Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Oxidoreductase
Patent
1994-11-01
1998-12-22
Prouty, Rebecca E.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Oxidoreductase
435471, 536232, 510374, 510392, C12N 908, C12N 1509, C12N 1553
Patent
active
058518118
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a national phase filing under 37 CFR 371 of PCT/DK93/00189 filed Jun. 1, 1993, which is incorporated herein by reference.
FIELD OF INVENTION
The present invention relates to a novel variant of peroxidase, and a bleaching agent or detergent composition comprising the peroxidase variant.
BACKGROUND OF THE INVENTION
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 or sold as constituents of a major part of the commercially available detergent compositions. Important conventional bleaching agents incorporated in detergent compositions are compounds which act as precursors of hydrogen peroxide formed in the course of the washing procedure. Perborates and percarbonates are the most important examples of compounds which are employed as bleaching agents and which exert a bleaching effect in this fashion. The detailed mechanism of bleaching by means of these bleaching agents is not known at present, but it is generally assumed that the hydrogen peroxide formed during washing converts coloured substances (responsible for stains on fabric) into non-coloured materials by oxidation and that some oxidation of the coloured substances may also take place due to their direct interaction with perborate or percarbonate.
One drawback of these commonly used bleaching agents is that they are not particularly efficient at the lower temperatures at which coloured fabrics are usually washed. Their efficiency may be enhanced by the use of activators (e.g. organic acid anhydrides, esters or imides) which give rise to the formation of peracids.
Apart from being employed for bleaching stains on fabric, such conventional bleaching agents have also been suggested for preventing surplus dyes from coloured fabrics which leach from the fabrics when these are washed from being deposited on other fabrics present in the same wash (this phenomenon is commonly known as dye transfer). The problem of dye transfer, of course, is most noticeable when white or light-coloured fabrics are washed together with fabrics of a darker colour from which dye is leached during washing.
It has been found that peroxidases utilizing hydrogen peroxide as their substrate are able to enhance the bleaching effect of hydrogen peroxide during washing. The use of peroxidase for bleaching stains on fabrics is described in WO 89/09813. It was also found that coloured substances leached from dyed fabrics could be bleached by means of peroxidases. The use of peroxidase for inhibiting the transfer of dye from a dyed fabric to another fabric during washing is described in WO 91/05839.
SUMMARY OF THE INVENTION
It has surprisingly been found that peroxidase variants with an improved stability towards hydrogen peroxide may be prepared by recombinant DNA techniques.
Accordingly, the present invention relates to a peroxidase variant with improved hydrogen peroxide stability, characterized by insertion, deletion or substitution of one or more amino acid residues located in or near the substrate channel of the parent peroxidase, near the heme group of the parent peroxidase or at or near the active site of the parent peroxidase.
Information about the three-dimensional structure of the parent peroxidase was obtained by aligning the amino acid sequence of the parent peroxidase to amino acid sequences of other known peroxidases (K. G. Welinder et al., "Structure and evolution of peroxidases" in Plant Peroxidase Biochemistry and Physiology, K. G. Welinder et al. (eds.), University of Copenhagen and Geneva 1993, in press). The sequence alignment showed that the parent peroxidase was homologous to yeast cytochrome c peroxidase (CCP) which has a known structure (J. Wang et al. , Biochemistry 29, 1990, p. 7160), and the overall structure of the parent peroxidase could therefore be inferred from the CCP structure. An example of how structural information for a homologous peroxidase can be inferred from the cry
REFERENCES:
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patent: 5286638 (1994-02-01), Tanaka et al.
patent: 5346823 (1994-09-01), Estell et al.
K.G. Welinder et al. "Srructure of Plant and Fungal Peroxidases", Biochem. Soc. Trans. 20:337-340. (May 1992).
M. Kjalke et al.,"Comparison of Ftructure and Activities of Peroxidases from Coprinus cinereus, Coprinus macrorhizus and Arthromyces romosus", Biocim. Biophys. Acta 1120(3)248-256 (Apr. 1992).
Andersen Morten Birket
Welinder Karen Gjesing
Gregg, Esq. Valeta
Novo Nordisk A S
Prouty Rebecca E.
Zelson Esq. Steve T.
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