Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1997-04-04
1998-11-03
Patterson, Jr., Charles L.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
510392, C12N 920, C11D 3386
Patent
active
058307362
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a 35 U.S.C. 371 national application of PCT/DK95/00425 filed Oct. 26, 1995 and claims priority under 35 U.S.C. 119 of Danish application 1235/94 filed Oct. 26, 1994, the contents of which are fully incorporated herein by reference.
TECHNICAL FIELD
The present invention relates to novel lipolytic enzymes. More specifically the invention provides novel lipolytic enzymes having the properties of a lipase native to the strain Fusarium culmorum CBS 513.94, or has immunochemical properties identical or partially identical to those of a lipase native to the strain Fusarium culmorum CBS 513.94.
BACKGROUND ART
Lipolytic enzymes find multiple industrial applications. Alkaline lipases are of particular interest for use in detergent compositions.
Alkaline lipases of microbial origin have been described, including lipases obtained from Fusarium. However, lipases obtained from Fusarium culmorum have never been disclosed.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide novel alkaline lipolytic enzymes (EC 3.1.1.3).
Accordingly, in its first aspect, the invention provides lipolytic enzymes having immunochemical properties identical or partially identical to those of a lipase obtained from the strain Fusarium culmorum CBS 513.94.
In its second aspect, the invention provides a process for the preparation of the lipolytic enzyme, which process comprises cultivation of a lipase producing strain of Fusarium culmorum in a suitable nutrient medium, containing carbon and nitrogen sources and other inorganic salts, followed by recovery of the lipolytic enzyme.
In its third aspect, the invention provides a process for the preparation of a lipolytic enzyme according to any of claims 1-6, which process comprises isolating a DNA fragment encoding the lipolytic enzyme; combining the DNA fragment with an appropriate expression signal in an appropriate plasmid vector; introducing the plasmid vector into an appropriate host either as an autonomously replicating plasmid or integrated into the chromosome; cultivating the host organism under conditions leading to expression of the lipolytic enzyme; and recovering of the enzyme from the culture medium.
In further aspects, the invention provides detergent compositions, as well as a detergent additives, comprising the lipolytic enzyme of the invention.
Finally, the invention provides a biologically pure culture of the strain Fusarium culmorum CBS 513.94.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 show the relation between pH and lipase activity of the lipolytic enzyme of the present invention determined at 30.degree. C. in the range of from pH 6 to pH 10.
DETAILED DISCLOSURE OF THE INVENTION
The present invention provides novel lipolytic enzymes having the properties of a lipase native to the strain Fusarium culmorum CBS 513.94.
The Microorganism
The invention provides lipolytic enzymes derived from a strain of the fungus Fusarium culmorum. Fusarium culmorum is a known species and strains of Fusarium culmorum have been deposited and are publicly available from depositary institutes, e.g. Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM), Germany, and American Type Culture Collection (ATCC), U.S.A.
In a preferred embodiment the invention provides a lipolytic enzyme derived from the strain Fusarium culmorum DSM 1094, Fusarium culmorum DSM Fusarium culmorum 62184, Fusarium culmorum DSM 62188, Fusarium culmorum DSM 62191, Fusarium culmorum DSM 62223, Fusarium culmorum ATCC 12656, Fusarium culmorum ATCC 15620, Fusarium culmorum ATCC 16430, Fusarium culmorum ATCC 16551, Fusarium culmorum ATCC 26556, Fusarium culmorum ATCC 34910, Fusarium culmorum ATCC 34913, Fusarium culmorum ATCC 36017, Fusarium culmorum ATCC 36879, Fusarium culmorum ATCC 36881, Fusarium culmorum ATCC 36886, Fusarium culmorum ATCC 44417, Fusarium culmorum ATCC 46040, Fusarium culmorum ATCC 56088, Fusarium culmorum ATCC 56089, Fusarium culmorum ATCC 60275, Fusarium culmorum ATCC 60362, Fusarium culmorum ATCC 62214
REFERENCES:
patent: 5439811 (1995-08-01), Yamashita et al.
Nagao, et al., J. Biochem, vol. 116, pp. 536-540 (Mar. 7, 1994).
Igumenov, et al., Prikl Biochem, vol. 18(5), pp. 652-658 (1982).
Lin, et al., Physiological Plant Pathology, vol. 17, pp. 1-15 (1980).
Soliday, et al., Archives of Biochemistry & Biophysics, vol. 176, pp. 334-343 (Feb. 25, 1976).
Ohkouchi et al., (1993) SEN-I Gakkaishi 49(4), 182-189.
Nakanishi et al., SEN-I Gakkaishi, vol. 49, No. 11, pp. 586-593 (1993).
P.E. Kolattukudy, Institute of Biological Chemistry and Biochemistry/Biophysics Program, From "Lipases", Borgstrom, et al, Editors (1984) pp. 471-504.
Nakanishi. S., et al. (1994) Chemical Abstracts 120:211249x.
Anant Patkar Shamkant
Borch Kim
Oxenb.o slashed.ll Karen Margrethe
Harrington James
Novo Nordisk A S
Patterson Jr. Charles L.
Zelson Steve T.
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