Part:BBa_K1484317
4-Hydroxybenzalacetone Generator
This part converts p-coumaric acid into p- or 4-hydroxybenzalacetone, which is often referred to simply as benzalacetone and can be reduced to raspberry ketone.
It consists of the enzymes 4-coumarate:coA ligase (from N. tabacum; BBa_K1484115) and benzalacetone synthase (from Rheum palmatum, Chinese rhubarb; BBa_K1484103), under the control of an arabinose inducible promoter.
To use, transform into E. coli and incubate in a growth medium of your choice with 50mM L-arabinose for maximum expression levels. 50 hour incubations with 3mM and 9mM p-coumaric acid in M9 minimal growth medium both showed significant amounts of benzalacetone production. See Experience page for spectra.
Beware that when E. coli were transformed to also express 'benzalacetone reductase' (BBa_M36705), raspberry ketone was NOT produced. This agrees with the findings of the lab that this gene was cloned by, led by Dr Jules Beekwilder in Wageningen, The Netherlands, that this protein was not shown to exhibit benzalacetone reductase activity either in vivo or in vitro.
The cds of the 4-coumarate:coA ligase was kindly sent by Jules Beekwilder and Esmer Jongedijk of Wageningen UR in the Netherlands, and that of benzalacetone synthase by Ikuro Abe and Takahiro Mori of The Graduate School of Pharmaceutical Sciences at the University of Tokyo.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 2350
Illegal BamHI site found at 2696
Illegal XhoI site found at 3978 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 3282
Illegal AgeI site found at 979 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2409
Illegal SapI site found at 961
| None |