Academic Journal
Molecular cloning, expression and biochemical characterization of periplasmic nitrate reductase from Campylobacter jejuni
| العنوان: | Molecular cloning, expression and biochemical characterization of periplasmic nitrate reductase from Campylobacter jejuni |
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| المؤلفون: | Mintmier, Breeanna, Mcgarry, Jennifer, Sparacino-Watkins, Courtney, Sallmen, Joseph, Fischer-Schrader, Katrin, Magalon, Axel, Mccormick, Joseph, Stolz, John, Schwarz, Günter, Bain, Daniel, Basu, Partha |
| المساهمون: | Indiana University - Purdue University Indianapolis (IUPUI), Indiana University System, University of Pittsburgh (PITT), Pennsylvania Commonwealth System of Higher Education (PCSHE), Duquesne University Pittsburgh, Universität zu Köln = University of Cologne, Laboratoire de chimie bactérienne (LCB), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS) |
| المصدر: | ISSN: 0378-1097. |
| بيانات النشر: | CCSD Wiley-Blackwell |
| سنة النشر: | 2018 |
| المجموعة: | Aix-Marseille Université: HAL |
| مصطلحات موضوعية: | Campylobacter, Heterologous Expression, NapA, Nitrate Reductase, Pathogenicity, Molybdenum, [SDV]Life Sciences [q-bio] |
| الوصف: | International audience ; Campylobacter jejuni, a human gastrointestinal pathogen, uses nitrate for growth under microaerophilic conditions using periplasmic nitrate reductase (Nap). The catalytic subunit, NapA, contains two prosthetic groups, an iron sulfur cluster and a molybdenum cofactor. Here we describe the cloning, expression, purification, Michaelis-Menten kinetics (kcat of 5.91 ± 0.18s - 1 and a KM (nitrate) of 3.40± 0.44 μM) in solution using methyl viologen as an electron donor. The data suggest that the high affinity of NapA for nitrate could support growth of C. jejunion nitrate in the gastrointestinal tract. Site-directed mutagenesis was used and the codon for the molybdenum coordinating residue, cysteine has been exchanged for serine. The resulting variant NapA is four fold less active than the native enzyme confirming the importance of this residue. The properties of the C. jejuni enzyme reported here represents the first isolation and characterization of an Epsilonproteobacterial NapA. Therefore, the fundamental knowledge of Nap has been expanded. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1093/femsle/fny151 |
| الاتاحة: | https://amu.hal.science/hal-01919478 https://amu.hal.science/hal-01919478v1/document https://amu.hal.science/hal-01919478v1/file/fny151.pdf https://doi.org/10.1093/femsle/fny151 |
| Rights: | http://creativecommons.org/licenses/by/ ; info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.A9A6E7E8 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1093/femsle/fny151 |
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