JPS58179439A - Animal feed additive or feed - Google Patents

Abstract

PURPOSE:A feed additive capable of improving extremely growth effect, feed efficiency, and digestibility, containing a variant of Bacillus natto having raised ability to produce an enzyme that is exerted by a microorganism, obtained by subjecting Bacillus natto to variation operation. CONSTITUTION:An animal feed additive containing a variant of Bacillus natto having raised ability to produce an enzyme (e.g., protease, amylase, cellulase, etc.), obtained by subjecting a wild strain of Bacillus natto to variation operation. The variant of Bacillus natto used is easily prepared by irradiation, treatment with a chemical substance (e.g., nitrosoguanidine, nitrous acid, etc.), method using the transformation ability of Bacillus subtilis, method by genetic engineering, etc. Growth effect, feed efficiency and digestibility can be extremely improved by adding a pharmaceutical preparation of the variant of Bacillus natto to animal feed.

Description

[Detailed description of the invention] The present invention relates to animal feed additives or feeds (/ζ).

More specifically, the present invention relates to a feed additive or feed 1 containing a mutant Bacillus natto with enhanced extracellular enzyme production ability as an active ingredient.

Currently, animal feed is undergoing a transition from conventional home-made feed to compound feed produced in modern factories, and the proportion of compound feed in all feed has become extremely high. It has become.

In addition to the main raw materials such as grains, oil cakes, fishmeal, oils and fats, these compounded feeds contain nutritional supplements, nutritional enhancement, improved digestion and absorption, growth promotion, and (b) divine purposes such as disease prevention. Amino acids, minerals, vitamins, antibiotics, enzymes, etc. are not used as auxiliary raw materials.However, among these, antibiotics cause allergies and changes in intestinal bacteria due to their residual transfer. Because of the risk of this, regulations regarding its use are being tightened, and the range of use is being limited and reduced.

Regarding enzymes, lipase agents, amylase agents, cellulase agents, protease agents, mixed enzyme agents, etc. are being studied, but their effects vary and it is currently not possible to expect consistent effects.

The present inventors have mutated the wild-type Bacillus natto strain to increase the ability to produce extracellular enzymes (grotease, amylase, cellulase, etc.). The present invention has been completed based on the discovery that it exhibits excellent effects such as improved efficiency and digestibility.5 That is, according to the present invention, (1) Obtained by mutating Bacillus natto;
A feed additive containing mutant Bacillus natto with increased ability to produce extracellular enzymes such as grotease, amylase, and cellulase.

and (2) obtained by mutating Bacillus natto.
Feed containing mutant natto bacteria with increased ability to produce extracellular enzymes such as grotease, amylase, and cellulase.

is provided.

The present invention will be explained in detail below.

Bacillus natto has been eaten for a long time, and it is well known that there is no problem with its safety. As is clear from the fact that it belongs to the same taxonomic and genetic species as Bacillus subtilis, it is one of the most stable bacteria in nature.

Currently, feed prepared from Bacillus natto spores is commercially available, but its mechanism of action is not clear and its effects are not sufficient.

The present invention uses Bacillus natto, which has a significantly higher extracellular enzyme production ability than wild-type Bacillus natto, and is more effective than conventional Bacillus natto preparations.

The animals targeted by the animal feed additive or feed of the present invention include livestock such as beef cattle, dairy cattle, calves, pigs, small pigs, sheep, goats, horses, rabbits, dogs, and cats; and Meat chickens, egg-laying hens, chicks,
Typical examples include poultry such as breeding chickens, ducks, geese, turkeys, quail, and foals.

The mutant Bacillus natto used in the present invention can be easily produced by normal mutation operations. For example, radiation (X-rays, r
Exocellular enzyme production can be achieved by any of the following methods: irradiation (rays, ultraviolet rays); treatment with chemicals such as trosoguanidine, nitrous acid, and base analogues; methods utilizing the transformation ability of Bacillus subtilis; and genetic manipulation methods. Even Bacillus natto with enhanced ability can be used in the present invention.

The mutant Bacillus natto may be cultured by liquid culture or solid culture according to the usual fermentation method.

As the medium, natural medium, semi-synthetic medium, synthetic medium, etc. can all be used. Composition of medium 1d (1) Carbon sources include glucose, sucrose, dextrin,
Maltose, glycerin, starch, molasses, etc. (11
) Nitrogen sources include peptone, meat extract, yeast extract,
Dry yeast, dog meal, urea, thiourea, ammonium salts, nitrates, and other organic or inorganic nitrogen compounds (i.
ii) As the inorganic salt, phosphates, nitrates, carbonates, chlorides, etc. of magne/rum, manganese, potassium, carunium, iron, etc. are used. In addition, amino acids, vitamins,
Nucleic acids and related compounds may also be added. The culture temperature is 10°C to 70°C in Nishinomiya, and 25°C to 50°C in Heavy. Further, the pH during culturing is usually pH 5 to pH 10, preferably pH 6 to pH 9. The culture period is long (1 hour ~
It contains in some form the mutant Bacillus natto obtained after 10 days, preferably 12 hours to 511 hours. For example, use the mutant Bacillus natto culture solution in a trench, or use flour,
It can be used by diluting it with an appropriate diluent such as starch, dextrin, etc., grains as raw materials for feed, rice bran, oil cake, etc.

Alternatively, only the bacterial cells may be separated from the culture solution, diluted in the same manner, and used.

The dosage of the animal feed additive or feed of the present invention cannot be unconditionally defined as it can vary depending on the target animal, the age of the target animal, and other components of the feed, but it is usually is 102 to 1015 pieces/day, preferably 1
04~1012 (solid/day, most preferably 105~
It is recommended to add 1010 pieces/day.

An example of a method for producing a mutant Bacillus natto and a method for producing a mutant Bacillus natto preparation according to the present invention will be shown below.

Method for producing mutant natto bacteria: Molasses 2y1 ammonium sulfate 12, K2HPO40-1? in a 500m7 Sakaguchi flask. , add yeast extract 05V, and add 100rrLt of tap water.

After adjusting the pH to 8.0 with 1N HCl and 1N NaOH, it is sterilized and inoculated with Bacillus natto isolated from commercially available natto. 3
Culture is carried out with shaking at 7°C, and bacterial cells in the logarithmic growth phase are taken out. This bacterial body was irradiated with 15 watts of ultraviolet light for a certain period of time, and
Heat treatment is performed at 0°C for 30 minutes.

This bacterial cell fluid was inoculated into a solid medium with the same medium composition as above,
After culturing at 37°C for 24 hours, the best growing strain was picked.

Among the 10 mutant strains thus obtained, we selected the 1L-1 strain, which has a high protease activity (casein degrading ability), which is about 120 times higher than the wild strain.

Method for producing a mutant Bacillus natto preparation: 1 ton of medium of the same group rJ-'i as above was placed in a jar fermentor, and after sterilization, the mutant Bacillus natto that had been precultured was inoculated and cultured for 241 hours to form spores. . After separating the bacterial cells from centrifugation (C), 1 kg of starch was added and dried to prepare a mutant Bacillus natto preparation.

An example of an animal administration test using the modified Y4 Bacillus natto spore preparation with high protease production ability obtained in this way is shown below.

Example 1 Using 120 broiler chickens (60 female each), each group:
11 groups of 30 birds (15 males and 15 females) were fed the basic feed shown in Table 1, a control group (starch added), a conventional natto bacterium preparation added, and a mutant natto SZ supplemented group in 2 groups. A total of 4 districts were established, and one feeding report examination was conducted over an 8-week period.

The results are shown in Table 2.

It showed remarkable efficacy in both body weight gain and feed conversion rate compared to conventional Bacillus natto preparations.

In addition, Table 5 shows the measurement results of digestibility in Table 5 (Table 1) Composition of this steel material) 4 Japanese compounded feed ■ Semi-test feed for SD broilers Early period SD broilers Late period 0 Crude protein (%) ≧23 ≧19 Crude fat (X)
≧ 5 ≧ 6 coarse fiber cone <X> ≦ 4
≦ 4 composition (%) ≦6.5 665M
1! (kc vomit/kg) ≧ 3000
≧ 3100 or more.As is clear from the results, adding the mutant Bacillus natto preparation to animal feed has the effect of increasing wages,
Feed efficiency and digestibility can be significantly improved.

patent applicant Mitsui Toatsu Chemical Co., Ltd.

Claims (6)

[Claims] (1) Obtained by mutating Bacillus natto,
A feed additive containing mutant natto bacteria with increased extracellular enzyme production ability. (2) The feed additive according to claim 1, which contains a mutant Bacillus natto with increased protease production ability. (3) The feed additive according to claim 1, which contains a mutant Bacillus natto with increased amylase production ability. (4) The feed additive according to claim 1, which contains a mutant Bacillus natto with increased cellulase production ability. (5) Obtained by mutating Bacillus natto,
Feed containing mutant natto bacteria with enhanced extracellular enzyme production ability. (6) Feed 1 according to claim 5 containing a mutant Bacillus natto with increased protease production ability; (7) Feed 1 comprising a mutant Bacillus natto with increased amylase production ability. (8) The feed according to claim 5, which contains a mutant reticulum sigma bacterium with increased cellulase production ability.