Aerobic purification of N5,N10-methylenetetrahydromethanopterin dehydrogenase, separated from N5,N10-methenyltetrahydromethanopterin cyclohydrolase, from Methanobacterium thermoautotrophicum strain Marburg

1989 ◽  
Vol 35 (4) ◽  
pp. 499-507 ◽  
Author(s):  
Biswarup Mukhopadhyay ◽  
Lacy Daniels
Keyword(s):  
Enzyme Activity ◽  
Ammonium Sulfate ◽  
Dehydrogenase Activity ◽  
Specific Activity ◽  
Methanobacterium Thermoautotrophicum ◽  
Anaerobic Conditions ◽  
Cell Extracts ◽  
First Time ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

The N5,N10-methylenetetrahydromethanopterin dehydrogenase from Methanobacterium thermoautotrophicum strain Marburg has been purified with reasonable yield and much higher specific activity than previously reported. For the first time it has been shown that both N5,N10-methylenetetrahydromethanopterin dehydrogenase and N5,N10-methenyltetrahydromethanopterin cyclohydrolase activities were stable under air and could be purified using aerobic operations. The dehydrogenase activity from Methanobacterium thermoautotrophicum Marburg was stable in phosphate buffer with or without glycerol or ammonium sulfate under both aerobic and anaerobic conditions. However, the presence of either 2-mercaptoethanol or dithiothreitol in the enzyme solution destroyed the enzyme activity during both aerobic and anaerobic incubations. Dehydrogenase was purified 62-fold using Phenyl-Sepharose and DEAE-Sephadex chromatography in succession under air. Both of these chromatographic methods separated dehydrogenase activity from N5,N10-methenyltetrahydromethanopterin cyclohydrolase; DEAE-Sephadex provided the best separation. Phenyl-Sepharose chromatography of the supernatant of cell extracts containing ammonium sulfate at 60% of saturation provided a 4.7-fold purification and 98% recovery of cyclohydrolase; this result established the air stability of N5,N10-methenyltetrahydromethanopterin cyclohydrolase from Methanobacterium thermoautotrophicum Marburg.Key words: methylenetetrahydromethanopterin dehydrogenase, methenyltetrahydromethanopterin cyclohydrolase, Methanobacterium, aerobic purification, oxygen stability.

scholarly journals Metabolism of dibenzo[1,4]dioxan by a Pseudomonas species

1979 ◽  
Vol 180 (3) ◽  
pp. 639-645 ◽  
Author(s):  
G M Klečka ◽  
D T Gibson
Keyword(s):  
Parental Strain ◽  
Culture Medium ◽  
Crystalline Form ◽  
Pseudomonas Sp ◽  
Anaerobic Conditions ◽  
Cell Extracts ◽  
Pseudomonas Species ◽  
Chemical Techniques ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Pseudomonas sp. N.C.I.B. 9816 strain 11, when grown on salicylate in the presence of dibenzo[1,4]dioxan, accumulated cis-1,2-dihydroxy-1,2-dihydrodibenzo[1,4]dioxan and 2-hydroxydibenzo[1,4]dioxan in the culture medium. Each metabolite was isolated in crystalline form and identified by a variety of conventional chemical techniques. Crude cell extracts prepared from the parental strain grown with naphthalene oxidized cis-1,2-dihydroxy-1,2-dihydrodibenzo[1,4]dioxan under both aerobic and anaerobic conditions to 1,2-dihydroxydibenzo[1,4]dioxan. Further degradation of this metabolite was not detected.

scholarly journals Calcite and vaterite biosynthesis by nitrate dissimilating bacteria in carbonatogenesis process under aerobic and anaerobic conditions

2019 ◽  
Author(s):  
Marwa Eltarahony ◽  
Sahar Zaki ◽  
Ayman Kamal ◽  
Desouky Abd-El-Haleem
Keyword(s):  
Nitrate Reduction ◽  
Reductase Activity ◽  
Specific Property ◽  
Bacterial Strains ◽  
Anaerobic Conditions ◽  
Lysinibacillus Sphaericus ◽  
Nucleation Sites ◽  
First Time ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Abstract. This study deals with 16S rDNA identified bacteria, Lysinibacillus sphaericus (71A), Raoultella planticola (VIP), and Streptomyces pluricolorescens (EM4) capable of precipitating CaCO3 through a nitrate reduction aerobically and anaerobically. The produced CaCO3 crystals were analyzed using XRD, EDX, and SEM. The results showed that the carbonatogenic bacteria served as nucleation sites for CaCO3 precipitation with distinct variation in polymorph and morphology; reflecting strain-specific property. Notably, the amount of precipitated CaCO3 recorded 3.27 (aerobic), 1.55 (anaerobic), 4.15 (aerobic), 3.75 (aerobic) and 1.87 (anaerobic) g/100 mL of strains 71A, EM4 and VIP, respectively, for 240 h of incubation. The study of changes in media chemistry during carbonatogenesis process revealed positive correlation between bacterial growth, nitrate reductase activity, pH, EC, amount of deposited CaCO3 and NO3− consumption. Therefore, the applications of these bacterial strains, which employed for the first time in carbonatogenesis process, are promising in the environmental, biomedical and civil engineering fields.

scholarly journals Reactivity of nitric oxide with the [4Fe–4S] cluster of dihydroxyacid dehydratase from Escherichia coli

2009 ◽  
Vol 417 (3) ◽  
pp. 783-789 ◽  
Author(s):  
Xuewu Duan ◽  
Juanjuan Yang ◽  
Binbin Ren ◽  
Guoqiang Tan ◽  
Huangen Ding
Keyword(s):  
Nitric Oxide ◽  
Escherichia Coli ◽  
Enzyme Activity ◽  
Anaerobic Conditions ◽  
Iron Sulfur Clusters ◽  
E Coli ◽  
Endonuclease Iii ◽  
Iron Sulfur ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Although the NO (nitric oxide)-mediated modification of iron–sulfur proteins has been well-documented in bacteria and mammalian cells, specific reactivity of NO with iron–sulfur proteins still remains elusive. In the present study, we report the first kinetic characterization of the reaction between NO and iron–sulfur clusters in protein using the Escherichia coli IlvD (dihydroxyacid dehydratase) [4Fe–4S] cluster as an example. Combining a sensitive NO electrode with EPR (electron paramagnetic resonance) spectroscopy and an enzyme activity assay, we demonstrate that NO is rapidly consumed by the IlvD [4Fe–4S] cluster with the concomitant formation of the IlvD-bound DNIC (dinitrosyl–iron complex) and inactivation of the enzyme activity under anaerobic conditions. The rate constant for the initial reaction between NO and the IlvD [4Fe–4S] cluster is estimated to be (7.0±2.0)×106 M−2·s−1 at 25 °C, which is approx. 2–3 times faster than that of the NO autoxidation by O2 in aqueous solution. Addition of GSH failed to prevent the NO-mediated modification of the IlvD [4Fe–4S] cluster regardless of the presence of O2 in the medium, further suggesting that NO is more reactive with the IlvD [4Fe–4S] cluster than with GSH or O2. Purified aconitase B [4Fe–4S] cluster from E. coli has an almost identical NO reactivity as the IlvD [4Fe–4S] cluster. However, the reaction between NO and the endonuclease III [4Fe–4S] cluster is relatively slow, apparently because the [4Fe–4S] cluster in endonuclease III is less accessible to solvent than those in IlvD and aconitase B. When E. coli cells containing recombinant IlvD, aconitase B or endonuclease III are exposed to NO using the Silastic tubing NO delivery system under aerobic and anaerobic conditions, the [4Fe–4S] clusters in IlvD and aconitase B, but not in endonuclease III, are efficiently modified forming the protein-bound DNICs, confirming that NO has a higher reactivity with the [4Fe–4S] clusters in IlvD and aconitase B than with O2 or GSH. The results suggest that the iron–sulfur clusters in proteins such as IlvD and aconitase B may constitute the primary targets of the NO cytotoxicity under both aerobic and anaerobic conditions.

Oxygen concentration affects frequency and range of transconjugants for the incompatibility (Inc) P-1 and P-7 plasmids pBP136 and pCAR1

2020 ◽  
Vol 85 (4) ◽  
pp. 1005-1015
Author(s):  
Kentaro Ochi ◽  
Maho Tokuda ◽  
Kosuke Yanagiya ◽  
Chiho Suzuki-Minakuchi ◽  
Hideaki Nojiri ◽  
...  
Keyword(s):  
Oxygen Concentration ◽  
Anaerobic Condition ◽  
Environmental Samples ◽  
Aerobic Condition ◽  
Recipient Strain ◽  
Donor Strain ◽  
Anaerobic Conditions ◽  
Filter Mating ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

ABSTRACT The frequency of transconjugants were compared for the incompatibility (Inc) P-1 and P-7 plasmids pBP136 and pCAR1 under aerobic and anaerobic conditions. Filter mating assays were performed with one donor strain and one recipient strain using different donors of Pseudomonas and recipient strains, including Pseudomonas, Pantoea, and Buttiauxella. Under anaerobic condition, frequencies of transconjugants for both plasmids were 101-103-fold lower than those under aerobic condition regardless of whether aerobically or anaerobically grown donors and recipients were used. To compare the transconjugant ranges under aerobic and anaerobic conditions, conjugation was performed between the donor of pBP136 and recipient bacteria extracted from environmental samples. Several transconjugants were uniquely obtained from each aerobic or anaerobic condition. Our findings indicate that a plasmid can differently spread among bacteria depending on the oxygen concentrations of the environment.

PRODUCTION AND PROPERTIES OF 2,3-BUTANEDIOL: VII. FERMENTATION OF WHEAT BY AEROBACILLUS POLYMYXA UNDER AEROBIC AND ANAEROBIC CONDITIONS

1946 ◽  
Vol 24f (1) ◽  
pp. 1-11 ◽  
Author(s):  
G. A. Adams
Keyword(s):  
Carbon Dioxide ◽  
Anaerobic Fermentation ◽  
Anaerobic Conditions ◽  
Mechanical Agitation ◽  
Fermentation Time ◽  
Aerobic Conditions ◽  
Ethanol Formation ◽  
Time Required ◽  
Aerobic And Anaerobic ◽  
Aerobic And Anaerobic Conditions

Aeration by mechanical agitation of 15% wheat mash fermented by Aerobacillus polymyxa inhibited the formation of 2,3-butanediol and particularly of ethanol. Aeration of similar mashes by passage of finely dispersed air or oxygen at the rate of 333 ml. per minute per litre of mash increased the rate of formation and yield of 2,3-butanediol but inhibited ethanol formation. However, the over-all time required for the completion of fermentation was not shortened from the usual 72 to 96 hr. required for unaerated mashes. There was no evidence of a shift from fermentative to oxidative dissimilation. Under aerobic conditions, the final butanediol–ethanol ratio was approximately 3:1. Anaerobic conditions, as produced by the passage of nitrogen or hydrogen through the mash, increased the rate of formation of both butanediol and ethanol and shortened the fermentation time to about 48 hr. Under these conditions, the butanediol–ethanol ratio was reduced to about 1.3:1.0. Carbon dioxide gave a butanediol–ethanol ratio resembling that of anaerobic fermentation but did not reduce fermentation time.

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