DNA encoding an enzyme with endoglucanase activity from Trichode

Details DNA encoding an enzyme with endoglucanase activity from Trichode DNA encoding an enzyme with endoglucanase activity from Trichode

1996-01-03

1998-10-06

Grimes, Eric

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

43525411, 4352543, 4353201, 435325, 536 232, C12N 510, C12N 924, C12N 1556, C12N 2102

Patent

active

058174990

DESCRIPTION:

BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of PCT/DK94/00275 filed Jul. 5, 1994, which is incorporated herein by reference.


FIELD OF INVENTION

The present invention relates to a DNA construct encoding an enzyme with endoglucanase activity, a method of producing the enzyme, an enzyme produced by the method, and an enzyme preparation containing the enzyme.


BACKGROUND OF THE INVENTION

Endoglucanases (EC no. 3.2.1.4) constitute a group of hydrolases, which catalyse endo hydrolysis of 1,4-.beta.-D-glycosidic linkages in cellulose, cellulose derivatives (such as carboxy methyl cellulose and hydroxy ethyl cellulose), lichenin, .beta.-1,4 bonds in mixed .beta.-1,3 glucans such as cereal .beta.-D-glucans or xyloglucans and other plant material containing cellulosic parts. The authorized name is endo-1,4-.beta.-D-glucan 4-glucano hydrolase, but the abbreviated term endoglucanase is used in the present specification. Reference can be made to R. F. Gould, "Cellulases and their Application", Advances in Chemistry Series 55, American Chemical Society (1969), T. M. Wood, "Properties and Mode of Action of Cellulases", in Biotechnology and Bioengineering Symposium, no. 5, John Wiley, 111-137 (1975), Y.-H. Lee and L. T. Fan, "Properties and Mode of Action of Cellulose", Advances in Biochemical engineering 17, 101-129 (1980), J. Goks.o slashed.yr and J. Eriksen, "Cellulases" in A. H. Rouse, Microbial Enzymes and Bioconversions, Academic Press, 283-330 (1980), T.-M. Enveri, "Microbial Cellulases" in W. M. Fogarty, Microbial Enzymes and Biotechnology, Applied Science Publishers, 183-224 (1983), Beguin, P. and Aubert, J-P., "The biological degradation of cellulose", FEMS Microbiology Reviews 13 (1994) 25-58. Celluloses are found in connection with many gums and they are components of cell walls in e.g. fruits, vegetables and cereals. Several cellulolytic genes from the genus Trichoderma have been cloned and characterized by Shoemaker et al. (1983), Teeri et al. (1983), Chen et al. (1987), Teeri et al. (1987), Penttila et al. (1986), van Arsdell et al. (1987), Saloheimo et al. (1988), Barnett et al. (1991) and Gonzalez et al. (1992).
The object of the invention is to provide a novel endoglucanase and a method for producing the endoglucanase in a better yield and higher purity than hitherto possible, as well as the use of the endoglucanase either alone or in combination with other enzymes for the degradation of plant cell wall tissue. Also it is the object of the invention to provide novel products, wherein the proportion of the endo-.beta.-1,4-glucanase is increased relative to the proportion in the original product.


SUMMARY OF THE INVENTION

The present inventors have now surprisingly isolated a novel DNA sequence from a strain of Trichoderma harzianum, which encodes an endoglucanase enzyme the deduced amino acid sequence of which has been found to be without homology to any of the above mentioned known endoglucanases or any other known enzymes.
Accordingly, in a first aspect the invention relates to a DNA construct comprising a DNA sequence encoding an enzyme with endoglucanase activity, which DNA sequence comprises the DNA sequence shown in SEQ ID No. 1 or an analogous sequence thereof being at least 70% homologous to the DNA sequence shown in SEQ ID No. 1.
The DNA sequence shown in SEQ ID No. 1 encodes an enzyme which in the following disclosure is referred to as Endoglucanase I.
In further aspects the invention relates to an expression vector harbouring a DNA construct of the invention, a cell comprising the DNA construct or expression vector and a method of producing an enzyme with endoglucanase activity which method comprises culturing said cell under conditions permitting the production of the enzyme, and recovering the enzyme from the culture.
In a still further aspect the invention relates to an enzyme with endoglucanase activity, which enzyme a) is encoded by a DNA construct of the invention as defined above, b) produced by the method of the invention, and/or c) is immunologica

REFERENCES:
Sidhu et al., Folia Microbiol. 31:293-302, 1986.
Dialog Information Services, File 351, Derwent WPI, Dialog accession No. 003541017, WPI accession No. 82-89009E/42, (AGEN) Agency of Sci Tech: "High Yield Cellulase Prodn. By Culturing Trichoderma harzianum In Medium Contg. Enzyme Hydrolysis Prod. Of Caseinas Cellulase Prodn. Accelerator", JP 57146579 A 820910 8242 (Basic) (1982).
Dialog Information Services, File 3512, Derwent WPI, Dialog accession No. 003541016, WPI, Dialog accession No. 82-89008E/42, (AGEN) Agency of Ind Sci Tech: "Cellulase Prodn. By Culturing Microorganizm Of Trichoderma Hazianum Genus", JP 57146578 A 82910 8242 (Basic) (1982).
Patent Abstracts of Japan, vol. 16, No. 257, C-949, abstract of Japan, A, 40-58889 (Aizo Matsushiro), 25 Feb. 1992 (25 Feb. 1992).

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