SU896071A1 - Lactobacillus acidophilus 578 strain as bacteriocin a-578 producent - Google Patents

Description

one

This invention relates to the microbiological industry and relates to a new strain producing bacteriocin.

A known strain of Lactobaci acidophiPus b74 is a bacteriocidal producer A-b74 1.

However, the known bacteriocin-producing strain is not active against cultures.

The purpose of the invention is a new strain of Lactobacieeus with claa ± from that producing bacteriocin active against LpCantarum cultures.

The proposed LactobaciC us acidophitus b78 strain was isolated from human saliva.

The strain Lactobaci & us acidophitua 578 (stored in the Museum of Cultures of Microorganisms of the Institute of Scientific Research Institute of Energetics under the number CMMP-1910) is a bacteriocin A-B78 producer.

Cultural and morphological signs. On a medium, the MPC-1 (48-hour culture) is polymorphic sticks 5-7 0.6-0.8 µm, arranged singly and in pairs sequentially with rounded ends. Volutin-grains do not form.

Gram-positive. Dispute and capsules do not have. Motionless.

Acetate Agar. After 72 hours of growth, carbon dioxide gas in the atmosphere will form up to 2 mm in diameter, round, grayish colonies, with a slightly convex, wrinkled, dull, dry surface, translucent, with a wavy edge, fibrous structure, and a dense texture.

Medium MPC-1. After 24 h of cultivation with uniform turbidity and a small amorphous precipitate.

1.5% agar mowed medium

15 MPC-1. After 48 hours of growth, with a grayish patina with a rough, dull, dry surface, a translucent, dense consistency, located on the surface of the medium.

20

Medium MPC-5. After 24 h, under cultivation at 37 ° C, a uniform turbidity of the medium and a small amorphous precipitate are observed.

Wednesday MPS-2. Does not grow at 50 and

25 Medium MPC-2 with 0.4% tipol. Growth is absent. Skimmed milk with 5% yeast autolysite and 1% glucose. Guards for 2-3 days.

Wednesday Davis. Staking and

30 yellowing of the medium for 2-3 days.

Milk. Guards for 4-5 days.

Physiological and biochemical signs. Optional anaerobic. The optimal growth at 32-37 C. Gelatin does not thin. Nitrates in nitrites do not restore. Catalase does not form.

On the Gibson and Abd-el-Malek medium, ferment glucose without generating gas. It ferments cellobiose, galactose, maltose, sucrose, mannose, lactose and salicin to form an acid. Does not cleave rhamnose, starch, sorbitol and mannitol.

Milk. After 14 days, the acid content reaches 0.38%.

Antagonistic properties. Bacteriocin A-b78 on 1.5 agar and liquid MPC-1 medium will be pleased.

Production of bacteriocin A-578 on 1.5% MPC-1 agar medium.

A daily culture of L.acidophifus 578 is inoculated on a plate with 1.5% MPC-1 agar medium and grown for 3–3 days at 37 ° C in a vacuum desiccator in which air is replaced by carbon dioxide. The grown macrocolonium is sterilized with chloroform vapor, after which the surface of the medium is poured with a second layer of 4 ml of 0.8 MPA-1 agar medium, melted and cooled, mixed with 0.5 ml of the 4-hour L.actis ATCC 800 culture culture. grown at 37 ° C for 2 days. Around the macrocolony, L.acidophiCus 578 is seen to be transparent with a not quite clear edge. Zones of growth inhibition of the indicator strain 16 mm in diameter. .....

Obtaining bacteriocin A-578 in liquid medium MPC-1.

Culture L.acidophi us 578 entre-. is watered in MPC-1 liquid medium and grown at 32 ° C for 2 days. For sterilization, 10% chloroform is added to the culture, mixed, and aged. for 30 minutes, after which the liquid is separated off with 6 tons of chloroform and centrifuged. Twice serial dilutions were prepared from the supernatant. One loop of each dilution is applied to the so-called cup surface with 1.5% MPC-J. Agar medium, seeded with 4 hours of L.bastig. ATCC 800 indicator culture. After growing for 2 days, bacteriocin A is counted -578 on liquid MPC-1 - 1; 4.

Spectrum of bacteriocin A-b78. .

Suppresses the growth of a number of cultures of L.acidophiCus, L.heCveficus, L.saBivarins L.

L.jugurti, J ... actis, L. Beichmannii, L.de6bineckii, L.casei, L.fermentum, L.brevis, L.bucheuri, a and L.pCantarum.

Bacteriocin A-578 is inactivated by trypsin, chymotrypsin, pepsin and papain, it is not destroyed by deoxyribonuclease, ribonuclease and lozicyma. Autoclaving at 1.5 atm is destroyed after 30 minutes. Diffuses through cellophane. Bacteriocin A-b78 replicates on the indicator strain L.Eactis ATCC 800.

The proposed strain L.acidophiCus 578 produces bacteriocin A-578 in comparison with the known ones, which can be used not only for bacteriocinotyping and bacteriocinogenotyping cultures of L.acidohiCus, L.heKveficus, L..saCivarins, L, jugurti, L.Pactis, L.bichmannii , L. deebmeckii, L. casei, L.fermentvim, L.brevis, and L.buchueri, but also of L. R. Claucher's cultures.

According to their sensitivity to bacteriocin A-578 (bacteriocinotyping), the cultures of the above-listed species of bacteriocotypes are divided into the following bacteriocinotypes: the first is sensitive to bacteriocin A-578 and the second is resistant to bacteriocin A-57

When bacteriocotyping 586 cultures of lactobacilus of different species, the following results were obtained.

Of the 187 cultures of L.acidophiCus, 155 belong to the first bacteriocin type and 32 cultures to the second. Of the 65 cultures of L.saBivarius 15, belong to the first bacteriocinotype and 50 to the third. Of the 16 L.jugiirti 15 cultures, it belongs to the first bacteriocotype and i culture to the second. Of the B4 cultures, L. fermentum 49 refers to the first bacteriocotype and 35 cultures to the second. Of the 13 L.burchueri cultures, 4 belong to the first bacteriocin type. And 9 cultures belong to the second. Of the 144 L.casci cultures, 82 belong to the first bacteriocotype and the B2 culture to the second, out of 23 cultures of L.p anterem 2 belong to the first bacteriocinotype and 21 cultures to the second.

4 cultures of L.heSveticus, 2 cultures of L.Eactis and L.6eichmannii, 1 Iculture Ir.aeCdmeckii and 43 cultures of L.brevis belong to the first bacteriocin type. 2 cultures of L.ceC obiosu are resistant to the action of bacteriocin A-578.

The use of the proposed bacteriocin A-578 producing strain makes it possible by genetic labels to carry out a further, deeper and more detailed intraspecific subdivision not only of cultures of L. acidophi6us, .leEveticus, L.saCivariVS ,, L jugrti, L, 6actis, L feichmannii, Jj, deCbmeckii, L.casei, L.fermeotvm, L.brevis and L.buchneri, but also L.pCanturum cultures with the same biochemical and physiological properties for bacteriocinotypes and bacteriocinogenotypes (bacteriocinotyping and bacteriocinogenotyping), also finds use in genetics research ziology of bacteriocinogenesis of lactobacilli, in the study of the chemical structure and mechanism of action of bacteriocin A-L78.

Claims (1)

1. USSR author's certificate on the loan 2685277, cl. C 12 D 9/22, 10 1978.