JP3268509B2 - Microbial carrier - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a carrier having a high alkaline and active microorganism fixability.

[0002]

2. Description of the Related Art Conventionally, as a carrier of microorganisms, natural materials such as crushed stone, lapilli, a mixture of ballas and sand, and the like are known.
Those having a large specific surface area and a high porosity are preferably used. In recent years, attempts have been made to artificially produce a porous ceramic body and use it as a carrier for microorganisms.

[0003]

However, the types of microorganisms are extremely large, the applications are diverse, the conditions such as culture conditions are complicated, and there is no universal support for all microorganisms. In particular, when trying to use microorganisms that are active under weakly alkaline conditions, there are examples in which shell fossils, coral lime, and the like are used as carriers, but they cannot be said to be perfect as microorganism carriers. An object of the present invention is to provide a good carrier for a microorganism which is alkaline and active.

[0004]

The microorganism carrier according to the present invention is composed of calcium carbonate having pores having a diameter of 1 to 10 μm in a single particle , or a powder of the calcium carbonate is used as a binder. And granulated and dried.

[0005] Limestone, which is a raw material of calcium carbonate satisfying this condition, is often produced in Southeast Asia. The present inventors have tried to use limestone and dolomite as a carrier of microorganisms, and as a result, among limestones produced naturally, soft limestone produced on a plateau at an altitude of about 300 m near Uonosari, south of Jogjagarta, Indonesia ( (Hereinafter referred to as soft limestone) has been found to have much better microbial activity than ordinary limestone, and has completed the present invention. When this soft limestone is observed with an electron microscope, calcium carbonate particles having a diameter of 0.5 to 50 μm exist in a single particle state or in a lightly aggregated state, and individual calcium carbonate particles having a diameter of 2 μm or more (single particles) One particle) has pores having a diameter of 1 to 10 μm. The origin of the vacancies is not clear.

[0006] This soft limestone is not a hard one usually produced in Japan from the Paleozoic Carboniferous to the Mesozoic, but a shallow marine calcareous sediment formed after the Cenozoic. Some are porous and have a water absorption of as much as 20%. In addition, since the degree of agglomeration is light, it is soft, has no corners, is rounded, and can be easily dispersed into single particles. The chemical composition is close to pure calcium carbonate and contains slight amounts of magnesium, silica, iron, aluminum and the like as impurities. On the other hand, such vacancies do not exist in single particles of domestic limestone, for example, kudzu limestone (dolomite) and Okinawan limestone (coral).

From the viewpoint of use, it is convenient to granulate and dry the above calcium carbonate particles using a binder. When the microorganism is added and fixed using the granulated material as a carrier, high fixability is exhibited. When the bioactivity was compared using this granulated product as a microorganism carrier, it showed much better bioactivity than ordinary limestone having the same particle size.

[0008] The binder used in the granulation includes starch, sodium alginate, gum arabic, casein,
Gelatin, methyl cellulose, polyvinyl alcohol,
It is possible to use hydrophilic binders such as sodium silicate and glycerin, and, depending on the application, hydrophobic binders such as paraffin. Binders also act as microbial elicitors. Granulation method, rolling granulation,
Any of pressing granulation and spray granulation may be used.

Hereinafter, the present invention will be described specifically with reference to Examples, but the present invention is not limited to the following Examples.

[0010]

Example 1 Indonesian limestone (soft limestone) in which single particles having pores having a diameter of 1 to 10 μm are lightly aggregated is sterilized at 140 ° C. for 2 hours.
It was adjusted to 5 mm or less, and the adjusted water content was set to 0.01%. On the other hand, microfabrication bacteria No. 12954 (FERM P-1295), which has been deposited at
4) was added to a liquid medium adjusted to pH 7.0 by adding 0.2% by weight of yeast extract and 0.5% by weight of peptone,
Shaking culture was performed at a rotation speed of 160 rpm for 168 hours. In this way, FERM P-12954 cultivated in a large amount to 1 × 10 ⁹ cells per 1 ml of the culture solution was added and fixed so that the number of cells per 1 g of the soft limestone became 1 × 10 ⁶ . The fixing method is as follows: 995 g of soft limestone is weighed and put into a stirrer while stirring.
1 ml of the culture solution containing FERM P-12954 cultured therein was diluted with 4 ml of sterilized water, and added using a spray. FERM P-12954 is a microorganism having an antibacterial activity against plant pathogenic fungi belonging to the genus Fusarium, and exhibits alkaline activity.

[0011]

Comparative Example 1 FERM P-1 mass-cultured in Example 1
2954 is sterilized at 140 ° C. for 2 hours, and the particle size is 0.5 m.
m and adjusted by the method of Example 1 so that the number of bacteria per g of Okinawan limestone (coral) adjusted to 1 m or less was 1 × 10 ⁶ .

[0012]

Comparative Example 2 Ferm P-1 Mass-cultured in Example 1
2954 is sterilized at 140 ° C. for 2 hours, and the particle size is 0.5 m.
m and adjusted by the method of Example 1 so that the number of bacteria per gram of limestone (dolomite) adjusted to 1 m or less was 1 × 10 ⁶ .

The microorganisms fixed on the carriers of Example 1 and Comparative Examples 1 and 2 were incubated at a temperature of 38 ° C. or -10 ° C., and the fixation rates after 10, 20, and 30 days were examined. . Table 1 shows the results. FERM of soft limestone used as carrier (Example 1)
The fixing rate of P-12954 showed a higher fixing rate than the other carriers (Comparative Examples 1 and 2) under the temperature condition of 38 ° C. or −10 ° C.

[Table 1]

[0014]

Example 2 The soft limestone used as a carrier in Example 1 was added to Microtechnological Laboratory No. 12955 (FERM P-1) deposited at the Institute of Microbial Industry and Technology, National Institute of Advanced Industrial Science and Technology.
2955) was cultured in the same manner as in Example 1. In this way, FERM P-12955, which had been cultured in a large amount to 1 × 10 ⁹ cells per 1 ml of the culture solution, was transformed into cells per 1 g of the soft limestone. Addition and fixing were performed in the same manner as in Example 1 so that the number became 1 × 10 ⁶ . FE
RM P-12955 is also a microorganism having an antibacterial activity against plant pathogenic fungi of the genus Fusarium, and is alkaline and active.

[0015]

[Comparative Example 3] FERM P-1 cultured in a large amount in Example 2
2955 was added and fixed to the same coral as used in Comparative Example 1 by the method of Example 1 so that the number of bacteria per gram was 1 × 10 ⁶ .

[0016]

[Comparative Example 4] FERM P-1 cultured in a large amount in Example 2
Example 1 was obtained by adding 2955 to dolomite similar to that used in Comparative Example 2 so that the number of bacteria per gram was 1 × 10 ^6.
And fixed by the method described above.

The microorganisms fixed on the carriers of Example 2 and Comparative Examples 3 and 4 were incubated at a temperature of 38 ° C. or -10 ° C., and the fixation rates after 10, 20, and 30 days were examined. . Table 2 shows the results. FERM of soft limestone used as carrier (Example 2)
The fixing rate of P-12955 showed a higher fixing rate than the other carriers (Comparative Examples 3 and 4) under the temperature condition of 38 ° C. or −10 ° C.

[Table 2]

[0018]

Example 3 Indonesian limestone (soft limestone) in which single particles having pores having a diameter of 1 to 10 μm are lightly agglomerated was ground through a 560 μm sieve at 140 ° C. for 2 hours.
The mixture was sterilized for an hour, added with 0.1% methylcellulose, granulated with a Roche granulator, dried at 110 ° C., and adjusted to a particle size of 2 to 4 mm. Then, FERM P-12954 cultivated in a large amount in Example 1 was added and fixed by the same method as in Example 1 so that the number of bacteria per 1 g of the granulated product was 1 × 10 ⁶ .

[0019]

Comparative Example 5 FERMP-12954 cultivated in large amounts in Example 1 was applied to corals granulated in the same manner as in Example 3 so that the number of bacteria per gram of coral granules was 1 × 10 ^6. The addition and fixing were carried out by the method of Example 1.

[0020]

Comparative Example 6 FERM P-12954 mass-cultured in Example 1 on dolomite granulated in the same manner as in Example 3
Was added and fixed by the method of Example 1 so that the number of bacteria per 1 g of dolomite granules was 1 × 10 ⁶ .

The microorganisms fixed on the carriers of Example 3 and Comparative Examples 5 and 6 were incubated at a temperature of 38 ° C. or -10 ° C., and the fixation rates after 10, 20, and 30 days were examined. . Table 3 shows the results. FERM of soft limestone used as carrier (Example 3)
The fixing rate of P-12954 showed a higher fixing rate than the other carriers (Comparative Examples 5 and 6) under the temperature conditions of 38 ° C. or −10 ° C.

[Table 3]

[0022]

Example 4 FER cultured in large amounts in Example 2 on the granules of soft limestone used as a support in Example 3
MP-12955 was added and fixed by the method of Example 1 so that the number of bacteria per 1 g of the granulated soft limestone was 1 × 10 ⁶ .

[0023]

Comparative Example 7 FERMP-129 cultured in large amounts in Example 2 was added to the coral granules used as a support in Comparative Example 5.
55 was added and fixed by the method of Example 1 so that the number of bacteria per g of coral granules was 1 × 10 ⁶ .

[0024]

Comparative Example 8 The FERM P-1 mass-cultured in Example 2 was added to the granulated dolomite used as a support in Comparative Example 6.
2955, the number of bacteria per 1 g of granulated dolomite was 1 × 1
0 by ^six to become as in Example 1. was added fixing.

The microorganisms fixed on the carriers of Example 4 and Comparative Examples 7 and 8 were incubated at a temperature of 38 ° C. or -10 ° C., and the fixation rates after 10, 20, and 30 days were examined. . Table 4 shows the results. FERM of soft limestone used as carrier (Example 4)
The fixing rate of P-12955 showed a higher fixing rate than the other carriers (Comparative Examples 7 and 8) under the temperature conditions of 38 ° C. or −10 ° C.

[Table 4]

[0026]

The microorganism carrier according to the present invention has a high fixability as a microorganism carrier.

──────────────────────────────────────────────────続 き Continued on front page (58) Field surveyed (Int. Cl. ⁷ , DB name) C12N 11/00-11/18

Claims (2)

(57) [Claims] 1. A microorganism carrier comprising a single particle of calcium carbonate having pores having a diameter of 1 to 10 μm . 2. A microorganism carrier comprising a calcium carbonate powder having pores having a diameter of 1 to 10 μm in a single particle, a binder added, granulated and dried.