A Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo- beta-Lactamase

[en] New Delhi metallo- beta-lactamase 1 (NDM-1) is a subclass B1 metallo-beta - lactamase that exhibits a broad spectrum of activity against beta -lactam antibiotics. Here we report the kinetic study of 6 Q119X variants obtained by site-directed mutagenesis of NDM-1. All Q119X variants were able to hydrolyze carbapenems, penicillins and first-, second-, third-, and fourth-generation cephalosporins very efficiently. In particular, Q119E, Q119Y, Q119V, and Q119K mutants showed improvements in kcat/Km values for penicillins, compared with NDM-1. The catalytic efficiencies of the Q119K variant for benzylpenicillin and carbenicillin were about 65- and 70-fold higher, respectively, than those of NDM-1. The Q119K and Q119Y enzymes had kcat/Km values for ceftazidime about 25- and 89-fold higher, respectively, than that of NDM-1.

Disciplines :

Biochemistry, biophysics & molecular biology

Author, co-author :

Marcoccia, Francesca

Mercuri, Paola ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques

Galleni, Moreno ; Université de Liège - ULiège > Département des sciences de la vie > Macromolécules biologiques

Celenza, Giuseppe

Amicosante, Gianfranco

Perilli, Mariagrazia

Language :

English

Title :

A Kinetic Study of the Replacement by Site Saturation Mutagenesis of Residue 119 in NDM-1 Metallo- beta-Lactamase

Publication date :

2018

Journal title :

Antimicrobial Agents and Chemotherapy

ISSN :

0066-4804

eISSN :

1098-6596

Publisher :

American Society for Microbiology, United States - District of Columbia

Volume :

Issue :

Peer reviewed :

Peer Reviewed verified by ORBi

Available on ORBi :

since 10 August 2018

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Bibliography

Groundwater PW, Xu S, Lai F, Váradi L, Tan J, Perry JD, Hibbs DE. 2016. New Delhi metallo--lactamase-1: structure, inhibitors and detection of producers. Future Med Chem 8:993–1012. https://doi.org/10.4155/fmc-2016-0015.
Wang LH, Liu PP, Wei DD, Liu Y, Wan LG, Xiang TX, Zhang YJ. 2016. Clinical isolates of uropathogenic Escherichia coli ST131 producing NDM-7 metallo--lactamase in China. Int J Antimicrob Agents 48:41–45. https://doi.org/10.1016/j.ijantimicag.2016.03.009.
Compain F, Vandenberghe A, Gominet M, Genel N, Lebeaux D, Rama-hefasolo A, Podglajen I, Decré D. 2017. Primary osteomyelitis caused by an NDM-1-producing K. pneumoniae strain of the highly virulent sequence type 23. Emerg Microbes Infect 6:e57. https://doi.org/10.1038/emi.2017.43.
Khan AU, Maryam L, Zarrilli R. 2017. Structure, genetics and worldwide spread of New Delhi metallo--lactamase (NDM): a threat to public health. BMC Microbiol 17:101–112. https://doi.org/10.1186/s12866-017-1012-8.
Toleman M, Karthikeyan K, Sharma M, Chaudhary U, Thirunarayan M, Krishnan P, Walsh T. 2010. Recent epidemic emergence of blaNDM-1 metallo--lactamase in enteric organisms from India is mostly linked to A/C plasmids. Clin Microbiol Infect 16(Suppl 2):S30.
Bush K. 2010. Alarming -lactamase-mediated resistance in multidrug-resistant Enterobacteriaceae. Curr Opin Microbiol 13:558 –564. https://doi.org/10.1016/j.mib.2010.09.006.
Yong D, Toleman MA, Giske CG, Cho HS, Sundman K, Lee K, Walsh TR. 2009. Characterization of a new metallo--lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother 53:5046–5054. https://doi.org/10.1128/AAC.00774-09.
King D, Strynadka N. 2011. Crystal structure of New Delhi metallo--lactamase reveals molecular basis for antibiotic resistance. Protein Sci 20:1484–1491. https://doi.org/10.1002/pro.697.
Zhang HM, Hau Q. 2011. Crystal structure of NDM-1 reveals a common -lactam hydrolysis mechanism. FASEB J 25:2574–2582. https://doi.org/10.1096/fj.11-184036.
Kim Y, Cunningham MA, Mire J, Tesar C, Sacchettini J, Joachimiak A. 2013. NDM-1, the ultimate promiscuous enzyme: substrate recognition and catalytic mechanism. FASEB J 27:1917–1927. https://doi.org/10 .1096/fj.12-224014.
Guo Y, Wang J, Niu G, Shui W, Sun Y, Zhou H, Zhang Y, Yang C, Lou Z, Rao Z. 2011. A structural view of the antibiotic degradation enzyme NDM-1 from a superbug. Prot Cell 2:384–394. https://doi.org/10.1007/s13238-011-1055-9.
Garau G, Garcia-Sàez I, Bebrone C, Anne C, Mercuri PS, Galleni M, Frère JM, Dideberg O. 2004. Update of the standard numbering scheme for class B -lactamases. Antimicrob Agents Chemother 48:2347–2349. https://doi.org/10.1128/AAC.48.7.2347-2349.2004.
Chen J, Chen H, Zhu T, Zhou D, Zhang F, Lao X, Zheng H. 2014. Asp120Asn mutation impairs the catalytic activity of NDM-1 metallo--lactamase: experimental and computational study. Phys Chem Chem Phys 16:6709–6716. https://doi.org/10.1039/c3cp55069a.
Ho SN, Hunt HD, Horton RM, Pullen JK, Pease LR. 1989. Site-directed mutagenesis by overlap extension using a polymerase chain reaction. Gene 77:51–59. https://doi.org/10.1016/0378-1119(89)90358-2.
Marcoccia F, Bottoni C, Sabatini A, Colapietro M, Mercuri PS, Galleni M, Kerff F, Matagne A, Celenza G, Amicosante G, Perilli M. 2016. Kinetic study of laboratory mutants of NDM-1 metallo--lactamase and the importance of an isoleucine at position 35. Antimicrob Agents Chemother 60:2366–2372. https://doi.org/10.1128/AAC.00531-15.
Clinical and Laboratory Standards Institute. 2012. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically; approved standard— 9th ed. Document M07-A9. Clinical and Laboratory Standards Institute, Wayne, PA.
Segel IH. 1976. Enzyme kinetics. Biochemical calculations: how to solve mathematical problems in general biochemistry, 2nd ed, p 236–241. John Wiley & Sons, New York, NY.
Chiou J, Leung TY, Chen S. 2014. Molecular mechanisms of substrate recognition and specificity of New Delhi metallo--lactamase. Antimicrob Agents Chemother 58:5372–5378. https://doi.org/10.1128/AAC.01977-13.
De Meester F, Joris B, Reckinger G. 1987. Automated analysis of enzyme inactivation phenomena: application to -lactamases and DD-peptidases. Biochem Pharmacol 36:2393–2403. https://doi.org/10.1016/0006-2952(87)90609-5.