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.
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This research was supported by the R&D Program of MKE/KEIT (No. 10035578, Development of 2,3-butanediol and derivative production technology for C-Zero bio-platform industry).
This work was supported by the Graduate School of Specialization for Biotechnology Program of the Ministry of Knowledge Economy (MKE).
Soojin Lee and Borim Kim contributed equally to this work.
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Lee, S., Kim, B., Oh, M. et al. Enhanced activity of meso-secondary alcohol dehydrogenase from Klebsiella species by codon optimization. Bioprocess Biosyst Eng 36, 1005–1010 (2013). https://doi.org/10.1007/s00449-012-0824-z
- Codon optimization
- Meso-secondary alcohol dehydrogenase
- Gene expression
- Escherichia coli