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1.
Activities of glycosidases ( n = 8), esterases ( n = 10), arylamidases ( n = 63), acid phosphatase, alkaline phosphatase and phosphoamidase were tested in 47 Borrelia strains. Forty-four were B. burgdorferi strains; 22 of which were isolated from human specimens (skin 13, cerebrospinal fluid six, and one each from blood, heart muscle and synovia), 13 were isolated from various organs of laboratory animals infected via tick bite or with human isolates, and nine from ticks. The remaining three were the relapsing fever strains B. coriaceae , B. hermsii , and B. turicatae. Strains were of low and high passage but the number of subcultures did not influence the enzyme patterns obtained by utilization of chromogenic substrates of constitutive enzymes. Glycosidase activity was absent in almost all strains tested. Esterase activity was high on molecules of chain length 9 carbons. 2-Naphthylamide derivatives were utilized by strains of human, rodent or tick origin in a range of 66.6 to 83.1%. Almost all strains utilized substrates for acid and alkaline phosphatase and phosphoamidase. Chymotrypsin activity was only found in three and two strains from specimens of human and rodent origin, respectively; and γ-glutamyltransferase activity only in three human skin isolates. No strain tested displayed trypsin activity. Overall, the specific activities of constitutive enzymes of the Borrelia strains tested are widely similar. Thus, the enzyme profiles did not discriminate between human, animal and tick isolates, or between human isolates of Borrelia whether cultivated from cerebrospinal fluid or from skin biopsy of Lyme borreliosis patients. 相似文献
2.
The vitamin requirements of 66 strains of Staphylococcus cohnii from various sources were determined using solidified and liquid chemically defined media. All the strains tested required nicotinic acid and thiamine. Biotin was either growth stimulatory or essential for 34 strains. Pantothenic acid was required by 21 of the 30 strains of known human origin but was not required by any of the 24 strains of murine origin. There was an association among Staph. cohnii strains to grow in the absence of pantothenic acid and biotin and the ability to produce acid from lactose and the possession of phosphatase activity (20. strains). The majority of strains requiring pantothenic acid and biotin were unable to ferment lactose and did not possess phosphatase activity (22 strains). 相似文献
3.
In this study, using the API-ZYM system, we have reported the enzyme profile of 42 soil strains and 2 clinical strains of
Nocardia asteroides isolated locally. Of the 19 enzymes tested, only 7 were demonstrable in over 90% of the soil isolates.
These included alkaline phosphatase, esterase lipase, leucine arylamidase, acid phosphatase, phosphohydrolase, α-glucosidase
and β-glucosidase. In addition, β-galactosidase activity was demonstrated in all the strains by the O-nitrophenyl-β-D-galactopyranoside
(ONPG) test. The enzymes which were not demonstrable in >95% of the strains included valine arylamidase, cystine arylamidase,
trypsin, chymotrypsin, α-galactosidase, β-glucoronidase, N-acetyl-β-glucosaminidase, α-mannosidase and α-fucosidase. With
the exception of valine arylamidase, which was lacking in all but one isolate, the enzyme profiles of the soil isolates were
comparable with the clinical isolates of N. asteroides reported in previous studies. The reasons for this difference in the
two sets of isolates is not clear. The study reinforces the view that specific differences in the enzymatic profiles of Nocardia
species could be used for their rapid identification. However, more extensive studies are needed to establish the reproducibility
of this method. To the best of our knowledge, this is the first study of the enzymatic profile of soil isolates of N. asteroides
originating from a single geographic region.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
Acid phosphatase activity was studied in the left and right ovaries of the chick during embryonic development. The cytochemical study indicated that local enzymatic activity is localized mainly in germ and somatic cells. From the results obtained in the study on the specific activity of this acid hydrolase, it can be inferred that the higher concentration of this enzyme in the right ovary would be determined by a decrease of total proteins in the total homogenate and in the cellular fractions of this organ. Besides, a decrease in the enzymatic activity of this ovary is not observed, but it occurs in the left ovary. This finding would indicate a lack of enzymatic segregation that could be related to the phenomenon of ovarian atrophy. Finally, the electrophoretic study indicated that it would apparently exist only one molecular form of the enzyme. Our results support the hypothesis that acid phosphatase would be involved in the atrophic processes of the right ovary during embryonary differentiation. 相似文献
5.
M C Garel V Joulin P Le Boulch M C Calvin M O Préhu N Arous R Longin R Rosa J Rosa M Cohen-Solal 《The Journal of biological chemistry》1989,264(32):18966-18972
Bisphosphoglycerate mutase is an erythrocyte-specific enzyme whose main function is to synthesize 2,3-diphosphoglycerate, the allosteric effector of hemoglobin. In addition to its main 2,3-diphosphoglycerate synthase activity, the enzyme displays phosphatase and mutase activities both involving 2,3-diphosphoglycerate in their reaction. The three activities have been demonstrated to be catalysed at a unique active site. To study the structure of such an active site we have developed a recombinant system producing mutants of human bisphosphoglycerate mutase in Escherichia coli, by site-directed mutagenesis. For this purpose the human bisphosphoglycerate mutase cDNA that we had previously cloned has been used to construct a procaryotic high level expression vector bearing the "tac" promoter. Human bisphosphoglycerate mutase produced in E. coli, a species which does not normally synthesize this enzyme, represented 8% of the total soluble bacterial protein and displayed the three catalytic activities (synthase, mutase, and phosphatase) characteristic of the enzyme. Since it has been suggested that the carboxyl-terminal region may be implicated in the catalytic activity of the enzyme, three variants deleted in this part of the protein were produced. Our results indicate that a minimal deletion of 7 amino acid residues in the carboxyl-terminal portion of the human bisphosphoglycerate mutase completely abolished the three catalytic activities of the enzyme. In contrast, the effects of the deletion of the last two lysine residues were limited to a 38% reduction in the synthase activity. These results show that the carboxyl-terminal amino acid residues are either directly or indirectly implicated in the three catalytic functions of the human bisphosphoglycerate mutase, and that the two terminal lysine residues are not essential for the major part of the enzymatic mechanism of the enzyme. 相似文献
6.
With the G?m?ri technique, lead precipitates have been found in thyroid follicle cells in unusual localizations such as apical hyaloplasm and microvilli; it has been established that they were actually significant for acid phosphatase activity: constant results in spite of repeated controls and several variations from the original cytochemical technique, allow to think that lead precipitates were not merely artefactual, but actually significant of enzymatic activity. However it is pointed to the fact that the origin of the enzyme has to be questioned; it is assumed that most likely acid phosphatase has diffused from its original lysosomal site. Such diffusion implies variations of the selective permeability of lysosomal membranes; inappropriate relation between the quantity of enzyme present in these organelles and the quantity of substrate used might also be considered, though changes in the amount (resp. concentration) of substrate remained ineffective and induced no modification in the localization of observed enzymatic activity. In addition, one point of interest is an obvious relation between the observed enzyme diffusion and the state of activity resp. rest of the cell; in the present state of investigations, this remains unexplained and likely related to factors escaping control during processing; moreover, no explanation can be provided for the fact that it revealed impossible to avoid such diffusion even by means of variations of the numerous parameters involved in the G?m?ri technique. So that it finally appears necessary to remain on a critical position regarding the results at the ultrastructural level of this standardized technique, and there is no doubt it would reveal useful that several assumptions in the literature about extra lysosomal acid phosphatase activity should be reinvestigated with a similar critical purpose. 相似文献
7.
Iu A Belaia E L Khasman L Iu Popova 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1984,(11):72-76
The overtime study of changes in the activity of cathepsin D, a lysosomal enzyme, in the splenocytes of CBA mice after their infection with virulent and avirulent Shigella strains of the same origin and with the same antigenic structure has been made. As the result of two months of observations, changes in the activity of this enzyme in the cytoplasmic and lysosomal cell fractions have been found to occur in phases. The activity of cathepsin D has been shown to depend on the virulence of Shigella strains used for inoculation. Virulent Shigella strains induce the pronounced and prolonged activation of the enzyme in the lysosomes, as well as in the cytoplasm. The latter phenomenon is probably indicative of the pathological labilization of the lysosomal membranes, induced by the virulent culture. Avirulent Shigella strains induce only the transient activity of the enzyme in the lysosomes without any essential changes in the permeability of their membranes. These data point to the possibility of differentiating virulent and avirulent Shigella strains by the determination of the enzymatic activity of splenocytes in infected animals. 相似文献
8.
Identification and characterization of thiamin repressible acid phosphatase in yeast 总被引:3,自引:0,他引:3
M E Schweingruber R Fluri K Maundrell A M Schweingruber E Dumermuth 《The Journal of biological chemistry》1986,261(34):15877-15882
We have identified a genetic locus, pho4, in Schizosaccharomyces pombe which encodes a minor expressed cell surface acid phosphatase that is repressed by low concentrations (0.5 microM) of thiamin. The enzyme was purified from a strain that overproduces the enzyme. It is an Asn-linked glycoprotein. Removal of the carbohydrates by endoglycosidase H does not abolish enzymatic activity. The molecular mass of deglycosylated and unglycosylated enzyme that accumulates in membranes when cells are grown in the presence of tunicamycin is 56 kDa as determined by sodium dodecyl sulfate-gel electrophoresis. Thiamin regulation, at least in part, operates by reducing the level of pho4-mRNA. Pho4 is not genetically linked to the phosphate repressible acid phosphatase gene pho1. Phosphate and thiamin repressible acid phosphatase differ in their substrate specificity. Their protein moieties are immunologically related. Pho4 and pho1 are the only genes in S. pombe that express cell surface acid phosphatases being enzymatically active with nitrophenyl phosphate as substrate. S. pombe is not unique in having a thiamin repressible acid phosphatase. In Saccharomyces cerevisiae this enzyme is encoded by PHO3. 相似文献
9.
The aims of the study were: the isolation of strains of black-pigmented Bacteroides from the gingival sulcus of different animals, their biochemical and immunological characterization and comparison of their properties for classification within the genus. A total of 104 strains, isolated from cats, dogs, racoons and a jaguar, were characterized on the basis of fermentation of carbohydrates, metabolic end products, haemagglutination studies, enzymatic activities, catalase production and indirect immunofluorescence. No differences were observed between the strains regardless of their animal origin. The strains did not ferment carbohydrates, produce phenylacetic acid, show an array of enzyme activities or agglutinate sheep red blood cells. They were catalase-positive and so differed from the human oral strains of Bact. gingivalis. Immunofluorescence microscopy revealed that the animal strains shared at least one major antigen with Bact. gingivalis but none with Bact. asaccharolyticus. Apart from their catalase activity, the animal strains isolated were similar to those of human Bact. gingivalis strains. 相似文献
10.
The aims of the study were: the isolation of strains of black-pigmented Bacteroides from the gingival sulcus of different animals, their biochemical and immunological characterization and comparison of their properties for classification within the genus. A total of 104 strains, isolated from cats, dogs, racoons and a jaguar, were characterized on the basis of fermentation of carbohydrates, metabolic end products, haemagglutination studies, enzymatic activities, catalase production and indirect immunofluorescence. No differences were observed between the strains regardless of their animal origin. The strains did not ferment carbohydrates, produce phenylacetic acid, show an array of enzyme activities or agglutinate sheep red blood cells. They were catalase-positive and so differed from the human oral strains of Bact. gingivalis. Immunofluorescence microscopy revealed that the animal strains shared at least one major antigen with Bact. gingivalis but none with Bact. asaccharolyticus. Apart from their catalase activity, the animal strains isolated were similar to those of human Bact. gingivalis strains. 相似文献
11.
Cytochemical study of macrophage lysosomal inorganic trimetaphosphatase and acid phosphatase 总被引:2,自引:0,他引:2
Cytochemical investigations have associated acid inorganic trimetaphosphatase (TMPase) activity with the lysosomes of certain cell types. We have used the modified staining technique of Berg to show that this enzyme activity is present in normal mononuclear phagocytes and macrophage cell lines. We have found this enzyme activity to be present in murine RAW264 macrophages, in human U937 macrophages, in normal human blood monocytes, and in guinea pig peritoneal macrophages. All of the RAW264 and U937 macrophages showed intense TMPase activity. Many of the human monocytes and most of the guinea pig macrophages were labeled by this method. The reaction product was associated with the lysosomes of these cell types. The lysosomal staining-pattern was similar to that of acid phosphatase. Differences with regard to Golgi staining were noted. This indicates that TMPase is a lysosomal enzyme of mammalian macrophages. The distinction between TMPase and acid phosphatase activity has been demonstrated by measuring the pH optimum of each enzyme. Using substrates identical to those of the ultrastructural cytochemistry, we show that the pH optimum of TMPase is 4.0 and that of acid phosphatase is 5.0. The enzymatic activities are therefore ultrastructurally and biochemically distinct. Following phagocytosis of latex, yeast (Saccharomyces cerevisiae), or Corynebacterium parvum, TMPase has been found to be associated with phagosomes. This enzyme may take part in the degradation of phagocytosed materials, particularly microorganisms which contain inorganic polyphosphates and metaphosphates. 相似文献
12.
Enzymatic profiles were determined by the API ZYM system for 15 strains of non 01 Vibrio cholerae, 4 strains of V. metschnikovii, 9 strains of V. anguillarum, 6 strains of Plesiomonas shigelloides and 115 strains motile Aeromonas sp. All of the tested strains produced alkaline phosphatase, leucine aminopeptidase and did not possess alpha-fucosidase and alpha-mannosidase. Some differences in enzymatic activities among the tested Vibrionaceae strains were noted. The strains of non 01 V. cholerae, V. metschnikovii, V. anguillarum and P. shigelloides did not produce trypsin, whereas all of the tested Vibrio sp. strains appeared to be positive for this enzyme. Only the strains of P. shigelloides produced BI-Phospho-hydrolase. The lack of acid phosphatase activity was observed among the strains of V. anguillarum. 相似文献
13.
In this paper evidences are presented strongly confirming that an extracellular 32P-phosphopeptide phosphatase activity of yeast is accounted for by acid phosphatase. Dephosphorylation of 32P phosphoseryl peptides was achieved with whole yeast cells, thus demonstrating extracellular location of protein phosphatase activity. The acid phosphatase and protein phosphatase activity copurified throughout purification procedure. Purified enzyme showed the same pH-profile and had the same Km value with phosphopeptide substrate as intact cells. Protein phosphatase activity is repressed by phosphate in the same manner as acid phosphatase activity, showing that not only repressible but also constitutive acid phosphatase displays protein phosphatase activity. Using mutant strains defective in acid phosphatase activity it was confirmed that acid phosphatase and protein phosphatase activities are the products of the same gene(s). 相似文献
14.
In a whole cell assay system with p-nitrophenyl phosphate as substrate, strains of Mycobacterium tuberculosis and M. bovis were identical in the pH-activity pattern of acid phosphatase. It was a one-peak curve with a pH optimum at 6.2 and sharp symmetrical slopes. The enzymatic activity did not reflect the virulence. When the cells were subjected to mechanical fractionation, the major part of the enzymatic activity was found in a particulate fraction and a minor portion in supernatant and cell walls, suggesting the location of the enzyme in the membrane. Exposure of the cells to free long-chain fatty acids, especially unsaturated ones, reduced the enzymatic activity in a dose-response manner with concomitant decrease in the viability. However, no causal relationship between these two effects was suggested from the collateral experiments. 相似文献
15.
E Kondo S Dejsirilert N Wejprasit D Chiewsilp K Kanai 《Japanese journal of medical science & biology》1991,44(2):51-62
In a whole cell assay system with p-nitrophenyl phosphate as substrate, strains of Pseudomonas pseudomallei showed a two-peak pattern in pH activity curve of acid phosphatase, suggesting the presence of two enzyme components different in pH optimum (4.2 and 5.2). The component of 5.2 pH optimum was detected in the outer membrane fraction and the activity was resistant to heating at 70 C for 30 min. The other component of 4.2 pH optimum was heat-labile. No substantial difference was observed in the enzymatic activity between R and S type colonies. 相似文献
16.
In two maize genotypes, the effect of exogenous phosphate deficiency on acid phosphatase activity of the apical part of primary
root was followed in dry and imbided grains. Higher acid phosphatase activity was found in the genotype LG 5. The enzyme activity
increased after 18 h grain imbibition in the two genotypes. After 48 h germination no differences were found between the genotypes.
After 24 h cultivation of root segments in a solution of 0.25 mM CaSO4.2H2O,0.1 mM MgSO4.7H2O, and further after 3,6 and 24 h cultivation in solutions with and without phosphate genotypic differences were found in
acid phosphatase activity as well as in dry mass production. An increase in enzymatic activity due to exogenous phosphate
deficiency was registered in the two genotypes only after 24 h cultivation. 相似文献
17.
A phosphotyrosyl-protein phosphatase activity associated with acid phosphatase from human prostate gland 总被引:6,自引:0,他引:6
Using [32P]P-Tyr-IgG and [32P]P-Tyr-casein phosphorylated by pp60v-src as substrates, studies on the phosphotyrosyl-protein phosphatase activity in human prostate gland indicate that it is associated with prostatic acid phosphatase. Evidence to support this conclusion include the following: (a) these two enzymatic activities co-purify to apparent homogeneity; (b) they co-migrated on polyacrylamide gel electrophoresis, ion-exchange and gel filtration chromatographies; (c) the exhibit identical thermostability; and (d) the phosphotyrosyl-protein phosphatase activity is sensitive to inhibition by p-nitrophenyl phosphate and by several classical inhibitors of prostatic acid phosphatase including L(+)-tartrate, molybdate, vanadate and NaF. The purified enzyme exhibits high specificity towards phosphotyrosyl-proteins with little activity towards several phosphoseryl-proteins and phosphothreonyl-proteins examined. The present findings indicate that prostatic acid phosphatase may function in vivo as a phosphotyrosyl-protein phosphatase. 相似文献
18.
Piotr M. Laidler Eulazio M. Taga Robert L. Van Etten 《Archives of biochemistry and biophysics》1982,216(2):512-521
The stoichiometry and the reactivity of the sulfhydryl groups of a human liver acid phosphatase have been studied. The smallest (Mr = 14,400) of the three molecular-weight forms of acid phosphatase from human liver, recently purified and characterized in our laboratory, was treated with various sulfhydryl group-specific reagents: p-hydroxymercuribenzoate, p-hydroxymercuriphenylsulfonate, fluorescein mercuriacetate, methyl methanethiosulfonate, p-nitrophenoxycarbonyl methyl disulfide, and thiosulfate. A total loss of enzymatic activity was obtained in each case. By spectrophotometric titration with 5,5′-dithiobis(2-nitrobenzoate) and p-hydroxymercuriphenylsulfonate it was shown that there are six free sulfhydryls per protein molecule, consistent with the amino acid analysis of this enzyme. The same number was deduced as a result of inactivation studies carried out with p-hydroxymercuribenzoate and p-hydroxymercuriphenylsulfonate. A total loss of activity was obtained at reagent to enzyme ratios of 6:1 in both cases. Similar results were obtained upon inactivation by p-nitrophenoxycarbonyl methyl disulfide, where the enzyme was found to possess only 10% residual activity at an inhibitor-to-enzyme ratio of 6:1. With fluorescein mercuriacetate as an inactivator, total loss of activity was found at a 2.5 times molar excess of this reagent over protein. Both the stoichiometry of inactivation and fluorescence titration experiments suggest that fluorescein mercuriacetate can function as a bifunctional sulfhydryl group reagent. The activity of a totally inactivated enzyme preparation obtained following reaction with excess of p-nitrophenoxycarbonyl methyl disulfide or with methyl methanethiolsulfonate could be almost completely restored upon treatment with dithiothreitol. These data are consistent with the interpretation that in each enzyme molecule, there are six free sulfhydryl groups of almost equal reactivity, at least one of which is essential for enzymatic activity. 相似文献
19.
T L Buraia A A Butakov V A Drozhennikov V N Nikulin B V Pinegin N D Iushchuk 《Zhurnal mikrobiologii, epidemiologii, i immunobiologii》1991,(10):52-55
In four series of experiments human peripheral blood neutrophils were found capable of synthetizing the active forms of such enzymes as myeloperoxidase and acid phosphatase after stimulation with opsonized zymosan, and the optimum conditions for testing the synthetizing activity of neutrophils were established. The examination of 39 practically healthy donors revealed an approximate equilibrium between the synthesis of the active forms of the enzyme and its excretion from the cell after this cell was activated. Considerable changes in the enzymatic activity of peripheral blood neutrophils were found in patients with yersiniosis, chronic herpes, chronic bronchitis and AIDS-associated complex. The possibility of a deficient enzymatic activity in some patients was suggested. 相似文献
20.
We previously reported the isolation from bovine liver of a novel 56-kDa inorganic pyrophosphatase named phospholysine phosphohistidine inorganic pyrophosphate phosphatase (LHPPase). It is a unique enzyme that hydrolyzes not only oxygen-phosphorus bonds in inorganic pyrophosphate but also nitrogen-phosphorus bonds in phospholysine, phosphohistidine and imidodiphosphate in vitro. In this study, we determined the partial amino acid sequence of the purified bovine LHPPase. To investigate whether humans have the same enzyme, we isolated a cDNA clone from a HeLa cell cDNA library that encodes for the human homologue of LHPPase. Although its sequence does not include the consensus sequence of a typical inorganic pyrophosphatase, it does contain a similar sequence of the active site in other phosphatases such as protein-tyrosine phosphatase, dual-specific phosphatase and low molecular weight acid phosphatase. Human LHPPase was highly expressed in the liver and kidney, and moderately in the brain. The recombinant protein was produced in E. coli. Its ability to hydrolyze oxygen-phosphorus bonds and nitrogen-phosphorus bonds was confirmed. The enzymatic characteristics of this human protein were similar to those of purified bovine LHPPase. Thus, we concluded that the cDNA encoded the human counterpart of bovine LHPPase. 相似文献