# US 5,705,375 A · Transgenic plants having a modified carbohydrate content

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- Number: US 5,705,375 A
- Document ID: US5705375A
- Status: Granted
- Kind: A
- Application number: 08/253,575
- Filed: 1994-06-03
- Granted: 1998-01-06
- Earliest claimed: 1990-09-13
- Assignees: Mogen International N.V.
- Inventors: Albert Johannes Joseph Van Ooyen; Krijn Rietveld; Wilhelmus Johannes Quax; Petrus Josephus Maria Van Den Elzen; Jan Pen; Andreas Hoekema; Peter Christiaan Sijmons
- Web page: https://patentgrep.com/patent/US5705375A

## CPC

- C12N 15/8245: {involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention provides plants with a modified taste, solids content and/or texture. The invention also provides methods of obtaining such plants via transformation with DNA constructs containing genes encoding enzymes capable of degrading plant polysaccharides and optionally additional genes encoding enzymes which are capable of further modifying the degradation products resulting from the first degradation step.

## Claims

### Claim 1

A method for modifying the carbohydrate composition of a plant or plant organ, wherein said method comprises growing a stably transformed, transgenic plant containing a recombinant DNA expression construct encoding a microbial alpha-amylase under conditions wherein said alpha-amylase-encoding construct is expressed and the carbohydrate composition of said plant or plant organ is modified.

### Claim 2 (depends on 1)

The method of claim 1, wherein said expression construct comprises a regulatory sequence operably linked to a nucleotide sequence encoding said alpha-amylase, which regulatory sequence directs expression of said alpha-amylase-encoding nucleotide sequence at a selected stage of development or maturity of the transgenic plant or plant organ.

### Claim 3 (depends on 1)

The method of claim 1 wherein said expression construct contains a regulatory sequence operably linked to a nucleotide sequence encoding said alpha-amylase, which regulatory sequence directs tissue-specific expression of said alpha-amylase-encoding nucleotide sequence in the transgenic plant or plant organ.

### Claim 4 (depends on 1)

The method of claim 1 wherein the DNA expression construct comprises a nucleotide sequence encoding a fusion protein consisting of a leader amino acid sequence which is fused to said alpha-amylase, and wherein the leader amino acid sequence targets the alpha-amylase to a cellular compartment or organelle.

### Claim 5 (depends on 1)

The method of claim 1 wherein said transgenic plant contains at least one expression cassette which contains a nucleotide sequence encoding a second microbial enzyme.

### Claim 6 (depends on 5)

The method of claim 5 wherein said second microbial enzyme is glucoamylase.

### Claim 7 (depends on 1)

The method of claim 1 wherein said .alpha.-amylase originates from Bacillus licheniformis.

### Claim 8

A recombinant DNA expression cassette comprising a regulatory sequence operably linked to a nucleotide sequence encoding a microbial enzyme selected from the group consisting of alpha-amylase and glucoamylase, which regulatory sequence directs expression of said enzyme-encoding nucleotide sequence at a selected stage of development or maturity of a transgenic plant or plant organ.

### Claim 9

A recombinant DNA expression cassette comprising a 35S CaMV promoter operably linked to a nucleotide sequence encoding a microbial enzyme selected from the group consisting of alpha-amylase and glucoamylase.

### Claim 10

A recombinant DNA expression cassette comprising a regulatory sequence operably linked to a nucleotide sequence encoding a microbial enzyme selected from the group consisting of alpha-amylase and glucoamylase, which regulatory sequence directs tissue-specific expression of said enzyme-encoding nucleotide sequence in a plant.

### Claim 11 (depends on 8, 9, 10)

A vector comprising an expression cassette according to any of the claims 8-10.

### Claim 12 (depends on 8, 9, 10)

A stably transformed, transgenic plant characterized in that said plant contains an expression cassette encoding a microbial alpha-amylase according to any one of claims 8-10.

### Claim 13 (depends on 11)

A bacterial strain characterized in that said bacterial strain contains a vector according to claim 11.

### Claim 14 (depends on 1, 5)

A stably transformed, transgenic plant or plant organ, characterized in that said plant or plant organ contains a modified carbohydrate composition, said plant or plant organ being made by the method of either claim 1 or claim 5.

### Claim 15 (depends on 5)

The method of claim 5 wherein said second microbial enzyme uses the degradation products resulting from the action of said first enzyme as a substrate.

### Claim 16 (depends on 10)

The method of claim 10 wherein the glucoamylase originates from Aspergillus niger.

### Claim 17 (depends on 1, 2, 3, 4, 5, 6, 7, 15, 16)

The method of any one of claims 1-4, 5-7 and 15-16, wherein said transgenic plant is selected from the group consisting of tomato, potato, corn, cassava carrot, lettuce, strawberry and tobacco.