Part:BBa_K398006:Design

Medium-Chain Aldehyde Dehydrogenase

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The original CDS can be found [http://www.ncbi.nlm.nih.gov/nuccore/AB047106.1 in this link], the file reports a cluster of proteins induced by alkanes. We took the ORF contained from 1213 to 2706. Nucleotide sequence optimized for expression in E.coli K12. We used [http://www.jcat.de/ jcat] for this purpose. Moreover, we deleted the restriction sites for EcoRI, PstI, SpeI and XbaI and we change the stop codon for a TAA.

The expected aminoacid sequence is:

                    MTMISAQTTEADLWIDGEWRPAASGERFDVIDPATGEVTARVAN
                    AGEDDVDAAVAIAEEAFSVRRWLAISPLERGRILRRIAELIRQHHCELAQLMTRENGM
                    PINLALFIEIPLADCFDFFASLVVKPQGEVLPFSVAGSAPDYMAWTMKEPIGVAGLIT
                    PWNFPLLMPTWKIAPALAAGCTMVVKPAPETPLTALKLAEICHEAGVPEGVINVLPGL
                    DEAGKALVRHPRVPKIAFTGETETGRHILQAAAPHIKRVTLELGGKSPNIIFADADLE
                    QAAKSALFGVFYNSGQVCQAGSRILVERTVYEPFVERLAERAKKLKVGPGTNPRSDLG
                    PVISREQYEKVLRYIEIGKQEGARLAAGGRALDGGGGGYFIEPTVFADVSPSMRIACE
                    EIFGPVAAVIPFADEEEAVRIANGTMYGLAAAVWTNDIKRALRLARRVKSGTVWLNTY
                    QVLSPTVPFGGYKQSGLGRELGMQALDAYLETKTVICDLNDRPMTLF

According to the authors [1], the purified protein has an activity of dodecanal dehydrogenase equal to 22.995 U/mg protein (4e-7 kat/mg protein).

Source

Genbank accession number: [http://www.ncbi.nlm.nih.gov/nuccore/AB047106 AB047106]

References

1. Kato T. et al. "Gene cloning and characterization of an aldehyde dehydrogenase from long-chain alkane-degrading Geobacillus thermoleovorans B23.", Extremophiles. 2010 Jan;14(1):33-9. Epub 2009 Sep 29.
2. National Center for Biotechnology Information [http://www.ncbi.nlm.nih.gov/]