# US 8,785,162 B2 · Aminotransferase and oxidoreductase nucleic acids and polypeptides and methods of using

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- Number: US 8,785,162 B2
- Document ID: US8785162B2
- Status: Granted
- Kind: B2
- Application number: 12/811,491
- Filed: 2008-12-31
- Published: 2011-01-27
- Granted: 2014-07-22
- Earliest claimed: 2008-01-03
- Assignees: Cargill, Incorporated
- Applicants: Mervyn L. De Souza; Paula M. Hicks; Sherry R. Kollmann; Jose Laplaza; Joshua M. Lundorff; Sara C. McFarlan; Erin Marasco; Wei Niu; Fernando A. Sanchez-Riera; Christopher Solheid; David P. Weiner; Peter Luginbuhl; Analia Bueno; Joslin Cuenca
- Inventors: Mervyn L. De Souza; Paula M. Hicks; Sherry R. Kollmann; Jose Laplaza; Joshua M. Lundorff; Sara C. McFarlan; Erin Marasco; Wei Niu; Fernando A. Sanchez-Riera; Christopher Solheid; David P. Weiner; Peter Luginbuhl; Analia Bueno; Joslin Cuenca
- Web page: https://patentgrep.com/patent/US8785162B2

## CPC

- C12P 13/227: {Tryptophan}
- C12N 9/0004: {Oxidoreductases (1.)}
- C12N 9/1096: {transferring nitrogenous groups (2.6)}
- C12P 17/10: Nitrogen as only ring hetero atom
- 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)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The invention provides for aminotransferase and oxidoreductase polypeptides and nucleic acids encoding such polypeptides. Also provided are methods of using such aminotransferase and oxidoreductase nucleic acids and polypeptides.

## Claims

### Claim 1

A method of converting tryptophan to indole-3-pyruvate or indole-3-pyruvate to tryptophan, comprising combining tryptophan or indole-3-pyruvate, respectively, with
a) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:219, wherein said nucleic acid molecule encodes the polypeptide comprising SEQ ID NO:220; or
b) a variant of a), wherein said variant comprises a variant nucleic acid molecule that encodes a variant polypeptide, wherein said variant polypeptide has at least 95% sequence identity to SEQ ID NO:220 and has aminotransferase activity.

### Claim 2 (depends on 1)

The method of claim 1, wherein said variant encodes a variant polypeptide that has at least 99% sequence identity to SEQ ID NO:220 and has aminotransferase activity.

### Claim 3 (depends on 1)

The method of claim 1, wherein said variant polypeptide is a mutant and the mutant is selected from one or more of the mutations shown in Table 43 or Table 52.

### Claim 4 (depends on 1)

The method of claim 1, wherein the variant nucleic acid molecule has been codon optimized.

### Claim 5 (depends on 1)

The method of claim 1, wherein the nucleic acid molecule or variant nucleic acid molecule is contained within an expression vector.

### Claim 6 (depends on 5)

The method of claim 5, wherein the nucleic acid molecule or variant nucleic acid molecule is overexpressed in an isolated transformed host cell.

### Claim 7 (depends on 1)

The method of claim 1, wherein the polypeptide or variant polypeptide is immobilized on a solid support.

### Claim 8 (depends on 1)

The method of claim 1, wherein the variant polypeptide is a chimeric polypeptide.

### Claim 9

A method of converting tryptophan to indole-3-pyruvate or indole-3-pyruvate to tryptophan, comprising combining tryptophan or indole-3-pyruvate, respectively, with a polypeptide comprising the amino acid sequence of SEQ ID NO:220.

### Claim 10 (depends on 9)

The method of claim 9, wherein the polypeptide is immobilized on a solid support.

### Claim 11

A method of converting tryptophan to indole-3-pyruvate or indole-3-pyruvate to tryptophan, comprising combining tryptophan or indole-3-pyruvate, respectively, with a variant polypeptide that has mat least 95% sequence identity to SEQ ID NO:220 and (ii) aminotransferase activity.

### Claim 12 (depends on 11)

The method of claim 11, wherein the variant polypeptide is a mutant selected from one or more mutations shown in Table 52.

### Claim 13 (depends on 11)

The method of claim 11, wherein said variant polypeptide has (i) at least 99% sequence identity to SEQ ID NO:220 and (ii) aminotransferase activity.

### Claim 14 (depends on 13)

The method of claim 13, wherein the variant polypeptide is a mutant selected from one or more mutations shown in Table 43.

### Claim 15 (depends on 11)

The method of claim 11, wherein the variant polypeptide is immobilized on a solid support.

### Claim 16 (depends on 11)

The method of claim 11, wherein the variant polypeptide is a chimeric polypeptide.