Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Oxidoreductase
Patent
1995-11-02
1998-09-01
Lankford, Jr., Leon B.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Oxidoreductase
435191, C12N 902, C12N 906
Patent
active
058010351
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a 371 of PCT/DK94/00168 filed Apr. 27, 1994, which is incorporated herein by reference.
TECHNICAL FIELD
The present invention relates to a novel L-Amino acid oxidase having a broad substrate specificity and being obtainable from the species Trichoderma harzianum. The enzyme of the invention may be used for in situ generation of hydrogen peroxide in various contexts and may be advantageously incorporated, together with a substrate, into detergent compositions which comprise also a peroxidase system for inhibiting the transfer of dye from dyed fabric to other fabrics during washing.
BACKGROUND ART
The use of bleaching agents in washing procedures and as constituents of detergent compositions is well known in the art. Thus, bleaching agents are incorporated in a large part of the commercially available detergent compositions or are sold as separate preparations designed for adding to a detergent when the wash is executed.
Bleaching agents incorporated into detergent compositions are most often precursors of hydrogen peroxide. Perborates and percarbonates are the most important examples of compounds which are employed as bleaching agents.
The original purposes of adding bleaching agents to detergents were to achieve and maintain general whiteness of garments and, more specifically, to bleach stains thereon, such as tea, coffee, fruit juices, red wine, etc.
Later, bleaching systems involving or derived from hydrogen peroxide have also been invoked for reducing dye transfer, i.e. for preventing the discoloration resulting from excess dye leaching from dyed fabrics and being deposited on other fabrics, but recently a system involving a hydrogen peroxide source and peroxidases (and possibly further components has been disclosed, vide e.g. International Patent Application WO 91/05839. One possibility for a hydrogen peroxide source taught by this patent application is an oxidase system, i.e. an oxidase together with a substrate for it. Such an oxidase must of course be compatible with the detergent matrix and, in particular, with the peroxidase system.
An L-amino acid oxidase from Trichoderma viride, having very high substrate-specificity to L-lysine has been reported in GB 2015532.
SUMMARY OF THE INVENTION
It has now been found that a novel L-amino acid oxidase with a very broad substrate-specificity is obtainable from Trichoderma harzianum. This novel enzyme possesses excellent properties for in situ generation of hydrogen peroxide.
Accordingly, the present invention provides an L-amino acid oxidase (E.C. 1.4.3.2) having a pH optimum in the 15 range of from pH 8.5 to 9.5 (determined after incubation for 20 minutes at 20.degree. C. in the presence of L-arginine), a pH stability of 80% or more relative to initial activity at pH 9.5 (determined after incubation for 1 hour at 40.degree. C. in the absence of substrate), and activity towards L-arginine, L-lysine, L-methionine, L-asparagine, L-phenylalanine, and L-leucine.
In another aspect, the invention provides a process for the preparation of an L-amino acid oxidase of the invention, which process comprises cultivation of an L-amino acid oxidase producing strain of Trichoderma harzianum, in a suitable nutrient medium, containing carbon and nitrogen sources and inorganic salts, followed by recovery of the desired enzyme.
In a further aspect, the invention provides a detergent additive comprising an L-amino acid oxidase of the invention, the additive being provided in the form of a granulate, preferably a non-dusting granulate, a liquid, in particular a stabilized liquid, a slurry, or a protected enzyme.
In a yet further aspect, the invention provides a detergent composition comprising an L-amino acid oxidase of the invention and a substrate for the oxidase.
MICROORGANISMS
A strain of Trichoderma harzianum A611 was deposited on 22 Feb. 1993 according to the Budapest Treaty on the International Recognition of the Deposits of Microorganisms for the Purpose of Patent Procedures, at Centraal Bureau vo
REFERENCES:
patent: 4234691 (1980-11-01), Kusakabe et al.
Hansen Svend Aage
Pedersen Anders Hjelholt
Schneider Palle
Lambiris Elias J.
Lankford , Jr. Leon B.
Novo Nordisk A S
Zelson Steve T.
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