Enhanced activity of meso-secondary alcohol dehydrogenase from Klebsiella species by codon optimization.
Meso-secondary alcohol dehydrogenases (meso-SADH) from Klebsiella oxytoca KCTC1686 and Klebsiella pneumoniae KCTC2242 were codon optimized and expressed in Escherichia coli W3110. The published gene data of K. pneumoniae NTUH-K2044 (NCBI accession number AP006725), K. pneumoniae 342 (NCBI accession number CP000964), and K. pneumoniae MGH 78578 (NCBI accession number CP000647), were compared with the meso-SADH sequences of each strain, respectively. Codon-optimized meso-SADH enzymes of K. oxytoca and K. pneumoniae showed approximately twofold to fivefold increased enzyme activities for acetoin reduction over native enzymes. The highest activities for each strain were obtained at 30-37 °C and pH 6-7 (yielding 203.1 U/mg of protein and 156.5 U/mg of protein, respectively). The increased enzyme activity of the codon-optimized enzymes indicated that these modified enzymes could convert acetoin into 2,3-butanediol with a high yield.
Department of Chemical and Biomolecular Engineering, Sogang University, Seoul 121-742, Korea.
Soojin
S
Lee
Borim
B
Kim
Minkyu
M
Oh
Youngrok
Y
Kim
Jinwon
J
Lee
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003062:Codon; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D007709:Klebsiella / Q000201:enzymology*; D008969:Molecular Sequence Data; D010957:Plasmids; D012689:Sequence Homology, Nucleic Acid
D003062:Codon; D017931:DNA Primers; D000426:Alcohol Dehydrogenase