RESUMEN
A novel bacterial strain of acetic acid bacteria capable of producing riboflavin was isolated from the soil sample collected in Wuhan, China. The isolated strain was identified as Gluconobacter oxydans FBFS97 based on several phenotype characteristics, biochemicals tests, and 16S rRNA gene sequence conducted. Furthermore, the complete genome sequencing of the isolated strain has showed that it contains a complete operon for the biosynthesis of riboflavin. In order to obtain the maximum concentration of riboflavin production, Gluconobacter oxydans FBFS97 was optimized in shake flask cultures through response surface methodology employing Plackett-Burman design (PBD), and Central composite design (CCD). The results of the pre-experiments displayed that fructose and tryptone were found to be the most suitable sources of carbon and nitrogen for riboflavin production. Then, PBD was conducted for initial screening of eleven minerals (FeSO4, FeCl3, KH2PO4, K2HPO4, MgSO4, ZnSO4, NaCl, CaCl2, KCl, ZnCl2, and AlCl3.6H2O) for their significances on riboflavin production by Gluconobacter oxydans strain FBFS97. The most significant variables affecting on riboflavin production are K2HPO4 and CaCl2, the interaction affects and levels of these variables were optimized by CCD. After optimization of the medium compositions for riboflavin production were determined as follows: fructose 25 g/L, tryptone 12.5 g/L, K2HPO4 9 g/L, and CaCl2 0.06 g/L with maximum riboflavin production 23.24 mg/L.
Asunto(s)
Ácido Acético/metabolismo , Gluconobacter oxydans/metabolismo , Modelos Estadísticos , Filogenia , ARN Bacteriano/análisis , Riboflavina/metabolismo , Medios de Cultivo , Genoma Bacteriano , Gluconobacter oxydans/genética , Gluconobacter oxydans/aislamiento & purificación , ARN Bacteriano/genética , ARN Ribosómico 16S/análisis , ARN Ribosómico 16S/genéticaRESUMEN
This paper considers the effect of multiwalled carbon nanotubes (MWCNTs) on the parameters of Gluconobacter oxydans microbial biosensors. MWCNTs were shown not to affect the structural integrity of microbial cells and their respiratory activity. The positive results from using MWCNTs were due to a decrease in the impedance of the electrode. The total impedance of the system decreased significantly, from 9000 kOhm (G. oxydans/chitosan composite) to 600 kOhm (G. oxydans/MWCNTs/chitosan). Modification of the amperometric biosensor with nanotubes led to an increase in the maximal signal from 65 to 869 nA for glucose and from 181 to 1048 nA for ethanol. The biosensor sensitivity also increased 4- and 5-fold, respectively, for each of the substrates. However, the addition of MWCNTs reduced the affinity of respiratory chain enzymes to their substrates (both sugars and alcohols). Moreover, the minimal detection limits were not reduced despite a sensitivity increase. The use of MWCNTs thus improved only some microbial biosensor parameters.
Asunto(s)
Técnicas Biosensibles , Técnicas Electroquímicas , Gluconobacter oxydans/aislamiento & purificación , Nanotubos de Carbono/química , CatálisisRESUMEN
Gluconobacter oxydans is known for its incomplete oxidation of a wide range of alcohols, sugars, and acids in a bioprocess. The corresponding oxidation products are secreted almost completely into the medium. Here, we present the high-quality draft genome sequence of G. oxydans WSH-003, an industrial strain with both high l-sorbose productivity and extreme tolerance to saccharides and alditols.
Asunto(s)
ADN Bacteriano/química , ADN Bacteriano/genética , Genoma Bacteriano , Gluconobacter oxydans/genética , Análisis de Secuencia de ADN , Antibacterianos/metabolismo , Metabolismo de los Hidratos de Carbono , Gluconobacter oxydans/efectos de los fármacos , Gluconobacter oxydans/aislamiento & purificación , Datos de Secuencia MolecularRESUMEN
A Real-Time PCR (RT-PCR) assay was developed using TaqMan minor groove binder (MGB) probes for the specific detection and quantification of five acetic acid bacteria (AAB) species (Acetobacter pasteurianus, Acetobacter aceti, Gluconacetobacter hansenii, Gluconacetobacter europaeus and Gluconobacter oxydans) in wine and vinegar. The primers and probes, designed from the 16S rRNA gene, showed good specificity with the target AAB species. The technique was tested on AAB grown in glucose medium (GY) and inoculated samples of red wine and wine vinegar. Standard curves were constructed with the five target species in all these matrices. Quantification was linear over at least 5 log units using both serial dilution of purified DNA and cells. When this technique was tested in GY medium and inoculated matrices, at least 10(2)-10(3) cells/ml were detected. To quantify low populations of AAB in microbiologically complex samples, a PCR enrichment including part of the 16S-23S rRNA gene ITS region was needed to increase the amount of target DNA compared to non-target DNA. The RT-PCR assay used in this study is a reliable, specific and fast method for quantifying these five AAB species in wine and vinegar.
Asunto(s)
Ácido Acético , Acetobacter/aislamiento & purificación , Gluconacetobacter/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Vino/microbiología , Ácido Acético/metabolismo , Acetobacter/genética , Sondas de ADN , Gluconacetobacter/genética , Gluconobacter oxydans/genética , Gluconobacter oxydans/aislamiento & purificaciónRESUMEN
Two types of bacterial biosensor were constructed by immobilization of Gluconobacter oxydans and Pseudomonas fluorescens cells on graphite electrodes modified with the conducting polymer; poly(1-(4-nitrophenyl)-2,5-di(2-thienyl)-1 H-pyrrole) [SNS(NO(2))]. The measurement was based on the respiratory activity of cells estimated by the oxygen consumption at -0.7 V due to the metabolic activity in the presence of substrate. As well as analytical characterization, the linear detection ranges, effects of electropolymerization time, pH and cell amount were examined by using glucose as the substrate. The linear relationships were observed in the range of 0.25-4.0 mM and 0.2-1.0 mM for G. oxydans and P. fluorescens based sensors, respectively.
Asunto(s)
Gluconobacter oxydans/aislamiento & purificación , Polímeros/química , Pseudomonas fluorescens/aislamiento & purificación , Pirroles/química , Adsorción , Técnicas Biosensibles , Células Inmovilizadas/química , Diálisis , Conductividad Eléctrica , Electroquímica , Electrodos , Gluconobacter oxydans/química , Gluconobacter oxydans/citología , Grafito/química , Concentración de Iones de Hidrógeno , Membranas Artificiales , Pseudomonas fluorescens/química , Pseudomonas fluorescens/citologíaRESUMEN
A bacterium isolated from patulin-contaminated apples was capable of degrading patulin to a less-toxic compound, ascladiol. The bacterium was identified as Gluconobacter oxydans by 16S rRNA gene sequencing, whereas ascladiol was identified by liquid chromatography-tandem mass spectrometry and proton and carbon nuclear magnetic resonance. Degradation of up to 96% of patulin was observed in apple juices containing up to 800 microg/ml of patulin and incubated with G. oxydans.