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Characterisation of a Recombinant NADP‐Dependent Glycerol Dehydrogenase from Gluconobacter oxydans and its Application in the Production of L ‐Glyceraldehyde
Author(s) -
Richter Nina,
Neumann Markus,
Liese Andreas,
Wohlgemuth Roland,
Eggert Thorsten,
Hummel Werner
Publication year - 2009
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.200900193
Subject(s) - hypocrea , glyceraldehyde , dehydrogenase , biochemistry , chemistry , oxidoreductase , enzyme , glyceraldehyde 3 phosphate dehydrogenase , glycerol , stereochemistry , enantiopure drug , catalysis , trichoderma reesei , enantioselective synthesis , cellulase
The acetic acid bacterium Gluconobacter oxydans has a high potential for oxidoreductases with a variety of different catalytic abilities. One putative oxidoreductase gene codes for an enzyme with a high similarity to the NADP + ‐dependent glycerol dehydrogenase (GlyDH) from Hypocrea jecorina . Due to this homology, the GlyDH (Gox1615) has been cloned, over‐expressed in Escherichia coli , purified and characterised. Gox1615 shows an apparent native molecular mass of 39 kDa, which corresponds well to the mass of 37.213 kDa calculated from the primary structure. From HPLC measurements, a monomeric structure can be deduced. Kinetic parameters and the dependence of the activity on temperature and pH were determined. The enzyme shows a broad substrate spectrum in the reduction of different aliphatic, branched and aromatic aldehydes. Additionally, the enzyme has been shown to oxidize a variety of different alcohols. The highest activities were observed for the conversion of D ‐glyceraldehyde in the reductive and L ‐arabitol in the oxidative direction. Since high enantioselectivities were observed for the reduction of glyceraldehyde, the kinetic resolution of glyceraldehyde was investigated and found to yield enantiopure L ‐glyceraldehyde on preparative scale.