Chemo-enzymatic process
The present disclosure relates to processes that combine microbial production of organic intermediates and subsequent synthetic transformation to provide compounds of industrial value, including compounds used in fragrances.
1. A chemo-enzymatic process for producing a macrolactone selected from the group consisting of: 16-hexadecalactone and 16-7(Z)-hexadecalactone, the method comprising:
culturing a recombinant Escherichia coli ( E. coli ) comprising a modified fatty acid biosynthetic pathway, wherein the modified fatty acid biosynthetic pathway comprises an exogenous thioesterase selected from the group consisting of: fatB, obtained from Arabidopsis thaliana , and fatB3, obtained from Cuphea hookeriana ; an endogenous mutated tesA; and an omega-hydroxylase or oxygenase, obtained from Myobacterium marinum M which are overexpressed,
wherein the omega-hydroxylase or oxygenase generates an omega-hydroxy fatty acid (ω-OH FA), wherein the ω-OH FA is 16-hydroxy-hexadecanoic acid or 16-hydroxy-7(Z)-hexadecenoic acid, and wherein the culturing is performed with a carbon-based feedstock; and
contacting the 16-hydroxy-hexadecanoic acid or the 16-hydroxy-7(Z)-hexadecenoic acid ex-vivo with a Lewis acid reagent under conditions sufficient to produce the macrolactone selected from the group consisting of 16-hexadecalactone and 16-7(Z)-hexadecelactone.
2. The chemo-enzymatic process of claim 1 , wherein said modified fatty acid biosynthetic pathway further comprises a desaturase which is overexpressed to increase unsaturated fatty acid derivatives produced by said recombinant E. coli.
3. The method of claim 1 , further comprising isolating the 16-hydroxy-hexadecanoic acid or 16-hydroxy-7(Z)-hexadecenoic acid prior to the contacting step.
4. The method of claim 1 , wherein the 16-hydroxy-hexadecanoic acid or 16-hydroxy-7(Z)-hexadecenoic acid is secreted from said recombinant E. coli and the contacting step is performed without isolating the 16-hydroxy-hexadecanoic acid or 16-hydroxy-7(Z)-hexadecenoic acid from the culturing step.
5. The method of claim 1 , wherein the contacting step comprises dehydration, lactonization, or combinations thereof.
6. The method of claim 1 , wherein said carbon-based feedstock comprises a simple carbon source.
7. The method of claim 1 , wherein said carbon-based feedstock is renewable.
8. The method of claim 1 , wherein the Lewis acid comprises a tertbutyl titanate Lewis acid.