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https://hdl.handle.net/2440/79704
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dc.contributor.author | Schumann, C. | - |
dc.contributor.author | Michlmayr, H. | - |
dc.contributor.author | del Hierro, A. | - |
dc.contributor.author | Kulbe, K. | - |
dc.contributor.author | Jiranek, V. | - |
dc.contributor.author | Eder, R. | - |
dc.contributor.author | Nguyen, T. | - |
dc.date.issued | 2013 | - |
dc.identifier.citation | Bioengineered, 2013; 4(3):147-152 | - |
dc.identifier.issn | 1949-1018 | - |
dc.identifier.issn | 1949-1026 | - |
dc.identifier.uri | http://hdl.handle.net/2440/79704 | - |
dc.description.abstract | Malolactic enzymes (MLE) are known to directly convert L-malic acid into L-lactic acid with a catalytical requirement of nicotinamide adenine dinucleotide (NAD (+) ) and Mn ( 2+) ; however, the reaction mechanism is still unclear. To study a MLE, the structural gene from Oenococcus oeni strain DSM 20255 was heterologously expressed in Escherichia coli, yielding 22.9 kU l (-1) fermentation broth. After affinity chromatography and removal of apparently inactive protein by precipitation, purified recombinant MLE had a specific activity of 280 U mg (-1) protein with a recovery of approximately 61%. The enzyme appears to be a homodimer with a molecular mass of 128 kDa consisting of two 64 kDa subunits. Characterization of the recombinant enzyme showed optimum activity at pH 6.0 and 45°C, and Km, Vmax and kcat values of 4.9 mM, 427 U mg (-1) and 456 sec (-1) for L-malic acid, 91.4 µM, 295 U mg (-1) and 315 sec (-1) for NAD (+) and 4.6 µM, 229 U mg (-1) and 244 sec (-1) for Mn ( 2+) , respectively. The recombinant MLE retained 95% of its activity after 3 mo at room temperature and 7 mo at 4°C. When using pyruvic acid as substrate, the enzyme showed the conversion of pyruvic acid with detectable L-lactate dehydrogenase (L-LDH) activity and oxidation of NADH. This interesting observation might explain that MLE catalyzes a redox reaction and hence, the requirements for NAD (+) and Mn ( 2+) during the conversion of L-malic to L-lactic acid. | - |
dc.description.statementofresponsibility | Christina Schümann, Herbert Michlmayr, Andrés M. del Hierro, Klaus D. Kulbe, Vladimir Jiranek, Reinhard Eder and Thu-Ha Nguyen | - |
dc.language.iso | en | - |
dc.publisher | Landes Bioscience | - |
dc.rights | © 2013 Landes Bioscience | - |
dc.source.uri | http://dx.doi.org/10.4161/bioe.22988 | - |
dc.subject | Escherichia coli | - |
dc.subject | Malates | - |
dc.subject | Lactic Acid | - |
dc.subject | Malate Dehydrogenase | - |
dc.subject | Bacterial Proteins | - |
dc.subject | Enzyme Stability | - |
dc.subject | Gene Expression | - |
dc.subject | Kinetics | - |
dc.subject | Molecular Weight | - |
dc.subject | Oenococcus | - |
dc.title | Malolactic enzyme from Oenococcus oeni: Heterologous expression in Escherichia coli and biochemical characterization | - |
dc.type | Journal article | - |
dc.identifier.doi | 10.4161/bioe.22988 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | Jiranek, V. [0000-0002-9775-8963] | - |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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