Arizona State University

David Nielsen

Rebekah McKenna

Shawn Pugh

Deirdre Meldrum

Weiwen Zhang

Bo Wang

Christopher Jones

Brian Thompson

Michael Machas

Compositions and methods for genetically modifying at least one nucleic acid sequence of interest in a polyploid organism.

Systems and methods for genetic engineering of a polyploid organism

Certain embodiments provide a method for preparing a biochemical product (e.g., phenol, catechol, or muconic acid, or a salt thereof). For example, such methods include contacting a recombinant host having two or more recombinant pathways with a fermentable carbon source and growing the recombinant cell for a time sufficient to synthesize the product. In certain embodiments, each recombinant pathway: 1) is capable of producing the same final biochemical product; 2) comprises at least one gene encoding a polypeptide; 3) is derived from a different endogenous metabolite as its immediate precursor; and 4) converges to the same final product or the same intermediate metabolite.

Synthetic metabolic funneling for biochemical production

Described herein are engineered metabolic pathways in recombinant microorganism host cells which result in the production of 2-phenylethanol or 2-phenylacetic acid. Also described herein are methods of using the recombinant microorganisms for the production of 2-phenylethanol or 2-phenylacetic acid.

Microbial production of 2-phenylethanol from renewable substrates

A method for the in vivo production of 1,3-butadiene from 2,4-pentadienoate is described (FIG.). Enzymes capable of decarboxylating 2,4-pentadienoate to 1,3-butadiene have been discovered. Recombinant expression of these newly discovered enzymes has resulted in the engineering of microorganisms capable of producing 1,3-butadiene when cultured in the presence of 2,4-pentadienoate. 1,3-butadienoate is an important monomer used in the manufacturing of rubbers and plastics. This invention will help to enable the biological production of 1,3-butadiene from renewable resources such as sugar, for example.

Enzymes and microorganisms for the production of 1,3-butadiene and other dienes

Construction and expression of synthetic pathways to produce (S) or (R)-3-hydroxybutyrate (3HB) as enantiomerically-pure products by genetically engineering cyanobacteriumsp. PCC 6803. Under optimized growth conditions, the pathway employing phaA and phaB fromwas the most effective, producing up to 533.4&#xb1;5.5 mg/l (R)-3HB after 21 days photosynthetic cultivation. For the first time, the feasibility and high efficiency of producing 3HB using solar energy and COas sole energy and carbon sources by engineered cyanobacteria is demonstrated.

Photosynthetic production of 3-hydroxybutyrate from carbon dioxide

Novel methods for the in vivo production of phenol from renewable substrates using a recombinant microorganism (FIG.). Additionally, methods for the in vivo production of catechol and cis,cis-muconic acid from renewable substrates using a recombinant microorganism are disclosed. A host cell expresses at least one gene encoding a polypeptide that possesses isochorismate synthase activity, at least one gene encoding a polypeptide that possesses isochorismate pyruvate lyase activity, and at least one gene encoding a polypeptide that possesses salicylic acid decarboxylase activity. In the case of catechol, the host cell must additionally express at least one gene encoding a polypeptide that possesses phenol 2-monooxygenase activity. In the case of cis,cis-muconic acid, the host cell must additionally express at least one gene encoding a polypeptide that possesses phenol 2-monooxygenase activity and at least one gene encoding a polypeptide that possesses catechol-1,2-dioxygenase activity.

Microorganisms engineered to produce phenol and its derivatives

Construction and expression of synthetic pathways to produce (5) or (R)-3-hydroxybutyrate (3HB) as enantiomerically-pure products by genetically engineering cyanobacteriumsp. PCC 6803. Under optimized growth conditions, the pathway employing phaA and phaB fromwas the most effective, producing up to 533.4&#xb1;5.5 mg/l (R)-3HB after 21 days photosynthetic cultivation. For the first time, the feasibility and high efficiency of producing 3HB using solar energy and COas sole energy and carbon sources by engineered cyanobacteria is demonstrated.

A method for the in vivo production of styrene from renewable substrates using a recombinant microorganism is disclosed. Additionally, a method for the in vivo production of styrene oxide from renewable substrates using a recombinant microorganism is also disclosed. In both cases, the host cell expresses at least one gene encoding a polypeptide that possesses phenylalanine ammonia lyase activity in addition to at least one gene encoding a polypeptide that possesses trans-cinnamic acid decarboxylase activity. In the case of styrene oxide, the host cell must additionally express at least one gene encoding a polypeptide that possesses styrene monooxygenase activity.

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Tempe, AZ, United States of America

David Nielsen