Biosynthesis of caffeic acid and caffeic acid derivatives by recombinant microorganisms
View Patent ↗Microorganisms are genetically engineered to synthesize caffeic acid from simple carbon sources via a tyrosine intermediate by means of a dual pathway that utilizes both endogenous and engineered enzymatic activities.
1. A genetically engineered cell comprising p-coumarate 3-hydroxylase (C3H) enzyme activity, wherein said cell has been metabolically engineered to express or overexpress a tyrosine ammonia lyase (TAL) and to overexpress a 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H) enzyme or enzyme complex.
2. The genetically engineered cell of claim 1 wherein the p-coumarate 3-hydroxylase activity is provided by an endogenous 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H) enzyme or enzyme complex.
3. The genetically engineered cell of claim 1 further comprising a metabolic pathway for the biosynthesis of tyrosine.
4. The genetically engineered cell of claim 3 which has been further metabolically engineered to overproduce tyrosine compared to a wild-type cell.
5. The genetically engineered cell of claim 4 which has been further metabolically engineered to reduce or eliminate feedback inhibition of tyrosine biosynthesis.
6. The genetically engineered cell of any of claim 4 or 5 which has been further metabolically engineered to redirect carbon flow toward tyrosine biosynthesis.
7. The genetically engineered cell of claim 1 which is a bacterial cell.
8. The genetically engineered cell of claim 7 which is an Escherichia coli cell.
9. The genetically engineered cell of claim 1 comprising at least one vector operably encoding the tyrosine ammonia lyase (TAL).
10. The genetically engineered cell of claim 9 comprising at least one vector operably encoding an enzyme having p-coumarate 3-hydroxylase (C3H) activity.
11. The genetically engineered cell of claim 10 wherein a single vector encodes both the tyrosine ammonia lyase (TAL) and the enzyme having p-coumarate 3-hydroxylase (C3H) activity.
12. The genetically engineered cell of claim 10 comprising a first vector encoding the tyrosine ammonia lyase (TAL) and a second vector encoding the enzyme having p-coumarate 3-hydroxylase (C3H) activity.