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11.
Christine Jacobs Alain Dubus Didier Monnaie Staffan Normark Jean-Marie Frère 《FEMS microbiology letters》1992,92(1):95-100
The involvement of the serine residue 318 in the specificity of a class C beta-lactamase was investigated. Multiple site-directed mutants at this position were generated using a polymerase chain reaction technique. These mutants were then probed for their activity towards various beta-lactam compounds. One mutant, S318G was further purified and its physico-chemical and catalytic properties determined. It was shown that the observed minimal inhibitory concentration values of this mutant could be correlated to its kinetic properties using a 'diffusion-hydrolysis' model. However, the data showed that residue 318 has little influence on the specificity of class C beta-lactamases. 相似文献
12.
Juana Magdalena Mats Forsman Mauro V. Lenzini Alain Brans Jean Dusart 《FEMS microbiology letters》1992,99(1):101-105
Two beta-lactamase genes called blaL and blaU have been cloned independently in Liège and in Ume?, from Streptomyces cacaoi. Genes blaL and blaU were found to differ largely in their nucleotide sequences, although the encoded proteins both belonged to the class A of beta-lactamases (active-site serine penicillinases). DNA-hybridization and polymerase chain reaction assays have now demonstrated that both blaL and blaU genes were present in the S. cacaoi strains used in Liège and in Ume?. 相似文献
13.
Purification and biochemical characterization of an endoxylanase from Aspergillus versicolor 总被引:1,自引:0,他引:1
Eleonora Cano Carmona Marcia Regina Brochetto-braga Aline Aparecida Pizzirani-kleiner João Atilio Jorge 《FEMS microbiology letters》1998,166(2):311-315
An endoxylanase (β-1,4-xylan xylanohydrolase, EC 3.2.1.8) was purified from the culture filtrate of a strain of Aspergillus versicolor grown on oat wheat. The enzyme was purified to homogeneity by chromatography on DEAE-cellulose and Sephadex G-75. The purified enzyme was a monomer of molecular mass estimated to be 19 kDa by SDS-PAGE and gel filtration. The enzyme was glycoprotein with 71% carbohydrate content and exhibited a pI of 5.4. The purified xylanase was specific for xylan hydrolysis. The enzyme had a K m of 6.5 mg ml−1 and a V max of 1440 U (mg protein)−1 . 相似文献
14.
Concepción González-Bello 《Bioorganic & medicinal chemistry letters》2017,27(18):4221-4228
Resistance to available antibiotics in pathogenic bacteria is currently a global challenge since the number of strains that are resistant to multiple types of antibiotics has increased dramatically each year and has spread worldwide. To unlock this problem, the use of an ‘antibiotic adjuvant’ in combination with an antibiotic is now being exploited. This approach enables us to prolong the lifespan of these life-saving drugs. This digests review provides an overview of the main types of antibiotic adjuvants, the basis of their operation and the remaining issues to be tackled in this field. Particular emphasis is placed on those compounds that are already in clinical development, namely β-lactamase inhibitors. 相似文献
15.
Cephalosporin was used to synthesize soluble and precipitating fluorogenic β-lactam substrates that demonstrated differential catalytic hydrolysis by three different subtypes of β-lactamase: TEM-1 (class A), p99 (class C), and a Bacillus cereus enzyme sold by Genzyme (class B). The most successful soluble substrate contained difluorofluorescein (Oregon Green 488) ligated to two cephalosporin moieties that, therefore, required two turnovers to produce the fluorescent Oregon Green 488 leaving group. The bis-cephalosporin modification was required so that the final reaction product was the Oregon Green 488 carboxylic acid rather than a less bright phenolic adduct of the dye. Hydrolysis in pH 5.5 Mes and pH 7.2 phosphate-buffered saline (PBS) buffers was similar, but in pH 8.0 Tris the hydrolysis rate nearly doubled. Activity of the β-lactamases on the various substrates was shown to depend highly on the linker between the cephalosporin and the fluorophore, with an allyl linker promoting faster turnover than a phenol ether linker. Measured Km values for dichlorofluorescein and difluorofluorescein cephalosporin substrates were approximately the same as Km values for penicillin G and ampicillin found in the literature (∼30–40 μM). 相似文献
16.
Anil D. Naik Marinela M. Dîrtu Bernard Tinant Jacqueline Marchand-Brynaert 《Inorganica chimica acta》2011,368(1):21-4043
Coordinating abilities of 4R-1,2,4-triazole derivatives (R = glycine ethyl ester (L1), glycine (L2), diethylamino malonate (L3), methionine (L4) and diethyl aminomethylphosphonate (L5)) towards ZnII ions have been studied in solution, in solid state and versus three zinc-β-lactamases. The crystal structure of [Zn3(L4)6(H2O)6] (6) is described; it is the first crystal structure involving a 1,2,4-triazole functionalized methionine. It forms a trinuclear complex with central zinc octahedrally coordinated by only L4, whereas terminal zinc ions coordination sphere is completed by three water molecules. L4 exhibits a dual functionality of a bridging bidentate ligand as well as an anion. A dense hydrogen bonding network connects these trinuclear entity into a 3D supramolecular network. The ZnII ions in 6 are held at equidistance (3.848 Å) which coincidently matches with the corresponding Zn?Zn distance in the binuclear zinc enzyme from Bacillus cereus (3.848 and 4.365 Å). Among L1-L5 screened for β-lactamase assay, L4 shows modest inhibition for BcII enzyme. 相似文献
17.
Christian Therrien Lakshmi P. Kotra Franois Sanschagrin Shahriar Mobashery Roger C. Levesque 《FEBS letters》2000,470(3)
Characterization of the biochemical steps in the inactivation chemistry of clavulanic acid, sulbactam and tazobactam with the carbenicillin-hydrolyzing β-lactamase PSE-4 from Pseudomonas aeruginosa is described. Although tazobactam showed the highest affinity to the enzyme, all three inactivators were excellent inhibitors for this enzyme. Transient inhibition was observed for the three inactivators before the onset of irreversible inactivation of the enzyme. Partition ratios (kcat/kinact) of 11, 41 and 131 were obtained with clavulanic acid, tazobactam and sulbactam, respectively. Furthermore, these values were found to be 14-fold, 3-fold and 80-fold lower, respectively, than the values obtained for the clinically important TEM-1 β-lactamase. The kinetic findings were put in perspective by determining the computational models for the pre-acylation complexes and the immediate acyl-enzyme intermediates for all three inactivators. A discussion of the pertinent structural factors is presented, with PSE-4 showing subtle differences in interactions with the three inhibitors compared to the TEM-1 enzyme. 相似文献
18.
19.
Abstract β-Lactamase genes of class-A ( Rtem ) and class-C ( ampC ) were placed under control of an inducible tac -promoter and expressed in Escherichia coli . Expression of RTEM had no observable effect on the growth properties of E. coli strains HB101 ( ampC + ) or MI1443 (Δ ampC ). E. coli MI1443 exhibited a decline in growth rate at mid-exponential phase which could be delayed by expression of AmpC at early-exponential phase. AmpC expression otherwise inhibited growth, particularly during the transition into exponential phase where growth was prevented altogether. We suggest that the AmpC β-lactamase, but not RTEM, may have an additional cellular function as a peptidoglycan hydrolase. 相似文献
20.
Abstract Aztreonam-resistant Klebsiella oxytoca strain SL7811 was selected on agar containing 1 μg of aztreonam per ml from a susceptible strain SL781. The MICs for the resistant mutant towards penicillins, aztreonam and ceftriaxone were much higher, to cefotaxime slightly higher and to ceftazidime unchanged. Synthesis of β-lactamase was 223-fold greater in the mutant compared with the susceptible strain. SL781 and its resistant mutant SL7811 produced β-lactamase with the same isoelectric point and substrate profile. The β-lactamase genes from SL781 and SL7811 were cloned in plasmid pBGS18 giving pBOF-1 and pBOF-4 respectively. The sequences of the two putative promoters indicated two modifications in the resistant plasmid pBOF-4: a transversion (G → T) in the first base of the − 10 consensus sequence and a deletion of one C residue four base pairs upstream of the − 10 hexamer. 相似文献