Title : The effect of high-frequency random mutagenesis on in vitro protein evolution: a study on TEM-1 beta-lactamase.

Pub. Date : 1999 Jan 15

PMID : 9878443






3 Functional Relationships(s)
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1 Here we report the results of an experiment in which we have hypermutated the bacterial enzyme TEM-1 beta-lactamase and selected small pools (<1.5x10(5)) of clones for enzymatic activity against the beta-lactam antibiotic cefotaxime. Cefotaxime CD248 molecule Homo sapiens
2 The experiment resulted in the isolation of a number of TEM-1 mutants with greatly improved activity against cefotaxime. Cefotaxime CD248 molecule Homo sapiens
3 Among these, clone 3D.5 (E104K:M182T:G238S) exhibited a minimum inhibitory concentration for cefotaxime 20,000-fold higher than wild-type TEM-1 and a catalytic efficiency (kcat/Km) 2383 times higher than the wild-type enzyme. Cefotaxime CD248 molecule Homo sapiens