Mutant transporters for bacterial uptake of terephthalic acid
The present disclosure relates to a non-naturally occurring microorganism that includes a gene encoding a MucK transporter protein, where the microorganism is capable of catabolizing terephthalic acid (TPA). In some embodiments of the present disclosure, the gene encoding the MucK transporter protein may contain at least one mutation, relative to a reference gene encoding a reference MucK transporter protein.
1. A non-naturally occurring microorganism comprising:
a gene encoding a MucK transporter protein; and
a deletion of an endogenous gene encoding a transcriptional regulator, wherein:
the microorganism is capable of catabolizing terephthalic acid (TPA).
2. The non-naturally occurring microorganism of claim 1 , wherein the gene encoding the MucK transporter protein contains at least one mutation, relative to a gene encoding a reference MucK transporter protein.
3. The non-naturally occurring microorganism of claim 2 , wherein the reference MucK transporter protein is at least 90% identical to SEQ ID NO: 2.
4. The non-naturally occurring microorganism of claim 2 , wherein the gene encoding the reference MucK transporter protein is at least 90% identical to SEQ ID NO: 1.
5. The non-naturally occurring microorganism of claim 1 , wherein the mutation to the MucK transporter protein comprises at least one point mutation.
6. The non-naturally occurring microorganism of claim 5 , wherein the point mutation is present at an amino acid located at at least one of positions 34, 53, 133, 341, or 342 on SEQ ID NO: 2.
7. The non-naturally occurring microorganism of claim 6 , wherein the point mutation comprises at least one of M34L, M34I, Y133C, T3421, or E53G.
8. The non-naturally occurring microorganism of claim 1 , further comprising a deletion of an endogenous gene encoding a MucK transporter protein.
9. The non-naturally occurring microorganism of claim 1 , wherein the microorganism is further capable of growing on TPA.
10. The non-naturally occurring microorganism of claim 9 , wherein the microorganism is characterized by a TPA consumption rate between greater than zero g TPA/L/hr and about 0.2 g/L/hr.
11. The non-naturally occurring microorganism of claim 9 , wherein the microorganism is capable of growth in a liquid media at a temperature between about 25° C. and about 35° C.
12. The non-naturally occurring microorganism of claim 11 , wherein the liquid media is maintained at pH between about 6 and about 7.
13. The non-naturally occurring microorganism of claim 1 , wherein the microorganism comprises at least one of a bacterium, a yeast, or a fungus.
14. The non-naturally occurring microorganism of claim 13 , wherein the microorganism is a bacterium.
15. The non-naturally occurring microorganism of claim 14 , wherein the bacterium comprises a strain from at least one of Acinetobacter baylyi, Pseudomonas putida, Pseudomonas fluorescens , or Pseudomonas stuzeri.
16. The non-naturally occurring microorganism of claim 15 , wherein the bacterium is Acinetobacter baylyi.
17. The non-naturally occurring microorganism of claim 16 , wherein the bacterium is Acinetobacter baylyi.
18. The non-naturally occurring microorganism of claim 1 , wherein the transcriptional regulator is a DcaS transcriptional regulator.