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3.5.1.B15: pyrazinamidase

This is an abbreviated version!
For detailed information about pyrazinamidase, go to the full flat file.

Word Map on EC 3.5.1.B15

Reaction

Pyrazinamide
+
H2O
=
pyrazinoic acid
 +
NH3

Synonyms

ASAC_0847, BsPncA, More, nicotinamidase/pyrazinamidase, PH0999, PncA, PolyNic, pyrazinamidase, pyrazinamidase/nicotinamidase, PZAase, PZAse

ECTree

     3 Hydrolases
         3.5 Acting on carbon-nitrogen bonds, other than peptide bonds
             3.5.1 In linear amides
                3.5.1.B15 pyrazinamidase

Cloned

Cloned on EC 3.5.1.B15 - pyrazinamidase

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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
-
expressed in a methionine auxotroph, Escherichia coli strain B834
expressed in Escherichia coli
gene pncA, DNA and amino acid sequence determination and analysis of enzyme mutants, genotyping
gene pncA, DNA and amino acid sequence determination and analysis, recombinant enzyme expression in Escherichia coli strain BL21(DE3)pLysS
gene pncA, genotyping, DNA and amino acid sequence determination and analysis of wild-type and mutants
gene pncA, genotyping, global statistics for 641 mutations in 171 (of 187) codons from 2760 resistant strains and 96 mutations from 3329 susceptible strains reported in 61 studies, overview
-
gene pncA, PCR-based in vitro expression, and recombinant expression of His-tagged mutant enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain TOP10
-
gene pncA, PCR-based in vitro expression, and recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain TOP10
gene pncA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)
gene pncA, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha
-
recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3)