Microbial growth on mercaptosuccinic acid

1968 ◽  
Vol 14 (5) ◽  
pp. 515-523 ◽  
Author(s):  
Mark R. Hall ◽  
Richard S. Berk
Keyword(s):  
Microbial Growth ◽  
Enrichment Culture ◽  
Basal Medium ◽  
Sole Source ◽  
Resting Cells ◽  
Metabolic Fate ◽  
Succinate Oxidation ◽  
Oxidase System ◽  
Mercaptosuccinic Acid ◽  
Inorganic Sulfate

Using enrichment culture technics a species of Alcaligenes (M1) was isolated from soil which was able to utilize mercaptosuccinic acid (MS) as a sole source of carbon, sulfur, and energy. Growth on a MS–salts basal medium was not significantly enhanced by single supplements of B-vitamins or by yeast extract. Comparative studies on succinate and MS oxidation by Alcaligenes and Pseudomonas aeruginosa indicated that MS was an inhibitor of succinate oxidation by resting cells of both microorganisms when they were grown in a medium lacking mercaptosuccinic acid such as a succinate–salts basal. However, when M1 was grown on MS, it was able to oxidize both succinate and MS, thereby indicating that the MS oxidase system was inducible. In addition, the MS oxidase system in cell-free extracts of M1 was relatively insensitive to 10–30 μmoles of malonate, whereas the succinoxidase system was inhibited 66% by 30 μmoles of the inhibitor. Cell-free extracts of succinate-grown cells of P. aeruginosa were unable to oxidize MS, indicating that the inactivity of resting cells was not due to a permease problem. Investigation of the metabolic fate of the sulfur moiety of MS by growing cells of M1 indicated that all of the available sulfur was liberated as inorganic sulfate, while no free sulfide was detected. Thiosulfate sulfurtransferase (rhodanese) was detected in extracts from cells grown both with and without mercaptosuccinic acid. However, growth in the MS medium enhanced the production of rhodanese approximately 40%. In addition, thiosulfate oxidase activity was also detected in resting cells and cell-free extracts prepared from MS-grown cells, but not from cells grown without mercaptosuccinic acid.

Biotransformation of nitrocellulose under methanogenic conditions

1996 ◽  
Vol 34 (5-6) ◽  
pp. 327-334 ◽  
Author(s):  
David L. Freedman ◽  
Bryan M. Caenepeel ◽  
Byung J. Kim
Keyword(s):  
Biological Treatment ◽  
Enrichment Culture ◽  
Basal Medium ◽  
Anaerobic Biodegradation ◽  
Anaerobic Treatment ◽  
Acid Digestion ◽  
Energetic Compounds ◽  
Nitrate Ester ◽  
Digestion Technique ◽  
Aerobic And Anaerobic

Treatment of wastewater containing nitrocellulose (NC) fines is a significant hazardous waste problem currently facing manufacturers of energetic compounds. Previous studies have ruled out the use of biological treatment, since NC has appeared to be resistant to aerobic and anaerobic biodegradation. The objective of this study was to examine NC biotransformation in a mixed methanogenic enrichment culture. A modified cold-acid digestion technique was used to measure the percentage of oxidized nitrogen (N) remaining on the NC. After 11 days of incubation in cultures amended with NC (10 g/L) and methanol (9.9 mM), the % N (w/w) on the NC decreased from 13.3% to 10.1%. The presence of NC also caused a 16% reduction in methane output. Assuming the nitrate ester on NC was reduced to N2, the decrease in CH4 represented almost exactly the amount of reducing equivalents needed for the observed decrease in oxidized N. An increase in the heat of combustion of the transformed NC correlated with the decrease in % N. There was no statistically significant decrease in % N when only NC was added to the culture, or in controls that contained only the sulfide-reduced basal medium. The biotransformed NC has a % N comparable to nonexplosive nitrated celluloses, suggesting that anaerobic treatment may be a technically feasible process for rendering NC nonhazardous.

Transformation of Alachlor by Microbial Communities

Weed Science ◽  
1990 ◽  
Vol 38 (4-5) ◽  
pp. 416-420 ◽  
Author(s):  
Hone L. Sun ◽  
Thomas J. Sheets ◽  
Frederick T. Corbin
Keyword(s):  
Carbon Source ◽  
Growth Medium ◽  
Basal Medium ◽  
Sole Source ◽  
Ring Cleavage ◽  
Side Chain ◽  
Microbial Culture ◽  
Mixed Microbial Culture ◽  
Alternative Carbon Source ◽  
Thin Layer Chromatographic Analysis

A mixed microbial culture able to transform alachlor at a concentration of 50 μg ml-1was obtained from alachlor-treated soil after an enrichment period of 84 days. The microbial community was composed of seven strains of bacteria. No single isolate was able to utilize alachlor as a sole source of carbon. There was no alachlor left in the enriched culture after a 14-day incubation, but only 12% of the14C-ring-labeled alachlor was converted to14CO2through ring cleavage during 14 days in the basal medium amended with alachlor as a sole carbon source. The presence of sucrose as an alternative carbon source decreased the mineralization potential of the enriched culture, but sucrose increased the mineralizing ability of a three-member mixed culture. Thin-layer chromatographic analysis showed that there were four unidentified metabolites of alachlor produced by the enriched culture. Sucrose decreased the amount of two of the four metabolites. The absence of a noticeable decline in radioactivity beyond the initial 12% suggested that the side chain of alachlor was utilized as carbon source by the enriched culture. Little difference in radioactivity between growth medium and cell-free supernatant of the growth medium suggested that the carbon in the ring was not incorporated into the cells of the degrading microorganisms.

Biochemical pathway and degradation of phthalate ester isomers by bacteria

2005 ◽  
Vol 52 (8) ◽  
pp. 241-248 ◽  
Author(s):  
J.-D. Gu ◽  
J. Li ◽  
Y. Wang
Keyword(s):  
Aromatic Ring ◽  
Enrichment Culture ◽  
Klebsiella Oxytoca ◽  
Sole Source ◽  
Phthalate Ester ◽  
Ring Cleavage ◽  
Mangrove Sediment ◽  
Individual Species ◽  
Dimethyl Phthalate ◽  
Dimethyl Phthalate Ester

Degradation of dimethyl isophthalate (DMI) and dimethyl phthalate ester (DMPE) was investigated using microorganisms isolated from mangrove sediment of Hong Kong Mai Po Nature Reserve. One enrichment culture was capable of utilizing DMI as the sole source of carbon and energy, but none of the bacteria in the enrichment culture was capable of degrading DMI alone. In co-culture of two bacteria, degradation was observed proceeding through monomethyl isophthalate (MMI) ester and isophthalic acid (IPA) before the aromatic ring opening. Using DMI as the sole carbon and energy source, Klebsiella oxytoca Sc and Methylobacterium mesophilicum Sr degraded DMI through the biochemical cooperation. The initial hydrolytic reaction of the ester bond was by K. oxytoca Sc and the next step of transformation was by M. mesophilicum Sr, and IPA was degraded by both of them. In another investigation, a novel bacterium, strain MPsc, was isolated for degradation of dimethyl phthalate ester (DMPE) also from the mangrove sediment. On the basis of phenotypic, biochemical and 16S rDNA gene sequence analyses, the strain MPsc should be considered as a new bacterium at the genus level (8% differences). This strain, together with a Rhodococcus zopfii isolated from the same mangrove sediment, was able to degrade DMPE aerobically. The consortium consisting of the two species degraded 450mg/l DMPE within 3 days as the sole source of carbon and energy, but none of the individual species alone was able to transform DMPE. Furthermore, the biochemical degradation pathway proceeded through monomethyl phthalate (MMP), phthalic acid (PA) and then protocatechuate before aromatic ring cleavage. Our results suggest that degradation of complex organic compounds including DMI and DMPE may be carried out by several members of microorganisms working together in the natural environments.

Metabolism of a monoterpene alcohol, linalool, by a soil pseudomonad

1977 ◽  
Vol 23 (3) ◽  
pp. 230-239 ◽  
Author(s):  
K. M. Madyastha ◽  
P. K. Bhattacharyya ◽  
C. S. Vaidyanathan
Keyword(s):  
Mineral Salt Medium ◽  
Enrichment Culture ◽  
Sole Source ◽  
Mineral Salt ◽  
Salt Medium ◽  
Culture Techniques ◽  
Unsaturated Lactone ◽  
Linalool Oxide

A microorganism of the genus Pseudomonas has been isolated from the soil by enrichment culture techniques with linalool(I) as the sole source of carbon and energy. The organism is also capable of utilizing limonene, citronellol, and geraniol as substrates but fails to grow on citral, citranellal, and 1,8-cineole. Fermentation of linalool by this bacterium in a mineral salt medium results in the formation of 10-hydroxylinalool(II), 10-carboxylinalool(III), oleuropeic acid(IX), 2-vinyl-2-methyl-5-hydroxyisopropyl-tetrahydrofuran(linalool oxide, V), 2-vinyl-2-methyl-tetrahydrofuran-5-one(unsaturated lactone, VI), and few unidentified minor metabolites. Probable pathways for the biodegradation of linalool are presented.

The metabolism of p-fluorophenylacetic acid by a Pseudomonas sp. I. Isolation and identification of intermediates in degradation

1971 ◽  
Vol 17 (5) ◽  
pp. 635-644 ◽  
Author(s):  
D. B. Harper ◽  
E. R. Blakley
Keyword(s):  
Sole Carbon Source ◽  
Culture Medium ◽  
Enrichment Culture ◽  
Culture Technique ◽  
Spectroscopic Techniques ◽  
Resting Cells ◽  
Pseudomonas Sp ◽  
Isolation And Identification ◽  
Hydroxyphenylacetic Acid ◽  
Organic Fluorine

A Pseudomonas sp. capable of growing on p-fluorophenylacetic acid as sole carbon source has been isolated using the enrichment culture technique. All the organic fluorine is released into the culture medium as fluoride ion during growth. A number of fluorinated intermediates have been isolated from the culture medium when resting cells were incubated with the substrate. Using infrared, nuclear magnetic resonance, and mass spectroscopic techniques together with chemical degradative procedures, these have been identified as D(+)-monofluorosuccinic acid, trans-3-fluoro-3-hexenedioic acid, (−)-4-carboxymethyl-4-fluorobutanolide, 4-fluoro-2-hydroxyphenylacetic acid, and 4-fluoro-3-hydroxyphenylacetic acid.

Degradation of (–)-Ephedrine by Pseudomonas putida Detection of (–)-Ephedrine: NAD+-oxidoreductase from Arthrobacter globiformis

1980 ◽  
Vol 35 (1-2) ◽  
pp. 80-87 ◽  
Author(s):  
E. Klamann ◽  
F. Lingens
Keyword(s):  
Pseudomonas Putida ◽  
Enrichment Culture ◽  
Culture Fluid ◽  
Sole Source ◽  
Culture Technique ◽  
Arthrobacter Globiformis ◽  
Muconic Acid ◽  
Ammonium Sulphate Precipitation ◽  
Wide Range ◽  
Nad Oxidoreductase

Abstract A bacterium utilizing the alkaloid (-)-ephedrine as its sole source of carbon was isolated by an enrichment-culture technique from soil supplemented with 4-benzoyl-1,3-oxazolidinon-(2). The bacterium was identified as Pseudomonasputida by morphological and physiological studies. The following metabolites were isolated from the culture fluid: methylamine, formaldehyde, methyl- benzoylcarbinol (2-hydroxy-1-oxo-1-phenylpropane), benzoid acid, pyrocatechol and cis, cis- muconic acid. A pathway for the degradation of (-)-ephedrine by Pseudomonas putida is proposed and compared with the degradative pathway in Arthrobacter globiformis.The enzyme, which is responsible for the first step in the catabolism of (-)-ephedrine could be demonstrated in extracts from Arthrobacter globiformis. This enzyme catalyses the dehydrogena- tion of (-)-ephedrine yielding phenylacetylcarbinol/methylbenzoylcarbinol and methylamine. It requires NAD+ as cofactor and exhibits optimal activity at pH 11 in 0.1 m glycine/NaOH buffer. The Km value for (-)-ephedrine is 0.02 mM and for NAD+ 0.11 mм, respectively. No remarkable loss of activity is observed following treatment with EDTA. The enzyme has been shown to react with a wide range of ethanolamines. A slight enrichment was obtained by ammonium sulphate precipitation. The name (-)-ephedrine: NAD+-oxidoreductase (deaminating) is proposed.

scholarly journals Lactate production is the major metabolic fate of glucose in splenocytes and is altered in spontaneously diabetic BB rats

1990 ◽  
Vol 272 (2) ◽  
pp. 445-452 ◽  
Author(s):  
C J Field ◽  
G Wu ◽  
M D Métroz-Dayer ◽  
M Montambault ◽  
E B Marliss
Keyword(s):  
Glucose Utilization ◽  
Lactate Production ◽  
Resting Cells ◽  
Metabolic Fate ◽  
Con A ◽  
Normal Cells ◽  
Immunological Activation ◽  
Proliferation Of Lymphocytes ◽  
Lactate And Pyruvate

Enhanced glucose metabolism is necessary to support the activation and proliferation of lymphocytes. To define further quantitatively the metabolic fates of glucose and assess glucose utilization both in normal cells and in an autoimmune disease with abnormal lymphocytes, [U-14C]glucose conversion into 14CO2 and the production of lactate and pyruvate were measured in splenocytes. Cells from non-diabetes-prone (BBn) and spontaneously diabetic (BBd) rats were studied both freshly isolated ‘resting’ and cultured for 96 h with and without concanavalin A (Con A) stimulation. (1) Lactate was confirmed to be the major end product in both freshly isolated (53% of utilized glucose) and unstimulated cultured (62% of utilized glucose) cells from BBn animals studied at (2-8) x 10(6) cells/ml concentration. The use of concentrations from 10 x 10(6) to 300 x 10(6) cells/ml resulted in progressively less lactate production per 10(6) splenocytes. (2) Cells from BBd animals after stimulation with Con A incorporated less [3H]thymidine and produced significantly less lactate (155 +/- 14 versus 305 +/- 24 nmol/2 h per 10(6) cells) than did BBn cells (P less than 0.05). (3) However, more lactate (101 +/- 8 versus 78 +/- 6 nmol/5 h per 10(6) cells) was produced by ‘resting’ cells from BBd animals compared with BBn (P less than 0.03), and this difference was sustained after 4 days in culture. (4) Significantly greater amounts of pyruvate were produced by BBd than by BBn cells, particularly when stimulated with Con A, suggesting an alteration in the availability of reducing equivalents in BBd cells. (5) These results are consistent with prior metabolic as well as immunological ‘activation’ of cells in vivo in the BB diabetic animals.

The degradation of p-toluenesulfonate by a Pseudomonas

1970 ◽  
Vol 16 (5) ◽  
pp. 309-316 ◽  
Author(s):  
D. D. Focht ◽  
F. D. Williams
Keyword(s):  
Carbon Dioxide ◽  
Oxygen Uptake ◽  
Aromatic Compounds ◽  
Carbon Sources ◽  
Sole Source ◽  
Carbon Chain ◽  
Ring Cleavage ◽  
Resting Cells ◽  
Resting Cell ◽  
Complete Mineralization

A Pseudomonas isolated from sewage was adapted to use p-toluenesulfonate as the sole source of both carbon and sulfur. Very few of over 30 aromatic compounds tested were used for growth as sole carbon sources. Significantly, sulfobenzoate, phenolsulfonates, and isomers of cresolsulfonates did not support growth. Respirometry studies with washed, resting cells showed similar results. In both studies, benzenesulfonate was always used more rapidly than p-toluenesulfonate. The degradation of p-toluenesulfonate was shown to be over 90% of the theoretical value required for complete mineralization to carbon dioxide, water, and sulfate. When resting cells were incubated with 35S-p-toluenesulfonate, the ratio of oxygen uptake to 35S-sulfate liberation remained constant during the complete degradation period. Radiochromatographic analysis showed no 35S-aromatic intermediates in resting-cell supernatants at any time. Resting cells previously incubated with 35S-p-toluenesulfonate liberated two 35S-labeled aromatic intermediates upon disruption. Resting cells incubated with 1-14C-p-toluenesulfonate produced labeled 3-methylcatechol, labeled acetate, and unlabeled pyruvate. The labeled intermediate, 3-methylcatechol, was degraded by cell-free extracts to labeled acetate. Hydroxylation, desulfonation, ring cleavage, and subsequent fissions of the carbon chain occurred in that order; all steps but the first were catalyzed by cell-free extracts.

scholarly journals The microbial metabolism of thiophen-2-carboxylate

1973 ◽  
Vol 134 (2) ◽  
pp. 353-366 ◽  
Author(s):  
Roger E. Cripps
Keyword(s):  
Methylene Blue ◽  
High Speed ◽  
Enrichment Culture ◽  
Specific Radioactivity ◽  
Microbial Metabolism ◽  
Sole Source ◽  
Cellular Protein ◽  
Sulphur Atom ◽  
Degradative Pathway ◽  
Anaerobic Reduction

1. An organism was isolated by enrichment culture that was capable of using thiophen-2-carboxylate as sole source of carbon, energy and sulphur for growth. 2. Analysis of the cellular protein after growth of the organism on thiophen-2-[14C]carboxylate showed that only glutamate, proline and arginine were labelled. All the radioactivity in the glutamate was confined to C-1. 3. In the presence of 2.1 mm-arsenite, suspensions of the organism converted thiophen-2-[14C]carboxylate into 14C-labelled 2-oxoglutarate which had the same specific radioactivity as the starting material. 4. Cell-free extracts of the organism catalysed the release of 14CO2 from thiophen-2-[14C]carboxylate. This activity was largely dependent on the presence of ATP and CoA and was stimulated by NAD+ and Mg2+. Inclusion of hydroxylamine resulted in the appearance of thiophen-2-carbohydroxamic acid, indicating that the ATP and CoA were involved in the formation of the CoA ester of thiophen-2-carboxylate. 5. High-speed centrifuging of cell-free extracts resulted in supernatants with decreased thiophen-2-carboxylate-degrading activity. Activity was restored by the addition of the high-speed pellet or by Methylene Blue. 6. The metabolism of the CoA ester of thiophen-2-carboxylate by cell-free extracts could be linked to the anaerobic reduction of Methylene Blue. 7. The sulphur atom of the thiophen nucleus was converted into sulphate by growing cultures and resting suspensions of the organism. 8. A degradative pathway is proposed involving the hydroxylation (at C-5) of the CoA ester of thiophen-2-carboxylate followed by further metabolism to 2-oxoglutarate and sulphate.

Isolation and Characterization of Phenol-Degrading Denitrifying Bacteria

1998 ◽  
Vol 64 (7) ◽  
pp. 2432-2438 ◽  
Author(s):  
Paula M. van Schie ◽  
L. Y. Young
Keyword(s):  
New York ◽  
Enrichment Culture ◽  
Phenol Degradation ◽  
Sole Source ◽  
New Members ◽  
Isolation And Characterization ◽  
East River ◽  
Pure Cultures ◽  
Orange Grove ◽  
Metabolic Properties

ABSTRACT Phenol is a man-made as well as a naturally occurring aromatic compound and an important intermediate in the biodegradation of natural and industrial aromatic compounds. Whereas many microorganisms that are capable of aerobic phenol degradation have been isolated, only a few phenol-degrading anaerobic organisms have been described to date. In this study, three novel nitrate-reducing microorganisms that are capable of using phenol as a sole source of carbon were isolated and characterized. Phenol-degrading denitrifying pure cultures were obtained by enrichment culture from anaerobic sediments obtained from three different geographic locations, the East River in New York, N.Y., a Florida orange grove, and a rain forest in Costa Rica. The three strains were shown to be different from each other based on physiologic and metabolic properties. Even though analysis of membrane fatty acids did not result in identification of the organisms, the fatty acid profiles were found to be similar to those of Azoarcusspecies. Sequence analysis of 16S ribosomal DNA also indicated that the phenol-degrading isolates were closely related to members of the genusAzoarcus. The results of this study add three new members to the genus Azoarcus, which previously comprised only nitrogen-fixing species associated with plant roots and denitrifying toluene degraders.

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