Granted Patent B2

US 12,703,875 · App. 17/681,434 · Granted Aug 11, 2026

Inventors: Bradley W. Biggs (Evanston, IL); Keith E.J. Tyo (Evanston, IL)

Assignee: Northwestern University

C12P7/24C12N1/20C12N9/0008C12N15/113C12Y102/01067

Abstract

Claims (21)

1 . An engineered Acinetobacter baylyi that is capable of production of vanillin-glucoside from carbon sources derived from lignin and that comprises a genome that is modified, wherein a catechol O-methyltransferase (COMT) gene and a UDP-glycosyltransferase (UGT) gene are each independently integrated into the genome of the engineered Acinetobacter baylyi , and wherein at least ten genes selected from ACIAD1725, ACIAD1430, ACIAD1429, ACIAD0503, ACIAD1577, ACIAD1578, ACIAD1009, ACIAD1716, ACIAD2018, ACIAD1879, ACIAD3339, ACIAD2774, ACIAD3612, ACIAD2015, ACIAD2929, ACIAD1743, ACIAD2542, ACIAD3616, ACIAD1950, and ACIAD3642 have been knocked out.

2 . The engineered Acinetobacter baylyi of claim 1 , wherein the Acinetobacter baylyi is ADP1.

3 . The engineered Acinetobacter baylyi of claim 1 , wherein the vanillin-glucoside is vanillin-4-O-D-glucoside.

4 . The engineered Acinetobacter baylyi of claim 1 , wherein the COMT gene is a human COMT gene and the UGT gene is Arabidopsis UGT72E2.

5 . The engineered Acinetobacter baylyi of claim 4 , wherein the human COMT gene is integrated into the genome of the engineered Acinetobacter baylyi at the vanAB locus and/or the Arabidopsis UGT72E2 gene is integrated into the genome of the engineered Acinetobacter baylyi at the pcaHG locus.

6 . The engineered Acinetobacter baylyi of claim 1 , wherein Car/Sfp is introduced into the engineered Acinetobacter baylyi.

7 . The engineered Acinetobacter baylyi of claim 6 , wherein Car/Sfp is introduced via a plasmid.

8 . The engineered Acinetobacter baylyi of claim 1 , wherein pcaH, pcaG, vanA, and/or vanB have been knocked out of the genome of the engineered Acinetobacter baylyi.

9 . The engineered Acinetobacter baylyi of claim 1 , wherein at least sixteen genes selected from ACIAD1725, ACIAD1430, ACIAD1429, ACIAD0503, ACIAD1577, ACIAD1578, ACIAD1009, ACIAD1716, ACIAD2018, ACIAD1879, ACIAD3339, ACIAD2774, ACIAD3612, ACIAD2015, ACIAD2929, ACIAD1743, ACIAD2542, ACIAD3616, ACIAD1950, and ACIAD3642 have been knocked out.

10 . The engineered Acinetobacter baylyi of claim 1 , wherein all of ACIAD1725, ACIAD1430, ACIAD1429, ACIAD0503, ACIAD1577, ACIAD1578, ACIAD1009, ACIAD1716, ACIAD2018, ACIAD1879, ACIAD3339, ACIAD2774, ACIAD3612, ACIAD2015, ACIAD2929, ACIAD1743, ACIAD2542, ACIAD3616, ACIAD1950, and ACIAD3642 have been knocked out.

11 . A method of producing a vanillin, comprising culturing an engineered Acinetobacter baylyi of claim 1 in the presence of a carbon source.

12 . The method of claim 11 , wherein the carbon source is a waste stream.

13 . The method of claim 11 , wherein the carbon source comprises a lignin.

14 . The method of claim 13 , wherein the lignin has undergone an alkali pretreatment.

15 . The method of claim 11 , wherein culturing occurs in a M9 medium.

16 . The method of claim 11 , wherein culturing occurs in the presence of trace elements.

17 . The method of claim 11 , wherein the carbon source is an alkali pretreated liquor lignin (APL).

18 . The method of claim 11 , wherein the vanillin is a vanillin-glucoside.

19 . The method of claim 18 , wherein the vanillin-glucoside is vanillin-4-O-D-glucoside.

20 . The method of claim 11 , wherein the vanillin is detectable after about 24 hours of culturing.

21 . The engineered Acinetobacter baylyi of claim 1 , wherein the COMT gene is integrated at a vanAB locus and the UGT gene is integrated at a pcaHG locus.

Assignments (2)

Continuity (2)

References Cited (16)