Microorganisms engineered for muconate production
Disclosed herein are the genetically modified Pseudomonas with improved tolerance to hydroxycinnamic acids.
1. A genetically modified Pseudomonas comprising:
a mutation of a gene encoding a cell membrane protein wherein:
the mutation improves tolerance to a hydroxycinnamic acid, and
the gene encodes for a protein that is at least 92% identical to SEQ ID NO: 2.
2. The genetically modified Pseudomonas of claim 1 capable of converting hydroxycinnamic acid to muconate.
3. The genetically modified Pseudomonas of claim 1 wherein the mutation of a gene encoding a cell membrane protein decreases the time that the Pseudomonas spends in a lag phase when exposed to a solution of hydroxycinnamic acid compared to a non-genetically modified Pseudomonas.
4. The genetically modified Pseudomonas of claim 1 wherein the mutation increases a growth rate of the Pseudomonas when exposed to a solution of hydroxycinnamic acid compared to a non-genetically modified Pseudomonas.
5. The genetically modified Pseudomonas of claim 1 , wherein the gene is at least 92% identical to SEQ ID NO: 1.
6. The genetically modified Pseudomonas of claim 1 , wherein the hydroxycinnamic acid is selected from the group consisting of coumarate and ferulate.
7. The genetically modified Pseudomonas of claim 1 , wherein the Pseudomonas is P. putida K2440.
8. The genetically modified Pseudomonas of claim 1 , wherein the Pseudomonas demonstrates at least a 3.3-fold increased tolerance to p-coumaric acid (pCA) compared to a non-genetically modified Pseudomonas.
9. The genetically modified Pseudomonas of claim 1 , wherein the Pseudomonas has at least a 37 hour decrease in lag phase when exposed to 20 g/L pCA compared to a non-genetically modified Pseudomonas.
10. The genetically modified Pseudomonas of claim 1 , wherein the Pseudomonas has at least a 2.4-fold increased growth rate when exposed to 20 g/L pCA compared to a non-genetically modified Pseudomonas.
11. A method for lignin valorization, the method comprising:
converting a hydroxycinnamic acid to muconate by contacting the hydroxycinnamic acid with a genetically modified Pseudomonas , wherein:
the Pseudomonas comprises a mutation of a gene encoding a cell membrane protein,
the modification increases the uptake of the hydroxycinnamic acid compared to a non-genetically modified Pseudomonas , and
the gene encodes for a protein that is at least 92% identical to SEQ ID NO: 2.
12. The method of claim 11 wherein the mutation decreases the time in a lag phase of the Pseudomonas when the Pseudomonas is exposed to a solution of hydroxycinnamic acid compared to a non-genetically modified Pseudomonas.
13. The method of claim 11 wherein the mutation increases a growth rate of the Pseudomonas when exposed to a solution of hydroxycinnamic acid compared to a non-genetically modified Pseudomonas.
14. The method of claim 11 wherein the mutation is to a gene that is at least 92% identical to SEQ ID NO: 1.
15. The method of claim 11 wherein the hydroxycinnamic acid is selected from the group consisting of coumarate, and ferulate.
16. The method of claim 11 wherein the Pseudomonas has at least a 3.3-fold increased tolerance to pCA, at least a 37 hour decrease in lag phase, and a 2.4-fold increased growth rate when exposed to 20 g/L pCA compared to a non-genetically modified Pseudomonas.
17. A genetically modified Pseudomonas comprising:
a deletion of a gene encoding a cell membrane protein, wherein:
the deletion improves tolerance to a hydroxycinnamic acid, and
the deleted gene encodes for the expression of protein PP_3350.
18. A method for lignin valorization, the method comprising:
converting a hydroxycinnamic acid to muconate by contacting the hydroxycinnamic acid with a genetically modified Pseudomonas , wherein:
the Pseudomonas comprises a deletion of a gene encoding a cell membrane protein,
the modification increases the uptake of the hydroxycinnamic acid compared to a non-genetically modified Pseudomonas , and
the deleted gene encodes for the expression of protein PP_3350.