Cloning, expression and characterization of thermostable YdaP from Bacillus licheniformis 9A

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dc.contributor.authorLako, Joseph D. Wani
dc.contributor.authorYengkopiong, Jada P.
dc.contributor.authorStafford, William H. L.
dc.contributor.authorTuffin, Marla
dc.contributor.authorCowan, Don A.
dc.date.accessioned2019-10-18T11:05:35Z
dc.date.available2019-10-18T11:05:35Z
dc.date.issued2017
dc.description.abstractThe Bacillus licheniformis ydaP gene encodes for a pyruvate oxidase that catalyses the oxidative decarboxylation of pyruvate to acetate and CO2. The YdaP form of this enzyme was purified about 48.6-folds to homogeneity in three steps. The enzyme was recovered in a soluble form and demonstrated significant activity on pyruvate using 2, 6-dichlorophenolindophenol (DCPIP) as an artificial electron acceptor. HPLC analysis of the YdaP-enzyme catalysed conversion of pyruvate showed acetate as the sole product, confirming the putative identity of pyruvate oxidase. Analysis of the substrate specificity showed that the YdaP enzyme demonstrated preference for short chain oxo acids; however, it was activated by 1% Triton X-100. The YdaP substrate-binding pocket from the YdaP protein differed substantially from the equivalent site in all of the so far characterized pyruvate oxidases, suggesting that the B. licheniformis YdaP might accept different substrates. This could allow more accessibility of large substrates into the active site of this enzyme. The thermostability and pH activity of the YdaP enzyme were determined, with optimums at 50ÂșC and pH 5.8, respectively. The amino acid residues forming the catalytic cavity were identified as Gln460 to Ala480.en_US
dc.identifier.citationLako, Joseph D. Wani et al. (2017). Cloning, expression and characterization of thermostable YdaP from Bacillus licheniformis 9A. Acta Biochimica Polonica, 65(1), 59-66. https://doi.org/10.18388/abp.2017_1499en_US
dc.identifier.urihttps://doi.org/10.18388/abp.2017_1499
dc.identifier.urihttp://hdl.handle.net/10566/5054
dc.language.isoenen_US
dc.publisherActa Biochimica Polonicaen_US
dc.subjectBacillus licheniformisen_US
dc.subjectPyruvate oxidaseen_US
dc.subjectYdaPen_US
dc.subjectThiamine diphosphate enzymeen_US
dc.titleCloning, expression and characterization of thermostable YdaP from Bacillus licheniformis 9Aen_US
dc.typeArticleen_US

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