In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect
Antibiotic resistance is a global threat. The Class D ?-lactamase OXA-143 has been described as an efficient penicillinase, oxacillinase, and carbapenemase. This Class of enzymes has an unusual lysine (K84) that in the carbamylated form acts as a general base. Recent studies performed in our laborat...
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| Formato: | Online |
| Idioma: | português |
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Universidade Estadual de Campinas
2019
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| _version_ | 1864359038038310912 |
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| author | Costa, Luiz Favaro, Denize Antunes, Víctor |
| author_facet | Costa, Luiz Favaro, Denize Antunes, Víctor |
| author_sort | Costa, Luiz |
| collection | Portal de Eventos Científicos da UNICAMP |
| container_reference | Revista dos Trabalhos de Iniciação Científica da UNICAMP; n. 27 (2019): Congresso de Iniciação Científica UNICAMP; 1-1 |
| description | Antibiotic resistance is a global threat. The Class D ?-lactamase OXA-143 has been described as an efficient penicillinase, oxacillinase, and carbapenemase. This Class of enzymes has an unusual lysine (K84) that in the carbamylated form acts as a general base. Recent studies performed in our laboratory showed that P227S is a positive mutation within the evolutionary pathway of the OXA-143 sub-family towards a more efficient carbapenemase. Once the mutant OXA-143(P227S) is an evolved version of OXA-143 it is important to analyze the effect of the buffer for the kinetic in vitro assays. Therefore, as the carboxylated lysine acts as a general base in the proposed covalent catalysis mechanism, in this study, we performed assays to investigate the effect of the pH in the structure and catalytic efficiency of the mutant OXA-143(P227S). Our results show a direct pH dependence on penicillinase activity, with an optimal range from pH 7 to 9. In addition, our findings also suggest the exchange of two hydrogens during the reaction with ampicillin. Furthermore, it was also observed a direct correlation between the catalytic efficiency in this pH range and the enzyme thermal stability. |
| first_indexed | 2025-10-08T13:57:00Z |
| format | Online |
| id | ojs-article-2296 |
| institution | Universidade Estadual de Campinas |
| issn | 2596-1969 |
| language | por |
| last_indexed | 2025-10-08T13:57:00Z |
| publishDate | 2019 |
| publisher | Universidade Estadual de Campinas |
| record_format | ojs |
| spelling | ojs-article-22962025-09-26T14:39:16Z In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect Costa, Luiz Favaro, Denize Antunes, Víctor Antibiotic resistance Carbapenem-hydrolyzing Class D β-lactamase Enzyme kinetics Antibiotic resistance is a global threat. The Class D ?-lactamase OXA-143 has been described as an efficient penicillinase, oxacillinase, and carbapenemase. This Class of enzymes has an unusual lysine (K84) that in the carbamylated form acts as a general base. Recent studies performed in our laboratory showed that P227S is a positive mutation within the evolutionary pathway of the OXA-143 sub-family towards a more efficient carbapenemase. Once the mutant OXA-143(P227S) is an evolved version of OXA-143 it is important to analyze the effect of the buffer for the kinetic in vitro assays. Therefore, as the carboxylated lysine acts as a general base in the proposed covalent catalysis mechanism, in this study, we performed assays to investigate the effect of the pH in the structure and catalytic efficiency of the mutant OXA-143(P227S). Our results show a direct pH dependence on penicillinase activity, with an optimal range from pH 7 to 9. In addition, our findings also suggest the exchange of two hydrogens during the reaction with ampicillin. Furthermore, it was also observed a direct correlation between the catalytic efficiency in this pH range and the enzyme thermal stability. Universidade Estadual de Campinas 2019-01-14 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artigo de convidado application/pdf https://econtents.sbu.unicamp.br/eventos/index.php/pibic/article/view/2296 10.20396/revpibic2720192296 Revista dos Trabalhos de Iniciação Científica da UNICAMP; n. 27 (2019): Congresso de Iniciação Científica UNICAMP; 1-1 2596-1969 por https://econtents.sbu.unicamp.br/eventos/index.php/pibic/article/view/2296/2346 Copyright (c) 2019 Revista dos Trabalhos de Iniciação Científica da UNICAMP http://creativecommons.org/licenses/by/4.0 |
| spellingShingle | Costa, Luiz Favaro, Denize Antunes, Víctor Antibiotic resistance Carbapenem-hydrolyzing Class D β-lactamase Enzyme kinetics In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect |
| title | In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect |
| title_full | In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect |
| title_fullStr | In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect |
| title_full_unstemmed | In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect |
| title_short | In vitro penicillinase activity of ?-lactamase OXA-143(P227S): pH effect |
| title_sort | in vitro penicillinase activity of ?-lactamase oxa-143(p227s): ph effect |
| topic | Antibiotic resistance Carbapenem-hydrolyzing Class D β-lactamase Enzyme kinetics |
| url | https://econtents.sbu.unicamp.br/eventos/index.php/pibic/article/view/2296 |
| work_keys_str_mv | AT costaluiz invitropenicillinaseactivityoflactamaseoxa143p227spheffect AT favarodenize invitropenicillinaseactivityoflactamaseoxa143p227spheffect AT antunesvictor invitropenicillinaseactivityoflactamaseoxa143p227spheffect |
