US 11,781,120 B2
Biosynthesis of polyketides
Ramon Gonzalez, Tampa, FL (US); Seokjung Cheong, Houston, TX (US); and James M. Clomburg, Houston, TX (US)
Assigned to Ramon Gonzalez, Tampa, FL (US)
Filed by Ramon Gonzalez, Tampa, FL (US)
Filed on Jul. 15, 2020, as Appl. No. 16/930,220.
Application 16/930,220 is a continuation in part of application No. 15/749,000, abandoned, previously published as PCT/US2016/045037, filed on Aug. 1, 2016.
Claims priority of provisional application 62/198,764, filed on Jul. 30, 2015.
Prior Publication US 2021/0054427 A1, Feb. 25, 2021
Int. Cl. C12N 9/10 (2006.01); C12P 15/00 (2006.01); C12P 17/06 (2006.01)
CPC C12N 9/1025 (2013.01) [C12P 17/06 (2013.01); C12Y 203/01174 (2013.01)] 13 Claims
OG exemplary drawing
 
1. A method of making a polyketide, comprising growing a genetically engineered microorganism in a nutrient broth for a time sufficient to produce a polyketide and isolating said polyketide or a spontaneously rearranged form of said polyketide or a derivative of said polyketide, said polyketide selected from triacetic acid lactone, dehydroacetic acid, olivetolic acid, orsellinic acid, or 6-methylsalicylic acid and said derivative of said polyketide is a prenylated aromatic, or a cannabinoid, or dehydroacetic acid, or olivetolic acid, or cannabigerolic acid, or orsellinic acid, or 6-methylsalicylic acid, wherein said microorganism has a polyketide-producing pathway comprising the following substrate(s) to product(s) conversions:
a) C(n)-acyl-CoA+acetyl-CoA→C(n+2)-ketoacyl-CoA catalyzed by a ketoacyl-CoA thiolase encoded by Streptomyces collinus fadA, Rhodococcus opacus pcaF, Pseudomonas putida pcaF, Streptomyces sp. pcaF, P. putida fadAx, P. putida fadA, Acinetobacter sp. ADP1 dcaF, or Ralstonia eutropha bktB;
b) C(n+2)-ketoacyl-CoA+acetyl-CoA→C(n+4)-polyketoacyl-CoA catalyzed by a polyketoacyl-CoA thiolase encoded by Streptomyces collinus fadA, Rhodococcus opacus pcaF, Pseudomonas putida pcaF, Streptomyces sp. pcaF, P. putida fadAx, P. putida fadA, Acinetobacter sp. ADP1 dcaF, or Ralstonia eutropha bktB;
c) optionally, C(n+4)-polyketoacyl-CoA→3-OH—C(n+4)-polyketoacyl-CoA;
d) optionally, 3-OH—C(n+4)-polyketoacyl-CoA→C(n+4)-polyketoenoyl-CoA; and
e) optionally, C(n+4)-polyketoenoyl-CoA→C(n+4)-α,β-saturated-polyketoacyl-CoA;
f) iterations of at least one of the reactions in steps b), c), d), and e) wherein said iterations are achieved by utilizing a product generated in reactions steps b), c), d), and e), respectively, as a substrates for condensation with acetyl-CoA to elongate said product by two carbons and add a beta-keto group; and
g) conversion of said product formed in steps b), c), d), e), or a product formed in step f) to a polyketide or a spontaneously rearranged form of said polyketide or a derivative of said polyketide;
wherein n>0 and <30.