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Results for “Lactoperoxidase”

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At least 19 recordsLinked to original sources

PubMed1979

Lactoperoxidase activity in guinea-pig milk and saliva: correlation in milk of lactoperoxidase with bactericidal activity against Escherichia coli.

The lactoperoxidase (LPO) activity in guinea-pig milk and saliva has been investigated in sows suckling normal young, and young orally infected with Escherichia coli. There was a 5-fold increase in activity in milk during the 3--4 weeks of lactation; infection of the young did not alter this. There was no comparable increase in lactoperoxidase activity of saliva during this same period, either in the infected or non-infected group. The antibacterial activity of milk from sows suckling normal young increased with the lactoperoxidase, and this bactericidal activity could be reversed by LPO inhibitors such as penicillamine and cysteine but not by addition of sufficient iron to saturate the lactoferrin. In milk from sows suckling infected young, bacteriostatic activity occurring in samples from about 14 days after infection needed iron or both iron and penicillamine (or cysteine) for reversal, indicating that both the antibody-lactoferrin system and the LPO system may be involved in the infected state.

Animals↗

PubMed1977

Enzyme activity of salivary lactoperoxidase adsorbed to human enamel.

Human saliva was incubated with human enamel powder, and the lactoperoxidase activity of the saliva was measured before and after incubation. Liquidphase lactoperoxidase activity was reduced in direct proportion to the weight of enamel powder added. Lactoperoxidase molecules were adsorbed to the enamel surface in an enzymatically active conformation, and this enamel-bound lactoperoxidase activity was also measured. The adsorption of lactoperoxidase was irreversible and produced at least a 40% increase in the concentration of lactoperoxidase in the enamel surface phase as compared with its concentration in the liquid phase. Enamel-bound lactoperoxidase, as well as the free enzyme, was capable of inactivating the key glycolytic enzyme hexokinase. The implications of the adsorption phenomenon for bacterial colonization are discussed.

Adsorption↗

PubMed1979

Lactoperoxidase binding to streptococci.

There have been conflicting reports regarding the binding of lactoperoxidase to bacterial cell surfaces. We describe here the effects of cell-bound lactoperoxidase on acid production by suspensions of Streptococcus mutans (NCTC 10449) in the presence of hydrogen peroxide and thiocyanate. Saline suspensions of log-phase bacteria were treated with 0.1 mg of lactoperoxidase per ml and were then washed thoroughly. The addition of hydrogen peroxide and thiocyanate markedly reduced the acid production of these lactoperoxidase-treated bacteria but had no effect on the acid production of untreated controls. After a 3-h incubation in saline, the lactoperoxidase-treated bacteria produced acid in the presence of hydrogen peroxide and thiocyanate at the same rate as untreated bacteria. These observations suggest that lactoperoxidase is initially bound to the cell surface in an enzymatically active form at a concentration sufficient to inhibit acid production. The lactoperoxidase is slowly degraded or desorbed as the bacteria stand in saline suspension.

Cell Membrane↗

PubMed1979

Interaction of lactoperoxidase with thiols and diiodotyrosine.

Glutathione and cysteine bind to the heme of lactoperoxidase, thereby causing a red shift of the Soret band which is reversed upon addition of iodide or guaiacol, two substrates for lactoperoxidase. The rate of formation of the enzyme-thiol complex is enhanced by diiodotyrosine. Binding of diiodotyrosine to lactoperoxidase does not cause a shift of the Soret band which indicates binding to the protein of the enzyme. At neutral pH and low ionic strength, lactoperoxidase is adsorbed on insolubilized diiodotyrosine (diiodotyrosine-agarose). It can be eluted at slightly increased ionic strength which shows that the binding is weak. In the presence of 5 X 10(-4) M glutathione, however, the binding of the enzyme to diiodotyrosine-agarose becomes much stronger so that a high salt concentration is required for elution. Lactoperoxidase is also adsorbed on insolubilized thiols (thiol-agarose). The presence of diiodotyrosine is not required for strong binding. A simple method for the preparation of lactoperoxidase from milk by affinity chromatography is based on the interactions of the enzyme with the two ligands, thiols and diiodotyrosine.

Cysteine↗

PubMed1979

Iodide binding and regulation of lactoperoxidase activity toward thyroid goitrogens.

The effects of the antithyroid goitrogens, methylthiouracil and methylmercaptoimidazole, on the oxidation of N-acetyltyrosylamide at pH 8.8 by lactoperoxidase have been evaluated in the presence and the absence of iodide for the purpose of elucidating the effects of iodide. At pH 8.8, iodine is not oxidized. In the absence of iodide, the two antithyroid drugs inactivate lactoperoxidase by a second order process. When iodide is added before methylthiouracil or methylmercaptoimidazole, enzyme inactivation does not occur as rapidly and both goitrogens are readily oxidized. The kinetics of the oxidation reactions have been analyzed in order to obtain the equilibrium constant of the iodide . lactoperoxidase complex. Essentially the same iodide dissociation constant, i.e. 2 x 10(-5) M, was found by studying its effects on the kinetics of oxidation of the two antithyroid drugs. A large difference absorption spectrum is observed in the Soret region between native lactoperoxidase and lactoperoxidase inactivated by methylthiouracil.

Iodides↗

PubMed1978

Degradation of aflatoxin by lactoperoxidase.

Three concentrations of lactoperoxidase, 5, 50, and 500 units/ml of reaction mixture, degraded aflatoxin in the presence of 225 micrometer NaCl and 50 micrometer H2O2 at 28 degrees C. Increasing the amount of lactoperoxidase from 50 to 500 units/ml of reaction mixture resulted in increasing the rate of degradation of aflatoxin B1 from 3.6 to 5.1%/24 h. When comparable amounts of lactoperoxidase were present, aflatoxin G1 was degraded approximately 1.5 times faster than was aflatoxin B1. At a given concentration of lactoperoxidase, aflatoxin degradation was independent of initial aflatoxin concentration. Derivatives that cochromatographed with aflatoxin B2a and derivatives that were water soluble were the major degradation products of aflatoxin B1. Similar derivatives, but in greater proportions, were noted as degradation products that resulted from activity of a blendure of mycelia of Aspergillus parasiticus.

Aflatoxins↗

PubMed1975

Immunological and biophysical properties of hepatitis B antigen labeled by the chloramine-T and by the lactoperoxidase methods.

Optimal conditions were sought for the radiolabeling of microgram quantities of hepatitis B surface antigen (HBs Ag) employing the chloramine-T or lactoperoxidase iodination procedures. Preparations of HBsAg labeled by these procedures are referred to as chloramine-T preparations and lactoperoxidase preparations, respectively. Labeled HBsAg having specific activities between 10-20 muCi/mug were found to display the greatest degree of sensitivity for unlabeled HBsAg and for anti-HBs using a double-antibody radioimmunoassay (RIA-DA). Increasing the specific activity above this level redulted in a decreased affinity of labeled 1251-HBs Ag for anti-HBs, indicating that soluble antigenic alterations had developed. At equivalent specific activities, chloramine-T preparations competed less effectively for unlabeled HBs Ag than lactoperoxidase preparations, and anti-HBs endpoint titers were slightly reduced, especially among preparations of high specific activity (greater than or equal to 65 muCi/mug). Chloramine-T preparations of HBs Ag (sp. act. 15--30 muCi/mug) showed essentially no antigenic deterioration over a 2-month period at minus 196 degrees C or minus 70 degrees C. Utilization of optimally labeled 1251-HBs Ag has increased the sensitivity of the RIA-DA for unlabeled HBs Ag 30-fold to a level below 1 ng/ml and enhanced antiamine-T method revealed that only the most acidic population was labeled (pH 3.75+/-0.5). In contrast, six antigenic components with distinct pI values ranging from 3.7 to 5.2 were detected by RIA-DA in both unlabeled HBs ag and in the chloramine-T preparation. This indicated that the chloramine-T method did not radically change the relative number or charge of each of the pI populations present in purified preparations of HBs Ag. Analysis of HBs Ag iodinated by the lactoperoxidase procedure revealed the presence of three of four populations of particles with pI values ranging from 3.9 to 4.5, suggesting that this procedure labels HBs Ag more uniformly.

Chloramines↗

PubMed1975

Molecular morphology of ribosomes. Iodination of Escherichia coli ribosomal proteins with solid-state lactoperoxidase.

Using either soluble or solid-state lactoperoxidase, a comparison was made between the enzymic iodination of ribosomal proteins iodinated as 30-S and 50-S subunits or as 70-S monosomes. Proteins S7, S11 and S12 of the 30-S subunit and proteins L2, L11, L26 and L28 of the 50-S subunit were labelled to a greater extent in isolated particles than in the 70-S ribosome. In contrast, proteins S4, S19 and S20 were labelled to a lesser extent in the isolated subunit. No significant differences were observed in the iodination patterns of ribosomes iodinated in the presence of soluble lactoperoxidase and those iodinated in the presence of lactoperoxidase bound to Sepharose 4B. It is suggested that the 30-S subunit undergoes a conformational change during its association with the 50-S subunit to form a 70-S monosome. Implications from results obtained with solid-state lactoperoxidase-catalyzed iodination of ribosomal proteins are also discussed.

Escherichia coli↗

PubMed1979

The effects of thioureylene compounds (goitrogens) on lactoperoxidase activity.

The rates of oxidation of several goitrogens by lactoperoxidase and the rates of inactivation of lactoperoxidase by the same goitrogens have been measured. The influence of iodide on both reactions has also been evaluated. It has been shown by us that iodide acts catalytically in regulating lactoperoxidase activity at pH 8.8. The rate data have been analyzed by a computer program which solves the differential equations for the above mentioned reactions. From this computer analysis we have been able to obtain binding constants of the goitrogens and inactivation rate constants of lactoperoxidase. Iodide was shown to inhibit goitrogenic activity either by increasing the rate of drug oxidation or by reducing the rate of enzyme inactivation, or both, depending on the particular drug. Iodide had little or no effect on the goitrogen-binding constants. We have also shown that the relative rates of enzyme inactivation can be correlated with the potency of the goitrogen as an antithyroid drug.

Antithyroid Agents↗

PubMed1979

Use of an iodide-specific electrode to study lactoperoxidase-catalyzed iodination of l-tyrosine.

An in vitro method employing an iodide-specific electrode for monitoring lactoperoxidase-catalyzed iodination is described. The method utilized lactoperoxidase, potassium iodide, and a glucose--glucose oxidase system for the generation of hydrogen peroxide and l-tyrosine. As iodination of l-tyrosine proceeded, the free iodide concentration in solution decreased and was monitored by an iodide-specific electrode. The iodide electrode was reliable when compared to a 131I-method for measuring free iodide changes in solution. Increasing concentrations of resorcinol, a well-known inhibitor of thyroid peroxidase-catalyzed iodination, in the reaction mixture resulted in graded inhibition of the initial rate of lactoperoxidase-catalyzed l-tyrosine iodination. This in vitro system can be used to assess inhibitory activity of various antithyroid substances.

Catalysis↗

PubMed1975

The transmembrane proteins in the plasma membrane of normal human erythrocytes. Evaluation employing lactoperoxidase and proteases.

The molecular architecture of the human erythrocyte membrane has been probed using lactoperoxidase-catalyzed iodination in conjunction with Pronase hydrolysis. Resealed, hemoglobin-free ghosts were labeled at the cytoplasmic surface and the external membrane surface was subsequently digested with Pronase. Changes in size of the components labeled at the cytoplasmic surface were readily detected by sodium dodecyl sulfate gel electrophoresis. The protein 3 molecular weight class labeled at the cytoplasmic surface was extensively hydrolyzed at the external surface to produce a major 65000 molecular weight fragment and a minor 45000 molecular weight fragment. When resealed membranes were labeled on the external surface the same 65000 molecular weight labeled component is produced. These results unequivocally demonstrate that the same polypeptides in the protein 3 molecular weight class that can be labeled by lactoperoxidase at the cytoplasmic membrane surface are digested by Pronase at the external surface and are, therefore, transmembrane components. Where it is possible to label one surface of a membrane with lactoperoxidase and reseal the membrane this procedure represents an alternate method for establishing transmembrane configuration of membrane proteins.

Blood Proteins↗

PubMed1978

Susceptibility of Escherichia coli to bactericidal action of lactoperoxidase, peroxide, and iodide or thiocyanate.

The bactericidal action that results from lactoperoxidase-catalyzed oxidation of iodide or thiocyanate was studied, using Escherichia coli as the test organism. The susceptibility of intact cells to bactericidal action was compared with that of cells with altered cell envelopes. Exposure to ethylenediaminetetraacetic acid, to lysozyme and ethylenediaminetetraacetic acid, or to osmotic shock were used to alter the cell envelope. Bactericidal action was greatly increased when the cells were exposed to the lactoperoxidase-peroxide-iodide system at low temperatures, low cell density, or after alteration of the cell envelope. When thiocyanate was substituted for iodide, bactericidal activity was observed only at low cell density or after osmotic shock. Low temperature and low cell density lowered the rate of destruction of peroxide by the bacteria. Therefore, competition for peroxide between the bacteria and lactoperoxidase may influence the extent of bactericidal action. Alteration of the cell envelope had only a small effect on the rate of destruction of peroxide. Instead, the increased susceptibility of these altered cells suggested that bactericidal action required permeation of a reagent through the cell envelope. In addition to altering the cell envelope, these procedures partly depleted cells of oxidizable substrates and sulfhydryl components. Adding an oxidizable substrate did not decrease the susceptibility of the altered cells. On the other hand, mild reducing agents such as sulfhydryl compounds did partly reverse bactericidal action when added after exposure of cells to the peroxidase systems. These studies indicate that alteration of the metabolism, structure, or composition of bacterial cells can greatly increase their susceptibility to peroxidase bactericidal action.

Escherichia coli↗

PubMed1975

Lactoperoxidase activity in human milk and in saliva of newborn infants.

Human milk and saliva from newborn infants were analyzed for their content of lactoperoxidase and thiocyanate. The activity of lactoperoxidase in infant saliva was variable but generally higher than that found in calf saliva. In contrast, the activity in human colostrum was low ( approximately 5%) compared with that found in cow's milk. The enzyme was resistant to gastric juice. Thiocyanate was demonstrated in infant saliva in concentrations about one-third of that in adult saliva. The amounts of lactoperoxidase and thiocyanate in infant saliva are quite sufficient to inhibit bacterial growth in in vitro systems. The importance of this system in vivo has not yet been demonstrated. The availability of this system to both newborn calves and humans (in calves provided largely by colostrum and in human babies by saliva) might be indirect evidence of its importance.

Adult↗

PubMed1978

Lactoperoxidase, peroxide, thiocyanate antimicrobial system: correlation of sulfhydryl oxidation with antimicrobial action.

The antimicrobial activity of the lactoperoxidase, peroxide, thiocyanate system against Escherichia coli was directly related to the oxidation of bacterial sulfhydryls. Lactoperoxidase catalyzed the oxidation of thiocyanate, which resulted in the accumulation of hypothiocyanite ion, OSCN-. A portion of the bacterial sulfhydryls were oxidized by OSCN- to yield sulfenic acid and sulfenyl thiocyanate derivatives. The remaining sulfhydryls were not oxidized, although OSCN- was present in large excess. The oxidation of sulfhydryls to sulfenyl derivatives inhibited bacterial respiration. This inhibition could be reversed by adding sulfhydryl compounds to reduce the sulfenyl derivatives and the excess OSCN-. Also, this inhibition could be reversed by washing the cells so as to remove the excess unreacted OSCN-. After washing, the bacteria underwent a time-dependent recovery of their sulfhydryl content. This recovery resulted in recovery of the ability to respire. The inhibited cells were viable if diluted and plated shortly after the incubation with the lactoperoxidase, peroxide, thiocyanate system. On the other hand, long-term incubation in the presence of the excess OSCN- resulted in loss of viability. Also, the inhibition of respiration became irreversible. During this long-term incubation, the excess OSCN- was consumed and the sulfenyl derivatives disappeared.

Dithiothreitol↗

PubMedDate not supplied

Xylitol-induced increase of lactoperoxidase activity.

Whole saliva samples of volunteers who ate a strict diet for two years with regard to the type of sweeteners used (sucrose, fructose, and xylitol) showed considerable differences in the lactoperoxidase activity. The consumption of a xylitol diet increased the activity of this enzyme fourfold to tenfold when compared to the other two test groups. Lactoperoxidase belongs to the natural defense mechanisms of the oral cavity. However, the consumption of a xylitol diet also leads to a strong reduction in the incidence of dental caries. It is suggested that the xylitol-induced elevation of the salivary lactoperoxidase activity and the cariostatic properties of xylitol are partly interrelated phenomena.

Adolescent↗

PubMed1979

Heterogeneity of chloramine T- and lactoperoxidase-radioiodinated human calcitonin.

Radioiodination reportedly damages peptides, but the nature of the damage has not been adequately examined. Utilizing isoelectric focusing, we examined the products of Chloramine T- and lactoperoxidase-directed radioiodinations of human calcitonin. Initially, the reaction products were purified by adsorption onto and elution from microfine silica (QUSO-G32). Radioiodination of the calcitonin by Chloramine T and lactoperoxidase produced a heterogeneous population of 125I-labeled peptides exhibiting apparent isoelectric points that were more acidic than that of unlabeled synthetic calcitonin. Variation in the products among radioiodinations and the inability of QUSO-G32 to resolve the components of the reaction mixture prompted our examination of alternative purification procedures. Anion-exchange chromatography on QAE-Sephadex effectively separated [125I]diiodotyrosine containing calcitonin from free iodine and [125I]iodolactoperoxidase. Our data indicate that: (a) radioiodination of human calcitonin by Chloramine T and lactoperoxidase induced alteration in the peptide as evidenced by isoelectric point, (b) specific [125I]iodopeptides vary in incidence and relative abundance among radioiodinations, (c) identification of the labeled amino acid in [125I]iodopeptides cannot ensure intergrity of the molecule, and (d) isoelectric focusing provides a method of comparing the products of peptide radioiodinations among laboratories.

Calcitonin↗

PubMed1976

An immobilized two-enzyme system for the activation of the lactoperoxidase antibacterial system in milk.

Lactoperoxidase catalyzes the oxidation of thiocyanate by hydrogen peroxide and an intermediary product is formed with antibacterial properties. The components of this system, with the exception of hydrogen peroxide, are present in milk. H2O2 may be introduced by means of enzymatic generation and thus make the system complete. A two-enzyme system consisting of beta-galactosidase and glucose oxidase has been developed for this purpose. The coupled enzyme reaction is shown to work with high efficiency at the neutral pH of milk although the enzymes as such, particularly lactases suitable for immobilization, have optimal activities at much lower pH values. The results indicate that the lactoperoxidase system may in this way be employed to inactivate bacteria present in milk.

Animals↗

PubMed1979

alpha-Amanitin: inactivation by bovine lactoperoxidase.

The principle amatoxin, alpha-amanitin, is found to be extremely sensitive toward lactoperoxidase catalyzed degradation, rather than iodination, of the indole nucleus. Extensive attenuation of inhibitor potency against eukaryotic DNA-dependent RNA polymerase II accompanies the treatment of alpha-amanitin with lactoperoxidase, iodide and hydrogen peroxide.

Amanitins↗