Escherichia coli K-12 substr. MG1655 Enzyme: NMN amidohydrolase

Gene: pncC Accession Numbers: G7409 (EcoCyc), b2700, ECK2695

Synonyms: ygaD

Regulation Summary Diagram

Regulation summary diagram for pncC

Subunit composition of NMN amidohydrolase = [PncC]2
         NMN aminohydrolase = PncC

NMN amidohydrolase takes part in the intracellular recycling of NAD, catalyzing the deamidation of NMN to NaMN [Hillyard81]. The enzyme is in the most active path (PNC IV) for recycling NAD in E. coli [Neidhardt96].

Until recently, the gene encoding NMN amidohydrolase was unknown. Aided by genomics-based reconstruction of NAD metabolism, the gene was finally identified by Galeazzi et al. [Galeazzi11].

Site-directed mutagenesis of conserved residues and kinetic analysis of the resulting mutant enzymes enabled the identification of residues involved in catalysis; the enzyme may employ a Ser/Lys dyad active site configuration. A detailed reaction mechanism was proposed [Sorci14].

PncC: "pyridine nucleotide cycle"

Citations: [SanchezCarron13]

Locations: cytosol

Map Position: [2,823,849 <- 2,824,346] (60.84 centisomes, 219°)
Length: 498 bp / 165 aa

Molecular Weight of Polypeptide: 17.582 kD (from nucleotide sequence), 20.0 kD (experimental) [Galeazzi11]

Molecular Weight of Multimer: 33.0 kD (experimental) [Hillyard81]

Unification Links: ASAP:ABE-0008882, DIP:DIP-48189N, EchoBASE:EB2560, EcoGene:EG12698, EcoliWiki:b2700, OU-Microarray:b2700, PortEco:ygaD, Pride:P0A6G3, Protein Model Portal:P0A6G3, RefSeq:NP_417180, RegulonDB:G7409, SMR:P0A6G3, String:511145.b2700, UniProt:P0A6G3

Relationship Links: InterPro:IN-FAMILY:IPR008136, Pfam:IN-FAMILY:PF02464

In Paralogous Gene Group: 311 (2 members)

Gene-Reaction Schematic

Gene-Reaction Schematic

GO Terms:
Biological Process:
Inferred by computational analysisGO:0019363 - pyridine nucleotide biosynthetic process [UniProtGOA11a]
Molecular Function:
Inferred from experimentInferred by computational analysisGO:0019159 - nicotinamide-nucleotide amidase activity [GOA01, Galeazzi11]
Inferred by computational analysisGO:0016787 - hydrolase activity [UniProtGOA11a]
Cellular Component:
Inferred by computational analysisGO:0005829 - cytosol [DiazMejia09]

MultiFun Terms: metabolismbiosynthesis of building blockscofactors, small molecule carriersnicotinamide adenine dinucleotide

Essentiality data for pncC knockouts:

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enrichedYes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB LennoxYes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerolYes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucoseYes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]

Created 30-Sep-2011 by Keseler I, SRI International
Last-Curated 12-Dec-2014 by Keseler I, SRI International

Enzymatic reaction of: NMN amidohydrolase

Inferred from experiment

Synonyms: nicotinamide mononucleotide amidohydrolase, nicotinamide-nucleotide amidase, NMN deamidase

EC Number:

β-nicotinamide D-ribonucleotide + H2O → ammonium + β-nicotinate D-ribonucleotide

The direction shown, i.e. which substrates are on the left and right sides, is in accordance with the Enzyme Commission system.

The reaction is favored in the direction shown.

Alternative Substrates [ Comment 4]:

In Pathways: NAD salvage pathway II

Kinetic Parameters:
Substrate Km (μM) kcat (sec-1) kcat/Km (sec-1 μM-1) Specific Activity (U/mg) Citations
β-nicotinamide D-ribonucleotide 14.0 [Hillyard81, BRENDA14]
β-nicotinamide D-ribonucleotide 7.0 4.1 0.59 [Sorci14]
β-nicotinamide D-ribonucleotide 6.0 3.3 0.55 10.8 [Galeazzi11]

Sequence Features

Protein sequence of NMN aminohydrolase with features indicated

Feature Class Location Citations Comment
Pfam PF02464 8 -> 160
Inferred by computational analysis[Finn14]
CinA : Competence-damaged protein [More...]

Gene Local Context (not to scale -- see Genome Browser for correct scale)

Transcription Unit

Transcription-unit diagram


Peter D. Karp on Thu Jan 16, 2003:
Predicted gene function revised as a result of E. coli genome reannotation by Serres et al. [Serres01].
Markus Krummenacker on Tue Oct 14, 1997:
Gene object created from Blattner lab Genbank (v. M52) entry.


Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

BRENDA14: BRENDA team (2014). Imported from BRENDA version existing on Aug 2014.

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Finn14: Finn RD, Bateman A, Clements J, Coggill P, Eberhardt RY, Eddy SR, Heger A, Hetherington K, Holm L, Mistry J, Sonnhammer EL, Tate J, Punta M (2014). "Pfam: the protein families database." Nucleic Acids Res 42(Database issue);D222-30. PMID: 24288371

Galeazzi11: Galeazzi L, Bocci P, Amici A, Brunetti L, Ruggieri S, Romine M, Reed S, Osterman AL, Rodionov DA, Sorci L, Raffaelli N (2011). "Identification of nicotinamide mononucleotide deamidase of the bacterial pyridine nucleotide cycle reveals a novel broadly conserved amidohydrolase family." J Biol Chem 286(46);40365-75. PMID: 21953451

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

Hillyard81: Hillyard D, Rechsteiner M, Manlapaz-Ramos P, Imperial JS, Cruz LJ, Olivera BM (1981). "The pyridine nucleotide cycle. Studies in Escherichia coli and the human cell line D98/AH2." J Biol Chem 1981;256(16);8491-7. PMID: 7021549

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Neidhardt96: Neidhardt FC, Curtiss III R, Ingraham JL, Lin ECC, Low Jr KB, Magasanik B, Reznikoff WS, Riley M, Schaechter M, Umbarger HE "Escherichia coli and Salmonella, Cellular and Molecular Biology, Second Edition." American Society for Microbiology, Washington, D.C., 1996.

SanchezCarron13: Sanchez-Carron G, Martinez-Monino AB, Sola-Carvajal A, Takami H, Garcia-Carmona F, Sanchez-Ferrer Á (2013). "New insights into the phylogeny and molecular classification of nicotinamide mononucleotide deamidases." PLoS One 8(12);e82705. PMID: 24340054

Serres01: Serres MH, Gopal S, Nahum LA, Liang P, Gaasterland T, Riley M (2001). "A functional update of the Escherichia coli K-12 genome." Genome Biol 2(9);RESEARCH0035. PMID: 11574054

Sorci14: Sorci L, Brunetti L, Cialabrini L, Mazzola F, Kazanov MD, D'Auria S, Ruggieri S, Raffaelli N (2014). "Characterization of bacterial NMN deamidase as a Ser/Lys hydrolase expands diversity of serine amidohydrolases." FEBS Lett 588(6);1016-23. PMID: 24530526

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

Other References Related to Gene Regulation

Nonaka06: Nonaka G, Blankschien M, Herman C, Gross CA, Rhodius VA (2006). "Regulon and promoter analysis of the E. coli heat-shock factor, sigma32, reveals a multifaceted cellular response to heat stress." Genes Dev 20(13);1776-89. PMID: 16818608

Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by Pathway Tools version 20.0 (software by SRI International) on Thu May 5, 2016, BIOCYC13A.