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1.
Article in English | MEDLINE | ID: mdl-33351739

ABSTRACT

A phylogenomic analysis based on 107 single-copy core genes revealed that three strains from sugar-rich environments, i.e. LMG 1728T, LMG 1731 and LMG 22058, represented a single, novel Gluconacetobacter lineage with Gluconacetobacter liquefaciens as nearest validly named neighbour. OrthoANIu and digital DNA-DNA hybridization analyses among these strains and Gluconacetobacter type strains confirmed that the three strains represented a novel Gluconacetobacter species. Biochemical characteristics and MALDI-TOF mass spectra allowed differentiation of this novel species from the type strains of G. liquefaciens and other closely related Gluconacetobacter species. We therefore propose to classify strains LMG 1728T, LMG 1731 and LMG 22058 in the novel species Gluconacetobacter dulcium sp. nov., with LMG 1728T (=CECT 30142T) as the type strain.


Subject(s)
Ananas/microbiology , Gluconacetobacter/classification , Phylogeny , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Genome Size , Gluconacetobacter/isolation & purification , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sugars
2.
Microbiol Res ; 244: 126651, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33383369

ABSTRACT

Gluconacetobacter diazotrophicus is a species of great agronomic potential due to its growth-promotion traits. Its colonization process in different plants has been reported. However, there have been no studies regarding its structural colonization in elephant grass. This is a fast-growing C4-Poaceae plant, and its application in Brazil is mainly aimed at feeding dairy cattle, due to its high nutritional value. Also, in the last decade, this grass has been applied in the production of biofuels. The present study aimed to monitor the colonization process of strain LP343 of G. diazotrophicus inoculated in elephant grass seedlings of PCEA genotype, by using a mCherry-tagged bacterium. Samples of roots and shoots collected at different periods were visualized by confocal laser-scanning microscopy. The colony-counting assay was used to compare the number of cells recovered in different niches and a qPCR was performed for the quantification of endophytic cells in root and shoot tissues. Results suggested that the strain LP343 quickly recognized the PCEA roots as host, attached to the elephant grass roots at 6 h, and 7 days after inoculation were able to colonize the xylem vessels of roots and shoots of elephant grass. This study advances our knowledge about the colonization process of G. diazotrophicus species in elephant grass, contributing to future studies involving the plant-bacteria interaction cultivated under gnotobiotic conditions.


Subject(s)
Gluconacetobacter/growth & development , Pennisetum/microbiology , Plant Roots/microbiology , Plant Shoots/microbiology , Brazil , Germ-Free Life , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Pennisetum/growth & development , Plant Roots/growth & development , Plant Shoots/growth & development , Seedlings/growth & development , Seedlings/microbiology
3.
J Mater Sci Mater Med ; 29(7): 90, 2018 Jun 25.
Article in English | MEDLINE | ID: mdl-29938364

ABSTRACT

Recent research was conducted to evaluate the healing efficiency of bacterial cellulose (BC) as a wound dressing in different pHs and its possibility of being a smart wound dressing that can indicate pHs. BC was produced by environmentally isolated bacterial strains. After washing the best achieved BC, it was floated in normal saline with different pHs with phenol red used as a pH indicator. Finally the wound healing effects of the acidic, neutral and alkaline BC membranes were evaluated in rat cutaneous wounds. Results showed that one of the isolates which its partial 16srRNA genome had 95% similarity with Gluconacetobacter intermedius, had the thickest layer. The microscopic and macroscopic evaluations showed that the acidic BC had the best healing activity. Although the color of the films remained unchanged during the experiments because they were transparent and thin, these changes could not be easily seen. This suggests the use of thicker films such as the ones which are cross linked with some materials (e.g., sterile gauze). In conclusion the pH can affect the healing ability of natural BC and acidic pH had the best wound healing efficiency. In future it is better to use the acidic BC instead of natural one for different wound healing purposes.


Subject(s)
Bandages , Cellulose/chemistry , Wound Healing , Animals , Biocompatible Materials/chemistry , Cellulose/biosynthesis , Cellulose/genetics , Cross-Linking Reagents , Female , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Gluconacetobacter/metabolism , Hydrogen-Ion Concentration , Materials Testing , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Rats , Rats, Wistar , Skin/injuries , Skin/pathology
4.
Appl Microbiol Biotechnol ; 101(3): 1003-1012, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27678116

ABSTRACT

Isolate B17 from Kombucha was estimated to be an efficient producer of bacterial cellulose (BC). The isolate was deposited under the number P 1463 and identified as Komagataeibacter rhaeticus by comparing a generated amplified fragment length polymorphism (AFLP™) DNA fingerprint against a reference database. Static cultivation of the K. rhaeticus strain P 1463 in Hestrin and Schramm (HS) medium resulted in 4.40 ± 0.22 g/L BC being produced, corresponding to a BC yield from glucose of 25.30 ± 1.78 %, when the inoculum was made with a modified HS medium containing 10 g/L glucose. Fermentations for 5 days using media containing apple juice with analogous carbon source concentrations resulted in 4.77 ± 0.24 g/L BC being synthesised, corresponding to a yield from the consumed sugars (glucose, fructose and sucrose) of 37.00 ± 2.61 %. The capacity of K. rhaeticus strain P 1463 to synthesise BC was found to be much higher than that of two reference strains for cellulose production, Komagataeibacter xylinus DSM 46604 and Komagataeibacter hansenii DSM 5602T, and was also considerably higher than that of K. hansenii strain B22, isolated from another Kombucha sample. The BC synthesised by K. rhaeticus strain P 1463 after 40 days of cultivation in HS medium with additional glucose supplemented to the cell culture during cultivation was shown to have a degree of polymerization of 3300.0 ± 122.1 glucose units, a tensile strength of 65.50 ± 3.27 MPa and a length at break of 16.50 ± 0.83 km. For the other strains, these properties did not exceed 25.60 ± 1.28 MPa and 15.20 ± 0.76 km.


Subject(s)
Cellulose/biosynthesis , Fermentation , Gluconacetobacter/metabolism , Kombucha Tea/microbiology , Amplified Fragment Length Polymorphism Analysis , Carbon/metabolism , Cellulose/metabolism , Culture Media/chemistry , Gluconacetobacter/classification , Gluconacetobacter/growth & development , Gluconacetobacter/isolation & purification , Glucose/metabolism
5.
Microb Ecol ; 72(1): 70-84, 2016 07.
Article in English | MEDLINE | ID: mdl-26944561

ABSTRACT

Greater Mexico City is one of the largest urban centers in the world, with an estimated population by 2010 of more than 20 million inhabitants. In urban areas like this, biological material is present at all atmospheric levels including live bacteria. We sampled the low atmosphere in several surveys at different points by the gravity method on LB and blood agar media during winter, spring, summer, and autumn seasons in the years 2008, 2010, 2011, and 2012. The colonial phenotype on blood agar showed α, ß, and γ hemolytic activities among the live collected bacteria. Genomic DNA was extracted and convenient V3 hypervariable region libraries of 16S rDNA gene were high-throughput sequenced. From the data analysis, Firmicutes, Proteobacteria, and Actinobacteria were the more abundant phyla in all surveys, while the genera from the family Enterobacteriaceae, in addition to Bacillus spp., Pseudomonas spp., Acinetobacter spp., Erwinia spp., Gluconacetobacter spp., Proteus spp., Exiguobacterium spp., and Staphylococcus spp. were also abundant. From this study, we conclude that it is possible to detect live airborne nonspore-forming bacteria in the low atmosphere of GMC, associated to the microbial cloud of its inhabitants.


Subject(s)
Air Microbiology , Bacteria/classification , Biodiversity , Phylogeny , Actinobacteria/genetics , Actinobacteria/isolation & purification , Bacillus/genetics , Bacillus/isolation & purification , Bacteria/isolation & purification , Cities , Culture Media , DNA, Bacterial/genetics , Genomics , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Mexico , Proteobacteria/genetics , Proteobacteria/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
6.
Rev Argent Microbiol ; 47(4): 335-43, 2015.
Article in English | MEDLINE | ID: mdl-26652262

ABSTRACT

It has been established that a decrease in the population of Gluconacetobacter diazotrophicus associated with sugarcane occurs after nitrogen fertilization. This fact could be due to a direct influence of NH(4)NO(3) on bacterial cells or to changes in plant physiology after fertilizer addition, affecting bacterial establishment. In this work, we observed that survival of G. diazotrophicus was directly influenced when 44.8mM of NH(4)NO(3) (640mgN/plant) was used for in vitro experiments. Furthermore, micropropagated sugarcane plantlets were inoculated with G. diazotrophicus and used for split root experiments, in which both ends of the system were fertilized with a basal level of NH(4)NO(3) (0.35mM; 10mgN/plant). Twenty days post inoculation (dpi) one half of the plants were fertilized with a high dose of NH(4)NO(3) (6.3mM; 180 mgN/plant) on one end of the system. This nitrogen level was lower than that directly affecting G. diazotrophicus cells; however, it caused a decrease in the bacterial population in comparison with control plants fertilized with basal nitrogen levels. The decrease in the population of G. diazotrophicus was higher in pots fertilized with a basal nitrogen level when compared with the corresponding end supplied with high levels of NH4NO3 (100dpi; 80 days post fertilization) of the same plant system. These observations suggest that the high nitrogen level added to the plants induce systemic physiological changes that affect the establishment of G. diazotrophicus.


Subject(s)
Gluconacetobacter/isolation & purification , Nitrogen/administration & dosage , Plant Physiological Phenomena , Plant Roots/microbiology , Saccharum/microbiology
7.
Appl Biochem Biotechnol ; 176(4): 1162-73, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25926011

ABSTRACT

Cellulose producing bacteria were isolated from fruit samples and kombucha tea (a fermented beverage) using CuSO4 solution in modified Watanabe and Yamanaka medium to inhibit yeasts and molds. Six bacterial strains showing cellulose production were isolated and identified by 16S rRNA gene sequencing as Gluconacetobacter xylinus strain DFBT, Ga. xylinus strain dfr-1, Gluconobacter oxydans strain dfr-2, G. oxydans strain dfr-3, Acetobacter orientalis strain dfr-4, and Gluconacetobacter intermedius strain dfr-5. All the cellulose-producing bacteria were checked for the cellulose yield. A potent cellulose-producing bacterium, i.e., Ga. xylinus strain DFBT based on yield (cellulose yield 5.6 g/L) was selected for further studies. Cellulose was also produced in non- conventional media such as pineapple juice medium and hydrolysed corn starch medium. A very high yield of 9.1 g/L cellulose was obtained in pineapple juice medium. Fourier transform infrared spectrometer (FT-IR) analysis of the bacterial cellulose showed the characteristic peaks. Soft cellulose with a very high water holding capacity was produced using limited aeration. Scanning electron microscopy (SEM) was used to analyze the surface characteristics of normal bacterial cellulose and soft cellulose. The structural analysis of the polymer was performed using (13)C solid-state nuclear magnetic resonance (NMR). More interfibrillar space was observed in the case of soft cellulose as compared to normal cellulose. This soft cellulose can find potential applications in the food industry as it can be swallowed easily without chewing.


Subject(s)
Acetobacter/metabolism , Fruit/microbiology , Genes, Bacterial , Gluconacetobacter xylinus/metabolism , Gluconacetobacter/metabolism , Kombucha Tea/microbiology , Acetobacter/classification , Acetobacter/genetics , Acetobacter/isolation & purification , Ananas/microbiology , Beverages , Bioreactors , Cellulose/metabolism , Cellulose/ultrastructure , Copper Sulfate/chemistry , Fermentation , Gluconacetobacter/classification , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Gluconacetobacter xylinus/classification , Gluconacetobacter xylinus/genetics , Gluconacetobacter xylinus/isolation & purification , Malus/microbiology , Musa/microbiology , Phylogeny , Polysaccharides, Bacterial/biosynthesis , Polysaccharides, Bacterial/ultrastructure , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Spectroscopy, Fourier Transform Infrared , Starch/metabolism , Zea mays/microbiology
8.
Int J Food Microbiol ; 204: 9-16, 2015 Jul 02.
Article in English | MEDLINE | ID: mdl-25828705

ABSTRACT

Culture-dependent and culture-independent methods were combined for the investigation of acetic acid bacteria (AAB) populations in traditionally produced vinegars and mother of vinegar samples obtained from apple and grape. The culture-independent denaturing gradient gel electrophoresis (DGGE) analysis, which targeted the V7-V8 regions of the 16S rRNA gene, showed that Komagataeibacter hansenii and Komagataeibacter europaeus/Komagataeibacter xylinus were the most dominant species in almost all of the samples analyzed directly. The culture-independent GTG5-rep PCR fingerprinting was used in the preliminary characterization of AAB isolates and species-level identification was carried out by sequencing of the 16S rRNA gene, 16S-23S rDNA internally transcribed to the spacer (ITS) region and tuf gene. Acetobacter okinawensis was frequently isolated from samples obtained from apple while K. europaeus was identified as the dominant species, followed by Acetobacter indonesiensis in the samples originating from grape. In addition to common molecular techniques, real-time PCR intercalating dye assays, including DNA melting temperature (Tm) and high resolution melting analysis (HRM), were applied to acetic acid bacterial isolates for the first time. The target sequence of ITS region generated species-specific HRM profiles and Tm values allowed discrimination at species level.


Subject(s)
Acetic Acid/metabolism , Acetobacter/genetics , Acetobacter/isolation & purification , Food Contamination/analysis , Gluconacetobacter/genetics , Base Sequence , DNA, Bacterial/genetics , DNA, Ribosomal Spacer/genetics , Denaturing Gradient Gel Electrophoresis , Gluconacetobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Real-Time Polymerase Chain Reaction , Sequence Analysis, DNA
9.
Appl Microbiol Biotechnol ; 98(3): 1065-75, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24190494

ABSTRACT

In this study, typical niches of acetic acid bacteria were screened for isolation of cellulose producer strains. Hestrin Schramm broth was used as enrichment and production media. Only nine out of 329 isolates formed thick biofilms on liquid surface and were identified as potential cellulose producers. Physiological and biochemical tests proved that all cellulose producers belonged to Gluconacetobacter genus. Most productive and mutation-resistant strain was subjected to 16S rRNA sequence analysis and identified as Gluconacetobacter hansenii P2A due to 99.8 % sequence similarity. X-ray diffraction analysis proved that the biofilm conformed to Cellulose I crystal structure, rich in Iα mass fraction. Static cultivation of G. hansenii P2A in HS medium resulted with 1.89 ± 0.08 g/l of bacterial cellulose production corresponding to 12.0 ± 0.3 % yield in terms of substrate consumption. Shaking and agitation at 120 rpm aided in enhancement of the amount and yield of produced cellulose. Productivity and yield reached up to 3.25 ± 0.11 g/l and 17.20 ± 0.14 % in agitated culture while a slight decrease from 78.7 % to 77.3 % was observed in the crystallinity index.


Subject(s)
Cellulose/metabolism , Gluconacetobacter/isolation & purification , Gluconacetobacter/metabolism , Bacterial Typing Techniques , Cluster Analysis , Culture Media/chemistry , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Gluconacetobacter/classification , Gluconacetobacter/growth & development , Molecular Sequence Data , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , X-Ray Diffraction
10.
Int J Syst Evol Microbiol ; 63(Pt 11): 3981-3988, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23793855

ABSTRACT

Ten strains of Gram-stain-negative, rod-shaped, non-spore-forming bacteria were isolated from the burial mound soil collected before the dismantling and samples collected during the dismantling work on the Takamatsuzuka Tumulus in Asuka village, Nara Prefecture, Japan in 2007. On the basis of the 16S rRNA gene sequence analysis of the isolates, they were accommodated in the genus Gluconacetobacter (class Alphaproteobacteria) and can be separated into four groups within the cluster containing the genus Gluconacetobacter. One of the groups demonstrated a phylogenetic position identical to that of Gluconacetobacter asukensis, which was isolated from small holes on plaster walls of the stone chamber interior of Kitora Tumulus in Asuka village, Nara Prefecture, Japan. The remaining three groups consisted of novel lineages within the genus Gluconacetobacter. A total of four isolates were selected from each group and carefully identified using a polyphasic approach. The isolates were characterized on the basis of their possessing Q-10 as the major ubiquinone system and C18 : 1ω7c (58.5-65.2 %) as the predominant fatty acid. A DNA-DNA hybridization test was used to determine that the three lineages represented novel species, for which the names Gluconacetobacter tumulisoli sp. nov., Gluconacetobacter takamatsuzukensis sp. nov. and Gluconacetobacter aggeris sp. nov. are proposed. The type strains are T611xx-1-4a(T) ( = JCM 19097(T) = NCIMB 14861(T)), T61213-20-1a(T) ( = JCM 19094(T) = NCIMB 14859(T)) and T6203-4-1a(T) ( = JCM 19092(T) = NCIMB 14860(T)), respectively.


Subject(s)
Gluconacetobacter/classification , Phylogeny , Soil Microbiology , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Japan , Molecular Sequence Data , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/analogs & derivatives , Ubiquinone/chemistry
11.
Carbohydr Polym ; 92(2): 2012-7, 2013 Feb 15.
Article in English | MEDLINE | ID: mdl-23399252

ABSTRACT

A strain producing bacterial cellulose (BC) screened from rotten mandarin fruit was identified as Gluconacetobacter intermedius CIs26 by the examination of general taxonomical characteristics and 16S rDNA sequence analysis. Furthermore, Fourier transform infrared (FT-IR) spectrum showed that pellicle produced by strain CIs26 was composed of glucan, and had the same functional group as a typical BC. X-ray diffractometry (XRD) analysis indicated that the BC was type I in structure with crystallinity index of 75%. BC yields of strain CIs26 in Hestrin-Schramn (HS), citrus waste modified HS (CMHS) and citrus waste solution (CWS) mediums were 2.1 g/L, 5.7 g/L, and 7.2 g/L, respectively. It was shown that citrus waste could stimulate BC production of strain CIs26 efficiently. Based on the ability of utilization of citrus waste, this strain appeared to have potential in BC manufacture on an industrial scale.


Subject(s)
Cellulose/biosynthesis , Gluconacetobacter/isolation & purification , Gluconacetobacter/metabolism , Citrus/microbiology , Fruit/microbiology
12.
Syst Appl Microbiol ; 36(1): 17-21, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23273842

ABSTRACT

Comparison of HaeIII- and HpaII-restriction profiles of PCR-amplified 16S-23S rDNA ITS regions of Gluconacetobacter sp. LMG 1529(T) and SKU 1109 with restriction profiles of reference strains of acetic acid bacteria described by Trcek and Teuber [34] revealed the same but unique restriction profiles for LMG 1529(T) and SKU 1109. Further analyses of nearly complete 16S rRNA gene sequences, nearly complete 16S-23S rDNA ITS sequences, as well as concatenated partial sequences of the housekeeping genes dnaK, groEL and rpoB, allocated both strains to a single phylogenetic cluster well separated from the other species of the genus Gluconacetobacter. DNA-DNA hybridizations confirmed their novel species identity by 73% DNA-DNA relatedness between both strains, and values below the species level (<70%) between SKU 1109 and the type strains of the closest phylogenetic neighbors. The classification of strains LMG 1529(T) and SKU 1109 into a single novel species was confirmed also by AFLP and (GTG)(5)-PCR DNA fingerprinting data, as well as by phenotypic data. Strains LMG 1529(T) and SKU 1109 can be differentiated from their closely related Gluconacetobacter species, Gluconacetobacter entanii and Gluconacetobacter hansenii, by their ability to form 2-keto-d-gluconic acid from d-glucose, their ability to use d-mannitol, d-gluconate and glycerol as carbon source and form acid from d-fructose, and their ability to grow without acetic acid. The major fatty acid of LMG 1529(T) and SKU 1109 is C(18:1ω7c) (60.2-64.8%). The DNA G+C content of LMG 1529(T) and SKU 1109 is 62.5 and 63.3mol% respectively. The name Gluconacetobacter maltaceti sp. nov. is proposed. The type strain is LMG 1529(T) (=NBRC 14815(T)=NCIMB 8752(T)).


Subject(s)
Acetic Acid/metabolism , Gluconacetobacter/classification , Gluconacetobacter/metabolism , Amplified Fragment Length Polymorphism Analysis , Bacterial Proteins/genetics , Bacterial Typing Techniques , Base Composition , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , DNA, Ribosomal Spacer/chemistry , DNA, Ribosomal Spacer/genetics , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Molecular Sequence Data , Nucleic Acid Hybridization , Phylogeny , Polymorphism, Restriction Fragment Length , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
13.
Int J Syst Evol Microbiol ; 63(Pt 3): 1119-1125, 2013 Mar.
Article in English | MEDLINE | ID: mdl-22729025

ABSTRACT

The phylogenetic position of a cellulose-producing acetic acid bacterium, strain ID13488, isolated from commercially available Colombian homemade fruit vinegar, was investigated. Analyses using nearly complete 16S rRNA gene sequences, nearly complete 16S-23S rRNA gene internal transcribed spacer (ITS) sequences, as well as concatenated partial sequences of the housekeeping genes dnaK, groEL and rpoB, allocated the micro-organism to the genus Gluconacetobacter, and more precisely to the Gluconacetobacter xylinus group. Moreover, the data suggested that the micro-organism belongs to a novel species in this genus, together with LMG 1693(T), a non-cellulose-producing strain isolated from vinegar by Kondo and previously classified as a strain of Gluconacetobacter xylinus. DNA-DNA hybridizations confirmed this finding, revealing a DNA-DNA relatedness value of 81 % between strains ID13488 and LMG 1693(T), and values <70 % between strain LMG 1693(T) and the type strains of the closest phylogenetic neighbours. Additionally, the classification of strains ID13488 and LMG 1693(T) into a single novel species was supported by amplified fragment length polymorphism (AFLP) and (GTG)5-PCR DNA fingerprinting data, as well as by phenotypic data. Strains ID13488 and LMG 1693(T) could be differentiated from closely related species of the genus Gluconacetobacter by their ability to produce 2- and 5-keto-d-gluconic acid from d-glucose, their ability to produce acid from sucrose, but not from 1-propanol, and their ability to grow on 3 % ethanol in the absence of acetic acid and on ethanol, d-ribose, d-xylose, sucrose, sorbitol, d-mannitol and d-gluconate as carbon sources. The DNA G+C content of strains ID13488 and LMG 1693(T) was 58.0 and 60.7 mol%, respectively. The major ubiquinone of LMG 1693(T) was Q-10. Taken together these data indicate that strains ID13488 and LMG 1693(T) represent a novel species of the genus Gluconacetobacter for which the name Gluconacetobacter medellinensis sp. nov. is proposed. The type strain is LMG 1693(T) ( = NBRC 3288(T) = Kondo 51(T)).


Subject(s)
Acetic Acid , Cellulose/biosynthesis , Gluconacetobacter/classification , Phylogeny , Amplified Fragment Length Polymorphism Analysis , Bacterial Typing Techniques , Base Composition , Colombia , DNA, Bacterial/genetics , Fatty Acids/analysis , Genes, Bacterial , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Molecular Sequence Data , Nucleic Acid Hybridization , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
14.
Appl Microbiol Biotechnol ; 96(6): 1479-87, 2012 Dec.
Article in English | MEDLINE | ID: mdl-22782249

ABSTRACT

Nowadays, bacterial cellulose has played more and more important role as new biological material for food industry and medical and industrial products based on its unique properties. However, it is still a difficult task to improve the production of bacterial cellulose, especially a large number of byproducts are produced in the metabolic biosynthesis processes. To improve bacterial cellulose production, ethanol and sodium citrate are added into the medium during the fermentation, and the activities of key enzymes and concentration of extracellular metabolites are measured to assess the changes of the metabolic flux of the hexose monophosphate pathway (HMP), the Embden-Meyerhof-Parnas pathway (EMP), and the tricarboxylic acid cycle (TCA). Our results indicate that ethanol functions as energy source for ATP generation at the early stage of the fermentation in the HMP pathway and the supplementation of ethanol significantly reduces glycerol generation (a major byproduct). While in the EMP pathway, sodium citrate plays a key role, and its supplementation results in the byproducts (mainly acetic acid and pyruvic acid) entering the gluconeogenesis pathway for cellulose synthesis. Furthermore, by adding ethanol and sodium citrate, the main byproduct citric acid in the TCA cycle is also reduced significantly. It is concluded that bacterial cellulose production can be improved by increasing energy metabolism and reducing the formation of metabolic byproducts through the metabolic regulations of the bypasses.


Subject(s)
Cellulose/biosynthesis , Citrates/metabolism , Ethanol/metabolism , Gluconacetobacter/metabolism , Metabolic Networks and Pathways , Actinidia/microbiology , Culture Media/chemistry , Culture Media/metabolism , Fermentation , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Industrial Microbiology , Sodium Citrate
15.
Food Microbiol ; 30(1): 98-104, 2012 May.
Article in English | MEDLINE | ID: mdl-22265289

ABSTRACT

Persimmon (Diospyros kaki) is a seasonal fruit with important health benefits. In this study, persimmon use in wine and condiment production was investigated using molecular methods to identify the yeast and acetic acid bacteria (AAB) isolated from the alcoholic fermentation and acetification of the fruit. Alcoholic fermentation was allowed to occur either spontaneously, or by inoculation with a commercial Saccharomyces cerevisiae wine strain, while acetification was always spontaneous; all these processes were performed in triplicates. Non-Saccharomyces yeast species were particularly abundant during the initial and mid-alcoholic fermentation stages, but S. cerevisiae became dominant toward the end of these processes. During spontaneous fermentation, S. cerevisiae Sc1 was the predominant strain isolated throughout, while the commercial strain of S. cerevisiae was the most common strain isolated from the inoculated fermentations. The main non-Saccharomyces strains isolated included Pichia guilliermondii, Hanseniaspora uvarum, Zygosaccharomyces florentinus and Cryptococcus sp. A distinct succession of AAB was observed during the acetification process. Acetobacter malorun was abundant during the initial and mid-stages, while Gluconacetobacter saccharivorans was the main species during the final stages of these acetifications. Four additional AAB species, Acetobacter pasteurianus, Acetobacter syzygii, Gluconacetobacter intermedius and Gluconacetobacter europaeus, were also detected. We observed 28 different AAB genotypes, though only 6 of these were present in high numbers (between 25%-60%), resulting in a high biodiversity index.


Subject(s)
Acetobacter/classification , Diospyros/microbiology , Fermentation , Food Handling/methods , Food Microbiology/methods , Yeasts/classification , Acetobacter/genetics , Acetobacter/isolation & purification , Bacterial Typing Techniques/methods , Colony Count, Microbial , DNA, Bacterial/genetics , Diospyros/metabolism , Fruit , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Mycological Typing Techniques/methods , Wine/analysis , Wine/microbiology , Yeasts/genetics , Yeasts/isolation & purification
16.
Int J Syst Evol Microbiol ; 62(Pt 8): 2032-2038, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22038998

ABSTRACT

Six Gram-negative, rod-shaped, non-spore-forming bacterial strains were isolated from small holes on plaster walls of the stone chamber interior of the Kitora Tumulus in Asuka village, Nara Prefecture, Japan. These were investigated by means of a polyphasic approach. All the isolates were strictly aerobic and motile by peritrichous flagella. Phylogenetic trees generated based on 16S rRNA gene sequences identified two novel lineages (comprising five isolates and one isolate, respectively) within the genus Gluconacetobacter. The isolates were characterized by having Q-10 as the major ubiquinone system and C(18:1)ω7c (58.7-63.1% of the total) as the predominant fatty acid. DNA-DNA hybridization experiments endorsed the species rank for the two lineages, for which the names Gluconacetobacter tumulicola sp. nov. (type strain K5929-2-1b(T) = JCM 17774(T) = NCIMB 14760(T)) and Gluconacetobacter asukensis sp. nov. (type strain K8617-1-1b(T) = JCM 17772(T) = NCIMB 14759(T)) are proposed.


Subject(s)
Gluconacetobacter/classification , Paintings , Phylogeny , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/analysis , Gluconacetobacter/genetics , Gluconacetobacter/isolation & purification , Japan , Molecular Sequence Data , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/analogs & derivatives , Ubiquinone/analysis
17.
Int J Syst Evol Microbiol ; 62(Pt 7): 1465-1469, 2012 Jul.
Article in English | MEDLINE | ID: mdl-21841006

ABSTRACT

Two novel acetic acid bacteria, strains G5-1(T) and I5-1, were isolated from traditional kaki vinegar (produced from fruits of kaki, Diospyros kaki Thunb.), collected in Kumamoto Prefecture, Japan. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains G5-1(T) and I5-1 formed a distinct subline in the genus Gluconacetobacter and were closely related to Gluconacetobacter swingsii DST GL01(T) (99.3% 16S rRNA gene sequence similarity). The isolates showed 96-100% DNA-DNA relatedness with each other, but <53% DNA-DNA relatedness with closely related members of the genus Gluconacetobacter. The isolates could be distinguished from closely related members of the genus Gluconacetobacter by not producing 2- and 5-ketogluconic acids from glucose, producing cellulose, growing without acetic acid and with 30% (w/v) d-glucose, and producing acid from sugars and alcohols. Furthermore, the genomic DNA G+C contents of strains G5-1(T) and I5-1 were a little higher than those of their closest phylogenetic neighbours. On the basis of the phenotypic characteristics and phylogenetic position, strains G5-1(T) and I5-1 are assigned to a novel species, for which the name Gluconacetobacter kakiaceti sp. nov. is proposed; the type strain is G5-1(T) (=JCM 25156(T)=NRIC 0798(T)=LMG 26206(T)).


Subject(s)
Acetic Acid/metabolism , Food Microbiology , Gluconacetobacter/classification , Gluconacetobacter/isolation & purification , Bacterial Typing Techniques , Base Composition , Cluster Analysis , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Fruit , Gluconacetobacter/genetics , Japan , Molecular Sequence Data , Nucleic Acid Hybridization , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
18.
Int J Food Microbiol ; 151(1): 105-12, 2011 Nov 15.
Article in English | MEDLINE | ID: mdl-21903289

ABSTRACT

The identification of acetic acid bacteria (AAB) from sound grapes from the Canary Islands is reported in the present study. No direct recovery of bacteria was possible in the most commonly used medium, so microvinifications were performed on grapes from Tenerife, La Palma and Lanzarote islands. Up to 396 AAB were isolated from those microvinifications and identified by 16S rRNA gene sequencing and phylogenetic analysis. With this method, Acetobacter pasteurianus, Acetobacter tropicalis, Gluconobacter japonicus and Gluconacetobacter saccharivorans were identified. However, no discrimination between the closely related species Acetobacter malorum and Acetobacter cerevisiae was possible. As previously described, 16S-23S rRNA gene internal transcribed spacer (ITS) region phylogenetic analysis was required to classify isolates as one of those species. These two species were the most frequently occurring, accounting for more than 60% of the isolates. For typing the AAB isolates, both the Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR and (GTG)5-PCR techniques gave similar resolution. A total of 60 profiles were identified. Thirteen of these profiles were found in more than one vineyard, and only one profile was found on two different islands (Tenerife and La Palma).


Subject(s)
Acetobacter/isolation & purification , Biodiversity , Gluconacetobacter/isolation & purification , Gluconobacter/isolation & purification , Vitis/microbiology , Acetobacter/classification , Acetobacter/genetics , Bacterial Typing Techniques , DNA, Ribosomal Spacer/genetics , Gluconacetobacter/classification , Gluconacetobacter/genetics , Gluconobacter/classification , Gluconobacter/genetics , Phylogeny , Polymerase Chain Reaction , RNA, Bacterial/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Spain
19.
J Bacteriol ; 193(13): 3395-6, 2011 Jul.
Article in English | MEDLINE | ID: mdl-21551293

ABSTRACT

Gluconacetobacter strains are prominent bacteria during traditional vinegar fermentation. Here, we report a draft genome sequence of Gluconacetobacter sp. strain SXCC-1. This strain was isolated from a fermentation starter (Daqu) used for commercial production of Shanxi vinegar, the best-known vinegar of China.


Subject(s)
DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Genome, Bacterial , Gluconacetobacter/genetics , Acetic Acid , China , Fermentation , Food Microbiology , Gluconacetobacter/isolation & purification , Molecular Sequence Data , Sequence Analysis, DNA
20.
J Bacteriol ; 193(10): 2670-1, 2011 May.
Article in English | MEDLINE | ID: mdl-21441523

ABSTRACT

Bacteria of the genus Gluconacetobacter are usually involved in the industrial production of vinegars with high acetic acid concentrations. We describe here the genome sequence of three Gluconacetobacter europaeus strains, a very common bacterial species from industrial fermentors, as well as of a Gluconacetobacter oboediens strain.


Subject(s)
DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Genome, Bacterial , Gluconacetobacter/genetics , Acetic Acid , Food Microbiology , Gluconacetobacter/isolation & purification , Industrial Microbiology , Molecular Sequence Data , Sequence Analysis, DNA
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