Literature summary for 3.5.1.B15 extracted from

Zapata-Perez, R.; Garcia-Saura, A.G.; Jebbar, M.; Golyshin, P.N.; Sanchez-Ferrer, A.
Combined whole-cell high-throughput functional screening for identification of new nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries (2016), Front. Microbiol., 7, 1915 .

Search for more literature for 3.5.1.B15

Cloned(Commentary)

Cloned (Comment)	Organism
combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward nicotinamidase activity (EC 3.5.1.19) and pyrazinamide (pyrazinamidase activity). Escherichia coli EPI300-T1 is used for the screening of the genomic libraries cloned in pCC1FOS fosmids library. Escherichia coli strain Rosetta2 (DE3) is used as the host for pET46Ek/LIC expression vector. The fosmid library contains inserts of mixed genomic DNA of 194 mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge, genotyping, DNA and amino acid sequence determination and analysis	uncultured bacterium

Cloned (Comment)

Organism

combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward nicotinamidase activity (EC 3.5.1.19) and pyrazinamide (pyrazinamidase activity). Escherichia coli EPI300-T1 is used for the screening of the genomic libraries cloned in pCC1FOS fosmids library. Escherichia coli strain Rosetta2 (DE3) is used as the host for pET46Ek/LIC expression vector. The fosmid library contains inserts of mixed genomic DNA of 194 mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge, genotyping, DNA and amino acid sequence determination and analysis

uncultured bacterium

Inhibitors

Inhibitors	Comment	Organism	Structure
5-bromo-nicotinaldehyde	-	uncultured bacterium
nicotinaldehyde	-	uncultured bacterium

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
pyrazinamide + H2O	uncultured bacterium	-	pyrazinoic acid + NH3	-	r

Organism

Organism	UniProt	Comment	Textmining
uncultured bacterium	-	mesophilic bacteria (MB) isolated from deep-sea hydrothermal vents sampled at various sites in Mid Atlantic Ridge	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
pyrazinamide + H2O	-	uncultured bacterium	pyrazinoic acid + NH3	-	r

Synonyms

Synonyms	Comment	Organism
More	cf. EC 3.5.1.19	uncultured bacterium
PolyNic	-	uncultured bacterium

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
37	-	assay at	uncultured bacterium

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.3	-	assay at	uncultured bacterium

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.000015	-	nicotinaldehyde	pH 7.3, 37°C	uncultured bacterium
0.00007	-	5-bromo-nicotinaldehyde	pH 7.3, 37°C	uncultured bacterium

General Information

General Information	Comment	Organism
additional information	combined whole-cell high-throughput functional screening for identification of nicotinamidases/pyrazinamidases in metagenomic/polygenomic libraries, screening of mesophilic marine bacteria (MB) polygenomic library. Development of two whole-cell methods using the chemical property of one of the products formed in the enzymatic reaction (pyrazinoic or NA) to form colored complexes with stable iron salts, such as ammonium ferrous sulfate or sodium nitroprusside (SNP), optimization of the assay. A fosmid polygenomic expression library obtained from deep-sea mesophilic bacteria is screened, discovering several positive clones with the ammonium ferrous sulfate method. Quantitative rescreening with the SNP method allowing the finding of the first nicotinamidase with balanced catalytic efficiency toward nicotinamidase activity (EC 3.5.1.19) and pyrazinamide (pyrazinamidase activity)	uncultured bacterium