Part:BBa_K4388002
Arabidopsis thaliana 4-Coumarate:CoA Ligase Mutant (At4CL L57I/L460H)
The Arabidopsis thaliana 4-coumarate:CoA Ligase (At4CL) is an enzyme that catalyses the activation of 4-coumaric acid with Coenzyme A to form 4-coumaroyl-CoA.
4CL is a key enzyme of the general phenylpropanoid pathway that has been used for the production of multiple polyphenols including naringenin, resveratrol, and pterostilbene. This version of the enzyme is a mutant with amino acid changes L57I and L460H. These mutations have been shown to increase catalytic efficiency from 6.72 (wild-type) to 31.76 μmol/h/g (Yan et al., 2021).
The ORF of this compound was obtained from the iGEM registry: BBa_K1033001. This sequence was then codon optimized for E. coli K12, mutated at the desired positions, and two stop codons (TAATAA) were added to terminate translation.
Yan, Z.-B., Liang, J.-L., Niu, F.-X., Shen, Y.-P., & Liu, J.-Z. (2021). Enhanced Production of Pterostilbene in Escherichia coli Through Directed Evolution and Host Strain Engineering. Frontiers in Microbiology, 12. doi:10.1186/s12934-017-0644-6
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1223
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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