Part:BBa_K4588035
Contents
4-hydroxy phenylacetate 3-monooxygenase reductase component C (E. coli), HpaC
This part encodes 4-hydroxy phenylacetate 3-monooxygenase reductase component C from Pasteurella multocida
Biology
The organism this gene is initially expressed in is Pasteurella multocida
HpaC is a coupling factor that enhances NADH oxidation for HpaB. This enzyme works with HpaB together to catalyze the oxidation of 4-hydroxyphenylacetate in the presence of NADH and FAD [1].
Design
This BioBrick contains a T7 promoter (BBa_I712074) for strong expression and a strong ribosome binding site (BBa_B0034) for optimal translation initiation. The composite BioBrick for HpaB consists of the coding region HpaC (BBa_K4588024) and is codon-optimized according to Escherichia coli codon bias for its expression in this bacteria. Following the coding region, a 6x His Tag (BBa_K1223006) was inserted for protein expression detection. The strong double terminator (BBa_B0015) was chosen to terminate gene transcription completely.
Usage
This enzyme is implemented in the synthesis pathway to produce salvianic acid A in an E. coli culture. This enzyme converts 4-hydroxyphenylpyruvate into 3,4-dihydroxy phenylpyruvate by adding a hydroxyl group to the benzene ring [2].
References
1. Xun, L., & Sandvik, E. R. (2000). Characterization of 4-hydroxyphenylacetate 3-hydroxylase (HpaB) of Escherichia coli as a reduced flavin adenine dinucleotide-utilizing monooxygenase. Applied and environmental microbiology, 66(2), 481–486. https://doi.org/10.1128/AEM.66.2.481-486.2000
2. Bloch, S. E., & Schmidt‐Dannert, C. (2014). Construction of a chimeric biosynthetic pathway for the de novo biosynthesis of rosmarinic acid in Escherichia coli. ChemBioChem, 15(16), 2393–2401. https://doi.org/10.1002/cbic.201402275
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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