Abstract
An SHV β-lactamase gene was amplified from a β-lactam resistant Klebsiella pneumoniae K-71 genomic DNA. After expression and purification, we demonstrated that peptide P1 could inhibit the hydrolysis activity of both TEM-1 and SHV β-lactamase in vitro. Three mutations were introduced into P1 in which the first residue S was replaced by F, the 18th residue V was mutated to Y, and the 15th residue Y was substituted with A, C, G, and R to obtain the mutants of P1-A, P1- C, P1-G, and P1-R, respectively. The mutant peptides were purified and their inhibitory constants against TEM-1 and SHV β-lactamase were determined. All these β-lactamase inhibitory peptides could inhibit the activity of both β-lactamases, while the mutant peptides showed stronger inhibitory activities against TEM-1 β-lactamase than against SHV β-lactamase. Inhibition data suggested that P1-A improved the β-lactamase inhibitory activity by over 3-fold compare to P1. When P1-A was incubated with K. pneumoniae K-71 in Luria-Bertani medium containing ampicillin, it showed a much stronger growth of inhibition ratio over P1. This study gives us a good candidate for development of novel β-lactamase inhibitors.
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Abbreviations
- BLIP:
-
β-lactamase inhibitory peptide
- IPTG:
-
isopropyl-β-D-thiogalactopyranoside
- MIC:
-
minimal inhibitory concentration
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Published in Russian in Biokhimiya, 2010, Vol. 75, No. 3, pp. 411–417.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM09-258, February 14, 2010.
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Xie, L., Xu, M., Yang, T. et al. Studies on amino acid replacement and inhibitory activity of a β-lactamase inhibitory peptide. Biochemistry Moscow 75, 336–341 (2010). https://doi.org/10.1134/S0006297910030107
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DOI: https://doi.org/10.1134/S0006297910030107