Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1994-10-05
1997-04-29
Wax, Robert A.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435 691, 4352523, 4352543, 4353201, 536 232, 536 2374, 935 14, 935 28, C12N 942, C12N 1556, C12N 1574, C12N 1580
Patent
active
056248350
DESCRIPTION:
BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of PCT/DK93/00108 filed Mar. 25, 1993, which is incorporated herein by reference.
The invention comprises an endo-.beta.-1,4-glucanase, a corresponding DNA sequence, a vector, a transformed host, a method for production of an endo-.beta.-1,4-glucanase, an enzyme preparation, and a use of the endo-/.beta.-1,4-glucanase.
The invention relates to genetic engineering and provides a partial amino acid sequence of an endo-.beta.-1,4-glucanase and partial DNA sequences.
Endo-.beta.-1,4-glucanases (EC no. 3.2.1.4) is a group of hydrolases, which catalyse endo hydrolysis of 1,4-.beta.-D-glycosidic linkages in cellulose, lichenin, cereal .beta.-D-glucans and other plant material containing cellulosic pans. The authorized name is endo-1,4-.beta.-D-glucan 4-glucano hydrolase, but the short term endo-.beta.-1,4-glucanase is used in this specification with claims. 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. Gokseyr 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). Celluloses are found in connection with many gums and they are components of cell walls in e.g. fruits, vegetables and cereals.
The above indicated partial amino acid sequence can be used for construction of DNA probes which can be used for screening a genomic library for organisms expressing such enzyme, or a cDNA library, thereby obtaining DNA sequences, which can be used either for an overproduction of endo-.beta.-1,4-glucanase, if inserted in the microorganism species, from which the parent DNA molecule originated, or for production of endo-.beta.-1,4-glucanase without accompanying closely related enzymes, if inserted in a host microorganism, which in its not-transformed condition does not produce any enzymes closely related to endo-.beta.-1,4-glucanase. The DNA sequences can be established otherwise, as will appear from the following.
Thus, the purpose of the invention is the provision of a new endo-.beta.-1,4-glucanase and of means and methods for production of endo-.beta.-1,4-glucanase in better yield and higher purity than hitherto possible, and of a use of endo-.beta.-1,4-glucanase either alone or in combination with other enzymes for degradation of plant cell wall tissue, more efficient than hitherto possible. Also it is the purpose of the invention to provide novel products, wherein the proportion of the endo-.beta.-1,4-glucanase is either increased or decreased in relation to the proportion in the original product.
The recombinant DNA sequence obtainable according to the invention comprises a DNA sequence coding for a polypeptide having endo-.beta.-1,4-glucanase activity, or a DNA sequence having substantial sequence homology to such endo-.beta.-1,4-glucanase coding sequence.
The endo-.beta.-1,4-glucanase according to the invention is characterized by the fact that it is immunologically reactive with an antibody raised against a purified endo-.beta.-1,4-glucanase derived from Aspergillus aculeatus, CBS 101.43.
In the present context the term "derived from" is intended not only to indicate an endo-.beta.-1,4-glucanase produced by strain CBS 101.43, but also an endo-.beta.-1,4-glucanase encoded by a DNA sequence isolated from strain CBS 101.43 and produced in a host organism transformed with said DNA sequence.
FIG. 1 shows the Ion Exchange Chromatograph (IEC) of the purification of the enzyme of the invention, depicting the endoglucanase fraction pooled from 0.2 m 0.25M NaCl (column: 7.5 * 20 cs, flow=75 ml/minute; cl
REFERENCES:
Torronen, A., et al., 1993, FEBS Letters, 321(2-3):135-139.
Dalb.o slashed.ge, H., et al., 1994, Molecular and General Genetics 243:253-260.
Williamson, G., et al., 1990, Carbohydrate Polymers, 13:387-397.
Ooi et al., Nucleic Acids Research, vol. 18, No. 19, p. 5884 (1990).
Murao et al., Agric. & Biological Chemistry, vol. 49, pp. 3511-3518 (1985).
Arai et al., Agric. & Biological Chemistry, vol. 51, pp. 627-633 (1987).
Ooi, T., et al., 1990, Current Genetics, 18:217-222.
Harkki, A., et al., 1991, Enzyme & Microbiol Technology, 13:227-233.
Sakamoto, R., et al., 1984, Journal of Fermentation Technology, 62(6):561-567.
Sakamoto, R., et al., 1985, Agricultural and Biological Chemistry, 49(5):1283-1290.
Yoshizumi, A., et al., 1987, Trends in Biotechnology 5(10):277-281.
Christensen Flemming M.
Dalb.o slashed.ge Henrik
Dorreich Kurt
Heldt-Hansen Hans P.
Mischler Marcel
Harrington, Esq. James
Moore William W.
Novo Nordisk A S
Wax Robert A.
Zelson Esq. Steve T.
LandOfFree
Endo-.beta.-1,4-glucanase and a DNA sequence does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Endo-.beta.-1,4-glucanase and a DNA sequence, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Endo-.beta.-1,4-glucanase and a DNA sequence will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-705090