JPS6236489B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to Lactobacillus, which has unique characteristics that were completely unknown in the past, and which the present inventors have successfully isolated and collected.
It relates to live bacterial preparations. The present inventors have already confirmed that a special strain of bacteria belonging to the Lactobacillus family can deodorize human-accumulated fecal odor, bad breath, and bad breath, and have applied for a patent in Japan and the United States for part of the deodorization (patent application). Showa 49-
134773, U.S. Pat. No. 3,957,974). Through subsequent research by the present inventors, the present inventors did not simply limit themselves to the strains discovered by the present inventors, but also conducted more detailed research on what kind of properties Lactobacillus can have and what kind of effects it can exert on deodorization. By studying those results, we finally arrived at the present invention. That is, the present inventors succeeded in isolating a large number of Lactobacilli that exhibit remarkable deodorizing properties, and when actually administered to human stock orally and by sprinkling on human stock feces, good deodorizing results were obtained in both cases. We not only obtained the results, but also investigated the correlation with the properties of the bacteria, and finally found out what properties Lactobacillus possesses.
We succeeded in elucidating what kind of deodorizing effect it exhibits. Furthermore, it has become clear that there are important issues regarding the handling of Lactobacillus, which has deodorizing properties, such as the need to use culture methods, subculture methods, and storage methods that are different from those of conventionally known Lactobacillus.
Regarding the above, the conclusions of the experimental results confirmed by the present inventor regarding Lactobacillus having deodorizing properties are briefly summarized as follows. That is, it has been found that in order for Lactobacillus to be recognized as having deodorizing properties in our noses, it must have the following properties. The essential conditions are: (1) bile resistance; (2) The nutritional requirement is significantly lower than that of known Lactobacillus. (3) Fast growth rate even in medium with low nutritional composition. (4) Single or multiple bacterial strains should have characteristics consistent with the SNC theory discovered by the present inventor, which will be explained in detail later. It is. Here, the SNC conditions advocated by the present inventor are S compounds, N compounds, and C compounds (lower fatty acids) (hereinafter SNC) among the odorous components contained in feces.
This means that compounds (called compounds) can be ingested as a nutritional source. In addition to the above, in order to better demonstrate the deodorizing effect, (5) it must contain enough antibiotics and lactic acid, (6) it must be resistant to bacteriostatic agents (including spices), was found to be an important condition. These will be explained in detail, but first, the parts related to the present invention regarding conventional Lactobacillus will be described. The classification of the genus Lactobacillus is defined as follows according to Mr. Mitsuoka. It is a Gram-positive, facultative anaerobic, non-spore-forming bacillus that exhibits coccobacillary, curved, coryneform, filamentous, etc. shapes depending on the species, but it does not exhibit a severely branched shape. They are usually non-motile, negative for catalase, and do not reduce nitrate. Also, it does not decompose gelatin and does not produce indole or hydrogen sulfide. Some species exhibit double-end staining, have very weak proteolytic and lipolytic properties, grow better in anaerobic or microaerobic conditions than in aerobic conditions, and have poor glycolytic properties. It is strong and acid resistant and produces lactic acid with a yield of over 50% through fermentation of glucose. It is not pathogenic to animals or plants. Next, it is also known that Lactobacillus, a group of bacteria with properties that meet this definition, cannot grow unless a medium with a fairly good nutritional composition is used. It requires amino acids, peptides, nucleic acid-related substances, vitamins, salts, fatty acids or their esters, and sugars. However, although the general properties of the Lactobacillus that the present inventor has recognized as having deodorizing properties match those of conventionally known Lactobacillus, it does not meet the above-mentioned points (2), (3), and (4). It turned out that there was a huge difference. The present invention provides the following features: 1. Lactobacillus live bacteria that can grow in either (a) Stephenson-Wesam medium, (b) Stephenson-Wesam + vitamin medium, or (c) Stephenson-Wesam + Casamino acid medium, as nutritional requirements, are sulfur-containing. A live bacteria preparation characterized by its coexistence with amino acids. 2. As a nutrient requirement, Lactobacillus can be grown in either (a) Stephenson-Wesum medium, (b) Stephenson-Westam + vitamin medium, or (c) Stephenson-Westam + Casamino acid medium, and the SNC compound is used. The Lactobacillus bacteria that are essential for growth or whose growth is promoted are treated with at least one compound selected from the group consisting of SNC compounds, sulfur-containing amino acids, glycine, glutamic acid, lysine, alanine, phenylalanine, arginine, asparagine, and aspartic acid. A live bacteria preparation characterized by the coexistence of bacteria. 3. The live bacteria preparation as described in 1 or 2 above, characterized in that Lactobacillus live bacteria is vacuum freeze-dried. 4. The live bacterial preparation according to any one of 1 to 3 above, wherein the Lactobacillus strain is resistant to various antibacterial agents including spices. 5. The viable bacterial preparation according to 4 above, wherein the Lactobacillus strain further has bile resistance. 6. The live bacterial preparation according to any one of 1 to 5 above, wherein the Lactobacillus strain further has an antibiotic-producing ability. Among these, the important points are detailed as follows. As mentioned above, the nutritional requirements of Lactobacillus, which are known in the art, require amino acids, peptides, nucleic acid-related substances, vitamins, salts, fatty acids or their esters, and sugars for growth, and are considered to be among the bacteria with relatively low nutritional requirements. It is considered. Therefore, in the past, good nutrient media such as Briggs and MRS had to be used heavily for culturing Lactobacillus. The Lactobacillus used in the present invention discovered by the present inventor (hereinafter referred to as Lactobacillus of the present invention unless otherwise specified) has properties completely different from conventionally known Lactobacillus. That is, the Lactobacillus of the present invention is
Of course, it grows well in a long nutrient composition such as Briggs' medium, but as shown in Table 1, it can also grow in a medium that does not have such a favorable composition, such as Stephenson-Whetham medium. Wessam medium (hereinafter referred to as S)
-W medium) has the following composition: KH ₂ PO ₄ 1 g; MgSO ₄ .7H ₂ O 0.7 g;
NaCl 1g; (NH ₄ ) ₂ HPO ₄ 4g; Glucose 5
g; Medium containing 0.03 g of FeSO ₄ .7H ₂ O in 1 water (hereinafter, the composition of the medium indicates the amount in 1 water).

【table】

[Table] The Lactobacillus of the present invention is as shown in Table 2.
The growth rate and final bacterial count in Briggs and MRS media are much higher than those of conventionally known Lactobacilli.

[Table] Furthermore, as is clear from the above nutritional requirements, for the conventionally known Lactobacillus, S.
-W medium, [(S-W) + vitamin medium] and [(S-W) + casamino acid]
Even the medium consisting of this is not a suitable medium for growth. The experimental results of the present inventors are also listed in Table 4. In contrast, in most cases, the Lactobacillus of the present invention not only grows in any of the following media, but also does not grow at a slow rate in such media. The final yield (number of bacteria/cc) is also not small. The present inventors have also confirmed that the Lactobacillus of the present invention, which cannot grow in the medium of (S-W)+vitamin+amino acid, has a deodorizing property. An example of the composition of the LC and (S-W) media described above is shown below. LC medium: Hepton
10g, meat extract 10g, NaCl 5g, K ₂ HPO ₄ 3g,
Glucose 10g, yeast extract 5g, _CaCO33
g, water 1, PH7.4, (S-W) medium: KH ₂ PO ₄ 1
g, _{MgSO47H2O0.7g} , _NaCl1g ,
(NH ₄ ) ₂ HPO ₄ 4 g, FeSO ₄ 7H ₂ O 0.03 g, glucose 5 g, water 1, Briggs medium: Briggs.M.
(1953). An improved medium for
By Lactobacilli.J.Dairy Res.20.36. MRS
Medium: DEMAN, JC., ROGOSA, M. &
SHARPE, M.E. (1960), J.appl.Bact.23(1),
According to 130-135. The above terms are explained in the conceptual table below (Table 3a,
It is summarized in b).

[Table] The growth rate and yield of the Lactobacillus of the present invention can be improved with a low nutritional composition like a medium.
The results are not much different from those obtained when conventional bacteria are cultured in a well-fed environment like Briggs. Part of the detailed experimental results of six representative Lactobacillus strains of the present invention with different nutritional requirements are shown below.

[Table] Conventionally known Lactobacilli are as mentioned above.
It is not known that the addition of SNC substances singly or in combination to the culture medium is essential for growth or that it promotes growth, and as shown in Table 6, this is not the case according to the research results of the present inventors. It is not allowed. On the contrary, the Lactobacillus of the present invention is SNC
When the substances or sulfur-containing amino acids were added to the culture medium of , growth essential or growth promotion was observed in all or any of , as shown in Table 6.

【table】

【table】

[Table] The SNC compounds proposed by the present inventors will be described in detail below. According to the inventors,
The extremely wide variety of odorous components contained in feces can be roughly divided into three groups: S compounds, N compounds, and C compounds. Among these compounds, S compounds include H ₂ S or As a Na ₂ S, N compound
NH ₃ , indole or skatole, and lower fatty acids are representative examples of C compounds, and it was found that research on the deodorization of odorous components in feces is primarily sufficient if only these were considered. . It should be noted that the P-compound is considered to be a secondary consideration. Also, these odorous components SNC
It has been revealed that when the Lactobacillus of the present invention is added to a suitably selected compound concentration and medium with low nutrition, the Lactobacillus exhibits a growth-essential or growth-promoting effect. Including these points, the present inventor calls the SNC theory regarding fecal odor deodorization and microorganisms. Next, Figure 1 shows the growth rate of the Lactobacillus of the present invention when LC medium was used as a representative nutrient and acetic acid or butyric acid was added thereto. The growth rates of the present strain 1 and the conventional strain 1' when 5 g of acetic acid were added, and 2 and 2' are the growth rates when 0.1 g to 1 g of acetic acid were added, respectively. As is clear from FIG. 1, during the logarithmic growth phase, the bacteria of the present invention also undergo growth promotion, but this is so slight that it is difficult to judge with the naked eye or by counting the number of viable bacteria. On the other hand, the susceptibility of conventional bacteria to the direction of growth promotion is very low. Similarly, using LC medium, Na ₂ S・9H ₂ O (2 g)
Figure 2 (Na ₂ S·9H ₂ O) and Figure 3 (NH 3 ) show the cases in which NH ₃ (2 g) and NH ₃ (2 g) were added. Conventional bacterium 1' suffers from growth inhibition at a lower concentration than inventive bacterium 1. In other words, it is sensitive to the direction of inhibition. 2 and 2′ are Na ₂ S・9H ₂ O and NH ₃ respectively.
A white test is shown where only 0.1g is present. Furthermore, the Lactobacillus of the present invention and conventionally known Lactobacillus were cultured using nutrient, mesotrophic, and low-nutrient media, and various concentrations of SNC substances were added during the logarithmic growth phase to determine the growth mode adopted by the bacteria. As shown in Figures 4 to 6,
In mesotrophic media, not only acetic acid but also Na ₂ S
When Na 2 S and NH ₃ are added, the Lactobacillus of the present invention has a concentration that promotes growth, but conventionally known Lactobacillus does not undergo growth promotion when Na ₂ S or NH ₃ is added. Furthermore, as shown in Figures 7 to 9, conventionally known Lactobacillus cannot grow in a low-nutrient medium, and cannot grow even when an SN compound is added thereto, whereas the Lactobacillus of the present invention does not grow in a low-nutrient medium. For example, bacteria that cannot grow in (S-W) medium are Na ₂ S
It becomes possible to grow by adding . That is, Na ₂ S is essential for the growth of the Lactobacillus of the present invention. The solid line in FIGS. 1 to 9 indicates Lactobacillus of the present invention, and the dotted line indicates conventionally known Lactobacillus. The above relationships are summarized in Table 7.

【table】

[Table] In addition to the above, in order to expect greater effects from the live Lactobacillus preparation of the present invention, it is more desirable to have the following properties. The first is that Lactobacillus has a great ability to form antibiotics. It has been found that some Lactobacilli of the present invention form antibiotics in large amounts, in small amounts, or not at all.
The second is to impart resistance to antibacterial agents (antibiotics and spices used). That is, the Lactobacillus of the present invention cannot fully function unless it is made resistant to the bacteriostatic agent used. This is a teaching in bacteriology, and the same was true for the Lactobacillus of the present invention. For example, it is clear from experimental results that the Lactobacillus of the present invention was actually used as an anti-inflammatory agent or a deodorizing agent, and Table 8 shows an example of its use as a deodorizing agent.

[Table] The Lactobacillus of the present invention was subcultivated in the presence of tetracycline to create a resistant strain, which was orally administered to humans and was further treated with tetracycline 4 times a day.
Tablets were administered. When a sensitive strain was used as a control, as shown in Table 8, the sensitive strain showed almost no deodorizing effect, whereas the resistant strain showed the expected results. Number three is bile resistance. Bile has a remarkable antibacterial effect, and there are records that it was used as a disinfectant in ancient times. When the Lactobacillus of the present invention is expected to proliferate in the intestines as a deodorizing agent, etc., it is essential to preserve bile resistance. The present inventors also confirmed this through actual administration experiments in human stock, an example of which is shown in Table 9.

[Table] While the Lactobacillus of the present invention made resistant to 2% bile powder exhibited a sufficient deodorizing effect, the bile-sensitive strain showed almost no deodorizing effect. Regarding the method for cultivating, passage, and enrichment of Lactobacillus of the present invention, the present inventors have confirmed that the deodorizing ability of the bacteria is significantly reduced depending on the medium composition during the passage and enrichment. Therefore, in order to further investigate this phenomenon, we categorized the culture medium into nutrient, meso-nutrient, and low-nutrient types as mentioned above, and added supplements with and without milk powder, which is said to be good for the growth of Lactobacillus, and with bile acid. Passaging and deodorization titer measurement experiments were carried out using culture media with and without treatment. Of these, the ones mentioned above are nutritional.
MRS, those based on LC medium, those based on milk powder, and mixtures thereof, and the fecal components include 0.5 g of Na ₂ S, 9H ₂ O, and skatole.
This is a combination of 0.5g and 1g of butyric acid (hereinafter referred to as component F). As for intermediate nutrients, the (S-W) medium is mainly supplemented with both casamino acids and vitamins, and is combined with F component, peptone, etc. Low nutritional content is based on (S-W) medium with amino acids or vitamins added, and F
Products with added ingredients and milk powder were also used in experiments. A part of the medium composition is shown below. (b) Low nutrient medium: (1), (S-W) + 1 g of casamino acids
+ bile end 10g, (2), (1) + F component (3), (1) + milk powder 30g
(4), (3) + F component (5), [S-W] + vitamin 0.1g
+ bile end 10g (6), (5) + F component (7), (5) + milk powder 30g
(8), (7) + F component (b) Mesotrophic medium: (9), (S-W) + Casamino acid 1g
+ 1g yeast extract + 10g bile end (10), (9) + F ingredient
(11), (9) + milk powder 30g (12), (11) + F component (13), peptone 2g + MgSO ₄ 7H ₂ O 0.5g + KH ₂ PO ₄ 0.5g
+ 1g of NaCl + 1.0g of bile ends (14), (13) + F component (15), 1/10 MRS medium (16), (15) + F component (17), 1/3 MRS (18), (17) + F component (c) Nutrient medium: (19), MRS medium (20), (19)
+F component (21), MRS + bile end 10g, (22),
(21) + F component (23), MRS + milk powder 30g, (24),
(23) + F component (25), MRS + milk powder 30g + bile powder 10
g (26), (25) + F component (27), milk powder medium (28), (27) + F component (29), LC medium (30),
(29) + F component (31), LC + bile end 10g (32),
(31) +F component Using such a medium, we carried out subculturing over a medium period and actually measured the deodorizing titer. Although there are various levels of growth, from weak to weak, deodorizing Lactobacillus can grow in any medium by adding the F component to the early, middle, and even all stages of growth. In the case of stimulation or subculturing to a certain extent in a low-nutrient medium, there is almost no change in the deodorizing ability of the bacteria when the F component is not added, with some exceptions, but with the addition of the F component, It was even observed that there was a slight increase in vitality over time. (ii) In the nutrient media (Nos. 19 to 32), the growth rate of bacteria was very good, and when the F component was added thereto, no growth stimulation could be detected by visual observation or by counting the number of viable bacteria. In addition, when subculturing is carried out in the above-mentioned nutrient medium, the deodorizing power of the bacteria is F
Regardless of whether ingredients were added or not, the bacterial titer tended to drop rapidly as the cells were passaged.
Although it does not seem to have any deodorizing power like one of the starter bacteria in commercially available lactic acid bacteria drinks, the growth and proliferation of bacteria that can be observed to grow even with moderate nutrition is actually inhibited by the addition of the F component from low to moderate nutrition. However, like the Lactobacillus of the present invention, it did not show any sensitivity in the nutrient medium. As is clear from the above, as a culture medium for enrichment that does not reduce deodorizing power, there is a medium based on a medium in which an F component is added to the base medium, and the addition stimulates the growth of bacteria. It was recognized that the medium to which F component was added was good. During the experiment on deodorizing Lactobacillus, the titer decreased. This occurs both during passaging and during storage.
What are the conditions for passage while maintaining the potency?
This has become one of the biggest problems. First, the inventors of the present invention took into consideration that when F component is added to a low-nutrient medium, the titer may slightly increase with passage, and that the titer decreases markedly when passage is carried out in a nutrient medium. We classified odorous fecal compounds as SNC compounds and conducted various experiments with or without adding them to the basal nutrient medium. Typical teaching examples are shown in Table 10.

[Table] In this example, Na ₂ S, NH ₃ and acetic acid are used as examples of additives to prevent the decrease in titer due to subculture, but another substance, which the inventors call SNC compound, is methyl sulfide. , mercaptan, skatole, indole, butyric acid, propionic acid, etc., it has become clear that no matter which combination is selected, as long as the SNC substances are sufficient in all cases, the same results can be obtained. Next, Table 11 (A) shows an example of the variation in titer due to passage when various amino acids and proteins were added to the basal medium.

[Table] As is clear from Table 11 (A), certain amino acids such as methionine, including cystine, sulfur-containing amino acids such as cysteine, as well as glutamic acid and lysine, are extremely effective for maintaining titer during passage. On the other hand, it has been shown that other types of amino acids, such as proline and tyrosine, rapidly reduce the deodorizing power. In this experiment, the present inventors discovered that some amino acids are effective in preserving the potency against deodorizing bacteria, while others, namely glycine, glutamic acid, and lysine, are slightly less effective, and others are effective against deodorizing bacteria. It was confirmed that different types can be roughly divided into three groups, each of which loses its effectiveness to a large extent. With this discovery, we were able to ascertain for the first time the true nature of the conventional experimental contradiction, namely, that improving nutrition in order to increase the growth of bacteria leads to a decrease in deodorizing potency, and we decided to resolve this contradiction. Successful. Furthermore, the SNC substance refers to an odorous component in feces, but it is extremely significant in that it has been shown that it can be added to the culture medium during passage in addition to being an odorous component and exhibits an effect that does not reduce the titer. is big. Furthermore, it has been found that the Lactobacillus of the present invention may contain the SNC element in a single compound in order to maintain its potency. In addition, this discovery becomes even more important from a practical point of view. Furthermore, as shown in Table 11 (B), good results are also shown by mixing the SNC compound with a specific (amino) acid. In addition, the SNC compound as used in the present invention refers to an S compound as an odorous component contained in feces, for example.
H ₂ S, Na ₂ S; Compounds such as NH ₃ , indole, skatole; C compounds such as lower fatty acids such as acetic acid, butyric acid, etc. Sulfur-containing amino acids include cystine, methionine and cysteine.

【table】

[Table] Tables 12 and 13 show the experimental results of the present inventors.

【table】

【table】

[Table] However, during vacuum freeze-drying or cryogenic storage, even if coated with milk, the titer will not decrease as much as the above results, and bacteria can be maintained in good condition. However, even in such cases, it is more desirable to mainly coat with sulfur-containing amino acids. As in the case of subculturing, good results were obtained in dry, semi-dry, and wet conditions when the SNC compound and amino acid were used together for storage. From Table 13, it was found that coating with amino acids such as cystine and methionine is effective in preserving the bacteria of the present invention during storage. When making a mold, it has the advantage that a wide range of methods can be considered in addition to vacuum freeze-drying and low-temperature storage. For example, when culturing with large amounts of Na ₂ S, NH ₃ and butyric acid added to the medium, if these have not been completely digested by the bacteria, the bacteria must be thoroughly washed. Therefore, the storage stability of the agent itself is not good, which means that cystine and methionine, which have a good storage stability for bacteria and are considered to be beneficial to living organisms, are used in the coating agent. The cultivation method and the like for the deodorizing Lactobacillus live bacterial preparation of the present invention have been described above in detail. The six deodorizing Lactobacillus strains of the present invention are listed in Table 10.
The measurement results in Tables 11 (A, B), 12, and 13 showed the same pattern, so Tables 10 and 11
Tables (A) and (B), Tables 12 and 13 respectively show the summarized results. Lactobacillus of the present invention (e.g., Microtechnical Institute Deposit No.
1946, 2742, 2779, 2780, 2081, and 2782), various activities ranging from biochemical properties to deodorizing ability are shown below. Biochemical properties are shown in Tables 14 a and b, and Tables 15 and 16
The table shows the relationship between nutritional requirements and growth. In addition, we conducted an experiment to reduce the odor of feces by adding live bacteria to the fresh feces of various animals or diluting them 5 times with AESE and culturing them. Table 17 shows this.

【table】

【table】

【table】

【table】

【table】

[Table] In Table 17, feces (%) 100%: 100% feces. 〃 20%: feces 20% water 80%. Live bacteria themselves: Take 3ase from the bacteria grown on the shear tray and add it to the above feces. Bacterial culture solution: Add 5 c.c. of culture solution in which bacteria have grown sufficiently in a test tube to the above feces. Fresh feces: Feces that have just been defecated. Next, six representative strains of Lactobacillus of the present invention were cultured in large quantities in the medium shown below, and Table 18 shows the deodorizing effects when each animal or human ingested them orally. Example of medium composition: KH ₂ PO ₄ , MgSO・7H ₂ O,
NaCg, ( _NH4 ) _2HPO , _FeSO4・_7H2O , starch, _CaCO3 , casamino acids, yeast extract,
After culturing the bacterial strain in a medium consisting of Na ₂ S・9H ₂ O, acetic acid, butyric acid, propionic acid, ammonia, indole, skatole, cystine, and vitamins, the bacteria were collected using a refrigerated centrifuge, and the bacterial cells were thoroughly washed. The bacteria were mixed with bread or butter in the form of a wet cake and 0.5 g/kg of bacterial cells were administered orally, and the degree of deodorization of excreted feces from the next day was examined. The degree of deodorization is as follows throughout this specification. 0: Completely odorless, 1: Slight odor that disappears immediately, 1': Smell that disappears after sniffing, 2: Emits a very weak but distinct odor, 2' : Weak odor, 3: Much weaker odor than control, 3': Slightly weaker odor than control, 4: Control odor.

[Table] The characteristics of the Lactobacillus of the present invention described in the present invention are as clear from the biological properties and nutritional requirements of each bacterium. They are very different from each other, as evidenced by the difference in their ability to grow in a (S-W) medium and another strain in a (S-W)+vitamin medium. All of the Lactobacilli of the present invention isolated by the present inventors have significantly lower nutritional requirements, faster growth rates, higher final yields, and special susceptibility to SNC compounds and higher sensitivity to various amino acids than conventional Lactobacilli. The present invention shows that they share fundamentally important points in microbiology, such as showing reactions not seen in conventional bacteria, and yet have unique characteristics that cannot be imagined based on conventional knowledge. It soon became clear. In other words, the important point of the present invention is that it has been made clear that even though the individual bacterial strains have different characteristics, there are extremely important commonalities from the viewpoint of deodorizing properties. There is. Therefore, the present inventor named the bacteria having these common characteristics as deodorizing Lactobacillus. The live bacteria preparation containing Lactobacillus of the present invention exhibits a remarkable deodorizing effect when administered orally or directly applied to feces.

[Brief explanation of the drawing]

1 to 9 show the growth rate curves of the Lactobacillus of the present invention and the conventional Lactobacillus in the logarithmic growth phase. Figure 1 shows LC medium with acetic acid added as a basal medium, Figure 2 shows Na ₂ S added to the same medium, and Figure 3 shows the same medium added with acetic acid.
NH ₃ is added, Figure 4 shows (S-W) + vitamin +
Figure 5 shows the same medium as in Figure 4 with addition of acetic acid, and Figure ₆ shows the same medium as in Figure 4 with addition of _NH3 . Fig. 7 shows the case where acetic acid was added to the low-nutrient medium, Fig. 8 shows the case where the same acetic acid was added, Na ₂ S.9H ₂ O, and Fig. 9 shows the case where the same NH ₃ was added. In the graph,
1 is when there is an additive of Lactobacillus of the present invention,
1' represents a white test of conventionally known Lactobacillus with additives, 2 represents a white test of Lactobacillus of the present invention without each additive, and 2' represents a white test of conventionally known Lactobacillus without each additive.

Claims (1)

[Claims] 1. As nutritional requirements, (a) Stephenson-Wesam medium, (b) Stephenson-Wesam + vitamin medium,
or (c) a live bacteria preparation characterized by the presence of live Lactobacillus bacteria that can grow in either a Stephenson-Wessam+casamino acid medium and a sulfur-containing amino acid. 2. The live bacteria preparation according to claim 1, characterized in that Lactobacillus live bacteria is vacuum freeze-dried. 3. The live bacterial preparation according to claim 1 or 2, wherein the Lactobacillus strain is resistant to various antibacterial agents including spices. 4. The live bacterial preparation according to claim 3, wherein the Lactobacillus strain further has bile resistance. 5. The live bacterial preparation according to any one of claims 1 to 4, wherein the Lactobacillus strain further has an antibiotic-producing ability. 6 As nutritional requirements, (a) Stephenson-Wessam medium, (b) Stephenson-Wessam + vitamin medium,
or (c) live Lactobacillus bacteria that can grow in either the Stephenson-Wessam + Casamino Acid medium, and whose growth is essential or growth-enhanced by the SNC compound, is treated with the SNC compound,
Sulfur-containing amino acids, glycine, glutamic acid, lysine, alanine, phenylalanine, arginine,
A live bacterial preparation characterized by coexisting with at least one compound selected from the group consisting of asparagine and aspartic acid. 7. The live bacteria preparation according to claim 6, characterized in that Lactobacillus live bacteria is vacuum freeze-dried. 8. The live bacterial preparation according to claim 6 or 7, wherein the Lactobacillus strain is resistant to various antibacterial agents including spices. 9. The live bacterial preparation according to claim 8, wherein the Lactobacillus strain further has bile resistance. 10. The live bacterial preparation according to any one of claims 6 to 9, wherein the Lactobacillus strain further has an antibiotic-producing ability.