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131.
N. Arfman E. M. Watling W. Clement R. J. van Oosterwijk G. E. de Vries W. Harder M. M. Attwood L. Dijkhuizen 《Archives of microbiology》1989,152(3):280-288
The enzymology of methanol utilization in thermotolerant methylotrophic Bacillus strains was investigated. In all strains an immunologically related NAD-dependent methanol dehydrogenase was involved in the initial oxidation of methanol. In cells of Bacillus sp. C1 grown under methanol-limiting conditions this enzyme constituted a high percentage of total soluble protein. The methanol dehydrogenase from this organism was purified to homogeneity and characterized. In cell-free extracts the enzyme displayed biphasic kinetics towards methanol, with apparent K
m values of 3.8 and 166 mM. Carbon assimilation was by way of the fructose-1,6-bisphosphate aldolase cleavage and transketolase/transaldolase rearrangement variant of the RuMP cycle of formaldehyde fixation. The key enzymes of the RuMP cycle, hexulose-6-phosphate synthase (HPS) and hexulose-6-phosphate isomerase (HPI), were present at very high levels of activity. Failure of whole cells to oxidize formate, and the absence of formaldehyde-and formate dehydrogenases indicated the operation of a non-linear oxidation sequence for formaldehyde via HPS. A comparison of the levels of methanol dehydrogenase and HPS in cells of Bacillus sp. C1 grown on methanol and glucose suggested that the synthesis of these enzymes is not under coordinate control.Abbreviations RuMP
ribulose monophosphate
- HPS
hexulose-6-phosphate synthase
- HPI
hexulose-6-phosphate isomerase
- MDH
methanol dehydrogenase
- ADH
acohol dehydrogenase
- PQQ
pyrroloquinoline, quinone
- DTT
dithiothreitol
- NBT
nitrobluetetrazolium
- PMS
phenazine methosulphate
- DCPIP
dichlorophenol indophenol 相似文献
132.
Methanogenium organophilum, a non-autotrophic methanogen able to use primary and secondary alcohols as hydrogen donors, was grown on ethanol. Per mol of methane formed, 2 mol of ethanol were oxidized to acetate. In crude extract, an NADP+-dependent alcohol dehydrogenase (ADH) with a pH optimum of about 10.0 catalyzed a rapid (5 mol/min·mg protein; 22°C) oxidation of ethanol to acetaldehyde; after prolonged incubation also acetate was detectable. With NAD+ only 2% of the activity was observed. F420 was not reduced. The crude extract also contained F420: NADP+ oxidoreductase (0.45 mol/min·mg protein) that was not active at the pH optimum of ADH. With added acetaldehyde no net reduction of various electron acceptors was measured. However, the acetaldehyde was dismutated to ethanol and acetate by the crude extract. The dismutation was stimulated by NADP+. These findings suggested that not only the dehydrogenation of alcohol but also of aldehyde to acid was coupled to NADP+ reduction. If the reaction was started with acetaldehyde, formed NADPH probably reduced excess aldehyde immediately to ethanol and in this way gave rise to the observed dismutation. Acetate thiokinase activity (0.11 mol/min·mg) but no acetate kinase or phosphotransacetylase activity was observed. It is concluded that during growth on ethanol further oxidation of acetaldehyde does not occur via acetylCoA and acetyl phosphate and hence is not associated with substrate level phosphorylation. The possibility exists that oxidation of both ethanol and acetaldehyde is catalyzed by ADH. Isolation of a Methanobacterium-like strain with ethanol showed that the ability to use primary alcohols also occurs in genera other than Methanogenium.Non-standard abbreviations ADH
alcohol dehydrogenase
- Ap5ALi3
P1,P5-Di(adenosine-5-)pentaphosphate
- DTE
dithioerythritol (2,3-dihydroxy-1,4-dithiolbutane)
- F420
N-(N-l-lactyl--l-glutamyl)-l-glutamic acid phosphodiester of 7,8-dimethyl-8-hydroxy-5-deazariboflavin-5-phosphate
-
Mg.
Methanogenium
- OD578
optical density at 578 nm
- PIPES
1,4-piperazine-diethanesulfonic acid
- TRICINE
N-(2-hydroxy-1,1-bis[hydroxymethyl]methyl)-glycine
- Tris
2-amino-2-hydroxy-methylpropane-1,3-diol
- U
unit (mol substrate/min) 相似文献
133.
Archaeoglobus fulgidus is an extremely thermophilic archaebacterium that can grow at the expense of lactate oxidation with sulfate to CO2 and H2S. The organism contains coenzyme F420, tetrahydromethanopterin, and methanofuran which are coenzymes previously thought to be unique for methanogenic bacteria. We report here that the bacterium contains methylenetetrahydromethanopterin: F420 oxidoreductase (20 U/mg), methenyltetrahydromethanopterin cyclohydrolase (0.9 U/mg), formyltetrahydromethanopterin: methanofuran formyltransferase (4.4 U/mg), and formylmethanofuran: benzyl viologen oxidoreductase (35 mU/mg). Besides these enzymes carbon monoxide: methyl viologen oxidoreductase (5 U/mg), pyruvate: methyl viologen oxidoreductase (0.7 U/mg), and membranebound lactate: dimethylnaphthoquinone oxidoreductase (0.1 U/mg) were found. 2-Oxoglutarate dehydrogenase, which is a key enzyme of the citric acid cycle, was not detectable. From the enzyme outfit it is concluded that in A. fulgidus lactate is oxidized to CO2 via a modified acetyl-CoA/carbon monoxide dehydrogenase pathway involving C1-intermediates otherwise only used by methanogenic bacteria.Non-standard abbreviations APS
adenosine 5-phosphosulfate
- BV
benzyl viologen
- DCPIP
2,6-dichlorophenolindophenol
- DMN
2,3-dimethyl-1,4-naphthoquinone
- DTT
DL-1,4-dithiothreitol
- H4F
tetrahydrofolate
- H4MPT
tetrahydromethanopterin
- CH2
H4MPT, methylene-H4MPT
- CH
H4MPT, methenyl-H4MPT
- Mes
morpholinoethane sulfonic acid
- MFR
methanofuran
- Mops
morpholinopropane sulfonic acid
- MV
methyl viologen
- Tricine
N-tris(hydroxymethyl)-methylglycine
- U
mol product formed per min 相似文献
134.
Teresa Urbanik-Sypniewska Ulrich Seydel Michaela Greck Jürgen Weckesser Hubert Mayer 《Archives of microbiology》1989,152(6):527-532
The structure of the lipopolysaccharide from Rhizobium meliloti 10406, a derivative of the wild-type strain MVII-1, was examined. The compositional analysis of its polysaccharide moiety demonstrated lack of heptose(s), but high contents in glucose, galacturonic acid and 2-keto-3-deoxy-octonate (dOclA) as characteristic features. The lipid A moiety consisted of a -1,6 linked glucosamine disaccharide carrying ester (at C-4) and glycosidically (at C-1) linked phosphate residues, both present exclusively as monoester phosphates but not as phosphodiesters. Ester- and amidelinked 3-hydroxy fatty acids were mostly present as non-3-O-acylated residues. Laser desorption mass spectrometry (LD-MS) revealed heterogeneity in the fatty acid substitution, as was also indicated by the non-stoichiometric ratios obtained by quantitative fatty acid analysis. The predominating lipid A structure contained at the reducing glucosamine residue ester-linked 3-hydroxy-tetradecanoic acid (3-OH-14:0) and amide-linked 3-OH-18:0, or 3-OH-18:1, respectively. The distal (non-reducing) glucosamine carried ester-bound the recently discovered 27-hydroxyoctacosanoic acid and 3-OH-14:0 and, as amide-linked fatty acid, mostly 3-hydroxy-stearic acid (3-OH-18:0).The isolated lipopolysaccharide exhibited a high extent of lethal toxicity in galactosamine-treated mice, comparable to that of enterobacterial lipopolysaccharide. The structural relationship of LPS and lipid A of Rhizobium meliloti to other rhizobial lipopolysaccharides and lipid A's with respect to questions of taxonomy and of phylogenetic relationships will be discussed.Abbreviations LPS
lipopolysaccharide
- dOclA
3-deoxy-D-mannooctulosonic acid (KDO)
- GalA
galacturonic acid
- DOC
sodium deoxycholate
- PAGE
polyacrylamide gel electrophoresis
- LD-MS
laser desorption-mass spectrometry 相似文献
135.
P.-J. Charmillot 《Entomologia Experimentalis et Applicata》1989,51(1):59-69
Résumé La métamorphose des insectes est régie par un équilibre hormonal complexe dans lequel l'hormone juvénile (HJ) joue un rôle important. Au dernier stade larvaire, la teneur en HJ est particulièrement faible dans le corps de l'insecte. Si un régulateur de croissance d'insectes (RCI)-un mimétique de l'HJ-est appliqué à ce moment-là, la mue nymphale est pertubée provoquant des déformations morphogénétiques caractéristiques. La teneur en HJ est également très faible dans les ufs fraîchement pondus. Les traitements aux RCI peuvent par conséquent perturber le développement embryonnaire de certaines espèces et produire ainsi un effet ovicide. Depuis quelques années deux RCI-le fenoxycarb et le CGA 45 128-ont été testés pour leur activité morphogénétique sur le dernier stade larvaire de quelques ravageurs tels qu'Adoxophyes orana F.v.R., ainsi que pour leur activité ovicide sur les ufs frais de Cydia pomonella L. et Grapholita funebrana Tr. Après quelques années d'expérimentation et de commercialisation des RCI dans les vergers européens, il s'avère que l'utilisation de ces produits peu toxiques, sélectifs et peu nocifs pour la faune utile, constitue une amélioration considérable pour l'aménagement de la lutte intégrée.
Insect growth regulators (IGR), mimics of juvenile hormone, as morphological and ovicidal means of control against orchard tortricids
Summary Metamorphosis is regulated by a complex hormonal balance in which juvenile hormone (JH) plays an important part. At the last larval instar the content of JH is particularly low in the insect body. If an insect growth regulator (IGR) — a mimic of JH-is applied at this time, the pupal moult may be disturbed with the characteristic morphogenetical deformations. The JH content is also very low in freshly laid eggs. Therefore IGR treatments may disturb the embryonic development of some species and produce an ovicidal activity. During a few years two IGR-fenoxycarb and CGA 45128-were evaluated for their morphogenetical effect on the last larval instar of Adoxophyes orana F.v.R. and their ovicidal effect on freshly laid eggs of Cydia pomonella L. and Grapholita funebrana Tr. After a few years of experimentation with both compounds and of commercialisation of fenoxycarb in European orchards, IGR confirmed to present a considerable improvement in integrated pest management due to selectivity, and low mammal toxicity.相似文献
136.
137.
Receptors for α2-macroglobulin-proteinase complexes have been characterized in rat and human liver membranes. The affinity for binding of 125I-labelled α2-macroglobulin · trypsin to rat liver membranes was markedly pH-dependent in the physiological range with maximum binding at pH 7.8–9.0. The half-time for association was about 5 min at 37°C in contrast to about 5 h at 4°C. The half-saturation constant was about 100 pM at 4°C and 1 nM at 37°C (pH 7.8). The binding capacity was approx. 300 pmol per g protein for rat liver membranes and about 100 pmol per g for human membranes. Radiation inactivation studies showed a target size of 466 ± 71 kDa (S.D., n = 7) for α2-macroglobulin · trypsin binding activity. Affinity cross-linking to rat and human membranes of 125I-labelled rat α1-inhibitor-3 · chymotrypsin, a 210 kDa analogue which binds to the α2-macroglobulin receptors in hepatocytes (Gliemann, J. and Sottrup-Jensen, L. (1987) FEBS Lett. 221, 55–60), followed by SDS-polyacrylamide gel electrophoresis, revealed radioactivity in a band not distinguishable from that of cross-linked α2-macroglobulin (720 kDa). This radioactivity was absent when membranes with bound 125I-α1-inhibitor-3 complex were treated with EDTA before cross-linking and when incubation and cross-linking were carried out in the presence of a saturating concentration of unlabelled complex. The saturable binding activity was maintained when membranes were solubilized in the detergent 3-[(3-cholamidopropyl)dimethylammonio]profane sulfonate (CHAPS) and the size of the receptor as estimated by cross-linking experiments was shown to be similar to that determined in the membranes. It is concluded that liver membranes contain high concentrations of an approx. 400–500 kDa α2-macroglobulin receptor soluble in CHAPS. The soluble preparation should provide a suitable material for purification and further characterization of the receptor. 相似文献
138.
Pedro J. N. Silva Richard K. Koehn Walter J. Diehl III Robin P. Ertl Elaine B. Winshell Mauro Santos 《Biochemical genetics》1989,27(7-8):451-467
Four samples of the musselMytilus edulis were taken between 1984 and 1987 from Stony Brook, New York, and used to study the glucose-6-phosphate isomerase (GPI) polymorphism
in this species.In vitro specific activity andin vivo flux measured in the same animals were found to be significantly correlated. A significant effect of GPI genotype on flux
was observed in one of the samples; overall, significant evidence of effect of genotype on enzyme activity was also obtained.
GPI activities of common genotypes tend to deviate less from the population mean than those of rare (frequency less than 5%)
genotypes. This suggests the possibility that rare GPI genotypes are rare as a consequence of having biochemical properties
that deviate from an optimum level and, therefore, having a lower fitness. In support of this hypothesis, we found in one
of our samples that shell length is a concave function of GPI activity with an intermediate optimum activity level.
The financial support provided to P.J.N.S. by the Luso-American Educational Commission (Fulbright Program), the Instituto
Nacional de Investigacao Científica (Portugal), and the Faculdade de Ciências da Universidade de Lisboa during several stages
of this research is gratefully acknowledged. Financial support from the Ministerio de Educatión y Ciencia (Spain) in the form
of a postdoctoral Fulbright/MEC fellowship to M.S. is also gratefully acknowledged. Research was supported by National Science
Foundation Grant BSR-8415060 to R.K.K. This is contribution No. 736 from the Program in Ecology and Evolution, State University
of New York at Stony Brook.
On leave from Departamento de Biologia Vegetal, Faculdade de Ciências, Universidade de Lisboa, Campo Grande C2, Lisboa, Portugal. 相似文献
139.
140.
Izumi Tabata Yoriko Atomi Hiroaki Kanehisa Mitsumasa Miyashita 《European journal of applied physiology and occupational physiology》1990,60(4):254-258
The purpose of this study was to determine the effects of high-intensity endurance training on isokinetic muscle power. Six male students majoring in physical-education participated in high intensity endurance training on a cycle ergometer at 90% of maximal oxygen uptake (VO2max) for 7 weeks. The duration of the daily exercise session was set so that the energy expenditure equalled 42 kJ.kg-1 of lean body mass. Peak knee extension power was measured at six different speeds (30 degrees, 60 degrees, 120 degrees, 180 degrees, 240 degrees, and 300 degrees.s-1) with an isokinetic dynamometer. After training, VO2max increased significantly from mean values of 51.2 ml.kg-1.min-1, SD 6.5 to 56.3 ml.kg-1.min-1, SD 5.3 (P less than 0.05). Isokinetic peak power at the lower test speeds (30 degrees, 60 degrees and 120 degrees.s-1) increased significantly (P less than 0.05). However, no significant differences in muscle peak power were found at the faster velocities of 180 degrees, 240 degrees, and 300 degrees.s-1. The percentage improvement was dependent on the initial muscle peak power of each subject and the training stimulus (intensity of cycle ergometer exercise). 相似文献