Production and separation of 3-hydroxypropionic acid
The disclosure provides methods and apparatus for producing 3-hydroxypropionic acid or a salt thereof, for removing 3-hydroxypropionic acid from aqueous solution (e.g., aqueous broth), and for using it to make various chemicals.
1. An expression system, comprising a first promoter that is either constitutive or inducible by a small molecule, a second promoter that is either constitutive or induced by a small molecule, a first gene encoding a protein involved in the synthesis of 3-HP, a salt of 3-HP, or coenzyme B12, a modified UTR, and a native sequence encoding up to 20 amino acids from the N-terminus of a native Pseudomonas denitrificans protein, wherein the native sequence is operably linked to the 3′-terminus of the modified UTR and operably linked to the 5′-terminus of the first gene, the first and second promoters are operably linked in tandem upstream of the modified UTR, and the modified UTR comprises a sequence selected from the group consisting of SEQ ID NO: 22-28 and 64 (UTR0-6).
2. The expression system of claim 1 , further comprising a third promoter; and a second gene encoding a transcriptional regulator configured to regulate expression of the first gene, wherein the third promoter is operably connected to the second gene, and the third promoter comprises a sequence that is 95% identical to the sequence selected from the group consisting of SEQ ID NO: 7-10 and 52-63.
3. The expression system of claim 1 , wherein the first promoter is operably linked to the 5′-terminus of the second promoter, the second promoter is operably linked to the 5′-terminus of the modified UTR, and the modified UTR is operably linked to the 5′terminus of the fusion gene.