Part:BBa_K5175032:Design
T7 promoter-fucO-aldA-T7 terminator
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 241
Illegal PstI site found at 1989
Illegal PstI site found at 2702 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2760
Illegal PstI site found at 241
Illegal PstI site found at 1989
Illegal PstI site found at 2702 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1527
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 241
Illegal PstI site found at 1989
Illegal PstI site found at 2702 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 241
Illegal PstI site found at 1989
Illegal PstI site found at 2702
Illegal AgeI site found at 957
Illegal AgeI site found at 1158
Illegal AgeI site found at 1799 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
It is a composite component consisting of the T7 promoter, lac operator, target genes fucO, aldA. It is responsible for enabling E.coli to increase E.coli 's ability to efficiently utilise EG.
We chose fucO as the gene for L-1,2-propanediol oxidoreductase and aldA as the gene for aldehyde dehydrogenase A. L-1,2-propanediol oxidoreductase is an iron-dependent group III dehydrogenase, and aldehyde dehydrogenase A is an enzyme with a relatively broad substrate specificity for small hydroxyaldehyde substrates. EG is first converted in E.coli to glycolaldehyde (GLA) by L-1,2 -propylene glycol oxidoreductase, which is subsequently converted to glycolic acid (GA) by aldehyde dehydrogenase A. GA can be metabolised by condensation with acetyl coenzyme A via the glyoxalate shunt to form malic acid.
Source
fucO, aldA are from E.coli.
References
[1] MONTELLA C, BELLSOLELL L, PéREZ-LUQUE R, et al. Crystal structure of an iron-dependent group III dehydrogenase that interconverts L-lactaldehyde and L-1,2-propanediol in Escherichia coli [J]. J Bacteriol, 2005, 187(14): 4957-66.
[2] ZHU Y, LIN E C. Loss of aldehyde dehydrogenase in an Escherichia coli mutant selected for growth on the rare sugar L-galactose [J]. J Bacteriol, 1987, 169(2): 785-9.