Animal feed additives

Details Animal feed additives Animal feed additives

1998-07-15

2001-06-12

Hobbs, Lisa J. (Department: 1652)

Chemistry: molecular biology and microbiology

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

Hydrolase

C435S254110, C435S006120, C426S053000, C426S623000, C426S635000, C536S023200

Reexamination Certificate

active

06245546

ABSTRACT:

CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation of serial no. PCT/DK96/00046 filed Jan. 26, 1996 and claims priority under 35 U.S.C. 119 of Danish application serial no. 0094/95 filed Jan. 26, 1995, the contents of which are fully incorporated herein by reference.
1. Technical Field
The present invention relates to animal feed additives, which additives comprise a monocomponent xylanase derived from a strain of Byssochlamus, Chaetomium, Humicola, Malbranchea, Mucor, Myceliophthora, Paecilomiyces, Talaromyces, Thermoascus, or Thielavia. In other aspects, the invention relates to monocomponent xylanase preparations, DNA constructs, recombinant expression vectors, host cells, and methods of producing monocomponent xylanase preparations.
2. Background Art
The types and amount of plant raw materials which can be used as components in animal feeds will often be limited by the ability of the animals to digest them. Feed enhancing enzymes are enzymes, usually of microbial origin, that by improving feed digestibility are able to increase the efficiency of its utilization.
Xylanolytic enzymes (EC 3.2.1.8) are well known as feed enhancing enzymes. Xylanases obtained from strains of Bacillus, Aspergillus, Trichoderna, Acremonium have been reported. Moreover, an enzyme preparation obtained by submerged fermentation of
Humicola insolens
have been marketed (Bio-Feed™Plus, available from Novo Nordisk A/S, Denmark).
Xylanase preparations obtained from strains of the fungus Thermomyces lanuginosus (Syn. Humicola lanuginosa) have been described [cf. Lischnig T, Purkarthofer H and Steiner W;
Biotechnology Letters
1993 15 (4) 411-414; Gomes J, Purkarthofer H, Hayn M, Kapplmüller J, Sinner M, and Steiner W,
Appl. Microbiol. Biotechnol
. 1993 39 700-707].
However, the use of a
Thermomyces lanuginosus
xylanase as a feed enhancing enzyme has never been disclosed.
Moreover, the xylanase preparations described in the prior art all relates to complex enzyme preparations comprising multiple enzyme components. Monocomponent xylanase preparations derived from Thermomyces by use of recombinant DNA technology have never been disclosed.
For many applications, the use of complex enzyme preparations is considered beneficial due to a synergistic effect arising from the co-operative action of multiple components. For some applications, e.g. the conversion of lignocellulose into liquid feedstocks or fuel, the processing of foods, and in particular for increasing digestibility of animal feed, a mixture of xylanolytic and cellulytic enzymes is regarded having optimal performance [Alam M, Gomes I, Mohiuddin G, & Hoq M M;
Enzyme Microb. Technol
. 1994 16 298-302].
SUMMARY OF THE INVENTION
According to the present invention it has now been found that when compared to conventional feed enhancing enzymes, the xylanase derived from
Thermomyces lanuginosus
is an excellent feed enhancing enzyme which shows significant improvement of the feed utilization when added to animal feed. Moreover, owing to an excellent thermostability, the xylanase preparation derived from
Thermomyces lanuginosus
is particularly well suited for being processed into feed additives at conditions preventing microbial infections, in particular Salmonella infection. It has also been found that the xylanase derived from
Thermomyces lanuginosus
exerts a significant reduction of digesta viscosity, which indicates a significant improvement in the chicken feed conversion efficiency.
Finally it has surprisingly been found that the recombinantly produced Thermomyces xylanase is significantly more thermotstable than the native xylnase, which makes the recombinantly produced xylanase particularly well suited for being processed into feed additives at conditions preventing microbial infections, in particular Salmonella infection.
Therefore it is an object of the present invention to provide a monocomponent xylanase preparation, which xylanase component is obtained by recombinant DNA techniques from a strain of Thermomzyces or a related genus.
Accordingly, in its first aspect, the present invention provides an animal feed additive, which additive comprises a monocomponent xylanase derived from a strain of Byssochlamus, Chaetomium, Humicola, Malbranchea, Mucor, Myceliophthora, Paecilomnyces, Talaromyces, Thermoascus, or Thielavia.
In another aspect, the present invention provides a monocomponent xylanase preparation, in which preparation the xylanase component is derived from a strain of Byssochlamus, Chaetomium, Humicola, Malbranchea, Mucor, Myceliophthora, Paecilomyces, Talaromyces, Thermoascus, or Thielavia.
In a further aspect, the invention relates to a DNA construct comprising a DNA sequence encoding a xylanase component, which DNA sequence comprises:
a) the xylanase encoding part of the DNA sequence presented as SEQ ID NO:1, or the DNA sequence obtainable from the plasmid in the strain
Saccharomyces cerevisiae
DSM 10133; or
b) a DNA sequence analogue to the xylanase encoding part of the DNA sequence presented as SEQ ID NO:1, or to the DNA sequence obtainable from the plasmid in the strain
Saccharomyces cerevisiae
DSM 10133, which analog DNA sequence either
i) is homologous to the xylanase encoding part of the DNA sequence presented as SEQ ID NO:1, or to the DNA sequence obtainable from the plasmid in the strain
Saccharomyces cerevisiae
DSM 10133; or
ii) hybridizes with the same oligonucleotide probe as the xylanase encoding part of the DNA sequence presented as SEQ ID NO:1, or with the DNA sequence obtainable from the plasmid in the strain
Saccharomyces cerevisiae
DSM 10133; or
iii) encodes a polypeptide which is at least 70% homologous to the polypeptide encoded by the DNA sequence presented as SEQ ID NO:1, or to the DNA sequence obtainable from the plasmid in the strain
Saccharomyces cerevisiae
DSM 10133; or
iv) encodes a polypeptide which is immunologically reactive with an antibody raised against the purified xylanase derived from the strain
Thermomyces lanuginosus
, DSM 4109, or encoded by the DNA sequence presented as SEQ ID NO:1, or the DNA sequence obtainable from the plasmid in the strain
Saccharomyces cerevisiae
DSM 10133.
In yet further aspects, the invention relates to an expression vector harbouring a DNA construct of the invention, a host cell comprising the DNA construct or expression vector, and a method of producing a mono component xylanase preparation of the invention, which method comprises culturing said host cell under conditions permitting the production of the enzyme, and recovering the enzyme from the culture.


REFERENCES:
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patent: WO 93/24621 (1993-12-01), None
patent: WO 94/21785 (1994-09-01), None
Gomes, et al., Appl. Microbiol Biotechnol, vol. 39, p. 700-707 (1993).
Lischnig T., et al., Biotechnology Letters, vol. 15, No. 4, p. 411-414, (Apr. 1993).
Mustafa Alam, et al., Enzyme Microb., vol. 16, p. 298-302, (Apr. 1994).
Kitpreechavanich V, et al., Dialog Info. Serv., accession No. 4460997 (1994).
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Ganju, et al., Canadian Jounal of Microbiology, vol. 35, No. 9 p. 836-842 (1989).
Anand et al., “Purification and Properties of Xylanase From the Thermophilic Fungus, Humicola Lanuginosa”, Archives of Biochemistry and Bipphysics, vol. 276, No. 2, Feb. 1, pp. 546-553, 1990.
Torronen, et al., Biotechnology, vol. 10, p. 1461-1465 (Nov. 1992).
Apel, et al., Molecular Plant Microbe Interactions, vol. 6, No. 4, p. 467-472 (Apr. 1993).

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