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Chemical Structure| 491-72-5 Chemical Structure| 491-72-5
Chemical Structure| 491-72-5

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Synonyms: OA; Olivetolcarboxylic Acid; Olivetolate

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Schäfer, Kilan J ; Chalwatzis, Lukas ; Feka, Athanasia M ; Aras, Marco ; Boles, Eckhard ; Kayser, Oliver

Abstract: annabinoids comprise a large class of bioactive compounds found primarily in the plant species Cannabis sativa and are of interest due to their pharmacological and therapeutic potential. The aromatic polyketide, olivetolic acid (OA), is a major precursor in the cannabinoid biosynthesis pathway and is derived from hexanoyl-CoA and malonyl-CoA by the action of olivetol synthase (OLS) and olivetolic acid cyclase (OAC). To date, most microbial cannabinoid production systems rely on the external supplementation of hexanoic acid together with the overexpression of acyl-activating enzyme 1 from C. sativa (CsAAE1) to provide hexanoyl-CoA. Here, we implement a heterologous OA biosynthesis pathway into the yeast Saccharomyces cerevisiae and describe various biochemical and metabolic engineering strategies to overcome the need for external hexanoic acid supplementation. We ensured a sufficient endogenous supply of hexanoyl-CoA and further optimized OA production by enhancing precursor supply. Moreover, we present a mutant phenylacetate-CoA ligase derived from Penicillium chrysogenum (PcPCL-K) which displayed superior hexanoyl-CoA ligase activity to CsAAE1. Together, these strategies enabled the production of 180 mg/L OA using shake flask cultures. Our results provide details of key metabolic engineering steps required for the biosynthetic production of cannabinoids and their precursors in S. cerevisiae.

Keywords: cannabinoids ; hexanoyl-CoA ; hexanoyl-CoA ligase ; malonyl-CoA ; olivetolic acid ; reverse β-oxidation pathway ; S. cerevisiae

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Schäfer, Kilan J ; Aras, Marco ; Boles, Eckhard ; Kayser, Oliver ;

Abstract: Medium chain fatty acids (MCFAs) are valuable platform compounds for the production of biotechnologically relevant chemicals such as biofuels and biochemicals. Two distinct pathways have been implemented in the yeast Saccharomyces cerevisiae for the biosynthetic production of MCFAs: (i) the mutant fatty acid biosynthesis (FAB) pathway in which the fatty acid synthase (FAS) complex is mutated and (ii) a heterologous multispecies-derived reverse β-oxidation (rBOX) pathway. Hexanoic acid has become of great interest as its acyl-CoA ester, hexanoyl-CoA, is required for the biosynthesis of olivetolic acid (OA), a cannabinoid precursor. Due to insufcient endogenous synthesis of hexanoyl-CoA, recombinant microbial systems to date require exogenous supplementation of cultures with hexanoate along with the overexpression of an acyl-CoA ligase to allow cannabinoid biosynthesis. Here, we engineer a recombinant S. cerevisiae strain which was metabolically optimized for the production of hexanoic acid via the FAB and rBOX pathways and we combine both pathways in a single strain to achieve titers of up to 120 mg L−1. Moreover, we demonstrate the biosynthesis of up to 15 mg L−1 OA from glucose using hexanoyl-CoA derived from the rBOX pathway

Keywords: Hexanoic acid ; Hexanoyl-CoA ; Medium-chain fatty acids ; Fatty acid synthase ; Reverse beta oxidation pathway ; Olivetolic acid ; Cannabinoids ; Pantothenate kinase

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Product Details of Olivetolic Acid

CAS No. :491-72-5
Formula : C12H16O4
M.W : 224.25
SMILES Code : O=C(O)C1=C(CCCCC)C=C(O)C=C1O
Synonyms :
OA; Olivetolcarboxylic Acid; Olivetolate
MDL No. :MFCD28383644
InChI Key :SXFKFRRXJUJGSS-UHFFFAOYSA-N
Pubchem ID :2826719

Safety of Olivetolic Acid

GHS Pictogram:
Signal Word:Warning
Hazard Statements:H302-H312-H332
Precautionary Statements:P261-P264-P270-P271-P280-P301+P312-P302+P352-P304+P340-P330-P363-P501
 

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