Structural characterization of pellicle polysaccharides of Acetobacter tropicalis SKU1100 wild type and mutant strains

Ali, Ibnaof Ali Ibnaof; Akakabe, Yoshihiko; Moonmangmee, Somporn; Deeraksa, Arpaporn; Matsutani, Minenosuke; Yakushi, Toshiharu; Yamada, Mamoru; Matsushita, Kazunobu

doi:10.1016/j.carbpol.2011.05.055

Cited by 19 publications

(12 citation statements)

References 28 publications

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“…A second mechanism in acid tolerance probably involves the presence of an acetic acid resistance ABC transporter ( aatA ; APA386B_103), an efflux pump in the cytoplasmic membrane capable of exporting acetic acid [60]. Thirdly, A. pasteurianus 386B contained the gene cluster involved in pellicle polysaccharide formation ( polABCDE ; APA386B_1394 - APA386B_1398), preventing the diffusion of acetic acid into the cytoplasm [46,61,62]. Fourthly, the genes coding for urease ( ureDABCEFG ; APA386B_1179 - APA386B_1184), an urea transporter ( urtABCDE ; APA386B_1640 - APA386B_1644), an allophanate hydrolase (APA386B_936 - APA386B_937), and an urea carboxylase (APA386B_218) were present, indicating the ability to transport urea and convert it into ammonia, which may contribute to the survival of A. pasteurianus 386B in acidic environments, such as the cocoa pulp-bean mass (pH 3.5 – 4.5).…”

Section: Resultsmentioning

confidence: 99%

Complete genome sequence and comparative analysis of Acetobacter pasteurianus 386B, a strain well-adapted to the cocoa bean fermentation ecosystem

2013

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BackgroundAcetobacter pasteurianus 386B, an acetic acid bacterium originating from a spontaneous cocoa bean heap fermentation, proved to be an ideal functional starter culture for coca bean fermentations. It is able to dominate the fermentation process, thereby resisting high acetic acid concentrations and temperatures. However, the molecular mechanisms underlying its metabolic capabilities and niche adaptations are unknown. In this study, whole-genome sequencing and comparative genome analysis was used to investigate this strain’s mechanisms to dominate the cocoa bean fermentation process.ResultsThe genome sequence of A. pasteurianus 386B is composed of a 2.8-Mb chromosome and seven plasmids. The annotation of 2875 protein-coding sequences revealed important characteristics, including several metabolic pathways, the occurrence of strain-specific genes such as an endopolygalacturonase, and the presence of mechanisms involved in tolerance towards various stress conditions. Furthermore, the low number of transposases in the genome and the absence of complete phage genomes indicate that this strain might be more genetically stable compared with other A. pasteurianus strains, which is an important advantage for the use of this strain as a functional starter culture. Comparative genome analysis with other members of the Acetobacteraceae confirmed the functional properties of A. pasteurianus 386B, such as its thermotolerant nature and unique genetic composition.ConclusionsGenome analysis of A. pasteurianus 386B provided detailed insights into the underlying mechanisms of its metabolic features, niche adaptations, and tolerance towards stress conditions. Combination of these data with previous experimental knowledge enabled an integrated, global overview of the functional characteristics of this strain. This knowledge will enable improved fermentation strategies and selection of appropriate acetic acid bacteria strains as functional starter culture for cocoa bean fermentation processes.

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Section: Resultsmentioning

confidence: 99%

Complete genome sequence and comparative analysis of Acetobacter pasteurianus 386B, a strain well-adapted to the cocoa bean fermentation ecosystem

2013

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“…Morphotype variations, including the transition from a mucoid to a non-mucoid phenotype, are common events within the family of Acetobacteraceae , especially for Acetobacter species. This phenotypic change is often connected with the ability of the bacteria to form a pellicle on the medium surface [ 10 , 11 ]. The pellicle is an assemblage of cells that permits them to float on the medium surface during static cultivation and ensures a high state of aeration.…”

Section: Discussionmentioning

confidence: 99%

“…A disruption of the polE gene in A. tropicalis SKU1100. however, leads to a defect in pellicle formation, thereby giving it a central role, either as rhamnosyl-transferase [ 35 ], or as galactosyl-transferase [ 11 ], which connects the CPS to the cell surface. Moreover, the mutant Pel − cells secreted EPS into the culture medium.…”

Section: Discussionmentioning

confidence: 99%

“…Briefly, 10% of the seed culture was inoculated to 30 mL NaG medium and incubated statically at 30 °C for 14 days. The pellicle EPS was purified from the culture and cells were separated from EPS via ultra-sonification (10 min) and mechanical disruption, related to the method of IAI Ali, Y Akakabe, S Moonmangmee, A Deeraksa, M Matsutani, T Yakushi, M Yamada and K Matsushita [ 11 ]. The culture was centrifuged (10 min, 10.000 g) and the supernatant was saved in another flask.…”

Section: Methodsmentioning

confidence: 99%

“…The downstream located polE gene showed only low similarity to known glycosyltransferases, whereas a transposon-induced disruption of the polE gene resulted in a non pellicle forming strain, due to the absence of CPS production. Instead of the CPS, however, the Pel − strain showed a smooth-surfaced colony and the HePS was now secreted into the medium, which had the same composition as the capsular pellicle polysaccharide [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

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Environmentally triggered genomic plasticity and capsular polysaccharide formation are involved in increased ethanol and acetic acid tolerance in Kozakia baliensis NBRC 16680

et al. 2017

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Background Kozakia baliensis NBRC 16680 secretes a gum-cluster derived heteropolysaccharide and forms a surface pellicle composed of polysaccharides during static cultivation. Furthermore, this strain exhibits two colony types on agar plates; smooth wild-type (S) and rough mutant colonies (R). This switch is caused by a spontaneous transposon insertion into the gumD gene of the gum-cluster, resulting in a heteropolysaccharide secretion deficient, rough phenotype. To elucidate, whether this is a directed switch triggered by environmental factors, we checked the number of R and S colonies under different growth conditions including ethanol and acetic acid supplementation. Furthermore, we investigated the tolerance of R and S strains against ethanol and acetic acid in shaking and static growth experiments. To get new insights into the composition and function of the pellicle polysaccharide, the polE gene of the R strain was additionally deleted, as it was reported to be involved in pellicle formation in other acetic acid bacteria.ResultsThe number of R colonies was significantly increased upon growth on acetic acid and especially ethanol. The morphological change from K. baliensis NBRC 16680 S to R strain was accompanied by changes in the sugar contents of the produced pellicle EPS. The R:ΔpolE mutant strain was not able to form a regular pellicle anymore, but secreted an EPS into the medium, which exhibited a similar sugar monomer composition as the pellicle polysaccharide isolated from the R strain. The R strain had a markedly increased tolerance towards acetic acid and ethanol compared to the other NBRC 16680 strains (S, R:ΔpolE). A relatively high intrinsic acetic acid tolerance was also observable for K. baliensis DSM 14400T, which might indicate diverse adaptation mechanisms of different K. baliensis strains in altering natural habitats.ConclusionThe results suggest that the genetically triggered R phenotype formation is directly related to increased acetic acid and ethanol tolerance. The polE gene turned out to be involved in the formation of a cell-associated, capsular polysaccharide, which seems to be essential for increased ethanol/acetic tolerance in contrast to the secreted gum-cluster derived heteropolysaccharide. The genetic and morphological switch could represent an adaptive evolutionary step during the development of K. baliensis NBRC 16680 in course of changing environmental conditions.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-017-1070-y) contains supplementary material, which is available to authorized users.

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The Genomes of Acetic Acid Bacteria

Ehrenreich¹,

Liebl²

2017

Biology of Microorganisms on Grapes, in Must and in Wine

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Structural characterization of pellicle polysaccharides of Acetobacter tropicalis SKU1100 wild type and mutant strains

Cited by 19 publications

References 28 publications

Complete genome sequence and comparative analysis of Acetobacter pasteurianus 386B, a strain well-adapted to the cocoa bean fermentation ecosystem

Complete genome sequence and comparative analysis of Acetobacter pasteurianus 386B, a strain well-adapted to the cocoa bean fermentation ecosystem

Environmentally triggered genomic plasticity and capsular polysaccharide formation are involved in increased ethanol and acetic acid tolerance in Kozakia baliensis NBRC 16680

The Genomes of Acetic Acid Bacteria

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