title: Biosynthesis of cis,cis-muconic acid and its aromatic precursors, catechol and protocatechuic acid, from renewable feedstocks by Saccharomyces cerevisiae.
abstract: Adipic acid is a high-value compound used primarily as a precursor for the synthesis of nylon, coatings, and plastics. Today it is produced mainly in chemical processes from petrochemicals like benzene. Because of the strong environmental impact of the production processes and the dependence on fossil resources, biotechnological production processes would provide an interesting alternative. Here we describe the first engineered Saccharomyces cerevisiae strain expressing a heterologous biosynthetic pathway converting the intermediate 3-dehydroshikimate of the aromatic amino acid biosynthesis pathway via protocatechuic acid and catechol into cis,cis-muconic acid, which can be chemically dehydrogenated to adipic acid. The pathway consists of three heterologous microbial enzymes, 3-dehydroshikimate dehydratase, protocatechuic acid decarboxylase composed of three different subunits, and catechol 1,2-dioxygenase. For each heterologous reaction step, we analyzed several potential candidates for their expression and activity in yeast to compose a functional cis,cis-muconic acid synthesis pathway. Carbon flow into the heterologous pathway was optimized by increasing the flux through selected steps of the common aromatic amino acid biosynthesis pathway and by blocking the conversion of 3-dehydroshikimate into shikimate. The recombinant yeast cells finally produced about 1.56 mg/liter cis,cis-muconic acid.
authors.affiliation: Institute of Molecular Biosciences, Goethe University Frankfurt, Frankfurt am Main, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Weber
authors.forename: Christine
authors.initials: C
authors.lastname: Brückner
authors.forename: Sheila
authors.initials: S
authors.lastname: Weinreb
authors.forename: Claudia
authors.initials: C
authors.lastname: Lehr
authors.forename: Christine
authors.initials: C
authors.lastname: Essl
authors.forename: Eckhard
authors.initials: E
authors.lastname: Boles
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics; D001711:Biotransformation; D002244:Carbon / Q000378:metabolism; D002396:Catechols / Q000378:metabolism*; D015870:Gene Expression; D062385:Hydroxybenzoates / Q000378:metabolism*; D060847:Metabolic Engineering; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D012765:Shikimic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D013011:Sorbic Acid / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D002396:Catechols; D062385:Hydroxybenzoates; D011994:Recombinant Proteins; C096387:3-dehydroshikimate; D012765:Shikimic Acid; C009091:protocatechuic acid; C005939:muconic acid; D002244:Carbon; C034221:catechol; D013011:Sorbic Acid