Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “PENICILLINASE”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 145 records · Page 8Linked to original sources

Effects of the recipient strain and ultraviolet irradiation on transduction kinetics of the penicillinase plasmid of Staphylococcus aureus.

When the penicillinase plasmid of Staphylococcus aureus PS 81(P(81))(T(81)) was transferred to its cured derivative of PS 81(N(P))(T(81)), there was a fivefold increase in the transduction frequency of penicillinase plasmid markers after ultraviolet (UV) irradiation of the phage instead of the expected decrease typical for plasmid-borne markers. These results were independent of the transducing phage, the donor, and the method of curing the recipient and were also obtained with a cured derivative of PS 80(PI(80)). With PS 52, a naturally occurring penicillin-sensitive strain, and a cured transductant of PS 52 as the recipients, typical plasmid kinetics were observed. The plasmid location of penicillinase plasmid markers in transductants was confirmed by their instability in ethidium bromide (EB). In a cross between isogenic plasmids (PI(258)penZ cad x PI(258)penI asa ero), transductants were doubly selected for cadmium and erythromycin resistances. There was a twofold increase in transduction frequency after UV irradiation of the transducing phage and an increase in the proportion of recombinant type transductants. CsCl-EB density centrifugation revealed that plasmid deoxyribonucleic acid (DNA) was present in PS 81(P(81))(N(T)) and its cured derivative [PS 81(N(P))(N(T))], but not in PS 52. Sucrose gradient analysis of plasmid DNA showed that the penicillinase plasmid of PS 81(P(81))(N(T)) was larger than the plasmid in its cured derivative. Thus, the cured derivative contains plasmid DNA which appears to recombine with the incoming plasmid, causing the rise in transduction frequency noted after UV irradiation of transducing phage.

Arsenic↗

Phenotypic suppression of methicillin resistance in Staphylococcus aureus by mutant noninducible penicillinase plasmids.

Methicillin (intrinsic) resistance of Staphylococcus aureus was suppressed almost completely by regulatory gene (penI(1)) mutations of penicillinase plasmids that made penicillinase production strictly noninducible. Methicillin resistance was restored by secondary regulatory gene mutations that altered the noninducible phenotype or by complementation with a compatible plasmid that did not bear the noninducible mutation. No evidence was obtained for genetic linkage between a penicillinase plasmid and the gene for methicillin resistance. We suggest, therefore, that the mutant noninducible repressor acted in trans by binding to a site on the methicillin resistance determinant. This hypothesis would imply an appreciable degree of homology between penicillinase plasmids and methicillin resistance genes.

Anti-Bacterial Agents↗

Immunoelectron microscopic double labeling of alkaline phosphatase and penicillinase with colloidal gold in frozen thin sections of Bacillus licheniformis 749/C.

The subcellular distribution of alkaline phosphatase and penicillinase was determined by double labeling frozen thin sections of Bacillus licheniformis 749/C with colloidal gold-immunoglobulin G (IgG). Antipenicillinase and anti-alkaline phosphatase antibodies were used to prepare complexes with 5- and 15-nm colloidal gold particles, respectively. The character of the labeling of membrane-bound alkaline phosphatase and penicillinase was different: the immunolabels for alkaline phosphatase (15-nm particles) were bound to a few sites at the inner surface of the plasma membrane, and the gold particles formed clusters of various sizes at the binding sites; the immunolabels for penicillinase (5-nm particles), on the other hand, were bound to the plasma membrane in a dispersed and random fashion. In the cytoplasm, immunolabels for both proteins were distributed randomly, and the character of their binding was similar. The labeling was specific: pretreating the frozen thin sections with different concentrations of anti-alkaline phosphatase or penicillinase blocked the binding of the immunolabel prepared with the same antibody. Binding could be fully blocked by pretreatment with 800 micrograms of either antibody per ml.

Alkaline Phosphatase↗

Identification and penicillinase testing of Neisseria gonorrhoeae from primary isolation cultures on modified New York City medium.

The fluorescent-antibody test, rapid carbohydrate utilization test (RCUT), and a test for penicillinase production were performed on the bacterial growth from primary cultures on modified New York City medium. Of 134 gonococcal infections in men, 88.8% were diagnosed by the fluorescent-antibody test and 70.9% by the RCUT after incubation for 24 h; the corresponding figures for 75 infections in women were 86.7 and 54.7%, respectively. After incubation for 48 h, 100% of infections were diagnosed by the fluorescent-antibody test, wherease 88.8% of infected males and 86.7% of females were also diagnosed by the RCUT. Primary isolation cultures on modified New York City medium proved suitable for determining the ability of strains to produce penicillinase by a modified RCUT procedure. The method of isolation and identification by using primary cultures from modified New York City medium is both rapid and economical. The rapidity of diagnosis provided by the RCUT makes this a very useful diagnostic method, particularly in laboratories lacking immunofluorescence equipment. Since the penicillinase production test forms part of a routine identification procedure, no extra culture media or subcultures are required. The rapidity of this test should make it of value in tracing contacts of patients infected with penicillinase-producing strains.

Bacteriological Techniques↗

Penicillinase-producing Neisseria gonorrhoeae in Great Britain, 1977-81: alarming increase in incidence and recent development of endemic transmission.

Since penicillinase-producing Neisseria gonorrhoeae appeared five years ago in West Africa and South-east Asia reported cases have doubled annually in Great Britain, primarily as a result of increasing importation. Importation of penicillinase-producing Neisseria gonorrhoeae has increased exponentially because dramatic expansion of these strains in their regions of origin has led to increasing infection of male air travellers. From 1977 to 1980 infections acquired in Great Britain played only a minor part in the exponential increase. During 1981 the number of indigenous cases increased much more rapidly than imported cases, indicating that these strains have become truly endemic in Great Britain. Currently, identification of patients at high risk and initial treatment with penicillinase-resistant antibiotics offers the best hope of containing the strains. The emergence and rapid spread of penicillinase-producing Neisseria gonorrhoeae shows the international consequences of the abuse of antibiotics.

Adolescent↗

Simple assay and extraction of periplasmic penicillinase in Escherichia coli.

Benzylpenicillin was clearly separated from benzylpenicilloic acid by ascending chromatography on a diethylaminoethyl cellulose paper using 0.1 M ammonium acetate as a solvent. Using this chromatographic system, penicillinase was assayed by measuring the formation of [14C]benzylpenicilloic acid from [14C]benzylpenicillin. This assay remedies the lack of specificity of the commonly used iodometric assays. Periplasmic penicillinase was released from Escherichia coli by suspension in a mixture of 1% phenethyl alcohol and 5 mM ethylenediaminetetraacetate (pH 7.0). This simple extraction method not only facilitates the assay of penicillinase in an E. coli culture, but will also be useful for large-scale purification of periplasmic penicillinase.

Cytoplasm↗

Accumulation of alkaliphilic Bacillus penicillinase cleaved within the signal sequence in cytoplasm of Escherichia coli.

Alkaliphilic Bacillus penicillinase produced by Escherichia coli is distributed in several subcellular compartments according to cultivation conditions. The penicillinase that accumulated in particular subcellular fractions of E. coli grown under different conditions was purified and characterized. Periplasmic or extracellular penicillinase (24 kDa) was mature protein, indicating that the putative precursor (27 kDa) was processed at the correct amino acid residue, probably by signal peptidase I. Cytoplasmic penicillinase contained two unusual proteins (25 kDa) that are produced by proteolytic cleavage of the precursor within its signal sequence.

Amino Acid Sequence↗

Gonorrhoea caused by a mixture of penicillinase-positive and -negative gonococci: a case report.

A 23-year-old unmarried man was infected with gonorrhoea from a prostitute in Fukuoka City and was treated with ampicillin which resulted in failure. By a rapid iodometric test it was found that the isolates before the treatment were penicillinase-negative. After the ampicillin treatment, however, the isolates turned to penicillinase-positive but were, in fact, a mixture of penicillinase-positive and penicillinase-negative gonococcal strains. Treatment by spectinomycin and doxycycline resulted in failure but the gonorrhoea was cured by ribostamycin.

Adult↗

A study of synergism between cloxacillin and gentamicin on resistant staphylococci (penicillinase producing and gentamicin resistant).

Twelve strains of Staphylococcus aureus were studied. All the strains were tested for penicillinase production by the iodometric and starch-paper methods. Ten out of 12 strains produced penicillinase. A strain which was gentamicin resistant and penicillinase producer was tested for synergism using the killing growth method. Synergism was observed between cloxacillin and gentamicin. These two antibiotics can now be used to treat infection by Staphylococcus aureus especially those that produce penicillinase and gentamicin resistant.

Cloxacillin↗

Modification and processing of Bacillus licheniformis prepenicillinase in Escherichia coli. Fate of mutant penicillinase lacking lipoprotein modification site.

We have previously shown that Bacillus licheniformis prepenicillinase is modified and processed to form membrane-bound penicillinase in Escherichia coli which contains N-acylglyceride-cysteine27 at the NH2 terminus. In the present study, we have constructed, by in vitro site-directed mutagenesis, two mutant penicillinase genes in which the modification site (the 27th cysteine residue in prepenicillinase) is either converted into serine (penPSer27) or is deleted along with the preceding four residues (Ala23 to Cys27, delta penP2327). The modification, processing, and subcellular localization of these two mutant penicillinases in E. coli cells were studied. Our results indicate that the delta penP2327 deletion mutant prepenicillinase is largely metabolically inert and the unmodified and uncleaved form is associated with the membrane fraction; a small fraction (about 7-9%) appears to contain glyceride-modified prepenicillinase (presumably at the Cys-21 position) which is not cleaved. In contrast, the Cys-27 in equilibrium Ser-27 point mutant prepenicillinase is processed into two forms which contain Asn-29 and Ser-35 at their NH2 termini, respectively, and the bulk of the processed penicillinase appears to be located in the peri-plasm. These results are discussed in terms of the substrate specificities of signal peptidases in E. coli.

Amino Acid Sequence↗

[Is methicillin better than cloxacillin in serious infections caused by strong penicillinase-producing staphylococci (phage-type 94/96)?].

An intravenous drug addict was treated with cloxacillin for septicaemia with Staphylococcus aureus because of pneumonia and suspected endocarditis. After 51 days of treatment Staphylococcus aureus was still found in blood and expectorate despite continued treatment with intravenous cloxacillin 1 g three and later four times daily and oral rifampicin. The staphylococcal isolates were all of phage type 94/96. Investigations have shown that Staphylococci aurei of phage type 94/96 produce large amounts of penicillinase, and that methicillin is the most penicillinase-resistant of the penicillinase-resistant penicillins followed by dicloxacillin and cloxacillin. The penicillinase production of the patient's Staphylococcus aureus strain was 304-362 units per mg bacteria which is high compared to typical values of 50-200. After 50 days of cloxacillin treatment, the treatment was changed to methicillin 2 g four times daily. Within a week the staphylococci had disappeared from the expectorate, and were never again recovered from the blood. It is suggested that methicillin should have superior efficiency in serious infections with Staphylococcus aureus of phage type 94/96.

Cloxacillin↗

Penicillinase as a potential enzyme marker for enzyme linked immunosorbent assay and its application in immunodiagnostics.

The present review article deals with the application of penicillinase in enzyme linked immunosorbent assay (ELISA) and describes the various characteristics of penicillinase. Using penicillinase as an enzyme marker, few kits have been developed to detect physiologically active proteins, infectious diseases and toxins. The review also highlights briefly the source of penicillinase, its production, isolation, purification and certain molecular properties.

Amino Acid Sequence↗

Penicillinase-resistant penicillin/gentamicin synergism. Effect in patients with Staphylococcus aureus bacteremia.

Gentamicin sulfate has been shown to enhance the effects of penicillinase-resistant penicillins against clinical isolates of Staphylococcus aureus in vitro, but the relevance of this observation to bacteremic patients is unclear. Therefore, serum samples from 14 patients with staphylococcal bacteremia were tested for growth inhibitory and bactericidal effects against the patients' own pathogen while they received a penicillinase-resistant penicillin alone, and again when gentamicin was added to the therapeutic regimen. Addition of gentamicin in vivo was associated with slight improvement in growth inhibitory activity but it caused profound increases in serum bactericidal activity. This effect could not be attributed to higher serum levels of penicillinase-resistant penicillin during gentamicin administration. Addition of gentamicin to penicillinase-resistant penicillin can lead to marked improvement in a patient's serum bactericidal activity against his own staphylococcal pathogen, and should be considered for any patient who does not respond to more conventional therapy.

Blood Bactericidal Activity↗

PURIFICATION AND PROPERTIES OF PENICILLINASES FROM TWO STRAINS OF BACILLUS LICHENIFORMIS: A CHEMICAL, PHYSICOCHEMICAL AND PHYSIOLOGICAL COMPARISON.

1. The penicillinases formed by penicillinase-constitutive mutant strains from two closely related varieties (749 and 6346) of Bacillus licheniformis have been isolated, characterized and compared. They are chemically, physicochemically and immunologically very similar, but differ enzymologically in absolute and relative activity on, and affinity for, different penicillins and cephalosporins. 2. The molecular weights of both types are approx. 23000. Neither enzyme contains any cyst(e)ine. However, in most other respects they show little resemblance to any of the other penicillinases so far isolated. 3. Their properties, whether isolated from cells (to which approx. 50% of the activity is normally bound) or from the culture supernatant, appear to be similar. However, the molecular weight of a preparation of enzyme from strain 749/C obtained from the culture supernatant was found to be significantly (over 20%) higher than that obtained from cells alone. 4. With benzylpenicillin, the enzyme from strain 749 has V(max.) approx. 6 times higher than that of the enzyme from strain 6346, but this difference is ;compensated' by its affinity being 6 times lower. Thus, at the very low biologically effective concentrations of penicillin met with under natural conditions, where neither type of enzyme is more than a fraction saturated with its substrate, the antibiotic is hydrolysed at the same rate by both. As expected, the penicillin-sensitivities of single cells from the two strains were found to be identical. 5. It is suggested that the concept of ;physiological efficiency' (defined as V(max.) divided by K(m)), applied to enzymes acting naturally under conditions of poor saturation with their substrates, may be useful for expressing their biological function in vivo.

Amino Acids↗

DRUG RESISTANCE OF STAPHYLOCOCCI. II. JOINT ELIMINATION AND JOINT TRANSDUCTION OF THE DETERMINANTS OF PENICILLINASE PRODUCTION AND RESISTANCE TO MACROLIDE ANTIBIOTICS.

Mitsuhashi, Susumu (Gunma University, Maebashi, Japan), Hajime Hashimoto, Megumi Kono, and Masato Morimura. Drug resistance of staphylococci. II. Joint elimination and joint transduction of the determinants of penicillinase production and resistance to macrolide antibiotics. J. Bacteriol. 89:988-992. 1965.-Strains of Staphylococcus aureus, which show high resistance to macrolide antibiotics (erythromycin, oleandomycin, leucomycin, and spiramycin) and the capacity to produce penicillinase, have been isolated from clinical sources. The determinants of penicillinase production (PCase(+)) and resistance to macrolide antibiotics (MAC(r)) of these strains were irreversibly eliminated by treatment with acridine or with ultraviolet light. Among the 18 strains tested, PCase(+) and MAC(r) were eliminated from all strains except one, which lost only PCase(+) but not MAC(r). The characters PCase(+) and MAC(r) were jointly transduced with the aid of phage lysates, obtained from the resistant donors by ultraviolet irradiation, into staphylococcal strains sensitive to PC and MAC. Segregation of PCase(+) and MAC(r) was rarely observed after transduction. From these results, it is suggested that the determinants of both PCase(+) and MAC(r) of staphylococci are located close together on a single genetic element, i.e., a plasmid (or episome), which exists extrachromosomally.

Anti-Bacterial Agents↗

Preparation and properties of an immobilized derivative of penicillinase.

Penicillinase (beta-lactamase I, EC 3.5.2.6) secreted by Bacillus cereus, strain 569/H, was covalently attached to aminoethyl cellulose via glutaraldehyde. The immobilized derivative shows increased thermostability and decreased susceptibility to conformational changes induced by certain substrates of penicillinase. The decline in the rate of hydrolysis of such substrates was consequently suppressed by immobilization. A marked increase in Km was observed with all substrates except for the unsubstituted 6-aminopenicillanic acid. The altered properties of the new derivative are attributed to the constraint imposed by immobilization on the conformational flexibility of the enzyme molecule. Thus, apart from obvious technological interest, immobilized penicillinase provides a useful model for the study of the role of flexibility in the function of an enzyme.

Bacillus cereus↗

Vesicle penicillinase of Bacillus licheniformis 749/C. Apparent identity with the plasma membrane enzyme.

A substantial fraction of the total membrane penicillinase of Bacillus licheniformis 749/C is attached to the vesicles released during conversion of the cells to protoplasts. This enzyme was purified since there was indirect evidence that it differed from the enzyme that remained with the protoplast. The purified vesicle penicillinase has the same molecular weight and general properties as the plasma membrane (protoplast) enzyme and, similarly, contains a covalently linked phosphatidylserine residue. Treatment of the two enzymes with trypsin produced phosphatidylserine-containing peptides which could not be distinguished by gel or paper electrophoresis. The two membrane penicillinases are very similar, if not identical.

Bacillus↗

A sensitive enzyme-linked immunosorbent assay (ELISA) for testosterone: use of a novel heterologous hapten conjugated to penicillinase.

A microplate enzyme-linked immunosorbent assay (ELISA) has been developed for the measurement of testosterone in plasma. The assay uses a heterologous system consisting of a novel hapten 4-(17 beta-hydroxy-3-oxoestra-4,9-dien-11 beta-yl)butanoic acid (1) conjugated to penicillinase (beta-lactamase). The key reaction in the synthesis of the hapten was the cuprate-mediated 1,4-conjugate addition on 3,3,17,17-bis-ethylenedioxy-5 alpha,10 alpha-oxido-estr-9(11)-ene by the Grignard reagent derived from trimethyl 4-bromoorthobutyrate; this regiospecifically introduces the 11 beta-butanoate function. The hapten-penicillinase conjugate was used in the assay in conjunction with the immunoglobulin G (IgG) fraction derived from a previously characterized, highly specific, antitestosterone serum raised against a testosterone-19-O-carboxymethyl ether-bovine serum albumin (T-19-O-CME-BSA) conjugate. This unique system represents one incorporating three elements of structural heterology: bridge, site, and ring heterology between the antigen hapten and enzyme-linked hapten. The limit of detection was 10 pg of testosterone with a sensitivity range between 15 and 1,000 pg. A low level of cross-reactivity with 5 alpha-dihydrotestosterone (6.17%) and 11 beta-hydroxytestosterone (1.03%) was noted. No interference was noted with other common androgens, estradiol, or progesterone. The sensitivity and selectivity observed in the assay may be attributable to the selection of penicillinase as the enzyme marker and the elements of conformational heterology between the antigen-linked and enzyme-conjugated steroid haptens.

Enzyme-Linked Immunosorbent Assay↗