scholarly journals Measurement in vivo of hydrogenase-catalysed hydrogen evolution in the presence of nitrogenase enzyme in cyanobacteria

1979 ◽  
Vol 177 (1) ◽  
pp. 139-144 ◽  
Author(s):  
A Daday ◽  
G R Lambert ◽  
G D Smith
Keyword(s):  
Gas Phase ◽  
Methyl Viologen ◽  
Nitrogenase Activity ◽  
Conclusive Evidence ◽  
Hydrogenase Activity ◽  
H2 Evolution ◽  
Total Inhibition ◽  
Low Concentrations ◽  
Cyanobacterium Anabaena

A method was devised that allows measurement in vivo of hydrogenase-catalysed H2 evolution from the cyanobacterium Anabaena cylindrica, independent of nitrogenase activity, which is also present. Addition of low concentrations of reduced Methyl Viologen (1-10mM) to intact heterocystous filaments of the organism resulted in H2 evolution, but produced conditions giving total inhibition of nitrogenase (acetylene-reducing and H2-evolving) activity. That the H2 formed under these conditions was not contributed to by nitrogenase was also supported by the observation that its rate of formation was similar in the dark or with Ar replaced by N2 in the gas phase, and also in view of the pattern of H2 evolution at very low Methyl Viologen concentrations. Conclusive evidence that the H2 formed in the presence of Methyl Viologen was solely hydrogenase-mediated was its evolution even from nitrogenase-free (non-heterocystous) cultures; by contrast ‘uptake’ hydrogenase activity in such cultures was greatly decreased. The hydrogenase activity was inhibited by CO and little affected by acetylene. Finally the hydrogenase activity was shown to be relatively constant at different stages during the batch growth of the organism, as opposed to nitrogenase activity, which varied.

Hydrogen Evolution Catalyzed by Hydrogenase in Cultures of Cyanobacteria

1981 ◽  
Vol 36 (1-2) ◽  
pp. 87-92 ◽  
Author(s):  
Patrick C. Hallenbeck ◽  
Leon V. Kochian ◽  
John R. Benemann
Keyword(s):  
Hydrogen Production ◽  
Hydrogen Evolution ◽  
High Frequency ◽  
Nitrogenase Activity ◽  
Inhibitory Effect ◽  
Hydrogenase Activity ◽  
Oxygen Inhibition ◽  
Oxygen Sensitive

Abstract Cultures of Anabaena cylindrica, grown on media containing 5 mᴍ NH4Cl (which represses heterocyst formation), evolved hydrogen after a period of dark incubation under an argon atmosphere. This hydrogen production was not due to nitrogenase activity, which was nearly undetectable, but was due to a hydrogenase. Cultures grown on media with tungsten substituted for molybdenum had a high frequency of heterocysts (15%) and inactive nitrogenase after nitrogen starvation. The hydrogenase activity of these cultures was three-fold greater than the activity of non-heterocystous cultures. The effects of oxygen inhibition on hydrogen evolution by hetero-cystous cultures suggest that two pools of hydrogenase activity exist - an oxygen sensitive hydrogen evolution in vegetative cells and a relatively oxygen-resistent hydrogen evolution in heterocysts. In either case, inhibition by oxygen was reversible. Light had an inhibitory effect on net hydrogen evolution. Hydrogen production in vitro was much higher than in vivo, indicating that in vivo hydrogenase activity is limited by endogenous reductant supply.

Differentiation of Rhizobium japonicum, I. Enzymatic Comparison of Nitrogenase Repressed and Derepressed Free Living Cells and of Bacteroids

1978 ◽  
Vol 33 (3-4) ◽  
pp. 245-252 ◽  
Author(s):  
D. Werner ◽  
R. Stripf
Keyword(s):  
Gas Phase ◽  
Liquid Culture ◽  
Nitrogenase Activity ◽  
Living Cells ◽  
Maximum Activity ◽  
Rhizobium Japonicum ◽  
Free Living ◽  
Low Concentrations ◽  
Specific Activities ◽  
Japonicum Strain

Derepressed free living cells of Rhizobium japonicum strain 61-A-101 with leucine as single nitrogen source develop a maximum nitrogenase activity of 180 nmol C2H4 · mg protein -1·h-1 in liquid culture under 2% O2 in the gas phase. Only 10% of this activity is found with no oxygen in the gas phase during a 90 min incubation period. The maximum activity under 2% oxygen in the gas phase is unaffected by addition of 1 -100 mM NH4* and by addition of low concentrations of glutamine (0.36 - 1.44 mᴍ).Specific activities of alanine dehydrogenase (E.C. 1.4.1.1.) aspartate aminotransferase (E.C. 2.6.1.1.) and, with much lower activities, of GOGAT (E.C. 1.4.1.13) in nitrogenase active free living cells are more similar to bacteroids than to nitrogenase repressed free living cells from liquid culture. The activities in nitrogenase repressed cells were about 50% lower. Glutamine synthetase (E.C. 6.3.1.2.) activity in bacteroids and in nitrogenase active cells were also similar, but only about 25 - 30% of that found in nitrogenase repressed Rhizobium japonicum cells.

Hydrogen metabolism of Azospirillum brasilense in nitrogen-free medium

1980 ◽  
Vol 26 (9) ◽  
pp. 1126-1131 ◽  
Author(s):  
Y. K. Chan ◽  
L. M. Nelson ◽  
R. Knowles
Keyword(s):  
Gas Phase ◽  
Cyclic Gmp ◽  
Azospirillum Brasilense ◽  
Nitrogenase Activity ◽  
Hydrogenase Activity ◽  
Similar Response ◽  
Uptake Hydrogenase ◽  
Hydrogen Metabolism ◽  
Free Medium ◽  
Batch Cultures

Production of H2 by Azospirillum brasilense under N2-fixing conditions was studied in continuous and batch cultures. Net H2 production was consistently observed only when the gas phase contained CO. Nitrogenase activity (C2H2 reduction) and H2 evolution (in the presence of 5% CO) showed a similar response to O2 and were highest at 0.75% dissolved O2. Uptake hydrogenase activity, ranging from 0.3 to 2.5 μmol H2/mg protein per hour was observed in batch cultures under N2. Such rates were more than sufficient to recycle nitrogenase-produced H2. Tritium-exchange assay showed that H2 uptake was higher under Ar than under N2. Uptake hydrogenase was strongly inhibited by CO and C2H2. Cyclic GMP inhibited both nitrogenase and uptake hydrogenase activities.

The hydrogenases of Thiocapsa roseopersicina

2005 ◽  
Vol 33 (1) ◽  
pp. 61-63 ◽  
Author(s):  
K.L. Kovács ◽  
Á.T. Kovács ◽  
G. Maróti ◽  
L.S. Mészáros ◽  
J. Balogh ◽  
...  
Keyword(s):  
Nitrogenase Activity ◽  
Electron Flow ◽  
Random Mutagenesis ◽  
Hydrogenase Activity ◽  
Transcriptional Level ◽  
Photosynthetic Membrane ◽  
Practical Applications ◽  
Thiocapsa Roseopersicina ◽  
In The Wild

The purple sulphur phototrophic bacterium, Thiocapsa roseopersicina BBS, contains several NiFe hydrogenases. One of these enzymes (HynSL) is membrane associated, remarkably stable and can be used for practical applications. HupSL is also located in the photosynthetic membrane, its properties are similar to other known Hup-type NiFe hydrogenases. A third hydrogenase activity was located in the soluble fraction and was analogous to the NAD-reducing hydrogenases of cyanobacteria. The hoxEFUYH genes are transcribed together. HoxE is needed for the in vivo electron flow to and from the soluble hydrogenase. Some of the accessory genes were identified using random mutagenesis, and sequencing of the T. roseopersicina genome is in progress. The HupD, HynD and HoxW gene products corresponded to the proteases processing the C-termini of the three NiFe hydrogenases respectively. HypF and HupK mutants displayed significant in vivo H2 evolution, which could be linked to the nitrogenase activity for the ΔhypF and to the bidirectional Hox activity in the ΔhupK strain. Both HypC proteins are needed for the biosynthesis of each NiFe hydrogenase. The hydrogenase expression is regulated at the transcriptional level through distinct mechanisms. The expression of hynSL is up-regulated under anaerobic conditions with the participation of an FNR (fumarate and nitrate reduction regulator)-type protein, FnrT. Although the genes encoding a typical H2 sensor (hupUV) and a two-component regulator (hupR and hupT) are present in T. roseopersicina, the system is cryptic in the wild-type BBS strain. The hupR gene was identified in the gene cluster downstream from hupSL. Introduction of actively expressed hupT repressed the hupSL gene expression as expected by analogy with other bacteria.

scholarly journals Catabolic Function of Compartmentalized Alanine Dehydrogenase in the Heterocyst-Forming Cyanobacterium Anabaena sp. Strain PCC 7120

2010 ◽  
Vol 192 (19) ◽  
pp. 5165-5172 ◽  
Author(s):  
Rafael Pernil ◽  
Antonia Herrero ◽  
Enrique Flores
Keyword(s):  
Fluorescent Protein ◽  
Nitrogenase Activity ◽  
Vegetative Cells ◽  
Reading Frame ◽  
Alanine Dehydrogenase ◽  
Cyanobacterium Anabaena ◽  
Green Fluorescent ◽  
Pcc 7120 ◽  
Diazotrophic Growth

ABSTRACT In the diazotrophic filaments of heterocyst-forming cyanobacteria, an exchange of metabolites takes place between vegetative cells and heterocysts that results in a net transfer of reduced carbon to the heterocysts and of fixed nitrogen to the vegetative cells. Open reading frame alr2355 of the genome of Anabaena sp. strain PCC 7120 is the ald gene encoding alanine dehydrogenase. A strain carrying a green fluorescent protein (GFP) fusion to the N terminus of Ald (Ald-N-GFP) showed that the ald gene is expressed in differentiating and mature heterocysts. Inactivation of ald resulted in a lack of alanine dehydrogenase activity, a substantially decreased nitrogenase activity, and a 50% reduction in the rate of diazotrophic growth. Whereas production of alanine was not affected in the ald mutant, in vivo labeling with [14C]alanine (in whole filaments and isolated heterocysts) or [14C]pyruvate (in whole filaments) showed that alanine catabolism was hampered. Thus, alanine catabolism in the heterocysts is needed for normal diazotrophic growth. Our results extend the significance of a previous work that suggested that alanine is transported from vegetative cells into heterocysts in the diazotrophic Anabaena filament.

Kinetics of Various 99mTc-Sn-Pyrophosphate Compounds in the Rat

1977 ◽  
Vol 16 (04) ◽  
pp. 157-162 ◽  
Author(s):  
C. Schümichen ◽  
B. Mackenbrock ◽  
G. Hoffmann
Keyword(s):  
Normal Saline ◽  
Half Life ◽  
Compound A ◽  
Short Half Life ◽  
Low Concentrations ◽  
The Stability ◽  
Kinetics Of ◽  
In Vitro Stability

SummaryThe bone-seeking 99mTc-Sn-pyrophosphate compound (compound A) was diluted both in vitro and in vivo and proved to be unstable both in vitro and in vivo. However, stability was much better in vivo than in vitro and thus the in vitro stability of compound A after dilution in various mediums could be followed up by a consecutive evaluation of the in vivo distribution in the rat. After dilution in neutral normal saline compound A is metastable and after a short half-life it is transformed into the other 99mTc-Sn-pyrophosphate compound A is metastable and after a short half-life in bone but in the kidneys. After dilution in normal saline of low pH and in buffering solutions the stability of compound A is increased. In human plasma compound A is relatively stable but not in plasma water. When compound B is formed in a buffering solution, uptake in the kidneys and excretion in urine is lowered and blood concentration increased.It is assumed that the association of protons to compound A will increase its stability at low concentrations while that to compound B will lead to a strong protein bond in plasma. It is concluded that compound A will not be stable in vivo because of a lack of stability in the extravascular space, and that the protein bond in plasma will be a measure of its in vivo stability.

99mTc-Sn-Pyrophosphate Complexes, an Investigation of its Possible Structures

1974 ◽  
Vol 13 (03) ◽  
pp. 252-257 ◽  
Author(s):  
K. Rörvik - Schümichen ◽  
G. Hoffmann ◽  
C. Schümichen
Keyword(s):  
In Vivo Distribution ◽  
Low Concentrations ◽  
Distribution Studies

SummaryAt least two different 99mTc-Sn-pyrophosphate complexes are formed, as it is shown by comparative in vivo distribution studies: A 2 : 2 Sn : pyrophosphate complex is predominant at higher concentrations. Only this complex shows bone seeking properties. A 2 : 1 Sn : pyrophosphate complex exists only at low concentrations. This complex shows no deposition in bone but in the kidneys. Which complex is predominant depends on the pyrophosphate concentration in the equilibrium. Both complexes are rapidly excreted by the kidneys.

scholarly journals Poor Mitochondrial Health and Systemic Inflammation? Testing of a Classical Hypothesis

2020 ◽  
Vol 4 (Supplement_1) ◽  
pp. 126-127
Author(s):  
Marta Zampino ◽  
Luigi Ferrucci ◽  
Richard Spencer ◽  
Kenneth Fishbein ◽  
Eleanor Simonsick ◽  
...  
Keyword(s):  
Mitochondrial Function ◽  
Fat Body ◽  
Reference Group ◽  
Linear Regression Analysis ◽  
Oxidative Capacity ◽  
Conclusive Evidence ◽  
Resonance Spectroscopy ◽  
Low Grade ◽  
Severe Impairment

Abstract Chronic low-grade inflammation often occurs with aging and has been associated with negative health outcomes. Despite extensive research on the origins of “inflammaging”, the causative mechanisms remain unclear. However, a connection between poor mitochondrial health and chronic inflammation has been hypothesized, with decreasing mitochondrial function occurring with age and precipitating an increase in reactive oxygen species and other pro-inflammatory macromolecules such as mitochondrial DNA. We tested this hypothesis on a population of 619 subjects from the Baltimore Longitudinal Study of Aging, measuring muscle mitochondrial oxidative capacity in vivo by phosphorus magnetic resonance spectroscopy (P-MRS), and plasma interleukin (IL)-6, the most widely used biomarker of inflammaging. The P-MRS-derived post-exercise phosphocreatine recovery time constant tau-PCr, a measure of oxidative capacity, was expressed as a categorical variable through assignment to quintiles. Participants in the first quintile of tau-PCr (best mitochondrial function) were taken as reference and compared to the others using linear regression analysis adjusted for sex, age, lean and fat body mass, and physical activity. Those participants with the lowest oxidative capacity had significantly higher log(IL-6) levels as compared to the reference group. However, data from the other quintiles was not significantly different from the reference values. In conclusion, severe impairment of oxidative capacity is associated with increased inflammation. This study design does not provide conclusive evidence of whether increased inflammation and impaired bioenergetic recovery are both caused by underlying poor health status, or whether mitochondrial deficits lead directly to the observed inflammation; we anticipate addressing this important question with longitudinal studies.

scholarly journals Fighting Bisphenol A-Induced Male Infertility: The Power of Antioxidants

Antioxidants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 289
Author(s):  
Joana Santiago ◽  
Joana V. Silva ◽  
Manuel A. S. Santos ◽  
Margarida Fardilha
Keyword(s):  
Male Infertility ◽  
Bisphenol A ◽  
Sperm Concentration ◽  
In Vivo Studies ◽  
Sperm Dna Damage ◽  
Testicular Cells ◽  
Low Concentrations ◽  
Sperm Dna ◽  
Male Reproductive Health

Bisphenol A (BPA), a well-known endocrine disruptor present in epoxy resins and polycarbonate plastics, negatively disturbs the male reproductive system affecting male fertility. In vivo studies showed that BPA exposure has deleterious effects on spermatogenesis by disturbing the hypothalamic–pituitary–gonadal axis and inducing oxidative stress in testis. This compound seems to disrupt hormone signalling even at low concentrations, modifying the levels of inhibin B, oestradiol, and testosterone. The adverse effects on seminal parameters are mainly supported by studies based on urinary BPA concentration, showing a negative association between BPA levels and sperm concentration, motility, and sperm DNA damage. Recent studies explored potential approaches to treat or prevent BPA-induced testicular toxicity and male infertility. Since the effect of BPA on testicular cells and spermatozoa is associated with an increased production of reactive oxygen species, most of the pharmacological approaches are based on the use of natural or synthetic antioxidants. In this review, we briefly describe the effects of BPA on male reproductive health and discuss the use of antioxidants to prevent or revert the BPA-induced toxicity and infertility in men.

scholarly journals In Bacillus subtilis, the Sirtuin Protein Deacetylase, Encoded by the srtN Gene (Formerly yhdZ), and Functions Encoded by the acuABC Genes Control the Activity of Acetyl Coenzyme A Synthetase

2009 ◽  
Vol 191 (6) ◽  
pp. 1749-1755 ◽  
Author(s):  
Jeffrey G. Gardner ◽  
Jorge C. Escalante-Semerena
Keyword(s):  
Bacillus Subtilis ◽  
Coenzyme A ◽  
Acetyl Coenzyme A ◽  
Acetyl Coenzyme ◽  
New Information ◽  
Low Concentrations ◽  
Dependent Protein ◽  
Protein Deacetylase

ABSTRACT This report provides in vivo evidence for the posttranslational control of the acetyl coenzyme A (Ac-CoA) synthetase (AcsA) enzyme of Bacillus subtilis by the acuA and acuC gene products. In addition, both in vivo and in vitro data presented support the conclusion that the yhdZ gene of B. subtilis encodes a NAD+-dependent protein deacetylase homologous to the yeast Sir2 protein (also known as sirtuin). On the basis of this new information, a change in gene nomenclature, from yhdZ to srtN (for sirtuin), is proposed to reflect the activity associated with the YdhZ protein. In vivo control of B. subtilis AcsA function required the combined activities of AcuC and SrtN. Inactivation of acuC or srtN resulted in slower growth and cell yield under low-acetate conditions than those of the wild-type strain, and the acuC srtN strain grew under low-acetate conditions as poorly as the acsA strain. Our interpretation of the latter result was that both deacetylases (AcuC and SrtN) are needed to maintain AcsA as active (i.e., deacetylated) so the cell can grow with low concentrations of acetate. Growth of an acuA acuC srtN strain on acetate was improved over that of the acuA + acuC srtN strain, indicating that the AcuA acetyltransferase enzyme modifies (i.e., inactivates) AcsA in vivo, a result consistent with previously reported in vitro evidence that AcsA is a substrate of AcuA.

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