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Abstract
Data revealed that only six isolates had high level of MIC ranging from 1500 to 7000 µg/ml & had the ability to produce B-lactamase activity but they differ in their levels of activities. There was one isolate which gave the highest level of resistance and this isolate produced the highest ẞ-lactamase activity
The B-lactamase enzyme of the selected isolate (S. aureus) precipitated by using ammonium sulphate (10 + (- 90)%) , purified by using DEAE- cellulose & sephadex G- 200 columns. The purified enzyme was subjected to SDS -PAGE to determined its molecular weight. The enzyme contained different amino acids and is rich in cysteine & alanine.
The genetic analysis of this isolate was studied using specific primers & PCR. Data revealed that genes responsible for resistance are blaZ & meca, localized on chromosome
Bacterial pathogens resistant to various ẞ-lactam antibiotics caused an important problem in the clinical flied. Therefore our study aimed to overcome this problem by screening for streptomycetes isolates which have the ability to produce B-lactamase inhibitory protein. Streptomycetes isolates ( Fs - 3 . Fs - 6 , Fs - 18 Fs - 20 proved to be the most active organisms to the other isolates.
Data revealed that maximum yield of active protein from these isolates was achieved by growing it on fishmeal extract medium incubated at 28 °C for five days under submerged condition. The isolate which produced the highest inhibitory protein was selected and the active protein was then purified.
SDS -PAGE revealed that its molecular weight is 48 KDa and MIC of the tested organism decreased from 7000 µg/ml to 1125 µg/ ml.