Enzymes with xylanase activity from Aspergillus aculeatus

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

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4352523, 4352541, 4352542, 4352543, 4353201, 536 232, 536 2374, C12N 924, C12N 122, C12N 1500, C07H 2104

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active

056935188

DESCRIPTION:

BRIEF SUMMARY
CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of PCT/DK94/00088 filed Mar. 2, 1994, which is incorporated herein by reference.


FIELD OF INVENTION

The present invention relates to an enzyme with xylanase activity, a method of producing the enzyme, an enzyme preparation containing the enzyme, and use of the enzyme for various industrial purposes.


BACKGROUND OF THE INVENTION

Xylan, a major component of plant hemicellulose, is a polymer of D-xylose linked by beta-1,4-xylosidic bonds. Xylan can be degraded to xylose and xylo-oligomers by acid or enzymatic hydrolysis. Enzymatic hydrolysis of xylan produces free sugars without the by-products formed with acid (e.g. furans).
Enzymes which are capable of degrading xylan and other plant cell wall polysaccharides are important for the food industry, primarily for baking and in fruit and vegetable processing such as fruit juice production or wine making, where their ability to catalyse the degradation of the backbone or side chains of the plant cell wall polysaccharide is utilised (Visser et al., Xylans and Xylanases, 1991).
Other applications for xylanases are enzymatic breakdown of agricultural wastes for production of alcohol fuels, enzymatic treatment of animal feeds for hydrolysis of pentosans, manufacturing of dissolving pulps Organic Chemicals from Biomass, (CRC Press, Boca Raton, Fla., 1981) 19-41.; Paice, M. G., and L. Jurasek., J. Wood Chem. Technol. 4: 187-198.; Pommier, J. C., J. L. Fuentes, G. Goma., Tappi Journal (1989): 187-191.; Senior, D. J., et al., Biotechnol. Letters 10 (1988):907-912!.
WO 92/17573 discloses a substantially pure xylanase derived from the fungal species H. insolens and recombinant DNA encoding said xylanase. The xylanase is stated to be useful as a baking agent, a feed additive, and in the preparation of paper and pulp.
WO 92/01793 discloses a xylanase derived from the fungal species Aspergillus tubigensis. It is mentioned, but not shown that related xylanases may be derived from other filamentous fungi, examples of which are Aspergillus, Disporotrichum, Penicillium, Neurospora, Fusarium and Trichoderma. The xylanases are stated to be useful in the preparation of bread or animal feed, in brewing and in reducing viscosity or improving filterability of cereal starch.
Shei et al., 1985, and Fournier et al., 1985 describe purification and characterization of endoxylanases isolated from A. niger.
WO 91/19782 and EP 463 706 discloses xylanase derived from Aspergillus niger origin and the recombinant production thereof. The xylanase is stated to be useful for baking, brewing, in the paper making industry, and in the treatment of agricultural waste, etc.


SUMMARY OF THE INVENTION

It is an object of the present invention to prepare single-component xylanases.
Accordingly, the present invention relates to an enzyme exhibiting xylanase activity, which enzyme is immunologically reactive with an antibody raised against a purified xylanase derived from Aspergillus aculeatus, CBS 101.43.
In the present context, the term "derived from" is intended not only to indicate a xylanase produced by strain CBS 101.43, but also a xylanase encoded by a DNA sequence isolated from strain CBS 101.43 and produced in a host organism transformed with said DNA sequence.
In another aspect, the invention relates to an enzyme exhibiting xylanase activity, which enzyme is encoded by a DNA sequence comprising at least one of the following partial sequences


______________________________________ (a) CATCAACATT CATTCATTCA (SEQ ID No. 7) (b) TTTAATTCAT TCCTCAAGCT (SEQ ID No. 8) (c) CAAGAGCAGT CATCCCTTCT (SEQ ID No. 9) (d) TTCCAACATG GTTCAAATCA (SEQ ID No. 10) (e) AAGCAGCTGC TCTGGCTGTC (SEQ ID No. 11) (f) CTTTTCGCCA GCAATGTGCT (SEQ ID No. 12) (g) CTCCAACCCC ATCGAGCCCCG (SEQ ID No. 13) (h) CCAGGCCTCG GTGAGCATCGA (SEQ ID No. 14) (i) TGCCAAATTA CAAGGCGCACG (SEQ ID No. 15) (j) CAAGAAGTAC CTGGGCACCAT (SEQ ID No. 16) (k) GAACCCCCAC AATCACGCAA (SEQ ID No. 17) (1) AAATGGTCGG ACTGCTTTCA (SEQ ID No. 18) (m) ATCACCGCGG CGC

REFERENCES:
Paice et al., J. of Wood Chem. and Tech., vol. 4 (2), p. 187-198, 1984.
Pommier et al., Tappi J., p. 187-191, 1989.
Senior et al., Biotech. Letters, vol. 10, No. 12, p. 907-912, 1988.
Shei et al., Biotech. and Bioeng., vol. XXVII, p. 533-538, 1984.
Murao et al., J. Ferment Tech., vol. 57, No. 3, p. 151-156, 1979.
Kiyoshi et al., Biosci. Biotech. Biochem., vol. 56 (6), p. 906-912, 1992.
Haas et al., Biochimica et Biophysica Acta, 1117, p. 279-286, 1992.
Grepinet et al., J. of Bacteriology, pp. 4576-4581, 4582-4588, 1988.
Shareck et al., Gene, vol. 197, p. 75-82, 1991.
Ito et al., Biosci. Biotech., Biochem., vol. 56 (8), p. 1338-1340, 1992.
Detroym R.W, CRC Press, p. 19-41, 1981.

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