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

ABSTRACT: 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.

SUBMITTER: Fitzgerald JW

PROVIDER: S-EPMC1176600 | biostudies-other | 1971 Feb

REPOSITORIES: biostudies-other

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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.

Project description: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.

| S-EPMC1176776 | biostudies-other

Differential expression of Pseudomonas fluorescens SBW25 during growth using glucose or MHPP as carbon sources

Project description:Identify genes involved in catabolism of the compound Methyl 3-(4-hydroxyphenyl)propionate

2024-03-22 | GSE228022 | GEO

Genetic analysis of the histidine utilization (hut) genes in Pseudomonas fluorescens SBW25.

Project description:The histidine utilization (hut) locus of Pseudomonas fluorescens SBW25 confers the ability to utilize histidine as a sole carbon and nitrogen source. Genetic analysis using a combination of site-directed mutagenesis and chromosomally integrated lacZ fusions showed the hut locus to be composed of 13 genes organized in 3 transcriptional units: hutF, hutCD, and 10 genes from hutU to hutG (which includes 2 copies of hutH, 1 of which is nonfunctional). Inactivation of hutF eliminated the ability to grow on histidine, indicating that SBW25 degrades histidine by the five-step enzymatic pathway. The 3 hut operons are negatively regulated by the HutC repressor with urocanate (the first intermediate of the histidine degradation pathway) as the physiological inducer. 5'-RACE analysis of transcriptional start sites revealed involvement of both sigma(54) (for the hutU-G operon) and sigma(70) (for hutF); the involvement of sigma(54) was experimentally demonstrated. CbrB (an enhancer binding protein for sigma(54) recruitment) was required for bacterial growth on histidine, indicating positive control of hut gene expression by CbrB. Recognition that a gene (named hutD) encoding a widely distributed conserved hypothetical protein is transcribed along with hutC led to analysis of its role. Mutational and gene fusion studies showed that HutD functions independently of HutC. Growth and fitness assays in laboratory media and on sugar beet seedlings suggest that HutD acts as a governor that sets an upper bound to the level of hut activity.

| S-EPMC1950622 | biostudies-literature

The Pseudomonas fluorescens Crc affects the length of the transition phase during diauxic growth

Project description:Pseudomonads, unlike enteric bacteria, prefer to utilize tricarboxylic acid cycle intermediates over glucose. The crc gene has been implicated to impart repression on glucose and other metabolic enzymes in various Pseudomonas species when grown in a mixture of glucose and various organic acids. The crc gene of P. fluorescens strain MB101 has been identified and a crc deletion derivative was constructed. Unlike a crc mutant of P. aeruginosa described earlier, the P. fluorescens deletion mutant crc did not begin to catabolize glucose until most of the succinate has been utilized in a medium containing both carbon sources. The transition phase during diauxic growth in succinate/glucose media was found to be shorter in a strain lacking crc and considerably longer in a strain overproducing Crc as compared to the wild type. DNA microarray analyses were performed with wild-type and crc mutant strains during the transition phase from succinate to glucose. In the wild-type strain, high expression signals were obtained for genes normally induced in cells that approach or are in stationary phase. By contrast, deletion mutant crc cells showed higher expression signals for genes associated with rapid growth than the wild-type strain. Taken together, this data indicates that Crc influences the length of the lag during the transition phase from succinate to glucose by directly or indirectly affecting the mRNA levels of target genes. Keywords: time course strain comparison

2008-09-11 | GSE5829 | GEO

Use of ammonium sulphate as a sulphur fertilizer: Implications for ammonia volatilization.

Project description:Ammonium sulphate is widely used as a sulphur (S) fertilizer, constituting about 50% of global S use. Within nitrogen (N) management, it is well known that ammonium-based fertilizers are subject to ammonia (NH3) volatilization in soils with pH > 7, but this has been overlooked in decision making on S fertilization. We reviewed 41 publications reporting measurements of NH3 loss from ammonium sulphate in 16 countries covering a wide range of soil types and climates. In field experiments, loss was mostly <5% of applied N in soils with pH (in water) <7.0. In soils with pH > 7.0, there was a wide range of losses (0%-66%), with many in the 20%-40% range and some indication of increased loss (ca. 5%-15%) in soils with pH 6.5-7.0. We estimate that replacing ammonium sulphate with a different form of S for arable crops could decrease NH3 emissions from this source by 90%, even taking account of likely emissions from alternative fertilizers to replace the N, but chosen for low NH3 emission. For every kt of ammonium sulphate replaced on soils of pH > 7.0 in temperate regions, NH3 emission would decrease from 35.7 to 3.6 t NH3. Other readily available sources of S include single superphosphate, potassium sulphate, magnesium sulphate, calcium sulphate dihydrate (gypsum), and polyhalite (Polysulphate). In view of the large areas of high pH soils globally, this change of S fertilizer selection would make a significant contribution to decreasing NH3 emissions worldwide, contributing to necessary cuts to meet agreed ceilings under the Gothenburg Convention.

| S-EPMC9290479 | biostudies-literature

Differential protein expression during growth on model and commercial mixtures of naphthenic acids in Pseudomonas fluorescens Pf-5.

Project description:Naphthenic acids (NAs) are carboxylic acids with the formula (Cn H2n + Z O2 ) and are among the most toxic, persistent constituents of oil sands process-affected waters (OSPW), produced during oil sands extraction. Currently, the proteins and mechanisms involved in NA biodegradation are unknown. Using LC-MS/MS shotgun proteomics, we identified proteins overexpressed during the growth of Pseudomonas fluorescens Pf-5 on a model NA (4'-n-butylphenyl)-4-butanoic acid (n-BPBA) and commercial NA mixture (Acros). By day 11, >95% of n-BPBA was degraded. With Acros, a 17% reduction in intensity occurred with 10-18 carbon compounds of the Z family -2 to -14 (major NA species in this mixture). A total of 554 proteins (n-BPBA) and 631 proteins (Acros) were overexpressed during growth on NAs, including several transporters (e.g., ABC transporters), suggesting a cellular protective response from NA toxicity. Several proteins associated with fatty acid, lipid, and amino acid metabolism were also overexpressed, including acyl-CoA dehydrogenase and acyl-CoA thioesterase II, which catalyze part of the fatty acid beta-oxidation pathway. Indeed, multiple enzymes involved in the fatty acid oxidation pathway were upregulated. Given the presumed structural similarity between alkyl-carboxylic acid side chains and fatty acids, we postulate that P. fluorescens Pf-5 was using existing fatty acid catabolic pathways (among others) during NA degradation.

| S-EPMC8289671 | biostudies-literature

The Pseudomonas fluorescens Crc affects the length of the transition phase during diauxic growth

Project description:Pseudomonads, unlike enteric bacteria, prefer to utilize tricarboxylic acid cycle intermediates over glucose. The crc gene has been implicated to impart repression on glucose and other metabolic enzymes in various Pseudomonas species when grown in a mixture of glucose and various organic acids. The crc gene of P. fluorescens strain MB101 has been identified and a crc deletion derivative was constructed. Unlike a crc mutant of P. aeruginosa described earlier, the P. fluorescens ïcrc did not begin to catabolize glucose until most of the succinate has been utilized in a medium containing both carbon sources. The transition phase during diauxic growth in succinate/glucose media was found to be shorter in a strain lacking crc and considerably longer in a strain overproducing Crc as compared to the wild type. DNA microarray analyses were performed with wild-type and crc mutant strains during the transition phase from succinate to glucose. In the wild-type strain, high expression signals were obtained for genes normally induced in cells that approach or are in stationary phase. By contrast, ïcrc cells showed higher expression signals for genes associated with rapid growth than the wild-type strain. Taken together, this data indicates that Crc influences the length of the lag during the transition phase from succinate to glucose by directly or indirectly affecting the mRNA levels of target genes. Keywords: time course strain comparison These are time course experiments to compare crc mutant strain to the wildtype MB101 strain at 5, 6, 7, 8, and 9 ht time points. Dye swaps were included in each time point comparison. There are 10 pairs in total.

2010-06-19 | E-GEOD-5829 | biostudies-arrayexpress

Genome Sequence of the Banana Plant Growth-Promoting Rhizobacterium Pseudomonas fluorescens PS006.

Project description:Pseudomonas fluorescens is a well-known plant growth-promoting rhizobacterium (PGPR). We report here the first whole-genome sequence of PGPR P. fluorescens evaluated in Colombian banana plants. The genome sequences contains genes involved in plant growth and defense, including bacteriocins, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, and genes that provide resistance to toxic compounds.

| S-EPMC4859179 | biostudies-literature

Complete Genome Sequence of the Plant Growth-Promoting Rhizobacterium Pseudomonas fluorescens LBUM677.

Project description:Pseudomonas fluorescens LBUM677 has shown the ability to increase plant biomass and seed oil yield in soybean, canola, and Buglossoides arvensis (corn gromwell) when inoculated in the rhizosphere. Here, we report a draft genome sequence of P. fluorescens LBUM677, with an estimated size of 6.14 Mb.

| S-EPMC6588369 | biostudies-literature

Dual involvement of CbrAB and NtrBC in the regulation of histidine utilization in Pseudomonas fluorescens SBW25.

Project description:Pseudomonas fluorescens SBW25 is capable of growing on histidine as a sole source of carbon and/or nitrogen. Previous work showed that the two-component regulatory system CbrAB is required for expression of the histidine utilization (hut) locus when histidine is the sole source of carbon and nitrogen. Here, using mutational analysis and transcriptional assays, we demonstrate involvement of a second two-component system, NtrBC. When histidine is the sole carbon source, transcription of the hutU operon is initiated from a sigma54-type promoter and requires CbrB (an enhancer binding protein for sigma54-recruitment). However, when histidine is the sole nitrogen source, the hutU operon is transcribed from a sigma70-type promoter and requires either CbrB or the nitrogen regulator, NtrC. No role was found for the SBW25 homolog of the nitrogen assimilation control protein (NAC). Biolog phenotypic microarray analysis of the ability of the three mutants (DeltacbrB, DeltantrC, and DeltacbrB DeltantrC) to utilize 190 carbon and 95 nitrogen substrates confirmed the central regulatory roles of CbrAB and NtrBC in cellular carbon and nitrogen catabolism: deletion of cbrB abolished growth on 20 carbon substrates; deletion of ntrC eliminated growth on 28 nitrogen substrates. A double cbrB-ntrC mutant was unable to utilize a further 14 nitrogen substrates (including histidine, proline, leucine, isoleucine, and valine). Our data show that CbrAB plays a role in regulation of both carbon and nitrogen catabolism and maintains activity of catabolic pathways under different C:N ratios.

| S-EPMC2206070 | biostudies-literature

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