Biomaterial Database

[Molecular characteristics of beta-lactamase from Klebsiella pneumoniae]. 2002

Yonglong Zhang, and Jiatai Li, and Mingwu Zhao

Institute of Clinical Pharmacology, Peking University First Hospital, Beijing 100083, China.

OBJECTIVE To study the molecular characteristics of beta-lactamase from Klebsiella pneumoniae. METHODS Beta-lactamase was prepared from strains of Klebsiella pneumoniae isolated clinically for which the minimal inhibitory concentration (MIC) of cefoperazone had been >or= 8 mg/L and then was reduced by 50% by the addition of sulbatam. The isoelectric points of different beta -lactamases were examined by LKB2117 Multiphor II Electrophoresis System. The molecular weights were examined by polyacrylamide gel electrophoresis. PCR was used to detect the TEM-type genes. The PCR products were sequenced by chemiluminescence. RESULTS All four strains of Klebsiella pneumoniae produced more than two kinds of beta-lactamase. TEM-type genes were amplified from all strains. 97% - 98% of the DNA sequence and amino acid sequence of the TEM-type enzymes from 2 strains of Klebsiella pneumoniae were identical with those of TEM-1 enzyme. The isoelectric points of the beta-lactamases ranged from 5.4 to 9.30 and the molecular weights were between 23.0 and 43.0 KD. The beta-lactamases from Klebsiella pneumoniae 99592 and K. pneumoniae 99607 hydrolysed ceftazidine, cefotaxime, and ceftriaxone. The beta-lactamase from K. pneumoniae 99595 had stronger activity against ceftazidime, ceftaxime, and ceftriaxone than that from K. pneumoniae 99607. CONCLUSIONS Klebsiella pneumoniae produces TEM-type extended spectrum beta -lactamase against ceftazidine, ceftaxime, and ceftriaxone.

UI	MeSH Term	Description	Entries
D007526	Isoelectric Point	The pH in solutions of proteins and related compounds at which the dipolar ions are at a maximum.	Isoelectric Points,Point, Isoelectric,Points, Isoelectric
D007711	Klebsiella pneumoniae	Gram-negative, non-motile, capsulated, gas-producing rods found widely in nature and associated with urinary and respiratory infections in humans.	Bacillus pneumoniae,Bacterium pneumoniae crouposae,Hyalococcus pneumoniae,Klebsiella pneumoniae aerogenes,Klebsiella rhinoscleromatis
D008970	Molecular Weight	The sum of the weight of all the atoms in a molecule.	Molecular Weights,Weight, Molecular,Weights, Molecular
D001618	beta-Lactamases	Enzymes found in many bacteria which catalyze the hydrolysis of the amide bond in the beta-lactam ring. Well known antibiotics destroyed by these enzymes are penicillins and cephalosporins.	beta-Lactamase,beta Lactamase,beta Lactamases
D016133	Polymerase Chain Reaction	In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.	Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain