title: New insights into the phylogeny and molecular classification of nicotinamide mononucleotide deamidases.
abstract: Nicotinamide mononucleotide (NMN) deamidase is one of the key enzymes of the bacterial pyridine nucleotide cycle (PNC). It catalyzes the conversion of NMN to nicotinic acid mononucleotide, which is later converted to NAD(+) by entering the Preiss-Handler pathway. However, very few biochemical data are available regarding this enzyme. This paper represents the first complete molecular characterization of a novel NMN deamidase from the halotolerant and alkaliphilic bacterium Oceanobacillus iheyensis (OiPncC). The enzyme was active over a broad pH range, with an optimum at pH 7.4, whilst maintaining 90 % activity at pH 10.0. Surprisingly, the enzyme was quite stable at such basic pH, maintaining 61 % activity after 21 days. As regard temperature, it had an optimum at 65 °C but its stability was better below 50 °C. OiPncC was a Michaelian enzyme towards its only substrate NMN, with a K m value of 0.18 mM and a kcat/K m of 2.1 mM(-1) s(-1). To further our understanding of these enzymes, a complete phylogenetic and structural analysis was carried out taking into account the two Pfam domains usually associated with them (MocF and CinA). This analysis sheds light on the evolution of NMN deamidases, and enables the classification of NMN deamidases into 12 different subgroups, pointing to a novel domain architecture never before described. Using a Logo representation, conserved blocks were determined, providing new insights on the crucial residues involved in the binding and catalysis of both CinA and MocF domains. The analysis of these conserved blocks within new protein sequences could permit the more efficient data curation of incoming NMN deamidases.
authors: Department of Biochemistry and Molecular Biology-A, Faculty of Biology, Regional Campus of International Excellence "Campus Mare Nostrum", University of Murcia, Campus Espinardo, Murcia, Spain.
authors: Guiomar
authors: G
authors: Sánchez-Carrón
authors: Ana Belén
authors: AB
authors: Martínez-Moñino
authors: Agustín
authors: A
authors: Sola-Carvajal
authors: Hideto
authors: H
authors: Takami
authors: Francisco
authors: F
authors: García-Carmona
authors: Álvaro
authors: Á
authors: Sánchez-Ferrer
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D008969:Molecular Sequence Data; D009535:Nicotinamidase / Q000737:chemistry / Q000145:classification* / Q000235:genetics*; D010802:Phylogeny; D017434:Protein Structure, Tertiary
chemical_list: D001426:Bacterial Proteins; D009535:Nicotinamidase