# US 7,745,184 B2 · Process for the biological production of 1,3-propanediol with high yield

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- Number: US 7,745,184 B2
- Document ID: US7745184B2
- Status: Granted
- Kind: B2
- Application number: 12/053,991
- Filed: 2008-03-24
- Published: 2008-07-24
- Granted: 2010-06-29
- Earliest claimed: 2003-10-06
- Assignees: E. I. Du Pont de Nemours and Company; Genencor International, Inc.
- Inventors: Marguerite A. Cervin; Philippe Soucaille; Fernando Valle
- Web page: https://patentgrep.com/patent/US7745184B2

## CPC

- C12P 7/18: polyhydric
- C12P 7/20: Glycerol
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}

## Abstract

The present invention provides a microorganism useful for biologically producing 1,3-propanediol from a fermentable carbon source at higher yield than was previously known. The complexity of the cofactor requirements necessitates the use of a whole cell catalyst for an industrial process that utilizes this reaction sequence to produce 1,3-propanediol. The invention provides a microorganism with disruptions in specified genes and alterations in the expression levels of specified genes that is useful in a higher yielding process to produce 1,3-propanediol.

## Claims

### Claim 1

A method for the bioproduction of 1,3-propanediol comprising contacting with a suitable carbon substrate under suitable conditions a E. coli strain comprising:
a) a disrupted endogenous phosphoenolpyruvate-glucose phosphotransferase system comprising one or more of:
i) a genetically disrupted endogenous ptsH gene preventing expression of active phosphocarrier protein;
ii) a genetically disrupted endogenous ptsl gene preventing expression of active phosphoenolpyruvate-protein phosphotransferase; and
iii) a genetically disrupted endogenous crr gene preventing expression of active glucose-specific IIA component;
b) a genetically up regulated endogenous galP gene encoding active galactose-proton symporter, said up regulation resulting in an increased galactose-proton symporter activity, wherein the up regulation is produced by:
i) introducing additional copies of said gene into said E. coli strain followed by integration; or
ii) replacing a native regulatory sequence with a strong non-native promoter or altered native promoter;
c) a genetically up regulated endogenous glk gene encoding active glucokinase, said up regulation resulting in an increased glucokinase activity, wherein the up regulation is produced by:
i) introducing additional copies of said gene into said E. coli strain followed by integration; or
ii) replacing a native regulatory sequence with a strong non-native promoter or altered native promoter; and
d) a genetically down regulated endogenous gapA gene encoding active glyceraldehyde 3-phosphate dehydrogenase, said down regulation resulting in a reduced glyceraldehyde 3-phosphate dehydrogenase activity;
whereby said E. coil strain is capable of bioconverting a suitable carbon source to 1,3-propanediol.

### Claim 2 (depends on 1)

The method of claim 1, wherein the E. coli strain comprises a genetically disrupted endogenous arcA gene preventing expression of active aerobic respiration control protein.

### Claim 3 (depends on 1)

The method of claim 1, wherein the E. coil strain further comprises:
(i) glycerol-3-phosphate dehydrogenase;
(ii) glycerol-3-phosphatase;
(iii) glycerol dehydratase or diol dehydratase; and
(iv) dehydratase reactivation factor.

### Claim 4 (depends on 2)

The method of claim 2, wherein the E. coil strain further comprises:
(i) glycerol-3-phosphate dehydrogenase;
(ii) glycerol-3-phosphatase;
(iii) glycerol dehydratase or diol dehydratase; and
(iv) dehydratase reactivation factor.