Cleaning compositions for solid surfaces – auxiliary compositions – Cleaning compositions or processes of preparing – For cleaning a specific substrate or removing a specific...
Reexamination Certificate
1998-12-23
2001-12-11
Delcotto, Gregory (Department: 1751)
Cleaning compositions for solid surfaces, auxiliary compositions
Cleaning compositions or processes of preparing
For cleaning a specific substrate or removing a specific...
C510S321000, C510S392000, C510S393000
Reexamination Certificate
active
06329332
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to phenol oxidizing enzymes, in particular, phenol oxidizing enzymes obtainable from strains of Pleurotus and methods for modifying colored compounds in textile, cleaning and pulp and paper applications at alkaline pH.
BACKGROUND OF THE INVENTION
Phenol oxidizing enzymes function by catalyzing redox reactions, i.e., the transfer of electrons from an electron donor (usually a phenolic compound) to molecular oxygen (which acts as an electron acceptor) which is reduced to H
2
O. While being capable of using a wide variety of different phenolic compounds as electron donors, phenol oxidizing enzymes are very specific for molecular oxygen as the electron acceptor.
Phenol oxidizing enzymes can be utilized for a wide variety of applications, including the detergent industry, the paper and pulp industry, the textile industry, the food industry and the wood processing industry. Most phenol oxidizing enzymes exhibit pH optima in the acidic pH range while being inactive in neutral or alkaline pHs.
Phenol oxidizing enzymes are known to be produced by a wide variety of fungi, including species of the genera Aspergillus, Neurospora, Podospora, Botytis, Pleurotus, Fomes, Phlebia, Trametes, Polyporus, Rhizoctonia and Lentinus. However, there remains a need to identify and isolate phenol oxidizing enzymes, and organisms capable of naturally-producing phenol oxidizing enzymes, which present pH optima in the alkaline range for use in pulp and paper applications and detergent washing methods.
SUMMARY OF THE INVENTION
The present invention relates to phenol oxidizing enzymes obtainable from
Pleurotus ostreatus
which are capable of modifying the color associated with dyes and colored compounds having different chemical structures, in particular at alkaline pH. Based on their color modifying ability,
Pleurotus ostreatus
phenol oxidizing enzymes of the present invention are used, for example, for pulp and paper bleaching at alkaline pH and for bleaching the color of stains on fabric at alkaline pH.
Accordingly, the present invention provides a method for modifying the color associated with a dye or colored compound in a sample comprising the step of contacting the sample with a composition comprising a purified phenol oxidizing enzyme at alkaline pH wherein, said purified phenol oxidizing enzyme is obtainable from
Pleurotus ostreatus
. In one aspect of the method, the pH is between about 7.5 and about 10.5 and in another aspect, the pH is between about 8 and about 10, and in another aspect, the pH is between about 8 and about 9. In another aspect of the method, said contacting takes place at a temperature between about 18° C. and up to about 50° C. and in another aspect between about 18° C. and up to about 60° C.
In one embodiment of the method, the phenol oxidizing enzyme is encoded by the genomic sequence as shown in
FIG. 1
(SEQ ID NO:1) and in another embodiment, the phenol oxidizing enzyme is encoded by the genomic sequence as shown in
FIG. 2
(SEQ ID NO: 2). In yet another embodiment of the method, the phenol oxidizing enzyme has the nucleic acid (SEQ ID NO:3) and amino acid (SEQ ID NO:4) as shown in FIG.
3
. The present invention also encompasses the use of phenol oxidizing enzymes that are variations of the amino acid sequence as shown in SEQ ID NO: 4 as long as the variation is able to modify the color associated with a dye or colored compound. Accordingly, in a further embodiment, the present invention encompasses the use of a phenol oxidizing enzyme obtainable from
Pleurotus ostreatus
and having at least about 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90% and at least 95% identity to the amino acid sequence disclosed in SEQ ID NO:4.
In one embodiment of the present invention, the phenol oxidizing enzymes are obtainable from strains of
Pleurotus ostreatus
, including, in particular,
Pleurotus ostreatus
having ATCC accession numbers 32783, 34672, 34673, 34674, 34675, 34676, 34677,44309, 58052, 58053 and 58054. In a preferred embodiment, the phenol oxidizing enzyme is obtainable from
Pleurotus ostreatus
ATCC accession number 32783.
Also provided herein are detergent compositions comprising a
Pleurotus ostreatus
phenol oxidizing enzyme of the present invention alone or in combination with an enhancer and other detergent ingredients, including proteases, amylases and/or cellulases.
Enhancers which can be used in detergent compositions of the present invention include but are not limited to phenothiazine-10-propionic acid (PPT), 10-methylphenothiazine (MPT), phenoxazine-10-propionic acid (PPO), 10-methylphenoxazine (MPO), 10-ethylphenothiazine-4-carboxylic acid (EPC) acetosyringone, syringaldehyde, methylsyringate, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonate (ABTS) and 4-Hydroxy-4-biphenyl-carboxylic acid or derivatives thereof.
The present invention also encompasses expression vectors and recombinant host cells comprising nucleic acid encoding a phenol oxidizing enzyme obtainable from
Pleurotus ostreatus
as well as methods for purifying the phenol oxidizing enzyme from such host cells. In a preferred embodiment, the host cell is an Aspergillus species. In another embodiment, the host cell is
Aspergillus niger
var.
awamori.
REFERENCES:
patent: WO 96/06930 A1 (1996-03-01), None
patent: WO 96/12846 A1 (1996-05-01), None
patent: WO 97/08325 (1997-03-01), None
patent: WO 97/11217 A1 (1997-03-01), None
patent: WO 98/27198 (1998-06-01), None
Copy of International Search Report for PCT/US99/30084, (7/01).
Giardina, P. et al., “The Gene Protein and Glycan Structures of Laccase fromPleurotus Osteatus,” Eur. J. Biochem., vol. 235, No. 3, Feb. 1, 1996, pp. 508-515, XP-000644722.
Giardina, P. et al., “The Gene Protein and Glycan Structures of Laccase fromPleurotus Osteatus,” EMBL Sequence Database, Apr. 20, 1995, XP-002136864.
Giardina, P. et al., “The Gene Protein and Glycan Structures of Laccase fromPleurotus Osteatus,” EMBL Sequence Database, Jun. 30, 1994, XP-002136865.
Giardina, P. et al., “The Gene Protein and Glycan Structures of Laccase fromPleurotus Osteatus,” EMBL Sequence Database, Nov. 1, 1997, XP-002136866.
Palmieri, G. et al., “Stability and activity of a phenol oxidase from the ligninolytic fungusPleurotus ostreatus”, Appl. Microbiol. Biotechnol., vol. 39, 1993, pp. 632-639, XP-000904759.
Bodie Elizabeth A.
Borneman William S.
Graycar Thomas P.
Winetzky Deborah S.
Delcotto Gregory
Genencor International Inc.
Ito Richard T.
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