Food or edible material: processes – compositions – and products – Fermentation processes – Of plant or plant derived material
Reexamination Certificate
2000-04-07
2003-03-18
Prouty, Rebecca E. (Department: 1652)
Food or edible material: processes, compositions, and products
Fermentation processes
Of plant or plant derived material
C435S254500, C435S183000, C435S203000, C435S209000, C435S197000, C435S256300, C426S656000, C426S018000, C426S044000
Reexamination Certificate
active
06534101
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a novel micro-organism, new enzymes and new enzymes mixture. In addition, the present invention relates to the composition of the enzymes mixture, its preparation and its use in feed, food and other industries including but not limited to the paper industry and the textile industry.
BACKGROUND OF THE INVENTION
Enzymes have been used for a long time for a variety of different industrial applications. Examples are known in the baking industry, in the wine and fruit juices industry (where enzymes are used to breakdown pectins and &bgr;-glucans), in the textile industry (where cellulases are used to obtain soft and smooth cellulosic fabrics) and also, which is not the least application, for animal feed. In this case the enzymes improve the digestibility of vegetable sources.
This last use enables the livestock to digest feed more efficiently. The value of a feed can be measured by the FCR (Feed Conversion Ratio), a nutritive ratio of the amount of feed consumed relative to the weight gain of the animal. A decrease in FCR, for a feed, indicates the animal gains proportionately more weight for a given quantity of feed ; i.e. the animal is able to utilize the feed more efficiently.
Poor digestibility of the feed components (starch, fat, protein/amino acids) is a noted feature of cereal-based feeds and, for example, particularly those containing a high barley or wheat content. In these cases it may be necessary to formulate the teed to contain higher levels of energy from other sources and other supplements such as amino acids. These enzymes increase the Apparent Metabolizable Energy value of the cereals incorporated into the Feed.
Another approach to resolve this problem has been to add enzyme supplements, cellulases, endo-1,3(4)-&bgr;-glucanases (&bgr;-glucanases), endo-1,4-&bgr;-xylanases (xylanases) etc., or mixtures of enzyme activities, to these cereal-based feeds. Enzyme supplements may have a specific use to hydrolyze the &bgr;-glucans, or to hydrolyze the arabinoxylans, found in the cereals (typically barley and wheat). The addition of enzymes has differents goals. One advantage which clearly proves the efficacy of feed enzyme supplements is the reduction in viscosity of materials in the gut of the animals which receive cereal-based feed containing the appropriate enzyme supplement. The higher viscosity is due, in part, to &bgr;-glucans and arabinoxylans found in barley and wheat. The lower viscosity, resulting from enzyme action, permits an easier absorption of nutritional components in the animal's gut. The other advantage is the release of nutrients entrapped by the cell walls of the cereals decreasing the requirement for other costly feed supplements. Overall the result is a significant reduction in the cost of the feed with a similar or beneficial effect as measured by the FCR.
Enzymes preparations originating from a range of different micro-organisms have been described to improve feed digestibility.
If we consider prior art related to the use of enzymes in the animal feed we can mention the European Patent No 0.699.762 which describes use of a phytase issued from
Schwanniomyces occidentalis
. This phytase is a phytase obtained from genetically modified organism obtained by including cloned gene that we would like to avoid in the present invention.
If we consider the WO 95/26398 patent application, again a modified cellulase is obtained by inclusion of foreign DNA sequence in an host cell which modifies the nature of the original strain which is chosen in the following list of micro-organisms: Bacillus, Streptomyces, Saccharomyces, Schizosaccharomyces, Aspergillus. In the present invention our main aim was to avoid foreign gene inclusion in the micro-organism which is the producer of the enzyme.
In the WO 96/05739 patent application, a mixture of enzymes (xylanase, protease and, optionally, &bgr;-glucanase) is obtained from different micro-organisms. The authors give example (page 5) of enzymes mixture with a ratio of xylanase activity to &bgr;-glucanase activity of the order of 1:5. It has been found that when a xylanase is included in a cereal-based diet at or around its optimum dosage level, the co-presence of enzymes possessing &bgr;-glucanase activity increase the FCR of the feed which is of course disadvantageous. Consequently the authors advise against the presence of &bgr;-glucanase, they recommend a maximum ratio of xylanase activity to &bgr;-glucanase activity of 1:0-0.25.
In some cases, in order to ensure all the enzyme activities relevant to the feed application are present, preparations are made up from preparations from more than one micro-organism. In a number of cases the enzyme preparations have been obtained from microorganisms subjected to genetic modification using recombinant DNA techniques.
We have discovered and developed a new micro-organism belonging to the class of
Penicillium funiculosum
, that contains new enzymes and a mixture of enzyme activities which can be used successfully to increase mainly the digestibility of cereal-based animal feeds.
SUMMARY OF THE INVENTION
Accordingly, the present invention relates to a new micro-organism derived from
Penicillium funiculosum
and a method for cultivating this micro-organism and for recovering the enzymes produced by this micro-organism.
In addition, in accordance with this invention, there are provided new enzymes issued from this micro-organism, nucleic acid sequences therefrom and new compositions containing those enzymes.
Further, in accordance with this invention, there is provided a method for improving the digestibility of aminoacids and cereal-based animal feeds and amino acids.
Another subject of the present invention is the reduction of phosphorus excretion and ammonia excretion from the battery where animals are fed.
DETAILED DESCRIPTION OF THE INVENTION
A. The New Strain
Penicillium funiculosum
This new strain of the fungus
Penicillium funiculosum
is deposited under the number IMI 378536 in a recognized International Depositary Authority under the Budapest Treaty (1977), the International Mycological Institute (IMI), Bakeham Lane, Englefield Green, Egham, Surrey, TW20 9TY, UK.
Filiation
The new strain has been obtained from
Penicillium funiculosum
IMI 134756 after successive UV and &bgr;-irradiations treatment of spores, including screening on selective medium. No genetic modification has been obtained by recombinant DNA techniques using inclusion of foreign DNA or RNA.
Identification and Typing
Penicillium funiculosum
IMI 378536 has been characterised by growth on Czapek Dox agar at 25° C. Colony characteristics and micro-morphology are typical for
Penicillium funiculosum
. The identification of the micro-organism as a
Penicillium funiculosum
has been confirmed at the International Mycological Institute, Bakeham Lane, Englefield Green, Egham, Surrey, TW20 9TY, UK. Growth is as a tough basal felt, with aerial growth, as ropes or bundles of hyphae (funiculose), mycelium is white with underlying red colouration in the substrate, margins are reverse pale but coloured red towards centres and may become deep red. This penicillium is typical, it shows conidiophores short mostly arising from funicles, biverticillate, acerose conidiogenous cells, conidia are elliptical and smooth.
The micro-organism used for the production of the enzyme preparation of this invention is grown under aerobic conditions in a medium which contains cellulose, corn steep liquor, calcium carbonate and ammonium sulphate.
B. Process of Fermentation
This new fungus is manufactured by fermentation of the deposited strain first on a seed medium preferably constituted of (in weight):
corn steep liquor
1% to 4%
antifoam
just to avoid foam
water
to 100%
NaOH
enough to adjust the pH to about pH 3.0 to
6.0 before sterilisation of the medium;
Temperature of incubation
27° C. to 36° C.
The production medium has preferably the following constitution (in weight):
corn steep liquor
0 to 4.0%
batched and fed cellulose
0.8 to 14%
Ca salt,
0 to 0.8%
Ammonium sul
Fish Neville Marshall
Haigh Nigel Paterson
Sabatier Alain
Aveatis Animal Nutrition S.A.
Baker & Botts LLP
Prouty Rebecca E.
Steadman David J.
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