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1. WO2016094604 - PHÉNYLPYRUVATE DÉCARBOXYLASE GÉNÉTIQUEMENT MODIFIÉE, PROCÉDÉS POUR LA PRÉPARER ET UTILISATIONS DE CETTE DERNIÈRE

Note: Texte fondé sur des processus automatiques de reconnaissance optique de caractères. Seule la version PDF a une valeur juridique

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CLAIMS

A process for genetically modifying a microorganism comprising

(A) selecting a microorganism that produces a C7-C11 2-ketoacid; and

(B) inserting a non-native nucleic acid sequence that encodes an amino acid sequence corresponding to SEQ. ID 4, 8, 14, 16, 18, 28, 30, 32, 34, 36, 38, 40, 42, 46, 52, 54, 56, 62, 64, 66, 68, or 76, or an amino acid sequence that is at least 90 percent homologous thereto;

such that a non-native phenylpyruvate decarboxylase is expressed in the microoorganism.

The process of Claim 1 wherein

(A) the microorganism is Escherichia coli;

(B) the non-native nucleic acid sequence is obtained from Azospirillum brasilense and optionally modified to correspond to the SEQ ID or is at least 90 percent homologous thereto prior to insertion; and

(C) the non-native phenylpyruvate decarboxylase takes part in a metabolic pathway that converts the C7-C11 2-ketoacids to C6-C10 aldehydes.

The process of Claim 2 wherein the metabolic pathway proceeds during anaerobic fermentation.

The genetically modified microorganism of any of Claims 1 to 3.

The microorganism of any of Claims 1 to 4 wherein

(A) the microorganism is Escherichia coli;

(B) the amino acid sequence is obtained from Azospirillum brasilense.

The microorganism of any of Claims 2 to 4 wherein

(A) the microorganism is a Clostridium species;

(B) the wild type metabolic pathway includes is a Wood-Ljungdahl pathway; and

(C) the amino acid sequence is obtained from Azospirillum brasilense.

A process to prepare a C6-C10 aldehyde, C6-C10 alcohol, a C6-C10 carboxylic acid, or a C6-C10 alkane, comprising the steps of

(A) contacting 2-ketobutyrate or 2-ketoisovalerate, isopropylmalate synthase, isopropylmalate isomerase, and isopropylmalate dehydrogenase,

under conditions such that the 2-ketobutyrate or 2-ketoisovalerate is converted to a C7-C11 2-ketoacid;

(B) contacting the C7-C11 2-ketoacid and a phenylpyruvate decarboxylase

which is expressed by a non-native nucleic acid sequence that encodes an amino acid sequence corresponding to SEQ ID 4, 8, 14, 16, 18, 28, 30, 32, 34, 36, 38, 40, 42, 46, 52, 54, 56, 62, 64, 66, 68, or 76, or an amino acid sequence that is at least 90 percent homologous thereto; under conditions such that the C7-C11 2-ketoacid is converted to a C6-C10 aldehyde having one less carbon atom than the C7-C11 2-ketoacid being converted; and

(C) optionally, contacting the C6-C10 aldehyde and

(1) an alcohol dehydrogenase under conditions to form a C6-C10 alcohol; or

(2) an aldehyde dehydrogenase under conditions to form a C6-C10 carboxylic acid; or

(3) a fatty aldehyde decarbonylase under conditions to form a C6-C10 alkane;

the process being carried out such that each step and substep occurs independently within or outside of a microbial organism

and under aerobic or anaerobic conditions.

The process of Claim 7 wherein the amino acid sequence is selected from the group consisting of SEQ ID 34, 36, 38, 40, 42, 46, 62, 68, and 76.

A polypeptide comprising an amino acid sequence corresponding to SEQ ID. 8, 14, 16, 18, 28, 30, 32, 34, 36, 38, 40, 42, 46, 52, 54, 56, 62, 64, 66, 68, or 76, or at least 90 percent homologous thereto.

The polypeptide of Claim 9 wherein the amino acid sequence is at least 95 percent homologous thereto.