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.

Pyridine degradation and heterocyclic nitrification by Bacillus coagulans

1997 ◽  
Vol 43 (6) ◽  
pp. 595-598 ◽  
Author(s):  
B. Uma ◽  
S. Sandhya
Keyword(s):  
Doubling Time ◽  
Enrichment Culture ◽  
Bacillus Coagulans ◽  
Sole Source ◽  
Culture Technique ◽  
Heterotrophic Nitrification ◽  
Aromatic Degradation ◽  
Pyridine Degradation ◽  
Unique Potential ◽  
Degrading Microorganism

A gram-positive, pyridine-degrading microorganism identified as Bacillus coagulans has been isolated from contaminated soil by enrichment culture technique. Pyridine was used as sole source of carbon, nitrogen, and energy. Bacillus coagulans has a unique potential to reduce nitrogen from aromatic ring to ammonia and subsequently heterotrophically to nitrite and nitrate. The maximum degradation of pyridine was 94.1% within 72 h at 30 °C with a 7.57-h doubling time. The study suggests possible existence of aromatic degradation and heterotrophic nitrification in Bacillus coagulans.Key words: pyridine degradation, heterotrophic nitrification, Bacillus coagulans.

scholarly journals The hydrolysis of algal galactans by enzymes from a Cytophaga species

1967 ◽  
Vol 105 (1) ◽  
pp. 311-316 ◽  
Author(s):  
J R Turvey ◽  
J. Christison
Keyword(s):  
Red Algae ◽  
Sea Water ◽  
Enrichment Culture ◽  
Hydrolytic Enzymes ◽  
Sole Source ◽  
Culture Technique ◽  
Partial Purification ◽  
Hydrolysis Of

1. Two bacteria were isolated from sea water by the enrichment culture technique, both of which could utilize the galactan sulphate, porphyran, as sole source of carbon. 2. From the cells of one bacterium, classified as a Cytophaga sp., hydrolytic enzymes were isolated. 3. Partial purification of the enzymes is described and some of the properties of the principal enzymes have been studied. 4. The action of the enzymes on several galactan sulphates of red algae suggests that an agarase is present in the mixture.

scholarly journals Characterization of a Novel CaCO3-Forming Alkali-Tolerant Rhodococcus erythreus S26 as a Filling Agent for Repairing Concrete Cracks

Molecules ◽  
2021 ◽  
Vol 26 (10) ◽  
pp. 2967
Author(s):  
Seunghoon Choi ◽  
Sungjin Park ◽  
Minjoo Park ◽  
Yerin Kim ◽  
Kwang Min Lee ◽  
...  
Keyword(s):  
Building Materials ◽  
Culture Fluid ◽  
Natural Phenomenon ◽  
Environment Friendly ◽  
Wide Range ◽  
Filling Agent ◽  
Calcium Source ◽  
Alkali Tolerance ◽  
High Alkali

Biomineralization, a well-known natural phenomenon associated with various microbial species, is being studied to protect and strengthen building materials such as concrete. We characterized Rhodococcus erythreus S26, a novel urease-producing bacterium exhibiting CaCO3-forming activity, and investigated its ability in repairing concrete cracks for the development of environment-friendly sealants. Strain S26 grown in solid medium formed spherical and polygonal CaCO3 crystals. The S26 cells grown in a urea-containing liquid medium caused culture fluid alkalinization and increased CaCO3 levels, indicating that ureolysis was responsible for CaCO3 formation. Urease activity and CaCO3 formation increased with incubation time, reaching a maximum of 2054 U/min/mL and 3.83 g/L, respectively, at day four. The maximum CaCO3 formation was achieved when calcium lactate was used as the calcium source, followed by calcium gluconate. Although cell growth was observed after the induction period at pH 10.5, strain S26 could grow at a wide range of pH 4–10.5, showing its high alkali tolerance. FESEM showed rhombohedral crystals of 20–60 µm in size. EDX analysis indicated the presence of calcium, carbon, and oxygen in the crystals. XRD confirmed these crystals as CaCO3 containing calcite and vaterite. Furthermore, R. erythreus S26 successfully repaired the artificially induced large cracks of 0.4–0.6 mm width.

Technology of production of biopesticides based on Bacillus thuringiensis

2021 ◽  
Vol 9 (16) ◽  
pp. 28-38
Author(s):  
Anatolii Bezusov ◽  
◽  
Valentyna Krutiakova ◽  
Olena Myroshnichenko ◽  
Nataliia Dotsenko ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Corn Oil ◽  
Raw Materials ◽  
Culture Fluid ◽  
Scientific Work ◽  
Insect Larvae ◽  
Effective Prevention ◽  
Research Results ◽  
Wide Range ◽  
Vegetable Raw Materials

Subject of research. Biopesticides are based on live cultures of specially selected beneficial microorganisms with controlled properties. They have a pronounced phytoprotective and stimulating effect, thus providing effective prevention and protection of plants from diseases. The obtained biological product increases productivity, improves the quality and structure of the crop, does not accumulate in plants, which allows to obtain environmentally friendly agricultural products and does not harm the environment. Among the large number of bacteria, as a source of microbiological insecticide is Bacillus thuringiensis, which infects lepidopteran pests and leads to their death. Preparations based on this strain are effective when used in low concentrations of solutions. The Bacillus thuringiensis strain produces several toxins with insecticidal action, including β-exotoxin and δ-endotoxin. Toxic effect is manifested and leads to paralysis of the intestinal tract of parasites. Preparations β-exotoxin and δ-endotoxin are obtained by culturing Bacillus thuringiensis bacteria in a liquid medium. The scientific work proposes a method of industrial production using by-products of vegetable raw materials, which makes it economically feasible to use such substrates. The purpose of the study is the development of technology, formulation of nutrient medium, process parameters for the cultivation of bacteria of the genus Bacillus thuringiensis and obtaining a culture fluid containing substances of the class of biopesticides. Methods. Standard and generally accepted methods of research of bioproducts in biotechnology. The formation of bioinsecticides was established by hydrolysis methods, followed by determination of the component of β-exotoxin – ribose, the formation of octagonal crystals of exotoxin – by microscopic method. Research results. Three variants of nutrient media, which include yeast-polysaccharide complex: corn flour, corn oil, yeast autolysate were developed. The parameters of the bacterial cultivation process were studied. The final product is a paste or powder with a titer of 35 .109 spores in 1 g of the bioproduct. Scope of research results. Microbiological preparations based on Bacillus thuringiensis are highly specific and act only on insect larvae from the classes Lepidoptera and Diptera. The resulting biopesticide can be used against pests of a wide range of vegetable and fruit crops.

scholarly journals Novel Pathway for the Degradation of 2-Chloro-4-Nitrobenzoic Acid by Acinetobacter sp. Strain RKJ12

2011 ◽  
Vol 77 (18) ◽  
pp. 6606-6613 ◽  
Author(s):  
Dhan Prakash ◽  
Ravi Kumar ◽  
R. K. Jain ◽  
B. N. Tiwary
Keyword(s):  
Salicylic Acid ◽  
Tricarboxylic Acid Cycle ◽  
Sole Source ◽  
Degradation Pathway ◽  
Ring Cleavage ◽  
Muconic Acid ◽  
Content Type ◽  
Nitrobenzoic Acid ◽  
Catabolic Genes ◽  
Catechol 1,2 Dioxygenase

ABSTRACTThe organismAcinetobactersp. RKJ12 is capable of utilizing 2-chloro-4-nitrobenzoic acid (2C4NBA) as a sole source of carbon, nitrogen, and energy. In the degradation of 2C4NBA by strain RKJ12, various metabolites were isolated and identified by a combination of chromatographic, spectroscopic, and enzymatic activities, revealing a novel assimilation pathway involving both oxidative and reductive catabolic mechanisms. The metabolism of 2C4NBA was initiated by oxidativeorthodehalogenation, leading to the formation of 2-hydroxy-4-nitrobenzoic acid (2H4NBA), which subsequently was metabolized into 2,4-dihydroxybenzoic acid (2,4-DHBA) by a mono-oxygenase with the concomitant release of chloride and nitrite ions. Stoichiometric analysis indicated the consumption of 1 mol O2per conversion of 2C4NBA to 2,4-DHBA, ruling out the possibility of two oxidative reactions. Experiments with labeled H218O and18O2indicated the involvement of mono-oxygenase-catalyzed initial hydrolytic dechlorination and oxidative denitration mechanisms. The further degradation of 2,4-DHBA then proceeds via reductive dehydroxylation involving the formation of salicylic acid. In the lower pathway, the organism transformed salicylic acid into catechol, which was mineralized by theorthoring cleavage catechol-1,2-dioxygenase tocis, cis-muconic acid, ultimately forming tricarboxylic acid cycle intermediates. Furthermore, the studies carried out on a 2C4NBA−derivative and a 2C4NBA+transconjugant demonstrated that the catabolic genes for the 2C4NBA degradation pathway possibly reside on the ∼55-kb transmissible plasmid present in RKJ12.

The metabolism of cyclohexanecarboxylic acid and 3-cyclohexenecarboxylic acid by Pseudomonas putida

1982 ◽  
Vol 28 (12) ◽  
pp. 1324-1329 ◽  
Author(s):  
E. R. Blakley ◽  
B. Papish
Keyword(s):  
Benzoic Acid ◽  
Pseudomonas Putida ◽  
Sole Source ◽  
Oxidation Pathway ◽  
Cyclohexanecarboxylic Acid

A strain of Pseudomonas putida grew rapidly on cyclohexanecarboxylic acid as a sole source of carbon. A CoA-mediated β-oxidation pathway was induced for the metabolism of the compound. The organism could not utilize 3-cyclohexenecarboxylic acid as a sole source of carbon for growth, but cells grown on gluconate in the presence of 3-cyclohexenecarboxylic acid were induced to metabolize cyclohexanecarboxylic acid, benzoic acid, and catechol. Evidence is presented that 3-cyclohexenecarboxylic acid was slowly metabolized by a β-oxidation pathway and by a pathway involving benzoic acid as an intermediate. For this strain of Pseudomonas putida, 3-cyclohexenecarboxylic acid acts as an oxidizable, nongrowth substrate and induces the metabolism of cyclohexanecarboxylic acid and benzoic acid.

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.

scholarly journals Time-Course Proteomic Analysis of Pseudomonas putida KT2440 during Mcl-Polyhydroxyalkanoate Synthesis under Nitrogen Deficiency

Polymers ◽  
2019 ◽  
Vol 11 (5) ◽  
pp. 748 ◽  
Author(s):  
Justyna Możejko-Ciesielska ◽  
Agnieszka Mostek
Keyword(s):  
Pseudomonas Putida ◽  
Proteomic Analysis ◽  
Cellular Response ◽  
Time Course ◽  
Molecular Mechanisms ◽  
Nitrogen Deficiency ◽  
Growth Stages ◽  
Bacterial Cells ◽  
Pseudomonas Putida Kt2440 ◽  
Wide Range

Medium-chain-length polyhydroxyalkanoates (mcl-PHAs) have gained great attention as a new green alternative to petrochemical-derived polymers. Due to their outstanding material properties they can be used in a wide range of applications. Pseudomonas putida KT2440 is a metabolically versatile producer of mcl-polyhydroxyalkanoates. Although the metabolism of polyhydroxyalkanoate synthesis by this bacterium has been extensively studied, the comparative proteome analysis from three growth stages of Pseudomonas putida KT2440 cultured with oleic acid during mcl-PHA synthesis has not yet been reported. Therefore; the aim of the study was to compare the proteome of Pseudomonas putida KT2440 at different time points of its cultivation using the 2D difference gel electrophoresis (2D-DIGE) technique. The analyses showed that low levels of a nitrogen source were beneficial for mcl-PHA synthesis. Proteomic analysis revealed that the proteins associated with carbon metabolism were affected by nitrogen starvation and mcl-PHA synthesis. Furthermore, the induction of proteins involved in nitrogen metabolism, ribosome synthesis, and transport was observed, which may be the cellular response to stress related to nitrogen deficiency and mcl-PHA content in bacterial cells. To sum up; this study enabled the investigators to acquire a better knowledge of the molecular mechanisms underlying the induction of polyhydroxyalkanoate synthesis and accumulation in Pseudomonas putida KT2440 that could lead to improved strategies for PHAs in industrial production.

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