stock ◽

Alexandra Wong ◽

Patricia S. Conville ◽

...

Keyword(s):

Acetic Acid ◽

Reca Protein ◽

Its Region ◽

Acetic Acid Bacteria ◽

Acetic Acid Bacterium ◽

Rrna Gene ◽

Optimum Temperature ◽

Dna Base Composition ◽

Dna Base ◽

The Family

A Gram-negative, aerobic, coccobacillus to rod-shaped bacterium was isolated from three patients with chronic granulomatous disease. The organism was subjected to a polyphasic taxonomic study. A multilocus phylogenetic analysis based on the 16S rRNA gene, the internal transcribed spacer (ITS) region and the RecA protein demonstrated that the organism belongs to a new sublineage within the acetic acid bacteria in the family Acetobacteraceae. Phenotypic features are summarized as follows: the organism grew at an optimum temperature of 35–37 °C and optimum pH of 5.0–6.5. It produced a yellow pigment, oxidized lactate and acetate, the latter weakly, produced little acetic acid from ethanol and could use methanol as a sole carbon source. The two major fatty acids were a straight-chain unsaturated acid (C18 : 1ω7c) and C16 : 0. The DNA base composition was 59.1 mol% G+C. The very weak production of acetic acid from ethanol, the ability to use methanol, the yellow pigmentation and high optimum temperature for growth distinguished this organism from other acetic acid bacteria. The unique phylogenetic and phenotypic characteristics suggest that the bacterium should be classified within a separate genus, for which the name Granulibacter bethesdensis gen. nov., sp. nov. is proposed. The type strain is CGDNIH1T (=ATCC BAA-1260T=DSM 17861T).

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Identification of new conserved and variable regions in the 16S rRNA gene of acetic acid bacteria and acetobacteraceae family

Molecular Biology ◽

10.1134/s0026893315050052 ◽

2015 ◽

Vol 49 (5) ◽

pp. 668-677 ◽

Cited By ~ 7

Author(s):

S. Chakravorty ◽

S. Sarkar ◽

R. Gachhui

Keyword(s):

Acetic Acid ◽

16S Rrna ◽

16S Rrna Gene ◽

Acetic Acid Bacteria ◽

Rrna Gene ◽

Variable Regions ◽

The 16S Rrna Gene

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A tunable l-arabinose-inducible expression plasmid for the acetic acid bacterium Gluconobacter oxydans

Applied Microbiology and Biotechnology ◽

10.1007/s00253-020-10905-4 ◽

2020 ◽

Vol 104 (21) ◽

pp. 9267-9282

Author(s):

Philipp Moritz Fricke ◽

Tobias Link ◽

Jochem Gätgens ◽

Christiane Sonntag ◽

Maike Otto ◽

...

Keyword(s):

Acetic Acid ◽

Stationary Phase ◽

Gene Knockout ◽

Glucose Dehydrogenase ◽

Gluconobacter Oxydans ◽

Acetic Acid Bacteria ◽

Acetic Acid Bacterium ◽

Basal Expression ◽

Fold Induction ◽

Membrane Bound

Abstract The acetic acid bacterium (AAB) Gluconobacter oxydans incompletely oxidizes a wide variety of carbohydrates and is therefore used industrially for oxidative biotransformations. For G. oxydans, no system was available that allows regulatable plasmid-based expression. We found that the l-arabinose-inducible PBAD promoter and the transcriptional regulator AraC from Escherichia coli MC4100 performed very well in G. oxydans. The respective pBBR1-based plasmids showed very low basal expression of the reporters β-glucuronidase and mNeonGreen, up to 480-fold induction with 1% l-arabinose, and tunability from 0.1 to 1% l-arabinose. In G. oxydans 621H, l-arabinose was oxidized by the membrane-bound glucose dehydrogenase, which is absent in the multi-deletion strain BP.6. Nevertheless, AraC-PBAD performed similar in both strains in the exponential phase, indicating that a gene knockout is not required for application of AraC-PBAD in wild-type G. oxydans strains. However, the oxidation product arabinonic acid strongly contributed to the acidification of the growth medium in 621H cultures during the stationary phase, which resulted in drastically decreased reporter activities in 621H (pH 3.3) but not in BP.6 cultures (pH 4.4). These activities could be strongly increased quickly solely by incubating stationary cells in d-mannitol-free medium adjusted to pH 6, indicating that the reporters were hardly degraded yet rather became inactive. In a pH-controlled bioreactor, these reporter activities remained high in the stationary phase (pH 6). Finally, we created a multiple cloning vector with araC-PBAD based on pBBR1MCS-5. Together, we demonstrated superior functionality and good tunability of an AraC-PBAD system in G. oxydans that could possibly also be used in other AAB. Key points • We found the AraC-PBADsystem from E. coli MC4100 was well tunable in G. oxydans. • In the absence of AraC orl-arabinose, expression from PBADwas extremely low. • This araC-PBADsystem could also be fully functional in other acetic acid bacteria.

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Dynamics and Biodiversity of Populations of Lactic Acid Bacteria and Acetic Acid Bacteria Involved in Spontaneous Heap Fermentation of Cocoa Beans in Ghana

Applied and Environmental Microbiology ◽

10.1128/aem.02189-06 ◽

2007 ◽

Vol 73 (6) ◽

pp. 1809-1824 ◽

Cited By ~ 187

Author(s):

Nicholas Camu ◽

Tom De Winter ◽

Kristof Verbrugghe ◽

Ilse Cleenwerck ◽

Peter Vandamme ◽

...

Keyword(s):

Acetic Acid ◽

Lactic Acid ◽

Lactic Acid Bacteria ◽

Fermentation Process ◽

Denaturing Gradient Gel Electrophoresis ◽

Acetic Acid Bacteria ◽

Cocoa Bean ◽

Rrna Gene ◽

Cocoa Bean Fermentation ◽

Leuconostoc Pseudomesenteroides

ABSTRACT The Ghanaian cocoa bean heap fermentation process was studied through a multiphasic approach, encompassing both microbiological and metabolite target analyses. A culture-dependent (plating and incubation, followed by repetitive-sequence-based PCR analyses of picked-up colonies) and culture-independent (denaturing gradient gel electrophoresis [DGGE] of 16S rRNA gene amplicons, PCR-DGGE) approach revealed a limited biodiversity and targeted population dynamics of both lactic acid bacteria (LAB) and acetic acid bacteria (AAB) during fermentation. Four main clusters were identified among the LAB isolated: Lactobacillus plantarum, Lactobacillus fermentum, Leuconostoc pseudomesenteroides, and Enterococcus casseliflavus. Other taxa encompassed, for instance, Weissella. Only four clusters were found among the AAB identified: Acetobacter pasteurianus, Acetobacter syzygii-like bacteria, and two small clusters of Acetobacter tropicalis-like bacteria. Particular strains of L. plantarum, L. fermentum, and A. pasteurianus, originating from the environment, were well adapted to the environmental conditions prevailing during Ghanaian cocoa bean heap fermentation and apparently played a significant role in the cocoa bean fermentation process. Yeasts produced ethanol from sugars, and LAB produced lactic acid, acetic acid, ethanol, and mannitol from sugars and/or citrate. Whereas L. plantarum strains were abundant in the beginning of the fermentation, L. fermentum strains converted fructose into mannitol upon prolonged fermentation. A. pasteurianus grew on ethanol, mannitol, and lactate and converted ethanol into acetic acid. A newly proposed Weissella sp., referred to as “Weissella ghanaensis,” was detected through PCR-DGGE analysis in some of the fermentations and was only occasionally picked up through culture-based isolation. Two new species of Acetobacter were found as well, namely, the species tentatively named“ Acetobacter senegalensis” (A. tropicalis-like) and “Acetobacter ghanaensis” (A. syzygii-like).

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Acetobacter senegalensis sp. nov., a thermotolerant acetic acid bacterium isolated in Senegal (sub-Saharan Africa) from mango fruit (Mangifera indica L.)

INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY ◽

10.1099/ijs.0.64678-0 ◽

2007 ◽

Vol 57 (7) ◽

pp. 1576-1581 ◽

Cited By ~ 34

Author(s):

Bassirou Ndoye ◽

Ilse Cleenwerck ◽

Katrien Engelbeen ◽

Robin Dubois-Dauphin ◽

Amadou Tidiane Guiro ◽

...

Keyword(s):

Acetic Acid ◽

Gluconic Acid ◽

Mangifera Indica ◽

Carbon Sources ◽

Acetic Acid Bacterium ◽

Sub Saharan Africa ◽

Rrna Gene ◽

Strictly Aerobic ◽

Mango Fruit ◽

Phenotypic Data

A thermotolerant acetic acid bacterium, designated strain CWBI-B418T, isolated in Senegal from mango fruit (Mangifera indica), was characterized in detail by means of genotypic and phenotypic methods. The novel strain was strictly aerobic and exhibited optimal growth on YGM medium at 35 °C. Cells were Gram-negative, motile and coccoid. The strain was assigned to the genus Acetobacter on the basis of 16S rRNA gene sequence analysis. DNA–DNA hybridization experiments with its phylogenetically closest relatives showed that strain CWBI-B418T represented a novel Acetobacter genospecies. The DNA G+C content of strain CWBI-B418T was 56.0 mol%. Phenotypic characteristics enabling the differentiation of strain CWBI-B418T from phylogenetically related Acetobacter species were: production of 2-keto-d-gluconic acid from d-glucose, but not 5-keto-d-gluconic acid, production of catalase but not oxidase, growth on yeast extract with 30 % d-glucose, growth with ammonium as sole nitrogen source with ethanol as carbon source, utilization of glycerol and ethanol but not maltose or methanol as carbon sources, and growth in the presence of 10 % ethanol. Based on the genotypic and phenotypic data presented, strain CWBI-B418T clearly represents a novel Acetobacter species, for which the name Acetobacter senegalensis sp. nov. is proposed. The type strain is CWBI-B418T (=LMG 23690T=DSM 18889T).

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Isolation of dihydroxyacetone-producing acetic acid bacteria in Vietnam

Science and Technology Development Journal ◽

10.32508/stdj.v19i4.625 ◽

2016 ◽

Vol 19 (4) ◽

pp. 31-38

Author(s):

Huong Thi Lan Vu ◽

Oanh Thi Kim Nguyen ◽

Van Thi Thu Bui ◽

Uyen Thi Tu Bui ◽

Nghiep Dai Ngo ◽

...

Keyword(s):

Acetic Acid ◽

Gluconobacter Oxydans ◽

Acetic Acid Bacteria ◽

Rrna Gene ◽

16S Rrna Gene Sequences ◽

Group Iii ◽

Group I ◽

Routine Identification ◽

Group Ii ◽

Potential Use

Sixty-six acetic acid bacteria (AAB) were isolated from fourty-five flowers and fruits collected in Hochiminh City, Vietnam. Of the sixty-six, thirty-one isolates were selected as dihydroxyacetone (DHA)-producing AAB based on the reaction with Fehling’s solution and grouped into three groups by routine identification with phenotypic features. Group I composed of fourteen isolates and was assigned to the genus Acetobacter, Group II composed of thirteen isolates and was assigned to the genus Gluconobacter and Group III was the remaining four isolates and was assigned to the genus Gluconacetobacter. Ten isolates among the thirteen isolates of Group II gave a larger amount of DHA (22.2–26.0 mg/mL) than Gluconobacter oxydans NBRC 14819T (19.8 mg/mL), promising for the potential use in producing DHA. In phylogenetic analysis based on 16S rRNA gene sequences, six isolates of the ten potential DHA producers were suggested to be candidates for new taxa in the genus Gluconobacter.

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