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1.
The specific Bchl a and c content of the vitamin B12-dependent Chlorobium limicola strain 1230 decreased strongly under vitamin B12 limitation. In comparison to a regularly grown culture (20 g vitamin B12/l) the specific Bchl c content of a B12-limited culture was reduced to 20% and the specific Bchl a content to 42%. By ultrathin sections it could be clearly demonstrated that B12-deficient cells contained no chlorosomes. After the addition of vitamin B12 to a deficient culture, chlorosomes were formed and the Bchl a and c content increased again to the level of regularly grown cells. The brown-colored Chlorobium phaeobacteroides strain 2430 (type strain) and the extremely low-light-adapted strain MN1 were compared with respect to the influence of light on the formation of chlorosomes and the Bchl e and carotenoid content. By ultrathin sections it could be demonstrated that strain MN1 produced two-fold larger chlorosomes. Chlorosome dimensions of strain MN1 decreased with increasing light intensities. The number of chlorosomes per cell in both strains did not change with different light intensities. Strain MN1 formed twice as much Bchl e as the type strain when grown at 30 or below 1 mol · m-2 · s-1. Under comparable light conditions strain MN1 formed 14–57% more carotenoids than the type strain. Low light intensities aaused the carotenoid content to increase by 25% in strain 2430 in comparison to high light intensity.  相似文献   

2.
Mercury resistance of Clostridium cochlearium T-2P was found to be controlled by a different mechanism from those reported so far since no mercury-reducing activity was detected in this strain. The H2S generating ability as well as the demethylating activity of this bacterium was eliminated by the treatment with acridine dye and recovered by the conjugation of the cured strain with the parent strain. In addition, the strain which lost their abilities to generate H2S and to decompose methylmercury, showed higher sensitivity to mercurials than the parent strain. From these results, the genes conferring both the activities seemed to reside on the plasmid and the mechanism of mercury resistance was probably based on a detoxification mechanism involving methylmercury decomposition and inactivation of the inorganic mercury with H2S.  相似文献   

3.
The vitamin B12 requirement of Rhodocyclus purpureus 6770, Rhodospirillum tenue 1/67, and Rhodopseudomonas palustris G 53/2 was determined. A wide variety of biogenetic precursors of the vitamin including cobinamide, cobyric acid, cobinic acid and several partially amidated cobyrinic acids showed growth-promoting activity in all three strains. In R. purpureus vitamin B12 could even be substituted by cobyrinic acid which is the first cobalt-containing precursor of vitamin B12 so far established. Neither methionine, deoxynucleosides, dimethylbenzimidazole nor increased amounts of cobalt could replace vitamin B12 as growth factor.Cupribalamin, which is a strong antimetabolite of vitamin B12 in Escherichia coli 113-3 and Lactobacillus leichmannii ATCC 7830, exhibited only a weak antagonistic effect on growth of R. purpureus and R. tenue. Growth of R. palustris was not inhibited by cupribalamin. The cells of all three strains were shown to contain metal-free corrinoids in addition to cobalt-containing corrinoids. The principal products were identified as 5-deoxyadenosylcobalamin and hydrogenobalamin, the metal free analogue of vitamin B12. The latter does not originate from the vitamin by removal of cobalt but is de novo biosynthesized as could be demonstrated in the case of R. purpureus by a labelling experiment with [13C] methyl-l-methionine.  相似文献   

4.
Radioactivity from [1-14C]riboflavin was incorporated into the 5,6-dimethylbenzimidazole moiety of Vitamin B12 in the aerobes Bacillus megaterium, Nocardia rugosa and Streptomyces sp. as well as in the aerotolerant anaerobe Propionibacterium freudenreichii, but not in the anaerobe Eubacterium limosum.As recently published for E. limosum, also in the anaerobe Clostridium barkeri radioactivity from [1-14C]glycine and [2-14C]glycine was found in the 5,6-dimethylbenzimidazole moiety, but not in the corrin moiety. The addition of l-[methyl-14C]methionine to C. barkeri led to the labeling of the corrin moiety and the 5,6-dimethylbenzimidazole moiety, showing that the seven extra methyl groups in the corrin ring as well as the two methyl groups of the base part originate from this precursor.In Clostridium thermoaceticum, forming the vitamin B12 analog 5-methoxybenzimidazolylcobamide, [1-14C]glycine and [2-14C]glycine were also incorporated into the 5-methoxybenzimidazole moiety, but not into the corrin ring.In E. limosum l-[U-14C]glutamate led to the labeling of the corrin ring of vitamin B12, but not of its base moiety.There results together with data from the literature indicate that a common biosynthetic pathway might exist for the corrinoid biosynthesis in aerobic microorganisms, and in those aerotolerant anaerobes like the Propionibacteria, which form the 5,6-dimethylbenzimidazole moiety of vitamin B12 only under aerobic conditions. They also show that this pathway differs from the pathway found in anaerobic bacteria.  相似文献   

5.
We examined 25 strains of Fusarium moniliforme from eight states known to be associated with equine leukoencephalomalacia, a disease caused by the mycotoxin fumonisin B1. We determined the mating population, mating type, and vegetative compatibility group to which each of these strains belonged. All 25 strains were in the A mating population; 12 were A+ and 13 were A. Seventeen of the 25 strains were female fertile; these strains also averaged higher levels of fumonisin B1 production than did the strains that were female sterile. Nitrate non-utilizing (nit) mutants were generated in all 25 strains and each strain was assigned to a unique vegetative compatibility group based on the inability of the derived nit mutants to form a prototrophic heterokaryon with complementary nit mutants derived from any of the other strains examined. From these data, we concluded that the production of fumonisin B1 is a general characteristic of strains from the A mating population of Gibberella fujikuroi associated with equine leukoencephalomalacia, since all 25 of the isolates that we examined were genetically distinct individuals.  相似文献   

6.
A biosorbent prepared by alkaline extraction of Aspergillus niger biomass was evaluated for its potential to remove mercury species – inorganic (Hg2+) and methyl mercury (CH3Hg+) – from aqueous solutions. Batch experiments were carried out to determine the pH and time profile of sorption for both species in the pH range 2–7. The Hg2+ exhibited more rapid sorption and higher capacity than the CH3Hg+. Further, removal of both mercury species from spiked ground water samples was efficient and not influenced by other ions. Sorption studies with esterified biosorbent indicated loss of binding of both mercury species (>80%), which was regained when the ester groups were removed by alkaline hydrolysis, suggesting the involvement of carboxyl groups in binding. Further, no interconversion of sorbed species occurred on the biomass. The biosorbent was reusable up to six cycles without serious loss of binding capacity. Our results suggest that the biosorbent from Aspergillus niger can be used for removal of mercury and methyl mercury ions from polluted aqueous effluents.  相似文献   

7.
Thirteen species of methanogenic bacteria were analyzed for corrinoids. Pseudo vitamin B12 (Co-[-(7-adenyl)]-cobamide) was the predominant cobamide of methanococcales and Methanoplanus. All other methanogens contained factor III (Co-[-(5-hydroxybenzimidazolyl)]-cobamide). Vitamin B12 (Co-[-(5,6-dimethylbenzimidazolyl)]-cobamide) was not detected in any of these archaebacteria. Their cobamide content was 100 to 1400 nmol per gram cell dry weight, indicating that abundant cobamides are essential for methanogens.  相似文献   

8.
Summary Transport of vitamin B12 across the cytoplamic membrane ofEscherichia coli requires the products ofbtuC andbtuD, two genes in thebtuCED operon. The role ofbtuE, the central gene of this operon, was examined. Deletions withinbtuE were constructed by removal of internal restriction fragments and were crossed onto the chromosome by allelic replacement. In-frame deletions that removed 20% or 82% of thebtuE coding region did not affect expression of the distalbtuD gene. These nonpolar deletions had little effect on vitamin B12 binding (whole cells or periplasmic fraction) and transport. They did not affect the utilization of vitamin B12 or other cobalamins for methionine biosynthesis, even in strains with decreased outer membrane transport of vitamin B12. ThebtuE mutations did not impair adenosyl-cobalamin dependent catabolism of ethanolamine or repression ofbtuB expression. Thus, despite its genetic location in the transport operon, thebtuE product plays no essential role in vitamin B12 transport.  相似文献   

9.
Growth yields and rates of 3 hyphomicrobia were improved by varying components of or adding compounds to medium 337. Methanol (0.5% v/v), and similarly methylamine·HCl (3.38g/l), were optimal among 22 C-sources tested; increasing the methylamine·HCl concentration to 5.07g/l gave higher Hyphomicrobium B-522 yields but also prolonged lag periods. Ten C-sources (organic acids, alcohols) stimulated growth slightly but significantly, even in subcultures. Sugar compounds were not utilized. Strains B-522 and ZV-580 were stimulated by l-lysine and gluconate, while NQ-521 gr was stimulated by aspartate.N-Sources tested were inorganic (3), organic (3), or complex (3). (NH4)2SO4 (0.5g/l) was optimal for strains ZV-580 and NQ-521 gr, but Hyphomicrobium B-522 grew best with urea-N. With NH 4 + , strain B-522 grew as homogeneous suspension, all other N-sources caused clumping and pellicle formation. Inorganic requirements (PO 4 3- , Mg, Ca, Fe, Mn, Mo) of strains B-522 and ZV-580 were optimized. Addition of Ni, Co, or Zn had no effect; metals 44 or Cu, resulted in growth inhibition.Vitamin B12 stimulated Hyphomicrobium B-522; 2.5g/l B12 decreased the doubling time from 9.3–10.8h to 5.4–5.8h. All combined single improvements resulted in a protein increase of 557% (B-522), 141% (NQ-521 gr), or 109% (ZV-580), respectively.  相似文献   

10.
We have shown that mercury affects energy transfer in Spirulina platensis. It inhibits energy transfer from phycocyanin to chlorophyll a by specifically bleaching the -84 chromophore of the chromo protein, phycocyanin (PC), in the cyanobacterium. This effect is observed during short-term exposure of cells to Hg2+ ions. Upon long-term (12 h) exposure, mercury at low concentrations (1–2.5 m) causes the gradual degradation of the polypeptide (22 kDa) of the PC of phycobilisomes in this cyanobacterium. The effect of mercury on this polypeptide is significant compared with the other phycobiliproteins.  相似文献   

11.
Summary Using an Escherichia coli lac deletion strain lysogenized with lambda phage carrying a metF-lacZ gene fusion (Flac), in which -galactosidase levels are dependent on metF gene expression, cis-acting mutations were isolated that affect regulation of the Salmonella typhimurium metF gene. The mutations were located in a region previously defined as the metF operator by its similarity to the E. coli metF operator sequence. Regulation of the metF gene was examined by measuring -galactosidase levels in E. coli strains lysogenized with the wild-type Flac phage and mutant Flac phage. The results suggest that the mutations disrupt the methionine control system mediated by the metJ gene product, but not the vitamin B12 control system mediated by the metH gene product. The results also demonstrate that negative control of the metF gene by the metH gene product and vitamin B12 is dependent on a functional metJ gene product.Abbreviations Ap ampicillin - dNTP deoxyribonucleoside triphosphates - GM glucose minimal - Km kanamycin - L-agar Luria agar - LM lactose minimal - SAM s-adenosyl-L-methionine - TPEG phenylethyl -D-thiogalactoside - X-gal 5-bromo-4-chloro-3-indolyl -D-galactopyranoside - [] designates plasmid-carrier state - :: novel joint  相似文献   

12.
Danilova  I. V.  Doronina  N. V.  Trotsenko  Yu. A.  Netrusov  A. I.  Ryzhkova  E. P. 《Microbiology》2004,73(2):134-138
The effect of vitamin B12 (cobalamin) on DNA biosynthesis in Methylobacterium dichloromethanicum was studied. When cultivated in media with methanol or dichloromethane, the bacterium produced approximately 10 g corrinoids per gram dry biomass, compared to about 7 g/g when cultivated on ethanol or succinate. Exogenous adenosylcobalamin (AdoCbl) stimulated DNA biosynthesis in M. dichloromethanicum cells grown under poor aeration, the effect being mediated by AdoCbl-dependent ribonucleotide reductase. In vitro studies showed that M. dichloromethanicumalso has AdoCbl-independent ribonucleotide reductase. Under good aeration, exogenous AdoCbl had no effect on DNA biosynthesis, while hydroxyurea suppressed it. These data suggest that AdoCbl-independent ribonucleotide reductase, which is likely to be activated by oxygen, plays an important part in DNA biosynthesis when M. dichloromethanicum is cultured with good aeration, whereas AdoCbl-dependent ribonucleotide reductase is active under conditions of poor aeration.  相似文献   

13.
Competitive immunoassay for analysis of vitamin B(12)   总被引:1,自引:0,他引:1  
In the current work, direct competitive enzyme-linked immunosorbent assay (ELISA) was developed for derivatized vitamin B12 by generating chicken egg yolk immunoglobulins (IgY) against derivatized vitamin B12 and purified using affinity chromatography. Checkerboard assay was performed with vitamin B12 antibody and vitamin B12–alkaline phosphatase conjugate followed by its conjugate characterization using ultraviolet (UV) spectroscopy and high-performance liquid chromatography (HPLC). The limit of detection was 10 ng/ml with a linear working range of 10 to 10,000 ng/ml. The affinity constant (Ka) of the vitamin B12 antibody was found to be 4.23 × 108 L/mol. Cross-reactivity with other water-soluble vitamins was found to be less than 0.01% except for analogs of vitamin B12 that showed 12% to 35%. The intra- and interassay coefficients of variation were found to be in the ranges from 0.0005% to 1.2% and 0.009% to 1.03%, respectively. The assay was validated with the HPLC method in terms of sensitivity, specificity, precision, and recovery of vitamin B12 with spiked multivitamin injections, tablets, capsules, and chocolates. The HPLC method had a detection limit of 500 ng/ml with a linear working range of 1000 to 10,000 ng/ml. After extraction of vitamin B12 using Amberlite XAD, the developed ELISA method correlated well with the established HPLC method with a correlation coefficient of 0.90.  相似文献   

14.
In this article, the three-dimensional structures of photosynthetic reaction centers (RCs) are presented mainly on the basis of the X-ray crystal structures of the RCs from the purple bacteria Rhodopseudomonas (Rp.) viridis and Rhodobacter (Rb.) sphaeroides. In contrast to earlier comparisons and on the basis of the best-defined Rb. sphaeroides structure, a number of the reported differences between the structures cannot be confirmed. However, there are small conformational differences which might provide a basis for the explanation of observed spectral and functional discrepancies between the two species.A particular focus in this review is on the binding site of the secondary quinone (QB), where electron transfer is coupled to the uptake of protons from the cytoplasm. For the discussion of the QB site, a number of newlydetermined coordinate sets of Rp. viridis RCs modified at the QB site have been included. In addition, chains of ordered water molecules are found leading from the cytoplasm to the QB site in the best-defined structures of both Rp. viridis and Rb. sphaeroides RCs.Abbreviations BA accessory bacteriochlorophyll in the active branch - BB accessory bacteriochlorophyll in the inactive branch - D primary electron donor (special pair) - DL special pair bacteriochorophyll bound by the L subunit - DM special pair bacteriochorophyll bound by the M subunit - QA primary electron acceptor quinone - QB secondary electron acceptor quinone - RC reaction center - Rb. Rhodobacter - Rp. Rhodopseudomonas - A bacteriopheophytin in the active branch - B bacteriopheophytin in the inactive branch  相似文献   

15.
W. Reisser 《Protoplasma》1981,105(3-4):273-284
Summary The greenStentor polymorphus harbours unicellular coccoid chlorophycean algae. They are located in food vacuoles, where they show various states of digestion, as well as in individual so-called perialgal vacuoles. According to their characteristic morphological properties the algae belong to the genusChlorella. They can be isolated from the ciliate and cultivated in mass cultures in a sterile defined inorganic medium supplemented with vitamins B1 and B12. The algae have no secondary carotenoids and excrete maltose by a pH-dependent mechanism. They thus show a conspicuous physiological similarity to the symbiotic chlorellae ofParamecium bursaria andHydra viridis, which also excrete maltose.A comparison of the properties of the chlorellae isolated fromStentor polymorphus and of the intactStentor polymorphus-Chlorella unit with the characteristic features of symbiotic chlorellae and with endosymbiotic systems containingChlorella sp. in general, lead to the conclusion that the greenStentor polymorphus is also a true endosymbiotic system.  相似文献   

16.
U. Heber  S. Neimanis  K. -J. Dietz 《Planta》1988,173(2):267-274
In order to obtain information on fractional control of photosynthesis by individual catalysts, catalytic activities in photosynthetic electron transport and carbon metabolism were modified by the addition of inhibitors, and the effect on photosynthetic flux was measured using chloroplasts of Spinacia oleracea L. In thylakoids with coupled electron transport, light-limited electron flow to ferricyanide was largely controlled by the QB protein of the electron-transport chain. Fractional control by the cytochrome f/b 6 complex was insignificant under these conditions. Control by the cytochrome f/b 6 complex dominated at high energy fluence rates where the contribution to control of the QB protein was very small. Uncoupling shifted control from the cytochrome f/b 6 complex to the QB protein. Control of electron flow was more complex in assimilating chloroplasts than in thylakoids. The contributions of the cytochrome f/b 6 complex and of the QB protein to control were smaller in intact chloroplasts than in thylakoids. Thus, even though the transit time for an electron through the electron-transport chain may be below 5 ms in leaves, oxidation of plastohydroquinone was only partially responsible for limiting photosynthesis under conditions of light and CO2 saturation. The energy fluence rate influenced control coefficients. Fractional control of photosynthesis by the ATP synthetase, the cytochrome f/b 6 complex and by ribulose-1,5-bisphosphate carboxylase increased with increasing fluence rates, whereas the contributions of the QB protein and of enzymes sensitive to SH-blocking agents decreased. The results show that the burdens of control are borne by several components of the photosynthetic apparatus, and that burdens are shifted as conditions for photosynthesis change.Abbreviations Chl chlorophyll - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DNP-INT 2,4-dinitro phenylether of 2-iodo-4-nitrothymol - pCMBS p-chloromercuribenzosulfonate  相似文献   

17.
Previous research has confirmed that cobalt ion and dimethylbenzimidazole (DMBI) are the precursors of vitamin B12 biosynthesis, and porphobilinogen synthase (PBG synthase) is a zinc-requiring enzyme. In this paper, the effects of Zn2+, Co2+ and DMBI on vitamin B12 production by Pseudomonas denitrificans in shake flasks were studied. Present experimental results demonstrated that the addition of the above mentioned three components to the fermentation medium could significantly stimulate the biosynthesis of vitamin B12. The concentrations of zinc sulphate, cobaltous chloride and DMBI in the fermentation medium were further optimized with rotatable orthogonal central composite design and statistical analysis by Data Processing System (DPS) software. As a result, vitamin B12 production was increased from 69.36 ± 0.66 to 78.23 ± 0.92 μg/ml.  相似文献   

18.
Novel vitamin B12 analogues in which the D-ribose moiety of the nucleotide loop was replaced by an oligomethylene group and a trimethylene analogue containing imidazole instead of 5,6-dimethylbenzimidazole as well as cobinamide methyl phosphate were tested for biological activities with Escherichia coli 215, a B12- or methionine-auxotroph, and Lactobacillus leichmannii ATCC 7830 as test organisms. A cyano form of 5,6-dimethylbenzimidazolyl tetramethylene, trimethylene and hexamethylene analogues supported the growth of L. leichmannii in this order. 5.6-Dimethylbenzimidazolyl dimethylene and imidazolyl trimethylene analogues did not show B12 activity and behaved as weak B12 antagonists when added together with cyanocobalamin. An adenosyl form of the biologically active analogues served as coenzymes for ribonucleotide reductase of this bacterium, whereas that of the inactive analogues did not. The latter acted as weak competitive inhibitors against adenosylcobalamin. ON the contrary, all the analogues did not support the growth of E. coli 215 at all by themselves and inhibited the growth when added with a suboptimum level of cyanocobalamin. A methyl form of the analogues also did not support the growth of E. coli 215, although they served as active coenzymes for methionine synthase of the bacterium. Since unlabeled analogues strongly inhibited the uptake of [3H]cyanocobalamin by this bacterium, it seems likely that the analogues exert their anti-B12 activity toward E. coli 215 by blocking the B12-transport systemAbbreviations AdoCbl adenosylcobalamin - MeCbl methylcobalamin - CN-Cbl cyanocobalamin or vitamin B12 - Cbl cobalamin - (CN, aq)Cbi cyanoaquacobinamide - MeCbi methylcobinamide - Cbi cobinamide - (CN, aq)Cbi-PMe cyanoaquacobinamide methyl phosphate - Cbi-PMe cobinamide methyl phosphate - DBI 5,6-dimethylbenzimidazole - DBIyl 5,6-dimethylbenzimidazolyl - FMNH2 fully reduced form of riboflavin 5-phosphate  相似文献   

19.
Electrochemically active composite film containing multiwalled carbon nanotubes (MWCNTs) and vitamin B12 was synthesized on glassy carbon, gold, and indium tin oxide electrodes by the potentiodynamic method. The presence of MWCNTs in the composite film (MWCNT–B12) modified electrode mediates vitamin B12’s redox reaction, whereas vitamin B12’s redox reaction does not occur at bare electrode. The electrochemical impedance spectroscopy studies reveal that MWCNTs present in MWCNT–B12 film enhance electron shuttling between the reactant and electrode surface. The surface morphology of bare electrode, MWCNT film. and MWCNT–B12 composite film was studied using atomic force microscopy, which reveals vitamin B12 incorporated with MWCNTs. The MWCNT–B12 composite film exhibits promising enhanced electrocatalysis toward hydrazine. The electrocatalysis response of hydrazine at MWCNT film and MWCNT–B12 composite film was measured using cyclic voltammetry and amperometric current–time (it) curve techniques. The linear concentration range of hydrazine obtained at MWCNT–B12 composite film using the it curve technique is 2.0 μM–1.95 mM. Similarly, the sensitivity of MWCNT–B12 composite film for hydrazine determination using the it curve technique is 1.32 mA mM−1 cm−2, and the hydrazine’s limit of detection at MWCNT–B12 composite film is 0.7 μM.  相似文献   

20.
The binding of inorganic mercury Hg(II) to phospholipid headgroups has been investigated by phosphorus-31 nuclear magnetic resonance of phosphatidylethanolamine (PE), phosphatidylserine (PS) and phosphatidylcholine (PC) in water micellar and multilamellar phases. HgCl2 triggers the aggregation of phospholipid micelles, leading to a lipid-mercury precipitate that is no longer detectable by high-resolution31P-NMR. The remaining signal area corresponds to micelles in the soluble fraction and is a non-linear function of the initial mercury-to-lipid molar ratio. Kinetics of micelle aggregation are exponential for the first 15 min and show a plateau tendency after 120 min. Apparent Hg(H) affinities for phospholipid headgroups are in the order: PE > PS > PC. The same binding specificity is observed when HgCl2 is added to (1:1) mixtures of different micelles (PE + PC; PS + PC). However, mercury binding to mixed micelles prepared with two lipids (PE/PC or PS/PC) induces the aggregation of both lipids. Hg(II) also leads to a31P-NMR chemical shift anisotropy decrease of PC, PS and mixed (1:1) PE/PC multilamellar vesicles and markedly broadens PS spectra. This indicates that HgCl2 binding forces phospholipid headgroups to reorient and that the concomitant network formation leads to a slowing down of PS membrane collective motions. Formation of a gel-like lamellar phase characterized by a broad NMR linewidth is also observed upon HgCl2 binding to PE samples both in fluid (L) or hexagonal (HII) phases. The PE hexagonal phase is no longer detected in the presence of HgCl2. Mixed PE/PC dispersions remain in the fluid phase upon mercury addition, indicating that no phase separation occurs. Addition of excess NaCl leads to the appearance of the non-reactive species HgCl inf4 sup2– and induces the reversal of all the above effects.Abbreviations A(t) time-dependence of peak area - A40 peak area at t=40 min - 1/ rate of peak area decrease - isotropic chemical shift - isotropic chemical shift change - chemical shift anisotropy - DPPC dipalmitoylphosphatidylcholine - Hg(II) inorganic mercury - NMR nuclear magnetic resonance - pCl –log [Cl] - PC phosphatidylcholine - PE phosphatidylethanolamine - PL phospholipid - PS phosphatidylserine - Ri mercury-to-lipid molar ratio - MLV multilamellar vesicles - SUV small unilamellar vesicles  相似文献   

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