# US 7,608,288 B2 · Caffeine fraction obtained from tea leaves and a method for inducing Agrobacterium tumefaciens-mediated genetic transformation in plants using said caffeine fraction

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 7,608,288 B2
- Document ID: US7608288B2
- Status: Granted
- Kind: B2
- Application number: 10/396,593
- Filed: 2003-03-25
- Published: 2004-01-15
- Granted: 2009-10-27
- Earliest claimed: 2003-03-25
- Assignees: Council of Scientific and Industrial Research
- Inventors: Indra Sandal; Ajay Kumar; Amita Bhattacharya; Ravindranath Srigiripuram Desikalhar; Ashu Gulati; Paramvir Singh Ahuja
- Web page: https://patentgrep.com/patent/US7608288B2

## CPC

- C12N 15/8205: {Agrobacterium mediated transformation}
- C07D 473/12: with methyl radicals in positions 1, 3, and 7, e.g. caffeine
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C07D: HETEROCYCLIC COMPOUNDS (macromolecular compounds C08)

## Abstract

The present invention relates to a thermolabile caffeine fraction useful for an efficient Agrobacterium-mediated genetic transformation in plant systems to develop desired traits in plants, and a method of preparing said fraction from tea leaves and also, an efficient and cost-effective method of introducing said Agrobacterium-mediated genetic transformation into plant systems using said caffeine fraction of tea leaves.

## Claims

### Claim 1

A thermolabile caffeine fraction produced by a method comprising:
a. extracting dried tea leaves with about 10-40 weight % aqueous acetone to obtain an aqueous acetone fraction,
b. extracting the aqueous acetone fraction of step (a) with n-hexane to obtain an aqueous layer and a lipid layer,
c. extracting the aqueous layer with petroleum ether and ethyl acetate to remove catechins from the aqueous layer, and
d. extracting the aqueous layer resulting from of step (c) with chloroform to obtain a chloroform fraction containing caffeine.

### Claim 2 (depends on 1)

The thermolabile caffeine fraction of claim 1, wherein the method further comprises concentrating the chloroform fraction to a lesser volume.