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Carbon and sulphur utilization during growth of Pseudomonas fluorescens on potassium D-glucose 6-O-sulphate as the sole sulphur source.

ABSTRACT: Pseudomonas fluorescens N.C.I.B. 8248, cultured on potassium d-glucose 6[(35)S]-O-sulphate as the sole sulphur source, liberated the 6-O-sulphate ester of d-gluconate into the culture medium. Extracts of bacteria grown under this cultural condition oxidized d-glucose 6-O-sulphate to yield the gluconate ester. Results suggest the involvement of a glucose dehydrogenase-like enzyme. The gluconate ester was apparently not oxidized further to any significant extent; however, it served as substrate for a desulphating enzyme found in extracts. Growth on d-glucose 6-O-sulphate as the sole source of sulphur was not associated with the appearance of a true glycosulphatase. Collectively, these results suggest that d-gluconate 6-O-sulphate, rather than the glucose ester, supplied the necessary sulphur for growth. Oxidative activities toward d-glucose 6-O-sulphate, d-glucose, d-gluconate 6-O-sulphate and d-gluconate found in extracts of P. fluorescens adapted to grow on d-glucose 6-O-sulphate as the sole source of carbon and sulphur are presented for comparative purposes.

SUBMITTER: Fitzgerald JW

PROVIDER: S-EPMC1176776 | biostudies-other | 1971 Apr

REPOSITORIES: biostudies-other

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Sulphur utilization during growth of pseudomonas fluorescens on potassium D-glucose 6-O-sulphate.

Project description:Pseudomonas fluorescens N.C.I.B. 8248 was adapted to grow on potassium d-glucose 6-O-sulphate as the sole carbon and sulphur source. Adapted bacteria grew optimally at 37 degrees C on 1.6% (w/v) sulphate ester and growth coincided with the disappearance of the ester from the culture medium at a rate of 2.4mg/h per ml. Three sulphated compounds were detected in the culture fluid at the termination of growth. One of these was present in traces only and has not been identified. The second was present in somewhat greater amounts and was identified as the 6-O-sulphate ester of d-gluconate, and the major metabolite was identified as d-glycerate 3-O-sulphate. Sulphur utilization by the organism was not associated with the appearance of a glycosulphatase enzyme in the cells. However, a novel enzyme system (or systems) was present that liberated inorganic (35)SO(4) (2-) ions from dipotassium d-gluconate 6[(35)S]-O-sulphate and from dipotassium dl-glycerate 3[(35)S]-O-sulphate. Activity towards the latter substrate could not be detected when the adapted or parent Pseudomonas strain was cultured on d-glucose and potassium sulphate as respective carbon and sulphur sources. Some properties of the enzyme acting on the glycerate ester are recorded.

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Project description:Chronic periodontitis is one of the most common forms of biofilm-induced diseases. Most of the recent studies were focus on the dental plaque microbial diversity and microbiomes. However, analyzing bacterial diversity at the taxonomic level alone limits deeper comprehension of the ecological relevance of the community. In this study, we compared the metabolic functional diversity of the microbial community in healthy subjects and periodontitis patients in a creative way--to assess the sole carbon source utilization using Biolog assay, which was first applied on oral micro-ecology assessment. Pattern analyses of 95-sole carbon sources catabolism provide a community-level phenotypic profile of the microbial community from different habitats. We found that the microbial community in the periodontitis group had greater metabolic activity compared to the microbial community in the healthy group. Differences in the metabolism of specific carbohydrates (e.g. ?-methyl-D-glucoside, stachyose, maltose, D-mannose, ?-methyl-D-glucoside and pyruvic acid) were observed between the healthy and periodontitis groups. Subjects from the healthy and periodontitis groups could be well distinguished by cluster and principle component analyses according to the utilization of discriminate carbon sources. Our results indicate significant difference in microbial functional diversity between healthy subjects and periodontitis patients. We also found Biolog technology is effective to further our understanding of community structure as a composite of functional abilities, and it enables the identification of ecologically relevant functional differences among oral microbial communities.

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Project description:Increasing lines of evidence indicate that while microbial profile might vary, community-level metabolic potential is often more stably correlated with healthy and diseased states. Here, we investigated the community-level metabolic diversity of dental plaque microbiota from caries-free (CF) and caries-affected (CA) children by measuring their sole carbon source utilization using a Biolog assay. The dietary habits of 32 CF and 31 CA children were recorded by a questionnaire. Supragingival plaque samples were collected and inoculated into Biolog AN Microplates to assess the metabolism of sole carbon sources by plaque bacteria. The results revealed significant differences in dietary habits between CF and CA children. Meanwhile, Biolog assay showed consistently higher, albeit not statistically significant, overall metabolic activity as measured by average well color development (AWCD) value in the plaque microbiota from CA group than CF group. Most importantly, the CA group had more than twice as many core-positive carbon sources (defined as being utilized by >90% of plaque microbiota from subjects within the group) as that of the CF group (31 vs. 14), including CA group-specific, cariogenic core-positive carbon sources such as sucrose, glucose and raffinose. Furthermore, CF and CA groups could be well distinguished by cluster and principle component analyses based on the types of sole carbon sources significantly differentially utilized by the two groups. Our results indicate that plaque communities associated with caries state are more metabolically versatile than those associated with healthy state, which could contribute to differential clinical caries states. Meanwhile, Biolog could be an effective tool in revealing the community-level physiological profiles of microbiota associated with different caries states.

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