Chemistry: natural resins or derivatives; peptides or proteins; – Peptides of 3 to 100 amino acid residues – 15 to 23 amino acid residues in defined sequence
Patent
1996-08-19
1999-01-19
Wax, Robert A.
Chemistry: natural resins or derivatives; peptides or proteins;
Peptides of 3 to 100 amino acid residues
15 to 23 amino acid residues in defined sequence
530300, 530350, 536 231, 536 236, 514 13, 4352523, 43525233, 435410, 435419, 4353201, C07K 708
Patent
active
058614800
DESCRIPTION:
BRIEF SUMMARY
This application claims benefit of international application PCT/GB95/00509, filed Mar. 9, 1995.
This invention relates to antimicrobial proteins, processes for their manufacture and use, and DNA sequences encoding them. In particular it relates to antimicrobial proteins capable of being isolated from seeds of Aralia or Impatiens.
In this context, antimicrobial proteins are defined as proteins or peptides possessing at least one of the following activities: antifungal activity (which may include anti-yeast activity); antibacterial activity. Activity includes a range of antagonistic effects such as partial inhibition or death. Antimicrobial proteins may be oligomeric or may be single peptide units.
The genus Aralia is part of the Araliaceae, a medium-sized plant family whose best known members are ivy and ginseng. Medicinal extracts have been obtained from some Aralia species, such as Aralia cordata.
The genus Impatiens is part of the Balsaminaceae plant family. There are five hundred to six hundred Impatiens species, many of which are commercially cultivated as greenhouse or pot plants.
Plants produce a wide array of antifungal compounds to combat potential invaders and over the last ten years it has become clear that proteins with antifungal activity form an important part of these defences. Several classes of such proteins have been described including thionins, beta-1,3-glucanases, ribosome-inactivating proteins, zeamatins, chitin-binding lectins and chitinases. . These proteins have gained considerable attention as they could potentially be used as biocontrol agents.
Antimicrobial proteins with activity against plant pathogenic fungi have been isolated from certain plant species. We have previously described the structural and antifungal properties of several such proteins, including: Biol Chem, 267:2228-2233; International Application Publication Number WO92/15691); 1992, Biochemistry, 37:4308-4314; International Application Publication Number WO92/21699); Delphinium, Cr-AFP from Catapodium, Ba-AFP from Bantisia and Ml-AFP from Microsensis (International Patent Application Publication Number WO94/11511); Biol Chem, 267:15301-13309) and related proteins such as Bn-AFP1 and Bn-AFP2 from Brassica napus, Br-AFP1 and Br-AFP2 from Brassica rapa, Sa-AFP1 and Sa-AFP2 from Sinapis alba, At-AFP1 from Arabidopsis thaliana, Dm-AMP1 and Dm-AMP2 from Dahlia merckii, Cb-AMP1 and Cb-AMP2 from Cnicus benedictus, Lc-AMP from Lathyrus cicera, Ct-AMP1 and Ct-AMP2 from Clitoria ternatea, Rs-nsLTP from Raphanus sativus (International Patent Application Publication Number WO93/05153).
These and other plant-derived antimicrobial proteins are useful as fungicides or antibiotics, particularly for agricultural purposes. The proteins may be applied to or around a plant or may be expressed within a plant.
We have now purified new potent antimicrobial proteins.
According to the present invention, there is provided an antimicrobial protein of about 3 kDa, capable of being isolated from seeds of Aralia or Impatiens.
We have purified two new antimicrobial proteins from seeds of Aralia chinensis, hereafter called Arc-AMP1 (Aralia chinensis--antimicrobial protein 1) and Arc-AMP2 (Aralia chinensis--antimicrobial protein 2).
We have also purified four new antimicrobial proteins from seeds of Impatiens balsamina, hereafter called Ib-AMP1 (Impatiens balsamina--antimicrobial protein 1), Ib-AMP2 (Impatiens balsamina--antimicrobial protein 2), Ib-AMP3 (Impatiens balsamina--antimicrobial protein 3) and Ib-AMP4 (Impatiens balsamina--antimicrobial protein 4).
An antimicrobial protein according to the invention is capable of being isolated from seeds of Aralia or Impatiens, and may also be capable of isolation from the seeds of both related and unrelated species, or may be produced or synthesised by any suitable method.
According to the present invention, there is further provided an antimicrobial protein having an amino acid sequence which is substantially homologous to a sequence selected from the group consisting of SEQ ID NO 10, SEQ ID NO 11,
REFERENCES:
Tailor et al. "A novel family of small cystein-rich antimicrobial peptides from seed of Impatiens balsamina . . . " J. Biol. Chem. 272 (39), 24480-24487, Sep. 26, 1997.
Attenborough Shelia
Broekaert Willem Frans
Osborn Rupert William
Ray John Anthony
Rees Sarah Bronwen
Nashed Nashaat T.
Thomson Marian T.
Wax Robert A.
Zeneca Limited
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