JP2012213396A - Microorganism selected using guaiacol-recognition as index, and guaiacol-containing composition having microorganism growth-promoting activity - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide: a method for selecting microorganisms recognizing a compound having a guaiacol skeleton such as ferulic acid existing in animal or plant living bodies and/or a compound having a 4-methoxyphenol skeleton, and promoting their growth; and a composition including a guaiacol skeleton and/or a compound having a 4-methoxyphenol skeleton having growth-promoting action of a probiotic lactic acid bacterium.SOLUTION: This invention is achieved by eagerly developing the method for selecting microorganisms recognizing a compound having a guaiacol skeleton and/or a compound having 4-methoxyphenol skeleton and promoting their growth. Thereby, the problem is solved by finding out microorganisms recognizing the compound having a guaiacol skeleton such as ferulic acid and/or compounds having a 4-methoxyphenol skeleton, and promoting their growth.

Description

  The present invention relates to a method for selecting a microorganism that recognizes and promotes growth of a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton present in a plant body, and ferulic acid having a growth promoting activity of probiotic lactic acid bacteria. A composition containing a compound having a guaiacol skeleton and / or a 4-methoxyphenol skeleton such as ferulic acid, a microorganism additive containing a microorganism that recognizes a compound having a guaiacol skeleton and / or a 4-methoxyphenol skeleton such as ferulic acid, and the like The present invention relates to a method for industrial effective utilization of the microorganisms and compounds, including fermented feed of food by-products using additives, forage crop / grass silage, TMR fermented roughage preparation method and food preparation method.

  Ferulic acid is a substance derived from plant cell walls and is a rich natural biomass material.

  Ferulic acid is mainly used for plants and seeds such as rice (Non-Patent Documents 1 and 2), wheat (Non-Patent Documents 3 and 4), vegetables (Non-Patent Document 5), and citrus fruits (Non-Patent Documents 6 and 7). It is a component existing in the cell wall, and is produced by metabolism of phenylalanine and tyrosine (Non-patent Document 8). Ferulic acid is a substance that has been confirmed to be safe, and has been secured in acute toxicity tests and mutagenicity tests (Non-patent Document 8), and mass purification techniques have already been established (Non-Patent Document 8).

  Lactic acid bacteria are extremely important beneficial microorganisms in the food and feed industry. Further, from the viewpoint of promoting the health of humans and livestock, the market value of the lactic acid bacteria growth promoting component itself is high. Application examples of lactic acid bacteria include prebiotics represented by indigestible carbohydrates (fructooligosaccharides, galactooligosaccharides, lactulose, etc.) (Non-patent Document 9).

  In recent years, the emergence of multidrug-resistant bacteria due to increased use of livestock antibiotics and concerns about environmental pollution have become a social issue, and the establishment of a reduction technology is an urgent issue. The reason for the increase in the amount of use is not only the promotion of the growth of livestock, but also the expectation for the improvement of the microflora aimed at inhibiting the growth of harmful microorganisms in the digestive tract (Non-patent Document 10).

  On the other hand, in humans, the microbiota in the digestive tract deteriorates due to various stresses such as mental stress, and many harmful microorganisms are detected (Non-patent Documents 11 and 12).

  However, there is no technology for improving the symbiotic bacterial flora balance that expects ferulic acid to promote probiotic growth.

  In the case of livestock feed, various antibiotics and their administration methods have been developed to control the gastrointestinal microflora (Non-patent Document 10). In addition, many techniques for improving microorganisms of the digestive tract using probiotic microorganisms, particularly lactic acid bacteria, have been proposed regardless of feed and food. Indigestible saccharides have been reported so far as functional components having an effect of promoting the growth of lactic acid bacteria, and many have been commercialized as so-called prebiotics.

Fujimaki M.F. , Et al. 1977, Agric. Biol. Chem. , 41, 1721-1725. Nishizawa Chieko et al., 1998. 45, 499-503. Japan Food Science Association Pushanaanawin V. , Et al. , 1988, J. Am. Agric. Food Chem. 36, 515-520. Wetzel D.H. L. , Et al. , 1988, Dev. Food Sci. , 17 409-428. Haimann W.H. , Hermann K. , 1971, Z. Lebensm. Unters. Fors. , 145, 20-26. Naim M.M. , Et al. , 1988, J. Am. Food Sci. 53, 500-512. Wheaton T. A. , Stewart I. , 1965, Nature, 206, 620-621. Masamoto Yamamoto, 2007, Bokkin Bobai, 35, 317-323. Robert fluid M.M. B. , 1998, Br. J. et al. Nutr. , 80, 197-202. Prescott J.M. F. , 2008, Animal Health Research Review, 9, 127-133. Holdman L. V. , Et al. , 1976, Appl. Environ. Microbiol. , 31, 359-375. Takatsuka H. et al. et al. , 2000, Int. J. et al. Hematol. , 71, 273-277.

  INDUSTRIAL APPLICABILITY The present invention exhibits a growth promoting effect on a microorganism selection method using as an index the recognizability of a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton, and lactic acid bacteria functioning as probiotics. Provided is a composition containing a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton. Specifically, by establishing a culture system using a low nutrient medium that reflects an in vivo digestive tract environment containing a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton, ferulic acid, etc. Strains belonging to Lactobacillus plantarum and Lactobacillus plantarum, Lactobacillus plantarum, Lactobacillus plantarum, Lactobacillus plantarum, Lactobacillus plantarum TO1002, and Lactobacillus plantarum Lactobacillus plantarum, Lactobacillus plantarum, and Lactobacillus plantarum I found several stocks. Until now, the inventor has not known the case where a lactic acid bacterium exhibiting a growth property with respect to a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton has been discovered. Therefore, the present invention provides a technique for improving a symbiotic bacterial flora balance that has been expected to promote probiotic growth promotion of a compound having a guaiacol skeleton and / or a 4-methoxyphenol skeleton such as ferulic acid, which has not been provided so far. I will provide a.

  In addition, a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton having a function of activating lactic acid bacteria has a chemical structure different from the conventionally reported indigestible saccharide components. As a biotic, a compound having a useful guaiacol skeleton and / or a 4-methoxyphenol skeleton is provided.

  Thus, for example, the present invention provides:

  In one aspect, a Lactobacillus genus lactic acid strain whose growth activity is increased by a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton is provided.

  In one embodiment, the present invention provides a Lactobacillus plantarum strain whose growth activity is increased by a compound having a guaiacol skeleton and / or a 4-methoxyphenol skeleton.

  In another embodiment, the present invention provides a Lactobacillus plantarum strain whose growth activity is increased by a compound having a compound having a guaiacol skeleton.

In one embodiment, the growth activity of the strain of the present invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH. _In a medium containing ₂ PO ₄ and 30% bovine serum (pH 6.5)
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4-fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride,
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride,
An increase of at least about 4.2 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 2.7 times in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
And at least about 1.8-fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 4 in the presence of 500 μM eugenol compared to the absence of eugenol. Have at least one feature selected from the group consisting of:

In one embodiment, the growth activity of the strain of the present invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH. _In a medium containing ₂ PO ₄ and 30% bovine serum (pH 6.5)
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4-fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride,
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 fold in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET) and compared to the absence of metanephrine hydrochloride Up to at least about 4.8 fold in the presence of 500 μM Metanephrine hydrochloride,
At least one feature selected from the group consisting of:

In one embodiment, the growth activity of the strain of the present invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH. _In a medium containing ₂ PO ₄ and 30% bovine serum (pH 6.5)
An increase of at least about 5.5 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4-fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride,
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 fold in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET) and compared to the absence of metanephrine hydrochloride Up to at least about 4.8 fold in the presence of 500 μM Metanephrine hydrochloride,
At least one feature selected from the group consisting of:

In a further embodiment, the growth activity of the strain of the invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. In a medium (pH 6.5) containing PO ₄ and 30% bovine serum,
An increase of at least about 7.9 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 3.4-fold in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 7 in the presence of 500 μM eugenol compared to the absence of eugenol. It has at least one feature selected from the group consisting of an increase of 8 times in addition to or independently from the above feature.

In a further embodiment, the growth activity of the strain of the invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. In a medium (pH 6.5) containing PO ₄ and 30% bovine serum,
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 2.7 times in the presence of 500 μM vanillin compared to the absence of vanillin,
An increase of at least about 3.8 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
An increase of at least about 5.8 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 2.7-fold in the presence of 500 μM silybin compared to the absence of silybin, and an increase of at least about 6.9-fold in the presence of 500 μM eugenol compared to the absence of eugenol At least one of the features selected from the group consisting of:

In a further embodiment, the growth activity of the strain of the invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. In a medium (pH 6.5) containing PO ₄ and 30% bovine serum,
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.2 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 2.7-fold in the presence of 500 μM silybin compared to the absence of silybin, and an increase of at least about 4.3-fold in the presence of 500 μM eugenol compared to the absence of eugenol At least one of the features selected from the group consisting of:

  In one embodiment, in the present invention, the compound is ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloric acid Salts, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine ( Metanephrine) hydrochloride and at least one selected from the group consisting of salts and solvates thereof.

  In another embodiment, in the present invention, the compound comprises ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride Salt, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), metanephrine (Metanephrine) ) Hydrochloride, guaiacol (2-methoxyphenol), 4-methoxyphenol, gingerol, shogaol, vanillic acid diethylamide, isoeugenol, eugenol, isovanillic acid, o-vanillin, trans Ferulic acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanic acid Ril alcohol, creosole, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, Coniferyl alcohol and at least one selected from the group consisting of salts and solvates thereof.

  In one embodiment, in the present invention, the compound is selected from the group consisting of ferulic acid, hydroxyferulic acid, vanillic acid and capsaicin.

  In another embodiment, the strain belongs to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei and Lactobacillus casei.

  In one embodiment, in the present invention, the strain is selected from Lactobacillus plantarum TO1000, Lactobacillus plantarum TO1001, Lactobacillus plantarum TO1002 or Lactobacillus plantarum TE1003 (I-PN-N95 Alternatively, the strain may be Lactobacillus paraplantarum LOOC 2020 (reception number: NITE AP-1308), Lactobacillus sake SG 171 (reception number: NITE AP-1309), Lactobacillus casei LO 2 (receipt number: NITE AP-1310), Lactobacillus casei PR143 (receipt number: NITE AP-1311) or Lactobacillus casei PR150 (receipt number: NITE AP-1312) may be.

  In another aspect, the present invention provides a composition for growth of Lactobacillus lactic acid strains comprising a compound having a guaiacol skeleton and / or a 4-methoxyphenol skeleton.

  In one embodiment, the present invention provides a composition for growth of Lactobacillus plantarum strains comprising a compound having a guaiacol backbone and / or a compound having a 4-methoxyphenol backbone.

  In another embodiment, the present invention provides a composition for growth of Lactobacillus plantarum comprising a compound having a compound having a guaiacol backbone.

  In one embodiment, in the present invention, the compound is ferulic acid or a derivative thereof.

  In one embodiment, in the present invention, the compound is ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine (Normetanephrine). Hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine (Metanephrine) hydrochloride and at least one selected from the group consisting of salts and solvates thereof.

  In another embodiment, in the present invention, the compound comprises ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride Salt, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), metanephrine (Metanephrine) ) Hydrochloride, guaiacol (2-methoxyphenol), 4-methoxyphenol, gingerol, shogaol, vanillic acid diethylamide, isoeugenol, eugenol, isovanillic acid, o-vanillin, trans Ferulic acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanic acid Ril alcohol, creosole, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, Coniferyl alcohol and at least one selected from the group consisting of salts and solvates thereof.

  In one embodiment, in the present invention, the compound is selected from the group consisting of ferulic acid, hydroxyferulic acid, vanillic acid, and capsaicin.

  In one embodiment, in the present invention, the compound comprises ferulic acid or a derivative thereof.

  In the embodiment, in the present invention, the strain belongs to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei, and Lactobacillus casei.

  In one embodiment, in the present invention, the strain is Lactobacillus plantarum TO1000 (Accession No. NITE P-958), Lactobacillus plantarum TO1001 (Accession No. NITE P-959), Lactobacillus planta No. Lactobacillus plantarum TO1003 (Accession number NITE P-961), Lactobacillus plantarum JCM1149, Lactobacillus paraplanum LOOC 2020 (Reception number: NITE AP1308), LactobaslTETE17TEL 309), Lactobacillus casei LOOC82 (receipt number: NITE AP-1310), Lactobacillus casei PR143 (receipt number: NITE AP-1311) or Lactobacillus casei PR150 (receipt number: a NITE AP-1312).

  In one embodiment, in the present invention, the strain is Lactobacillus plantarum TO1000 (Accession Number NITE P-958), TO1001 (Accession Number NITE P-959), TO1002 (Accession Number NITE P-960), TO1003 (Accession Number). NITE P-961) or JCM1149.

  In another aspect, the present invention provides a probiotic composition comprising the above strain of the present invention.

  In another aspect, the present invention provides a synbiotic composition obtained by combining the strain of the present invention with a compound having a guaiacol skeleton.

  In another aspect, the present invention provides a symbiotic composition obtained by combining the strain of the present invention with a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton.

In one embodiment, in the present invention, the compound is included at least 10 μM per 1 × 10 ⁶ colony forming units (cfu) of the strain when present in the body.

  In another aspect, the present invention provides a prebiotic composition comprising a compound having a guaiacol skeleton for use in combination with the above strain of the present invention.

  In another aspect, the present invention provides a prebiotic composition comprising a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton for use in combination with the above strain of the present invention.

In another aspect, the present invention is a method for selecting lactic acid bacteria whose proliferation activity is increased by a compound having a guaiacol skeleton, the method comprising:
A) a providing step of providing a sample containing lactic acid bacteria;
B) magnesium sulfate (MgSO _4), ammonium nitrate _(NH 4 _NO 3), potassium chloride (KCl), D - (+ ) - glucose, potassium dihydrogen phosphate _(KH 2 PO ₄₎ and in a medium containing calf serum Culturing the sample in the presence or absence of a compound having a guaiacol skeleton; and C) not growing in the absence of the compound and growing in the presence or the presence of the compound A method is provided that includes a separation step of separating a strain whose growth is promoted below that in the absence.

In still another aspect, the present invention is a method for selecting lactic acid bacteria whose proliferation activity is increased by a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, the method comprising:
A) a providing step of providing a sample containing lactic acid bacteria;
B) magnesium sulfate (MgSO _4), ammonium nitrate _(NH 4 _NO 3), potassium chloride (KCl), D - (+ ) - glucose, potassium dihydrogen phosphate _(KH 2 PO ₄₎ and in a medium containing calf serum A culturing step of culturing the sample in the presence or absence of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton; and C) non-proliferation and presence in the absence of the compound A method is provided that comprises a separation step of isolating a strain that has grown underneath, or has been promoted for growth in the presence of the compound than in the absence.

  In one embodiment, in the present invention, the magnesium sulfate is 1.16 to 1.74 mM, the ammonium nitrate is 8.02 to 9.81 mM, the potassium chloride is 3.83 to 5.74 mM, the D -(+)-Glucose is present in the medium at 3.96-39.64 mM, the potassium dihydrogen phosphate is 2.09-3.14 mM, and the bovine serum is 15-30%, Has a pH of 6.0 to 7.0.

  In one embodiment, in the present invention, the culturing step is performed for 32-48 hours.

  In one embodiment, in the present invention, the culturing step is performed at 25 to 40 ° C.

In one embodiment, in the present invention, the culturing step is performed under conditions of 0 to 5% CO ₂ and 1 to 20% O ₂ .

  In one embodiment, in the present invention, the lactic acid bacteria belong to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei, and Lactobacillus casei.

  In one embodiment, in the present invention, the lactic acid bacterium is Lactobacillus plantarum.

In one embodiment, in the present invention, the sample is selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplantum, Lactobacillus sakei and Lactobacillus between 5 × 10 ^{5 and} 5 × 10 ⁶ colony forming units / well. Contains bacteria.

In one embodiment, in the present invention, the sample comprises Lactobacillus plantarum between 5 × 10 ^{5 and} 5 × 10 ⁶ colony forming units / well.

  In various embodiments, the present invention includes any one or more features of the various embodiments described above.

  Since the microorganism of the present invention (for example, Lactobacillus plantarum TO1002) is derived from grass silage, it is expected to be widely used in the livestock field as a probiotic agent that is expected to have a probiotic effect on food residues, silage, and fermentation TMR. Is done. In addition, food residue, silage, and fermentation TMR also include “food / plant-derived ferulic acid”, and it cannot be overlooked that guaiacol skeleton-containing compounds such as ferulic acid can be recognized and promoted. Silage fermentation acceleration from a viewpoint different from conventional silage fermentation preparation technology by lactic acid bacteria such as sugar and moisture adjustment by focusing on compounds having guaiacol skeleton such as plant ferulic acid and / or compounds having 4-methoxyphenol skeleton The added value as an agent can also be greatly expected, and the technology transfer to feed manufacturers is expected as a “new lactic acid bacteria additive for silage preparation” and “lactic acid bacteria growth promoting additive”.

  Since compounds having a guaiacol skeleton such as ferulic acid and compounds having a 4-methoxyphenol skeleton are widely distributed in plant food materials as described above, the utility when safety is ensured is It is considered extremely high.

  In recent years, in the field of livestock feed, for the purpose of expanding domestic self-sufficient feed, feed rice, feed wheat or waste vegetables have been promoted. Guaiacol skeletons such as ferulic acid contained in a large amount thereof and / or 4 -There is interest in the functionality of compounds having a methoxyphenol skeleton that contribute to the healthy growth of livestock. That is, the application of the present invention is envisaged not only in foods but also in the feed field, and the target market scale that can be expected is large and the economic effect is high.

  The present invention improves the balance of intestinal bacteria and does not depend on antibacterial agents by the action of promoting the growth of lactic acid bacteria by a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton that can be easily applied to food and feed. It contributes to livestock health.

  In addition, the selection technique invented in this study enables screening of microorganisms derived from various environments. For example, by selecting probiotic lactic acid bacteria derived from human feces, safety derived from humans is expected, and in the future, development of functional foods for humans can be expected. In addition, by effectively using the present composition derived from plant resources (vegetables, fruits, seeds, etc.) that ensure food experience and safety, it can be used as a new functional ingredient that exerts prebiotic-like effects. I can expect enough. The indigestible carbohydrates of the preceding examples have many components derived from plants and seaweeds, and are relatively easily accepted by manufacturers and consumers.

  Antibiotics are being watched internationally as something to be reduced due to the emergence of multidrug-resistant bacteria and concerns about environmental pollution. In addition, a compound having a guaiacol skeleton such as ferulic acid and / or 4-methoxyphenol is expected in the expectation of an effect of improving the gut microbiota by a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton. Although there was no method for selecting a safe and beneficial microorganism using recognizability for a compound having a skeleton as an index, the present invention enabled application of a probiotic microorganism that actively utilizes this action.

  Therefore, the present invention provides establishment of a culture technique for promoting efficient growth and activity of lactic acid bacteria.

  Considering the recent market scale related to growth-promoting components of lactic acid bacteria, the needs for the growth-promoting components of beneficial microorganisms are extremely high, but they contain compounds having a guaiacol skeleton such as ferulic acid and / or compounds having a 4-methoxyphenol skeleton. There has been no report on the growth promoting effect of lactic acid bacteria by the composition in Japan and abroad, and the promoting action with such a substance was provided for the first time.

  In addition, an increase in patients with irritable bowel syndrome and an increase in various diseases such as allergies caused by deterioration in the intestinal microbiota balance is serious, and medical costs that continue to increase are a problem. The present invention provides means for solving such problems from the viewpoint of preventive medicine.

  Ferulic acid is known to have an “antibacterial effect” against lactic acid bacteria. And it is also known that antibacterial action becomes remarkable at pH 5 or less. Then, it is considered that the finding that ferulic acid is provided and the lactic acid bacteria grow so as to lower the pH, found in the present invention, is in the opposite direction to the conventional finding. Therefore, from the conventional knowledge, it is considered that the effect found in the present invention could not be predicted.

FIG. 1 shows the culture turbidity, which is an index of the growth promoting activity of ferulic acid against Lactobacillus plantarum TO1002 strain. As a reference showing the pH state after completion of the culture, the measurement results of the culture solution pH measured simultaneously are shown. No. TO1002 strains containing No. 0, 0.5, 1, 5, 10, 100 or 500 (μM) ferulic acid respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity (A) and culture solution pH (B) of the culture solution after the culture were measured. Significant differences when compared with the non-added medium are indicated by * (P <0.05), ** (P <0.01), *** (P <0.001). FIG. 2 shows the growth promoting activity of ferulic acid against Lactobacillus plantarum TO1000, TO1001, TO1003, and JCM1149 strains. These strains were designated as No. 1 containing 0, 0.5, 1, 5, 10, 100 or 500 (μM) ferulic acid, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. Significant differences when compared with the non-added medium are indicated by * (P <0.05) and *** (P <0.001). A represents TO1000 strain, B represents TO1001 strain, C represents TO1003 strain, and D represents JCM1149 strain. FIG. 3 shows the measurement results of the pH of the culture solution measured simultaneously with ferulic acid as a reference showing the pH status after cultivation with Lactobacillus plantarum TO1000, TO1001, TO1003, and JCM1149 strain. These strains were designated as No. 1 containing 0, 0.5, 1, 5, 10, 100 or 500 (μM) ferulic acid, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the culture solution pH after the culture was measured. Significant differences when compared with the non-added medium are indicated by * (P <0.05), ** (P <0.01), *** (P <0.001). A represents TO1000 strain, B represents TO1001 strain, C represents TO1003 strain, and D represents JCM1149 strain. FIG. 4 shows culture turbidity, which is an index of the growth promoting activity of vanillin, hydroxyferulic acid and capsaicin against Lactobacillus plantarum TO1002 strain. As a reference showing the pH state after completion of the culture, the measurement results of the culture solution pH measured simultaneously are shown. TO1002 strain is 0, 0.5, 1, 5, 10, 100 or 500 (μM) vanillin, or 0, 0.5, 1, 5, 10, 100 or 500 (μM) hydroxyferulic acid, or No. 0, 0.1, 0.25, 0.5, 0.75, 1 or 1.5 (mM) capsaicin. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity (A) and culture solution pH (B) of the culture solution after the culture were measured. Significant differences when compared with the non-added medium are indicated by * (P <0.05), ** (P <0.01), *** (P <0.001). FIG. 5 shows 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) The growth promotion activity with respect to Lactobacillus plantarum TO1002 strain of a hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine (Metanephrine) hydrochloride is shown. No. TO1002 strains containing No. 0, 0.5, 1, 5, 10, 100 or 500 (μM) MHPG, normetanephrine, VMA, 3MT, HVA, VA, MOPET or metanephrine, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. Significant differences when compared with the non-added medium are indicated by * (P <0.05), ** (P <0.01), *** (P <0.001). FIG. 6 shows 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine hydrochloride (Metanephrine) hydrochloride with Lactobacillus plantarum TO1002 The measurement result of the culture solution pH measured simultaneously is shown. No. TO1002 strains containing No. 0, 0.5, 1, 5, 10, 100 or 500 (μM) MHPG, normetanephrine, VMA, 3MT, HVA, VA, MOPET or metanephrine, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the culture solution pH after the culture was measured. Significant differences when compared with the non-added medium are indicated by * (P <0.05), ** (P <0.01), *** (P <0.001). FIG. 7 shows the growth-promoting activity of guaiacol, 3-methoxyphenol, 4-methoxyphenol, anisole, and phenol against Lactobacillus plantarum TO1002 strain. The TO1002 strain was obtained from No. 1 containing 10,100 or 500 (μM) of guaiacol, 3-methoxyphenol, 4-methoxyphenol, anisole and phenol, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. *** (P <0.001) indicates a significant difference when compared with the non-added medium. FIG. 8 shows the growth-promoting activity of Gingerol and Shogaol against Lactobacillus plantarum TO1002 strain. The TO1002 strains are No. 1 containing 0.1, 0.25, 0.5, 0.75, 1 or 1.5 (mM) gingerol and shogaol, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity (A, B) and culture solution pH (C, D) of the culture solution after the culture were measured. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). FIG. 9 shows vanillic acid diethylamide (a), isoeugenol (b), isovanillic acid (c), o-vanillin (d), transferuric acid (e), isoferulic acid (f), isocreole (g), 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one (h), hesperetin (i), 3- (4-hydroxy-3-methoxyphenyl) propionic acid (j), isovanillyl Alcohol (k), Creosol (l), o-vanillic acid (m), methyl vanillate (n), methyl-3- (4-hydroxy-3-methoxyphenyl) propionate (o), 5-nitro Guaiacol (p), scopoletin (q), silybin (r), vanillyl alcohol (s), ethyl vanillate (t), coniferyl alcohol (u) and eugeno Shows the growth promoting activity against Lactobacillus plantarum TO1002 strains of Le (v). TO1002 strains were each 0, 0.5, 1, 5, 10, 100 or 500 (μM) vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, creoleol, o-vanillic acid, vanillin No. containing methyl acid, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol or eugenol. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). FIG. 9 shows vanillic acid diethylamide (a), isoeugenol (b), isovanillic acid (c), o-vanillin (d), transferuric acid (e), isoferulic acid (f), isocreole (g), 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one (h), hesperetin (i), 3- (4-hydroxy-3-methoxyphenyl) propionic acid (j), isovanillyl Alcohol (k), Creosol (l), o-vanillic acid (m), methyl vanillate (n), methyl-3- (4-hydroxy-3-methoxyphenyl) propionate (o), 5-nitro Guaiacol (p), scopoletin (q), silybin (r), vanillyl alcohol (s), ethyl vanillate (t), coniferyl alcohol (u) and eugeno Shows the growth promoting activity against Lactobacillus plantarum TO1002 strains of Le (v). TO1002 strains were each 0, 0.5, 1, 5, 10, 100 or 500 (μM) vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, creoleol, o-vanillic acid, vanillin No. containing methyl acid, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol or eugenol. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). FIG. 10 shows vanillic acid diethylamide (a), isoeugenol (b), isovanillic acid (c), o-vanillin (d), transferuric acid (e), isoferulic acid (f), isocreole (g), 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one (h), hesperetin (i), 3- (4-hydroxy-3-methoxyphenyl) propionic acid (j), isovanillyl Alcohol (k), Creosol (l), o-vanillic acid (m), methyl vanillate (n), methyl-3- (4-hydroxy-3-methoxyphenyl) propionate (o), 5-nitro Guaiacol (p), scopoletin (q), silybin (r), vanillyl alcohol (s), ethyl vanillate (t), coniferyl alcohol (u) and Euge Shows the growth promoting activity against Lactobacillus plantarum TO1002 strain Lumpur (v). TO1002 strains were each 0, 0.5, 1, 5, 10, 100 or 500 (μM) vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, creoleol, o-vanillic acid, vanillin No. containing methyl acid, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol or eugenol. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the pH of the culture solution after the culture was measured. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). FIG. 10 shows vanillic acid diethylamide (a), isoeugenol (b), isovanillic acid (c), o-vanillin (d), transferuric acid (e), isoferulic acid (f), isocreole (g), 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one (h), hesperetin (i), 3- (4-hydroxy-3-methoxyphenyl) propionic acid (j), isovanillyl Alcohol (k), Creosol (l), o-vanillic acid (m), methyl vanillate (n), methyl-3- (4-hydroxy-3-methoxyphenyl) propionate (o), 5-nitro Guaiacol (p), scopoletin (q), silybin (r), vanillyl alcohol (s), ethyl vanillate (t), coniferyl alcohol (u) and Euge Shows the growth promoting activity against Lactobacillus plantarum TO1002 strain Lumpur (v). TO1002 strains were each 0, 0.5, 1, 5, 10, 100 or 500 (μM) vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, creoleol, o-vanillic acid, vanillin No. containing methyl acid, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol or eugenol. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the pH of the culture solution after the culture was measured. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). Fig. 11 shows Lactobacillus paraplantarum LOOC2020 strain, Lactobacillus sakei SG171 strain, and Lactobacillus casei strain LOOC82 that promotes Lactobacillus casei LOOC82 activity against capsaicin, vanillin, ferulic acid, silybin, guaiacol and eugenol. Each strain was designated as No. containing 500 (μM) capsaicin, vanillin, ferulic acid, silybin, guaiacol or eugenol. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05).

  The present invention will be described below. Throughout this specification, it should be understood that the singular forms also include the plural concept unless specifically stated otherwise. Thus, it should be understood that singular articles (eg, “a”, “an”, “the”, etc. in the case of English) also include the plural concept unless otherwise stated. Unless defined otherwise, it is understood that similar terms apply to variations thereof and other corresponding parts of speech. In addition, it is to be understood that the terms used in the present specification are used in the meaning normally used in the art unless otherwise specified. Thus, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

(Definition of terms)
In this specification, the following abbreviations are used as necessary.

L. : Lactobacillus
L. plantarum: Lactobacillus plantarum
L. paraplantarum: Lactobacillus paraplantarum
L. sakei: Lactobacillus sakei
L. casei: Lactobacillus casei
MHPG: 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt VMA: DL-4-hydroxy-3-methoxymandelic acid 3MT: 3-methoxytyramine hydrochloride HVA: homovanillic acid VA: vanillic acid MOPET: homovanillyl alcohol.

  The meaning of each term used in this specification will be described below. In the present specification, each term is used in a unified meaning, and is used in the same meaning when used alone or in combination with other terms.

  In the present specification, “a compound having (including) a guaiacol skeleton” or “a compound containing a guaiacol skeleton” means any compound having a guaiacol (2-methoxyphenol) skeleton. It is understood that the compound having a guaiacol skeleton may be any isomer (such as an optical isomer) and refers to both when there is a D / L isomer. Examples of such substances include guaiacol, ferulic acid, vanillin, creosole, 4-ethylguaiacol, hydroxyferulic acid, capsaicin, gingerol (gingerol), shogaol, 4-hydroxy-3-methoxyphenyl glycol hemipiperazi Nium salt, DL-normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid, 3-methoxytyramine hydrochloride, homovanillic acid, vanillic acid (o-vanillic acid), homovanillyl alcohol, metanephrine hydrochloride, coni Ferryl alcohol, vanillic acid diethylamide, isoeugenol, eugenol, peonidin chloride, 2-methoxyhydroquinone, 4′-hydroxy-3′-methoxyacetophenone, iso Nilin, o-vanillin, sodium 4-hydroxy-3-methoxybenzoate, trans-ferulic acid (transferulic acid), isoferulic acid, isocresol, 3-hydroxy-4-methoxycinnamic acid, 4-hydroxy-3 -Methoxybenzonitrile, 2-methoxy-5-methylphenol, 3-hydroxy-4-methoxyaniline, hesperetin, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, 3- (4 -Hydroxy-3-methoxyphenyl) propionic acid, isovanillic acid, isovanillyl alcohol, 2-methoxy-4-methylphenol, 3-methoxysalicylic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) ) Propionate, 3- (4-hydroxy-3-methoxy) Phenyl) propionate, 5-nitro guaiacol, mention may be made of scopoletin, silybin, vanillyl alcohol, ethyl vanillic acid and the like.

  Typical preferable examples of the compound having a guaiacol skeleton include ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine (Normetanephrine) Hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), metanephrine ( Metanephrine) hydrochloride, guaiacol (2-methoxyphenol), gingerol, shogaol, vanillic acid diethylamide, isoeugenol, eugenol, isovanillic acid, o-vanillin, transfer Acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl Alcohol, creosole, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coni Mention may be made of ferryl alcohol and their salts and solvates.

  Further preferred embodiments may include ferulic acid, hydroxyferulic acid, vanillic acid and capsaicin.

  In the present specification, the “catecholamine metabolite” of the compounds having a guaiacol skeleton refers to a catecholamine metabolite in vivo or a variant thereof among the compounds having a guaiacol skeleton.

  In the present specification, “a compound having (including) a 4-methoxyphenol skeleton” or “a compound containing a 4-methoxyphenol skeleton” means any compound having a 4-methoxyphenol skeleton. It is understood that the compound having a 4-methoxyphenol skeleton may be any isomer (such as an optical isomer) and refers to both when there is a D / L isomer. Examples of such substances include 4-methoxyphenol, 4-methoxy-3,5-dimethylphenol, 2,6-di-tert-butyl-4-methoxyphenol, and 3-t-butyl-5-methoxycatechol. 4-tert-butyl-5-methoxycatechol, 1- (2-hydroxy-4,5-dimethoxyphenyl) -1-butanone, 5-hydroxy-8-methoxy-3,4-dihydro-1 (2H)- Naphthalenone, butylhydroxyanisole, 2,5-bis (1,1-dimethylbutyl) hydroquinone monomethyl ether, 3-bromo-4-hydroxyanisole, 3-chloro-4-hydroxyanisole, 3.4-dimethoxyphenol, 2- Fluoro-4-methoxyphenol, 2'-hydroxy-5'-methoxyacetophenone, 2-hydroxy-5-methoxybenzaldehyde, 4-methoxy-1-naphtho , 5-methoxy salicylic acid, 4-methoxy-2-nitrophenol, methyl 5-methoxy salicylate, can be cited syringaldehyde alcohol, 3,4,5-trimethoxy phenol.

  In the present specification, “water-soluble” of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton is not only soluble in water but also soluble in water via a hydrophilic solvent (for example, ethanol). Includes possible properties. In the context of the present invention, compounds having a water-soluble “highly” guaiacol skeleton and / or compounds having a 4-methoxyphenol skeleton can be classified by their solubility in ethanol and the “water-soluble” guaiacol skeleton Means at least 50 mg / ml soluble in ethanol, but is not limited thereto, for example, at least 100 mg / ml, at least 90 mg / ml, at least 80 mg / ml, at least 70 mg / ml, at least 60 mg / ml, Alternatively, it means dissolving at least 40 mg / ml, at least 30 mg / ml, at least 20 mg / ml, at least 10 mg / ml.

  In the present specification, the “molecular weight” of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton is calculated by a method usually calculated in this field. In one embodiment, exemplary molecular weights that are suitable in the context of the present invention are about 1000 Da or less, about 900 Da or less, about 800 Da or less, about 700 Da or less, about 600 Da or less, about 400 Da or less, For example, it is not limited to about 350 Da or less. Since the molecular weight of guaiacol itself is 124.14, the minimum molecular weight is the molecular weight of this guaiacol.

  In the present specification, “ferulic acid” is an organic compound (CA537-98-4) present as a phytochemical on the cell wall of a plant,

It is represented by the structural formula of Ferulic acid is known to have an antioxidant action, and recently, research has also been conducted on antibacterial action. Ferulic acid exhibits antibacterial properties against various bacteria (including lactic acid bacteria), but is an acid-type antibacterial component that exhibits antibacterial properties on the acidic side, like sorbic acid. Although it is not desired to be bound by theory, typically, the optimum pH varies depending on the bacteria, but is set to pH 5.0 or less. Therefore, the present invention provides a new activity within the range where there is no antibacterial action. Attention is also paid to the points found.

In the present specification, “Lactobacillus plantarum” or “L. plantarum” is a gram-positive bacterium classified into the genus Lactobacillus and particularly refers to a strain having the following microbiological characteristics.
-By sequence analysis of the 16S rRNA gene sequence, It exhibits high homology such as 99% with a 16S rRNA sequence (DDBJ / EMBL / GenBank accession number, X52653) of a plantarum reference strain (for example, JCM1149T strain) ・ A phylogenetic tree analysis by MEGA etc. after alignment analysis by CLUSTALX L. A phylogenetically related positional relationship with the plantarum reference strain is observed. The recA gene is amplified by the multiplex PCR method targeting the recA gene. Show the same electrophoresis pattern as the byproduct of the plantarum reference strain.

In this specification, “Lactobacillus paraplantarum” or “L. paraplantarum” is a gram-positive bacterium classified into the genus Lactobacillus, and particularly refers to a strain having the following microbiological characteristics.
-By sequence analysis of the 16S rRNA gene sequence, It shows high homology such as 99% with 16S rRNA sequence (DDBJ / EMBL / GenBank accession number, AJ306297) of paraplantarum reference strain (eg DSM10667T) ・ By phylogenetic tree analysis by MEGA after alignment analysis by CLUSTALX etc. , L. A phylogenetically related positional relationship with the reference plant of paraplantarum is observed. A PCR amplification product of recA is obtained by L. Show the same electrophoretic pattern as the byproduct of the paraplantarum reference strain.

In the present specification, “Lactobacillus sakei” or “L.sakei” refers to a gram-positive bacterium classified into the genus Lactobacillus, and particularly refers to a strain having the following microbiological characteristics.
-By sequence analysis of the 16S rRNA gene sequence, A high homology such as 99% with a 16S rRNA sequence (DDBJ / EMBL / GenBank accession number, AM113784) of a sake reference strain (eg, DSM20017T). , L. A positional relationship closely related to the sakei reference strain is recognized.

In the present specification, “Lactobacillus casei” or “L. casei” is a Gram-positive bacterium classified into the genus Lactobacillus, and particularly refers to a strain having the following microbiological characteristics.
-By sequence analysis of the 16S rRNA gene sequence, Casei reference strain (eg, ATCC393T) 16S rRNA sequence (DDBJ / EMBL / GenBank accession number, AF469172) exhibits high homology such as 99%, etc. , L. A positional relationship closely related to the casei reference strain is recognized.

As used herein, “Lactobacillus genus” refers to a bacterial genus that produces a large amount of lactic acid, is a Gram-positive bacterium classified into the Lactobacillus genus, etc., and generally refers to a strain having the following microbiological characteristics.
• Cells are Gram positive.
-Cell shape is Neisseria gonorrhoeae or cocci.
-Catalase negative.
・ Does not form endogenous spores.
-Does not have mobility.
-Produces 50% or more lactic acid based on consumed glucose.
・ Require niacin from the vitamin B group.

In the present specification, “Lactobacillus lactic acid bacteria (strain)” refers to a strain of any species belonging to the genus Lactobacillus among lactic acid bacteria. Its characteristics are as follows: Lactobacilli that do not form spores.
-Absolute homo type fermentation, facultative hetero type fermentation or absolute hetero type fermentation type is shown.
Bacterial species described in Bactery's Manual of Systemic Bacteriology, Second edition, 2009, Volume Three (The Firmices), Lactobacillus (P465-P511).

  Examples of “Lactobacillus” include L. plantarum, as well as L. paraplantarum, L .; pentosus, L.M. sakei and L. casei may be mentioned, but is not limited thereto.

  For example, L. plantarum TO1000, TO1001, TO1002 or TO1003 (accession number NITE P-958, NITE P-959, NITE P-960 or NITE P-961 (reception number NITE AP-958, NITE AP-959, NITE AP-960 or NITE respectively) AP-961), Lactobacillus paraplantarum LOOC 2020 (reception number NITE AP-1308), Lactobacillus sakei SG171 (reception number NITE AP-1309), Lactobacillus casei LON 82L NITE AP-1311) PR14 Or Lactobacillus casei PR150 (receipt number NITE AP-1312) can be mentioned. These strains are about 2.3 to 6.3 times or more than the control group by adding a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton. It was found in the present invention that the proliferation activity was significantly increased. As another strain of Lactobacillus genus lactic acid bacteria, the JCM1149T strain can be mentioned. plantarum reference strain, which can be obtained from the same site. It should be noted that the L.A. The plantarum reference strain was also found to be reactive with ferulic acid according to the present invention (addition of 500 μM of the test compound was about 3.0 times that of the control group). Proliferative activity is increased.)

  L. plantarum and L. Casei is a member of the genus Lactobacillus in the "Probiotics Types and Microbiological Properties" section of 2006, "Probiotics / Prebiotics / Biogenics" (supervised by the Bifido Bacteria Center of Japan, edited by Tomochi Mitsuoka). (See also Ouwehand, A. C., et al., (2003). Bulletin of the IDF 380 (pp. 4-19)). . In addition, L. paraplantarum has also been reported as a bacterial species having a probiotic action (Martin R., et al., 2009, J. Dairy Res., 76, 418-425.).

In the present specification, “growth (activity)” (growth (activity)) refers to an increase in the number of individuals or cells of the microorganism, or the activity thereof. Proliferative activity can be measured with various parameters, such as pH, absorbance at a specific wavelength, etc., but in this specification 1.45 mM MgSO ₄ , 8.92 mM unless otherwise specified. Absorbance at 600 nm when cultured in medium (pH 6.5) containing NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum The turbidity of is used as a reference. Thus, the fold turbidity of the absorbance at 600 nm in the presence of that factor at that concentration is relative to the turbidity of the absorbance at 600 nm in the absence of that factor for a factor at that concentration. It can express with the magnification of.

  As used herein, “a composition for growth” refers to, for example, L. plantarum, L .; paraplantarum, L .; sakei and L. When a lactic acid bacterium such as a casei strain is mentioned, it means a composition for the growth of the bacterium. In the present invention, typically, a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton is included, and a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton is obtained by metabolism or the like. Substances such as composites can also be used in addition to or in place of compounds having a substantially guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton as components of a composition for growth Is understood.

  As used herein, “probiotic” or “probiotic composition” refers to a microorganism that has a positive effect on the human or animal body, or a product, food, or the like containing them. By increasing the number of so-called “good bacteria” (for example, lactic acid bacteria) in the body and improving the bacterial flora in the digestive tract, the balance of intestinal bacteria is maintained, the body's health is positively affected, and the body is less susceptible to disease. It works by For example, fermented milk (for example, yogurt etc.), lactic acid bacteria beverages, live bacteria preparations and the like are used as probiotics.

  As used herein, “prebiotic” or “prebiotic composition” refers to a component having an ability to increase microorganisms having a probiotic effect, such as a dietary component or a supplement. Non-digestible carbohydrates such as oligosaccharides that have a positive effect on intestinal bacteria can be exemplified.

  As used herein, “symbiotic” or “symbiotic composition” is defined to include probiotics (compositions) and synbiotics (compositions). Since prebiotics are defined as components that have an effect on a particular probiotic, synbiotics are prepared as appropriate by selecting and combining the appropriate prebiotics once a probiotic is selected. can do.

(Preferred embodiment)
Preferred embodiments of the present invention are listed below. The embodiments provided below are provided for a better understanding of the present invention, and the scope of the present invention should not be limited to the following description. Therefore, it is obvious that those skilled in the art can make appropriate modifications within the scope of the present invention with reference to the description in the present specification.

  In one aspect, the present invention relates to a Lactobacillus lactic acid strain whose growth activity is increased by a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton (for example, L. plantarum, L. paraplantarum, L. sakei and L. casei strain, etc.). The Lactobacillus genus lactic acid bacterial strain may be of any origin, but if it is derived from grass silage, it can be added to food residues, silage, and fermentation TMR to proliferate the probiotic effect. As such, it is expected to be widely used in the livestock field. By selecting probiotic lactic acid bacteria derived from human feces, safety derived from humans is expected more. In addition, the selection of probiotic lactic acid bacteria derived from fermented foods such as vegetables and pickles, which are experienced in food for humans and empirically ensured safety, may be harmful in the future in response to stress such as environmental changes. Development of functional foods that maintain the digestive tract environment while actively eliminating microorganisms can be expected. In an exemplary embodiment, the strain is provided as a strain belonging to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei, and Lactobacillus casei. Preferably, the compound in the present invention is a compound having a guaiacol skeleton. Without wishing to be bound by theory, this is because compounds have been found herein that have proliferative activity even below μM. However, when comparing guaiacol and 4-methoxyphenol, they have almost the same activity, and thus correspond to compounds having a guaiacol skeleton (that is, they have the same substituents except for the skeleton). A compound having a -methoxyphenol skeleton is expected to have the same activity.

  The compound having a guaiacol skeleton and / or the compound having a 4-methoxyphenol skeleton may be any compound and has a compound having a guaiacol skeleton and / or a 4-methoxyphenol skeleton used in the present invention. Examples of the compound include 4-methoxyphenol, guaiacol, ferulic acid, vanillin, creosole, 4-ethylguaiacol, hydroxyferulic acid, capsaicin, gingerol (gingerol), shogaol, and 4-hydroxy-3 described in the present specification. -Methoxyphenylglycol hemipiperazinium salt, DL-normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid, 3-methoxytyramine hydrochloride, homovanillic acid, vanillic acid (o-vanillic acid), Homovanillyl alcohol, metanephrine hydrochloride, coniferyl alcohol, vanillic acid diethylamide, isoeugenol, eugenol, peonidin chloride, 2-methoxyhydroquinone, 4′-hydroxy-3′-methoxyacetophenone, isovanillin, o-vanillin 4-sodium 4-hydroxy-3-methoxybenzoate, trans-ferulic acid (transferulic acid), isoferulic acid, isocreole, 3-hydroxy-4-methoxycinnamic acid, 4-hydroxy-3-methoxybenzonitrile, 2-methoxy-5-methylphenol, 3-hydroxy-4-methoxyaniline, hesperetin, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, 3- (4-hydroxy -3-methoxyphenyl) propionic acid, isovanillic acid, isovanillyl alcohol, 2-methoxy-4-methylphenol, 3-methoxysalicylic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propion Acid salt, methyl 3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, their salts, solvates (eg hydrates) In addition to any specific examples such as, other compounds having the same skeleton may be mentioned. Typical examples of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton that can be generally used include those having high water solubility (for example, those that dissolve in ethanol at 50 mg / ml). It is not limited to them. Alternatively, molecular weight can also be indexed (eg, about 500 daltons or less, about 400 daltons or less, about 350 daltons or less, etc.).

  Alternatively, catecholamine metabolites (eg, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloric acid Salt, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine hydrochloride (may be 3,4-dihydroxyphenylglycolaldehyde (MOPEGAL), etc.) It may correspond to. Without wishing to be bound by theory, the strains of the present invention comprise a hydroxy moiety at a specific position of the guaiacol skeleton and a moiety containing a methoxy group, or a hydroxy moiety at a specific position of the 4-methoxyphenol skeleton. It is considered that the proliferation activity is increased by recognizing a portion containing a methoxy group and receiving a stimulus. Thus, without wishing to be bound by theory, it is important that the compounds of the present invention are able to recognize the guaiacol skeleton and / or the 4-methoxyphenol skeleton for strains, and the guaiacol skeleton and / or 4 It is understood that the other part may have any structure as long as it is a compound capable of recognizing the methoxyphenol skeleton, and may be guaiacol or 4-methoxyphenol itself. In addition, any isomer may be used, and when there is a D / L isomer, it is understood that either the D isomer or the L isomer can be used in the present invention. In addition, compounds such as compounds and composites that produce a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton by metabolism or the like are also used as a component of the present invention. It is understood that it can be used in addition or alternatively to compounds having a 4-methoxyphenol skeleton. Although not wishing to be bound by theory, these compounds having a guaiacol skeleton and / or compounds having a 4-methoxyphenol skeleton have a specific structure that the microorganism of the present invention recognizes as a growth active ingredient. Can be cited as a preferable reason.

  The reason why ferulic acid is preferable is that it is not desired to be bound by theory, but lactic acid bacteria are beneficial. However, excessive growth in food may increase acidity and may be inappropriate in flavor. When fermenting milk near neutrality, it works as a substance that contributes to rapid fermentation, and when the pH drops, it is considered that the remaining substance will stop the reaction antibacterial at the same time it is decomposed .

In one embodiment, the growth activity of the strain of the present invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. It has at least one of the following activities in a medium (pH 6.5) containing PO ₄ and 30% bovine serum.

An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4-fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride,
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride,
An increase of at least about 4.2 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 2.7 times in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 4 in the presence of 500 μM eugenol compared to the absence of eugenol. It rises 3 times.

In one embodiment, the growth activity of the strain of the present invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO. _In a medium (pH 6.5) containing ₄ and 30% bovine serum, it has at least one of the following activities.

An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid;
An increase of at least about 4.4 fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid;
An increase of at least about 5.0-fold in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase;
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride;
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ;
An increase of at least about 5.8-fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride;
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA);
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride;
An increase of at least about 7.9 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 3.4-fold in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 4 in the presence of 500 μM eugenol compared to the absence of eugenol. It rises 3 times.

In another embodiment, the growth activity of the strain of the invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. It has at least one of the following activities in a medium (pH 6.5) containing PO ₄ and 30% bovine serum.

An increase of at least about 5.5 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid;
An increase of at least about 4.4 fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid;
An increase of at least about 5.0-fold in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase;
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride;
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ;
An increase of at least about 5.8-fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride;
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA);
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride,
An increase of at least about 7.9 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 3.4-fold in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 7 in the presence of 500 μM eugenol compared to the absence of eugenol. It rises 8 times.

In a further embodiment, the growth activity of the strain of the invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. In a medium (pH 6.5) containing PO ₄ and 30% bovine serum,
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 2.7 times in the presence of 500 μM vanillin compared to the absence of vanillin,
An increase of at least about 3.8 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
An increase of at least about 5.8 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 2.7-fold in the presence of 500 μM silybin compared to the absence of silybin, and an increase of at least about 6.9-fold in the presence of 500 μM eugenol compared to the absence of eugenol At least one of the features selected from the group consisting of:

In a further embodiment, the growth activity of the strain of the invention is 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH _2. In a medium (pH 6.5) containing PO ₄ and 30% bovine serum,
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.2 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 2.7-fold in the presence of 500 μM silybin compared to the absence of silybin, and an increase of at least about 4.3-fold in the presence of 500 μM eugenol compared to the absence of eugenol At least one of the features selected from the group consisting of:

  In one preferred embodiment, the compound having a guaiacol skeleton utilized in the present invention comprises a metabolite of catecholamine. Although not wishing to be bound by theory, catecholamines are neurotransmitters that exert physiological effects as hormones and neurotransmitters in animals and plants, and not only at the end of nerve fibers but also in the digestive tract. Because it is also present in tissues, digestive tract contents, feces, urine, and blood, the potential amount is abundant, and highly effective use as a prebiotic component can be fully expected. Can be used as probiotics, and since such a microorganism has not been found so far, it has a unique utility. Examples of such catecholamine metabolites include 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine ( 3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine hydrochloride, other examples include, for example, 3,4- Dihydroxyphenyl glycol aldehyde (MOPEGAL) can also be exemplified.

  Resistant carbohydrates (fructooligosaccharides, galactooligosaccharides, lactulose, etc.) have been studied as substances exhibiting a prebiotic effect against lactic acid bacteria, which are extremely important beneficial microorganisms in the food and feed industries. As a representative example of a compound having a guaiacol skeleton, the presence of pathogenic bacteria that recognize and promote the growth of catecholamine metabolites that increase in vivo concentration due to stress such as environmental changes has been shown. Utilizing a microorganism having a catecholamine metabolite as a proliferative active component as probiotics leads to competitive inhibition of pathogenic bacteria, and can be expected to be a new preventive or therapeutic method against stress.

  In one embodiment, the compounds used in the present invention are preferably ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL- Normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET) ), And metanephrine hydrochloride, guaiacol (2-methoxyphenol), 4-methoxyphenol, gingerol, shogaol, vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin , Transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid , Isovanillyl alcohol, creosole, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, vanillin Ethyl acid, coniferyl alcohol, eugenol and their salts and solvates, more preferably ferulic acid, hydroxyferulic acid, vanillic acid and capsaicin, most preferably ferulic acid or its derivatives are utilized. .

  Although not wishing to be bound by theory, the reason why these compounds are preferable is that these compounds have been confirmed to be effective as antioxidants or antibacterial agents for their activities on the living body, and they can cope with stress. This is because it is recognized as a substance of the nature and is expected to be effective as stress-responsive probiotics. Although not wishing to be bound by theory, some of these compounds not only function as antibiotics in the animal body, but also exist as physiologically active substances in plants and foods with abundant dietary experience. It has been confirmed. Plants and food ingredients such as vegetables containing these compounds were invented in this study. L. as probiotics when ingested with plantarum. This is because efficient growth promotion of plantarum is expected. Furthermore, although not wishing to be bound by theory, when ingesting plants and food ingredients such as vegetables, harmful fungi are removed while these compounds become probiotic ingredients that live in the living body. There is also a reason that it can be expected to promote the growth of biotics and contribute to improving the gut microbiota balance.

  In a preferred embodiment, the compound having a guaiacol skeleton used in the present invention includes ferulic acid or a derivative thereof. Although not wishing to be bound by theory, ferulic acid is known to have an antioxidant effect, and recently, research on antibacterial activity has also progressed, so it can be applied to foods and feeds near neutrality. However, lactic acid bacteria are beneficial, but excessive growth in food may increase acidity and may be inappropriate in flavor. When fermenting milk near neutrality, it works as a substance that contributes to rapid fermentation, and when pH drops, it is expected that the remaining substance will stop the reaction antibacterial at the same time it is decomposed. This is because that.

  In one embodiment, the strain of the invention is a strain belonging to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei and Lactobacillus casei, for example, in the exemplary embodiment L. plantarum TO1000, L. plantarum TO1001, L. plantarum TO1002 or L. plantarum TO1003 (Accession Number NITE P-958, NITE P-959, NITE P-960 or NITE P-961). Alternatively, Lactobacillus planetarum JCM1149, Lactobacillus paraplanum LOOC 2020 (reception number NITE AP-1308), Lactobacillus sakei SG171 (reception number NITE AP-1309), Lactacillus 82 AP-1311) or Lactobacillus casei PR150 (reception number NITE AP-1312) may also be used.

  In another aspect, the present invention provides a probiotic or probiotic composition comprising a strain of the present invention (eg, Lactobacillus lactic acid bacteria such as L. plantarum, Lactobacillus paraplanum, Lactobacillus sakei and Lactobacillus casei strains).

  In another aspect, the present invention includes a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton. Compositions for the growth of Lactobacillus genus lactic acid strains such as plantarum, Lactobacillus paraplantum, Lactobacillus sakei and Lactobacillus casei strains are provided. Such a composition can be used as prebiotics and is useful. Embodiments of the composition of the present invention include the following: foods, beverages, feeds, e.g. in the case of ingestion of plantarum, simultaneous intake or simultaneous intake as any additive, L. Fermented dairy products such as yogurt with plantarum, fruit (eg, banana) sauce, vegetable juice, fruit juice and the like. It is understood that the compounds that can be included in the composition of the present invention can include compounds having any guaiacol skeleton described herein, as well as compounds having any other same skeleton.

  In one embodiment, the compound included in the composition of the invention is preferably ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET) and metanephrine hydrochloride, guaiacol (2-methoxyphenol), 4-methoxyphenol, gingerol, shogaol, vanillic acid diethylamide, isoeugenol, eugenol, isovanillin Acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxy) Phenyl) propionic acid, isovanillyl alcohol, creosole, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vani Ril alcohol, ethyl vanillate, coniferyl alcohol and their salts and solvates, more preferably ferulic acid, hydroxyferulic acid, vanillic acid and capsaicin, most preferably ferulic acid or derivatives thereof Is done.

  In one embodiment, the subject strain of the composition of the present invention is L. plantarum TO1000, TO1001, TO1002 or TO1003 (Accession number NITE P-958, NITE P-959, NITE P-960 or NITE P-961), JCM1149, Lactobacillus paraplanum LOOC1ac-1 sap1 308 (Receive number NITE AP-1309), Lactobacillus casei LOOC82 (Receive number NITE AP-1310), Lactobacillus casei PR143 (Receive number NITE AP-1311) or Lactobacillus casei PR150 (Receive number NITE AP-1 It is a plurality of strain.

  In another aspect, the present invention provides a prebiotic or prebiotic composition comprising a compound having a guaiacol skeleton of the present invention and / or a compound having a 4-methoxyphenol skeleton for use in combination with the strain of the present invention. Offer things. Examples of the compound to be included in such prebiotics include any compound described above in this specification.

  In another aspect, the present invention provides a symbiotic or a symbiotic composition in which the strain of the present invention is combined with a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton. Examples of specific products and products as such synbiotics include, for example, 1) a compound having a guaiacol skeleton derived from food or plants and / or a compound having a 4-methoxyphenol skeleton and a lactic acid bacterium at the same time. Yogurt (solid, drink), fermented dairy products such as cheese; 2) Tablets designed to contain a compound having a guaiacol skeleton derived from food or plant and / or a compound having a 4-methoxyphenol skeleton and lactic acid bacteria at the same time Tablet-type supplement products such as 3) and fermented foods such as pickles (vegetables) fermented with plantarum and kimchi ”. That is, if the food form is such that the lactic acid bacteria and the compound exist naturally or at the same time in design, it is intended to correspond to the synbiotic of the present invention. In the present specification, it is understood that a case where a strain and a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton are used alone is a probiotic and a prebiotic. The

In one embodiment, the compounds utilized in the present invention, when present in the body, are at least 5 μM, at least 10 μM, more preferably at least 20 μM per 1 × 10 ⁶ colony forming units (cfu) of the strain of the present invention, More preferably at least 30 μM, more preferably at least 40 μM, more preferably at least 50 μM, more preferably at least 60 μM, more preferably at least 70 μM, more preferably at least 80 μM, more preferably at least 90 μM, more preferably at least 100 μM. Although it is preferable, it is understood that the present invention is not necessarily limited to this, and that it can work even at a lower concentration. Therefore, it should be understood by those skilled in the art that, in the present invention, if the number of bacteria is not more than the above cfu, it may be preferable even at a lower concentration (for example, at least 1 μM).

In one aspect, the present invention provides a method for selecting a lactic acid bacterium (for example, L. plantarum strain) whose proliferation activity is increased by a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton. This method is described in A) L. providing step provides a sample comprising plantarum bacteria; B) magnesium sulfate (MgSO _4), ammonium nitrate _(NH 4 _NO 3), potassium chloride (KCl), D - (+ ) - glucose, potassium dihydrogen phosphate (KH ₂ A culture step of culturing the sample in the presence or absence of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton in a medium containing PO ₄ ) and bovine serum; and C) of the compound A separation step is included that isolates a strain that does not grow in the absence and that has grown in the presence or that has been promoted to grow in the presence of the compound than in the absence.

  In one embodiment, magnesium sulfate contained in the medium used in the present invention is 1.16 to 1.74 mM, ammonium nitrate is 8.02 to 9.81 mM, and potassium chloride is 3.83 to 5.74 mM. The D-(+)-glucose is present in the medium at 3.96 to 39.64 mM, the potassium dihydrogen phosphate is 2.09 to 3.14 mM, and the bovine serum is 15 to 30%. The medium has a pH of 6.0 to 7.0, but is not limited to this, and any medium can be used as long as growth is observed even if there is more or less than these. Although not wishing to be bound by theory, the reason why these ranges are desirable is that the composition reflects the environment in the living body rather than a widely used medium for lactic acid bacteria (such as MRS medium for lactic acid bacteria). Since it contains serum as a component in the body, it is more desirable as an experimental system for selecting and identifying lactic acid bacteria that recognize compounds having a guaiacol skeleton such as ferulic acid and catecholamine metabolites and / or compounds having a 4-methoxyphenol skeleton. It becomes a composition. Although not wishing to be bound by theory, the reason why these ranges are desirable is that it does not contain any microorganism-derived components such as yeast extract contained in the medium for lactic acid bacteria. Is expected to prevent contamination of the culture medium, and it can be strongly expected to prevent misperception due to contamination of other microorganism-derived components with respect to the performance and properties of the microorganisms subjected to culture. . Although not wishing to be bound by theory, the reason why these ranges are desirable is also that the lactic acid bacteria can be cultured while maintaining the glucose concentration at a very low concentration compared to conventional media, For example, it is possible to set under low nutritional conditions assuming the large intestine and the like, and taking into account that the characteristics of the cells change dramatically depending on the nutritional conditions, one target is the guaiacol skeleton of living organisms found in the digestive tract etc. It can also be mentioned that it is extremely significant and characteristic in terms of the nature of the selection method using the recognition of a compound having a 4- and / or a compound having a 4-methoxyphenol skeleton as an index.

For example, preferred medium compositions include 1.44 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 7.93 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and Examples include, but are not limited to, those containing 30% bovine serum (pH 6.5).

  In one embodiment, the culturing step of the method of the invention is performed for 32-48 hours. Preferably, it is performed in 36 hours (for example, a range of about plus or minus 3 hours is allowed), and it can be determined within 48 hours, but it is not limited thereto. The determination at such time in the low nutrient condition medium is difficult in the conventional method, and is advantageous in that respect.

  In one embodiment, the culturing step of the method of the present invention is performed at 20 to 45 ° C., preferably 25 to 40 ° C. (eg, 30 to 37 ° C.), but is not limited thereto. While not wishing to be bound by theory, the reason why these temperature conditions are desirable is that they are usually Since it is a temperature range suitable for the growth of lactic acid bacteria such as plantarum, there is an advantage that a more efficient growth temperature condition is set as a background environment, and the strain of the present invention has an advantage of ferulic acid in vivo. From the recognition of a compound having a guaiacol skeleton such as a catecholamine metabolite and / or a compound having a 4-methoxyphenol skeleton, it is also meaningful in the sense that it is equivalent to a body temperature or an intestinal temperature.

In one embodiment, the culturing step of the method of the present invention is performed under conditions of 0-5% CO ₂ and 1-20% O ₂ , preferably 5% CO ₂ , 1% O ₂ . However, it is not limited to this. While not wishing to be bound by theory, it is preferred that these ranges are those of L. Lactobacillus such as plantarum can grow even under normal atmospheric pressure oxygen conditions, but by setting the oxygen condition to about 1%, it is more efficient as an original characteristic of the strain as anaerobic bacteria. It is possible to proliferate and to enable rapid determination as an evaluation system. In addition, although not wishing to be bound by theory, in addition to the advantages on the evaluation system, when considering the virtual intestinal tract, the intestinal tract is extremely anaerobic, so by making it hypoxic, One of the preferable reasons is that it can be expected to be established as a system reflecting the reaction in the intestine. On the other hand, although not wishing to be bound by theory, there is also an advantage that the upper gastrointestinal tract (including the oral cavity and esophagus) is relatively aerobic and can be studied in a wide gas range. Although not wishing to be bound by theory, the carbon dioxide concentration is set to an appropriate range as a tissue / cell culture model that reflects the in vivo environment, such as 5% during normal cell culture. There is also an advantage that can be made.

In one embodiment, the sample used in the method of the invention is between 5 × 10 ^{5 and} 5 × 10 ⁶ colony forming units / well, preferably 1 × 10 ⁶ colony forming units / well. Examples include, but are not limited to, lactic acid bacteria such as Lactobacillus genus lactic acid strains such as plantarum, Lactobacillus paraplantarum, Lactobacillus sakei, Lactobacillus casei. Although not wishing to be bound by theory, these ranges are preferable because it is possible to evaluate the reactivity of test compounds in an appropriate concentration range within 48 hours by setting the number of bacteria within the indicated range Because it becomes. It can be carried out even below this, and those skilled in the art can appropriately select the conditions based on the description of the present specification.

  As a culture vessel that can be used in the present invention, a general culture glass test tube, a plastic petri dish for culture, and the like can be used, and it is possible to test a large number of samples as a selection system. A 12- to 48-well plastic culture plate can also be used.

  It should be noted that in the selection method of the present invention, it is to be understood that a “selection system” for separating and selecting a microorganism to be tested from some environment is performed by a well-known technique.

  The method of the present invention examines microorganisms obtained by such a well-known method using the recognition of a compound having a guaiacol skeleton such as ferulic acid and / or a compound having a 4-methoxyphenol skeleton as an “selection system”. It should be noted.

  That is, using such a known method, the microorganism L. Lactic acid bacteria such as plantarum can be separated and selected relatively frequently mainly from plant environments. Therefore, the separation and selection method of the starting material itself does not require specially difficult techniques and equipment, and should be easy to handle and obtain with knowledge of microbial operation, and should have a certain degree of generality. .

  As such an exemplary method, for example, separation from timothy orchardgrass mixed grass silage is performed according to the following method, and timothy orchard grass mixed grass silage is sufficiently homogenized with sterilized water (1:10 w / v), and silage is obtained. The suspension supernatant is serially diluted with sterilized water, and then applied to Man Rogosa Sharpe (MRS) agar medium (Difco, Detroit, USA) and cultured for 48 hours (30 ° C. under anaerobic conditions). After culturing, the formed colonies can be purely cultured and stored at −80 ° C. in a nutrient broth (Difco) containing 10% glycerol until use.

  The microorganism used in the present invention was L. lactic acid bacteria derived from grass silage. is a plantarum and has already been reported in many cases as a bacterial species frequently isolated from fermented and stored feed for livestock feed (Rossi F. & Dellaglio F., Journal of Applied Microbiology (2007) Vol. 103, 1707-1715; Ennahar S. et al., Applied and Environmental Microbiology (2003) Vol.69, 444-451; Stevenson DM, et al., Applied microbiology 29 In either case, it is easily and frequently separated from each feed suspension by a medium for lactic acid bacteria.

  Examples of the medium that can be used for selection of starting materials include, but are not limited to, the technique described in, for example, page 18 of “Science and Technology of Lactic Acid Bacteria” (Academic Publishing Center, Inc.). In this document, the composition of the main medium used in the lactic acid bacteria described at the end of the book is made, and these documents are incorporated herein by reference.

  As the medium that can be used for selection of the starting material, any appropriate medium can be exemplified. For example, MRS medium, GYP medium, BL medium, BCP-added plate count agar medium, GAM can be mentioned as well-known medium. Examples thereof include a medium and a tomato juice medium. Looking at the composition of these media, all of them contain yeast extract and the like, and it is recognized that the glucose concentration is relatively high.

  On the other hand, the growth activity is increased by the compound having a guaiacol skeleton and / or the compound having a 4-methoxyphenol skeleton of the present invention. A method for selecting a lactic acid bacterium including Lactobacillus lactic acid strains such as plantarum, Lactobacillus paraplantarum, Lactobacillus sakei and Lactobacillus casei strains cannot be easily assumed from such well-known methods. The recognizability of a compound having a guaiacol skeleton such as ferulic acid and / or a compound having a 4-methoxyphenol skeleton is not a well-known fact. A compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton of a lactic acid strain such as lactobacillus lactic acid strains such as plantarum, lactobacillus paraplantarum, lactobacillus sakei and lactobacillus casei Remarkableness is also recognized in that it became.

  In addition, the medium used in the selection system of the present invention has different properties from those used in known techniques. For example, taking glucose as an example, the new medium used in the present invention is about 1/20 of the most widely used MRS medium. In addition, since yeast extract is used as a vitamin source and amino acid source and is contained in many culture media, it has a common recognition as an essential component in the field, and yeast extract is included as in the culture media of the present invention. The advantages such as absence and undernutrition are significant in the present invention.

  In one exemplary embodiment, the selection is as follows.

No. ₄ containing MgSO ₄ , NH ₄ NO ₃ , KCl, C ₆ H ₁₂ O ₆ , KH ₂ PO ₄ , bovine serum. Developed 189-091201 medium. Each component contained in this exemplary medium is preferably prepared at the following concentrations and pH (1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum, pH 6.5). The details of each recommended component are as follows, but there is no problem in the medium performance as long as it is an equivalent product.

D (+)-glucose (C ₆ H ₁₂ O ₆ ), Wako Pure Chemical Industries, Ltd. (reagent special grade, 041-00595)
Potassium dihydrogen phosphate (KH ₂ PO ₄ ), Wako Pure Chemical Industries, Ltd. (special reagent grade, 169-04245)
Ammonium nitrate (NH ₄ NO ₃ ), Wako Pure Chemical Industries, Ltd. (special grade reagent, 017-03235)
Potassium chloride (KCl), Wako Pure Chemical Industries, Ltd. (special grade reagent, 163-03545)
Magnesium sulfate heptahydrate, Wako Pure Chemical Industries, Ltd. (special reagent grade, 131-00405)
Bovine serum, adult, Sigma (B5433).

  The composition contained in the medium is newly prepared in the present invention and is not conventionally known. In addition, since it does not contain any microorganism-derived components such as yeast extract commonly used in microorganism culture media (for example, LB medium for Escherichia coli, MRS medium for lactic acid bacteria, etc.), It is expected to prevent contamination inside. Thereby, it can be strongly expected that the performance and properties of the microorganisms subjected to the culture are prevented from being misidentified due to contamination with other microorganism-derived components. In addition, the nutrient concentration is set to low nutritional conditions such as in the large intestine of the living body such that the lactic acid bacteria can be cultured while maintaining the glucose concentration at a very low concentration compared to the conventional medium. This is because the properties of the cells change dramatically depending on the nutrient conditions.

The test microorganism is pre-cultured in an appropriate medium (for example, MRS medium or the above-mentioned No. 189-091201 medium in the case of L. plantarum TO1002), sufficiently proliferated, and the cells are collected by centrifugation, and then cultured on the medium. Remove the supernatant and centrifuge thoroughly with fresh PBS or medium several times. No. used illustratively. The cells were resuspended in a 189-091201 medium, and each of the tested compounds having a guaiacol skeleton such as ferulic acid and / or a compound having a 4-methoxyphenol skeleton was subjected to anaerobic conditions (5% CO 2) in a 24-well plate. ₂ or 1% O ₂ ) or aerobic conditions (5% CO ₂ , 20% O ₂ ) (adjust the number of test microorganisms to about 1 × 10 ⁶ CFU / well. CFU) Are colony forming units). The cells are cultured for about 36 hours, and the growth of the bacterial cells is analyzed by measuring the number of colonies formed, the culture turbidity (OD600), or the culture solution pH.

  It is understood that these conditions are merely examples, and those skilled in the art can change based on the spirit of the present invention.

  The usefulness of the selection method of the present invention that can be expected includes the following. The selection technique found in the present invention enables screening of microorganisms derived from various environments. For example, for probiotic lactic acid bacteria derived from human or domestic animal feces, recognition of compounds having a guaiacol skeleton such as ferulic acid and / or compounds having a 4-methoxyphenol skeleton having antibacterial activity when the environment is inclined to an acidic type In the future, we will develop functional foods and feeds that maintain the gastrointestinal tract environment while ensuring the safety derived from the host and actively competing against harmful microorganisms when recognizing environmental changes. You can expect. Since food residues, silage, and fermentation TMR also contain “food / plant-derived ferulic acid”, when preparing livestock fermented feeds that effectively use these materials, lactic acid bacteria selected by the selection technique of the present invention Can be expected to be effectively used as a “lactic acid bacterium additive for feed preparation”. Furthermore, if harmful microorganisms such as stool-derived Escherichia coli are selected, it will lead to the identification of harmful microorganisms that grow rapidly upon recognition of environmental changes, and contribute to drug discovery that expects bactericidal and bacteriostatic effects against these microorganisms. Can be expected. It is also important that a plurality of bacterial cell samples can be examined simultaneously. That is, it is not desired to be bound by theory, but the culture conditions and the initial number of added bacteria may be important because the system cannot be established unless they are optimally set.

  The features of the present invention include the following. That is, in general, in order to grow a specific bacterium from a sample derived from a certain environment, a selective medium for preferential growth and selection of the bacterium is used. Currently, it is widely used in research on functions and properties and in selection tests. As a typical selective medium for lactic acid bacteria, MRS medium and the like are known. General features of these media include (1) containing a high concentration of glucose, (2) containing yeast extract as a vitamin and amino acid source, etc., and allowing the desired lactic acid bacteria to grow more efficiently and speedily. The ability is high, and it is very meaningful to use it when separating and selecting lactic acid bacteria from certain environmental samples, and it was widely used. There is one significance. In addition, L. is frequently observed from plant samples. It has been reported that lactic acid bacteria such as plantarum are separated and selected, and there are already many known literatures. In these well-known media, in general, low nutrients such as in the large intestine and plant samples after nutrient absorption. The growth conditions under conditions cannot be reflected. Moreover, since highly efficient growth is already achieved by the high nutrient components of the culture medium itself, it is difficult to determine its action when examining an unknown growth promoting factor. In addition, since lactic acid bacteria are known to have relatively high nutritional requirements, if the detailed nutritional conditions are set to low nutritional conditions without being studied, problems such as failure to establish culture may occur. It can be said that control is difficult.

  L. isolated in well-known MRS medium. When examining the new functionality of lactic acid bacteria such as Lactobacillus genus lactic acid strains such as plantarum, Lactobacillus paraplantarum, Lactobacillus sakei and Lactobacillus casei strains, it is not preferable to construct a selection method as it is in MRS medium. The development of designed culture conditions is considered to be extremely important. According to the present invention, it is considered that a growth selection test of lactic acid bacteria under conditions of low nutritional components is possible, and a selection system capable of further examining the growth ability has been constructed. In addition, it should not be overlooked that there is no research or technology for the purpose of “selecting a lactic acid bacterium that recognizes a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton”.

  In consideration of the fact that trials for adding a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton to lactic acid bacteria have not been carried out until now, the “new composition medium” addressed in the present invention, “Additional trial of compound having guaiacol skeleton and / or compound having 4-methoxyphenol skeleton” and “culture suitable for determining the action of compound having guaiacol skeleton and / or compound having 4-methoxyphenol skeleton” `` Recognizes compounds with guaiacol skeleton and / or compounds with 4-methoxyphenol skeleton, which were difficult to prove or find by conventional probiotic lactic acid bacteria culture test, function test, selection test by setting test conditions '' Proliferation and promotion Io selection of tick lactic acid bacteria "is thought to have become possible.

(General technology)
Biochemical techniques and microbiological techniques used in the present specification are well known and commonly used in the art, and examples thereof include the following. “Probiotics / Prebiotics / Biogenics” (supervised by the Nihon Bifidobacteria Center, edited by Tomokazu Mitsuoka); "Experiment Manual: From Separation to Identification" (Asakura Shoten, supervised by Michio Kosaki, Yasushi Uchimura and Sanae Okada); "Probiotics and Biogenics Scientific Basis and Future Development Prospects" (NTS Corporation, edited by Kuji Ito) Representative); “Health Function and Application of Lactic Acid Bacteria” (CMC Publishing Co., Ltd., supervised by Shuichi Uenogawa); robotics and Prebiotics (Wiley-Interscience, Seppo Salminen) and Handbook of Probiotics (Wiley-Interscience, Yuan-Kun, Koji Nomoto, Sepio Sep.

  References such as scientific literature, patents and patent applications cited herein are hereby incorporated by reference in their entirety to the same extent as if each was specifically described.

  The present invention has been described with reference to the preferred embodiments for easy understanding. In the following, the present invention will be described based on examples, but the above description and the following examples are provided only for the purpose of illustration, not for the purpose of limiting the present invention. Accordingly, the scope of the present invention is not limited to the embodiments or examples specifically described in the present specification, but is limited only by the scope of the claims.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, the technical scope of this invention is not limited by this Example. The reagents used in the following were commercially available unless otherwise specified.

(Example 1: Method for selecting a microorganism that recognizes a compound having a guaiacol skeleton such as ferulic acid)
In this example, a novel method for selecting a microorganism that recognizes a compound having a guaiacol skeleton such as ferulic acid was developed.

(Materials and methods)
(New selection method for microorganisms that recognize compounds with guaiacol skeleton)
(Culture medium)
1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% Adult
Using a medium containing bovine serum (pH 6.5, No. 189-091201 medium), the cells were cultured under microaerobic (5% CO ₂ , 1% O ₂ ) conditions.

In this example, the details of each recommended component are as follows, but it is expected that there is no problem in the medium performance if it is an equivalent product.
D (+)-glucose (C ₆ H ₁₂ O ₆ ), Wako Pure Chemical Industries, Ltd. (reagent special grade, 041-00595)
- potassium dihydrogen phosphate _(KH 2 _PO 4), Wako Pure Chemical Industries, Ltd. (special grade, 169-04245)
・ Ammonium nitrate (NH ₄ NO ₃ ), Wako Pure Chemical Industries, Ltd. (special grade reagent, 017-03235)
・ Potassium chloride (KCl), Wako Pure Chemical Industries, Ltd. (special grade reagent, 163-03545)
Magnesium sulfate heptahydrate, Wako Pure Chemical Industries, Ltd. (special grade, 131-00405) Bovine serum, Adult, Sigma (B5433).

  The composition of the medium of the present example was newly prepared in the present example, and was derived from microorganisms such as yeast extract widely used in microbial medium (for example, LB medium for Escherichia coli, MRS medium for lactic acid bacteria, etc.) Since no components are contained, it is expected that microbial components other than the microorganisms subjected to the culture are prevented from being contaminated in the medium. Thereby, it can be strongly expected that the performance and properties of the microorganisms subjected to the culture are prevented from being misidentified due to contamination with other microorganism-derived components.

  In addition, the nutrient concentration is set to low nutritional conditions such as in the large intestine of the living body such that the lactic acid bacteria can be cultured while maintaining the glucose concentration at a very low concentration compared to the conventional medium. Considering that the characteristics of bacterial cells change dramatically depending on nutritional conditions, a selection method using as an index the recognition of compounds with guaiacol skeletons such as ferulic acid known as antibiotics and antioxidants It is considered to be extremely meaningful and characteristic in terms of the nature of.

(experimental method)
The chemical structure of ferulic acid used in this study is shown below.

  The dose-dependent growth of the test lactic acid strain in the medium containing this compound was analyzed by measuring the culture turbidity.

All experiments were performed in triplicate and repeated three times as independent experiments. GraphPad
All data obtained were statistically analyzed by PRISM (Prism software, CA, USA).

Specifically, the test microorganisms were pre-cultured in an appropriate medium (for example, MRS medium or the above-mentioned No. 189-091201 medium in the case of L. plantarum TO1002) and allowed to grow sufficiently. The cells were collected by centrifugation, the medium supernatant was removed, and the cells were thoroughly washed by centrifugation several times with fresh PBS or medium. No. The cells were resuspended in a 189-091201 medium, and each test guaiacol skeleton-containing compound was mixed with a 24-well plate under anaerobic conditions (5% CO ₂ , 1% O ₂ ) or aerobic conditions (5% CO ₂ , 20% O ₂ ) (the number of test microorganisms was adjusted to about 1 × 10 ⁶ CFU / well. CFU is a colony forming unit). The cells were cultured for about 36 hours, and the growth of the bacterial cells was analyzed by measuring the number of colonies formed, the culture turbidity (OD600), or the culture solution pH.

(result)
The resulting strain was obtained from L. As a plantarum TO1000, TO1001, TO1002, or TO1003 (Accession number NITE P-958, NITE P-959, NITE P-960 or NITE P-961), the patent of the National Institute of Technology and Evaluation for Independent Administrative Agency on July 7, 2010 Deposited at the Microorganism Deposit Center (2-5-8 Kazusa Kamashika, Kisarazu City, Chiba Prefecture).

  By adding ferulic acid (anaerobic condition, culture for 36 hours), the dose-dependent L.P. proliferation of plantarum TO1002 was observed (FIG. 1A). In addition, a marked decrease in the pH of the culture solution was observed with the growth of TO1002 (FIG. 1B). From the above, ferulic acid recognition property plantarum TO1002 strain and ferulic acid having lactic acid bacteria growth promoting action are expected to be industrially useful and useful.

(Discussion)
From this example, the usefulness of this selection method that can be expected is as follows. The selection technique invented in this study enables screening of microorganisms derived from various environments. For example, for probiotic lactic acid bacteria derived from humans and livestock feces, by selecting as an index the recognition of ferulic acid known as an antibiotic or antioxidant, the safety derived from the host is ensured, and in the future, The development of functional foods and feeds that maintain the digestive tract environment while actively competing against harmful microorganisms when environmental changes are recognized can be expected. Since food residues, silage, and fermentation TMR also contain “food / plant-derived guaiacol skeletal compounds”, when preparing livestock fermented feeds that effectively use these materials, lactic acid bacteria selected by this selection technology It can be expected to be effectively used as a “lactic acid bacteria additive for feed preparation”. In addition, if harmful microorganisms such as Escherichia coli derived from stool are selected, it will lead to the identification of harmful microorganisms that grow rapidly upon recognition of environmental changes, and contribute to drug discovery that expects bactericidal and bacteriostatic effects against these microorganisms. There is expected.

(About microorganisms tested in this example)
The microorganism used in this example was L. lactic acid bacteria derived from grass silage. is a plantarum and has already been reported in many cases as a bacterial species frequently isolated from fermented and stored feed for livestock feed (Rossi F. & Dellaglio F., Journal of Applied Microbiology (2007) Vol. 103, 1707-1715; Ennahar S. et al., Applied and Environmental Microbiology (2003) Vol.69, 444-451; Stevenson DM, et al., Applied microbiology 29 In either case, it is easily and frequently separated from each feed suspension by a medium for lactic acid bacteria. Although it is a bacterial species that is easily separated, it is not a known fact about the recognizability of guaiacol skeletal compounds. It is considered that lactic acid strains such as plantarum can be evaluated for the ability to recognize guaiacol skeleton compounds such as ferulic acid.

(Example 2: Measurement of proliferation activation of other strains with ferulic acid)
Next, with respect to strains other than TO1002 isolated in Example 1, experiments were performed on the recognition ability of ferulic acid, which is a representative example of a guaiacol skeleton compound.

The method used is the same as the culture method, culture turbidity (OD600) measurement or culture solution pH measurement described in Example 1. Specifically, ferulic acid L.P. The growth promotion activity with respect to plantarum TO1000, TO1001, TO1003, and JCM1149 strain | stump | stock is shown. These strains were designated as No. 1 containing 0, 0.5, 1, 5, 10, 100 or 500 (μM) ferulic acid, respectively. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured.

(result)
L. of ferulic acid FIG. 2 shows the results of the growth promotion activity (plant turbidity after culture) for plantarum TO1000, TO1001, TO1003, and JCM1149, and FIG. 3 shows the pH of the medium. For ferulic acid, the same tendency for proliferation activation as that of TO1002 strain including the standard strain was shown. Moreover, although pH value fell when proliferation activity was recognized, all were pH5.0 or more.

(Example 3: Activity measurement of vanillin, hydroxyferulic acid and capsaicin)
Next, the representative strain TO1002 isolated in Example 1 was tested for its ability to recognize vanillin, hydroxyferulic acid and capsaicin as representative examples of guaiacol skeleton compounds other than ferulic acid.

The method used is the same as the culture method, culture turbidity (OD600) measurement or culture solution pH measurement described in Example 1. Specifically, the TO1002 strain is 0, 0.5, 1, 5, 10, 100 or 500 (μM) vanillin, or 0, 0.5, 1, 5, 10, 100 or 500 (μM) hydroxy, respectively. No. containing ferulic acid or 0, 0.1, 0.25, 0.5, 0.75, 1 or 1.5 (mM) capsaicin. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity (A) and culture solution pH (B) of the culture solution after the culture were measured.

(result)
L. vanillin, hydroxyferulic acid and capsaicin. FIG. 4A shows the turbidity and FIG. 4B shows the pH of the medium for the growth promoting activity against the plantarum TO1002 strain. Also for vanillin, hydroxyferulic acid and capsaicin, which are compounds other than ferulic acid having a guaiacol skeleton, the strain of the present invention showed the same growth activation tendency as that for ferulic acid. Moreover, although pH value fell when proliferation activity was recognized, all were pH5.0 or more.

(Example 4: Activity measurement of a compound having another guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton)
Next, for representative strain TO1002 isolated in Example 1, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL as a representative example of a guaiacol skeleton compound other than the compounds used in Examples 1 to 3 -Normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol ( MOPET), metanephrine hydrochloride, guaiacol, 3-methoxyphenol, 4-methoxyphenol, anisole, phenol, gingerol, shogaol, vanillic acid diethylamide, isoeugenol, eugenol , Isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3 -Methoxyphenyl) propionic acid, isovanillyl alcohol, creosole, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin , Vanillyl alcohol, ethyl vanillate, coniferyl alcohol, vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferulic acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-Butene- -One, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, cresol, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) ) Experiments on the recognition ability of propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol and eugenol were conducted.

The method used is the same as the culture method, culture turbidity (OD600) measurement or culture solution pH measurement described in Example 1. Specifically, the TO1002 strain is designated as No. containing 0, 0.5, 1, 5, 10, 100 or 500 (μM) MHPG, Normetanephine, VMA, 3MT, HVA, VA, MOPET or Metanephine. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity of the culture solution after the culture was measured. For guaiacol, 3-methoxyphenol, 4-methoxyphenol, anisole, and phenol, experiments were conducted at a concentration of 10,100 or 500 (μM), respectively. Gingerol and Shogaol were tested at concentrations of 0.1, 0.25, 0.5, 0.75, 1 or 1.5 (mM), respectively. Vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-Hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, cresol, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5- Nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate and coniferyl alcohol were tested at concentrations of 0, 0.5, 1, 5, 10, 100 or 500 (μM), respectively. Vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-Hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, cresol, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5- Nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol and eugenol were tested at concentrations of 0, 0.5, 1, 5, 10, 100 or 500 (μM), respectively.

(result)
4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride, DL-4-hydroxy-3-methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloric acid Salts of homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine hydrochloride. FIG. 5 shows turbidity and FIG. 6 shows the pH of the medium for the growth promoting activity against plantarum TO1002 strain. For many other compounds having a guaiacol skeleton other than the compounds demonstrated in Examples 1 to 3, the strains of the present invention showed the same growth activation tendency as for ferulic acid, vanillin, hydroxyferulic acid and capsaicin. Indicated. Moreover, although pH value fell when proliferation activity was recognized, all were pH5.0 or more.

  FIG. 7 shows the growth promoting activity of guaiacol (2-methoxyphenol), 3-methoxyphenol, 4-methoxyphenol, anisole, and phenol against Lactobacillus plantarum TO1002 strain. Guaiacol and its para isomer, 4-methoxyphenol, showed significant growth, whereas 3-methoxyphenol, anisole, and phenol did not grow significantly at the concentrations tested. Thus, for the purposes of the present invention, it is shown that the guaiacol skeleton can be substituted with the corresponding para isomer.

  Gingerol (also called gingerol; (S) -5-hydroxy-1- (4-hydroxy-3-methoxyphenyl) -3-decanone) and shogaol ((E) -1- (4-hydroxy-3-methoxyphenyl) FIG. 8 shows the growth-promoting activity and pH change of L) to Lactobacillus plantarum TO1002). The measured values of turbidity (A, B) and culture solution pH (C, D) of the culture solution after the culture are shown. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). Gingerol has a higher proliferation effect as the concentration is higher, whereas shogaol has a remarkable proliferation activity at a low concentration of around 0.25 mM, but shows a decrease in proliferation activity at a higher concentration.

  In FIG. 9 (FIGS. 9-1 and 9-2), vanillic acid diethylamide (a), isoeugenol (b), isovanillic acid (c), o-vanillin (d), transferuric acid (e), isoferulic acid (F), isoceosol (g), 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one (h), hesperetin (i), 3- (4-hydroxy-3-methoxy Phenyl) propionic acid (j), isovanillyl alcohol (k), creosole (l), o-vanillic acid (m), methyl vanillate (n), methyl-3- (4-hydroxy-3-methoxyphenyl) ) Propionate (o), 5-nitroguaiacol (p), scopoletin (q), silybin (r), vanillyl alcohol (s), ethyl vanillate (t), coniferyl Indicating the call (u) and eugenol (v) of Lactobacillus plantarum TO1002 lines to growth-promoting activity (turbidity). Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). Vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, cresol, o-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5- Nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol and eugenol were all shown to have proliferative activity at increasing concentrations.

  In FIG. 10 (FIGS. 10-1 and 10-2), vanillic acid diethylamide (a), isoeugenol (b), isovanillic acid (c), o-vanillin (d), transferuric acid (e), isoferulic acid (F), isoceosol (g), 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one (h), hesperetin (i), 3- (4-hydroxy-3-methoxy Phenyl) propionic acid (j), isovanillyl alcohol (k), creosole (l), o-vanillic acid (m), methyl vanillate (n), methyl-3- (4-hydroxy-3-methoxyphenyl) ) Propionate (o), 5-nitroguaiacol (p), scopoletin (q), silybin (r), vanillyl alcohol (s), ethyl vanillate (t), conifer Alcohol (u) and eugenol (v) growth promoting activity against Lactobacillus plantarum TO1002 strains (pH) is shown. Significant differences in comparison with the non-added medium are indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). Depending on the pH value, vanillic acid diethylamide, isoeugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid, isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-butene-2- ON, Hesperetin, 3- (4-Hydroxy-3-methoxyphenyl) propionic acid, Isovanillyl alcohol, Creosol, o-vanillic acid, Methyl vanillate, Methyl-3- (4-hydroxy-3-methoxyphenyl) Propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate and coniferyl alcohol were all shown to have proliferative activity at increasing concentrations.

(Example 5: Method for selecting a microorganism recognizing a compound having a 4-methoxyphenol skeleton)
In this example, a novel method for selecting a microorganism that recognizes a compound having a 4-methoxyphenol skeleton (for example, 4-methoxyphenol) was developed.

(Materials and methods)
(New selection method for microorganisms that recognize 4-methoxyphenol skeleton)

(Culture medium)
1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% Adult
Using a medium containing bovine serum (pH 6.5, No. 189-091201 medium), the cells were cultured under microaerobic (5% CO ₂ , 1% O ₂ ) conditions.

In this example, the details of each recommended component are as follows, but it is expected that there is no problem in the medium performance if it is an equivalent product.
D (+)-glucose (C ₆ H ₁₂ O ₆ ), Wako Pure Chemical Industries, Ltd. (reagent special grade, 041-00595)
- potassium dihydrogen phosphate _(KH 2 _PO 4), Wako Pure Chemical Industries, Ltd. (special grade, 169-04245)
・ Ammonium nitrate (NH ₄ NO ₃ ), Wako Pure Chemical Industries, Ltd. (special grade reagent, 017-03235)
・ Potassium chloride (KCl), Wako Pure Chemical Industries, Ltd. (special grade reagent, 163-03545)
Magnesium sulfate heptahydrate, Wako Pure Chemical Industries, Ltd. (special grade, 131-00405) Bovine serum, Adult, Sigma (B5433).

  The composition of the medium of the present example was newly prepared in the present example, and was derived from microorganisms such as yeast extract widely used in microbial medium (for example, LB medium for Escherichia coli, MRS medium for lactic acid bacteria, etc.) Since no components are contained, it is expected that microbial components other than the microorganisms subjected to the culture are prevented from being contaminated in the medium. Thereby, it can be strongly expected that the performance and properties of the microorganisms subjected to the culture are prevented from being misidentified due to contamination with other microorganism-derived components.

In addition, the nutrient concentration is set to low nutritional conditions such as in the large intestine of the living body such that the lactic acid bacteria can be cultured while maintaining the glucose concentration at a very low concentration compared to the conventional medium. Considering that the properties of bacterial cells change dramatically depending on nutritional conditions, as one target, the recognition of compounds having a 4-methoxyphenol skeleton such as 4-methoxyphenol known as antibiotics and antioxidants Due to the nature of the selection method used as an indicator, it is considered to be extremely meaningful and characteristic.
(experimental method)
The substance used in this study is 4-methoxyphenol.

  The dose-dependent growth of the test lactic acid strain in the medium containing this compound was analyzed by measuring the culture turbidity.

  All experiments were performed in triplicate and repeated three times as independent experiments. All data obtained were statistically analyzed by GraphPad PRISM (Prism software, CA, USA).

Specifically, the test microorganisms were pre-cultured in an appropriate medium (for example, MRS medium or the above-mentioned No. 189-091201 medium in the case of L. plantarum TO1002) and allowed to grow sufficiently. The cells were collected by centrifugation, the medium supernatant was removed, and the cells were thoroughly washed by centrifugation several times with fresh PBS or medium. No. The cells were resuspended in a 189-091201 medium, and each test 4-methoxyphenol skeleton-containing compound was combined with a 24-well plate under anaerobic conditions (5% CO ₂ , 1% O ₂ ) or aerobic conditions (5% (CO ₂ , 20% O ₂ ) (the number of test microorganisms was adjusted to about 1 × 10 ⁶ CFU / well. CFU is a colony forming unit). The cells were cultured for about 36 hours, and the growth of the bacterial cells was analyzed by measuring the number of colonies formed, the culture turbidity (OD600), or the culture solution pH.

(result)
The resulting strain was obtained from L. As a plantarum TO1000, TO1001, TO1002, or TO1003 (Accession number NITE P-958, NITE P-959, NITE P-960 or NITE P-961), the patent of the National Institute of Technology and Evaluation for Independent Administrative Agency on July 7, 2010 Deposited at the Microorganism Deposit Center (2-5-8 Kazusa Kamashika, Kisarazu City, Chiba Prefecture).

  By adding 4-methoxyphenol (anaerobic condition, culture for 36 hours), the dose-dependent L.P. The growth of plantarum TO1002 was observed (FIG. 7). From the above, 4-methoxyphenol recognition Industrial utility and utilization of plantarum TO1002 strain and 4-methoxyphenol skeleton-containing compounds such as 4-methoxyphenol having a lactic acid bacteria growth promoting action are expected.

(Discussion)
From this example, the usefulness of this selection method that can be expected is as follows. The selection technique invented in this study enables screening of microorganisms derived from various environments. For example, for probiotic lactic acid bacteria derived from human or livestock feces, the safety derived from the host is ensured by selecting the recognition ability of 4-methoxyphenol skeleton-containing compounds as an index. Development of functional foods and feeds that maintain the digestive tract environment while actively competing against harmful microorganisms can be expected. Since food residues, silage, and fermentation TMR also contain “food and plant-derived 4-methoxyphenol skeleton-containing compounds”, selection is made using this selection technique when preparing livestock fermented feeds that effectively use these materials. It can be expected that the produced lactic acid bacteria can be effectively used as “additives for lactic acid bacteria for feed preparation”. In addition, if harmful microorganisms such as Escherichia coli derived from stool are selected, it will lead to the identification of harmful microorganisms that grow rapidly upon recognition of environmental changes, and contribute to drug discovery that expects bactericidal and bacteriostatic effects against these microorganisms. There is expected.

  In addition to 4-methoxyphenol, 4-methoxy-3,5-dimethylphenol, 2,6-di-tert-butyl-4-methoxyphenol, 3-t-butyl-5-methoxycatechol, 4- tert-Butyl-5-methoxycatechol, 1- (2-hydroxy-4,5-dimethoxyphenyl) -1-butanone, 5-hydroxy-8-methoxy-3,4-dihydro-1 (2H) -naphthalenone, butyl Hydroxyanisole, 2,5-bis (1,1-dimethylbutyl) hydroquinone monomethyl ether, 3-bromo-4-hydroxyanisole, 3-chloro-4-hydroxyanisole, 3.4-dimethoxyphenol, 2-fluoro-4 -Methoxyphenol, 2'-hydroxy-5'-methoxyacetophenone, 2-hydroxy-5-methoxybenzaldehyde, 4-methoxy-1-naphthol, 5-metho Shisarichiru acid, 4-methoxy-2-nitrophenol, methyl 5-methoxy salicylate, a syringaldehyde alcohol, also 3,4,5-methoxyphenol available.

(About microorganisms tested in this example)
The microorganism used in this example was L. lactic acid bacteria derived from grass silage. is a plantarum and has already been reported in many cases as a bacterial species frequently isolated from fermented and stored feed for livestock feed (Rossi F. & Dellaglio F., Journal of Applied Microbiology (2007) Vol. 103, 1707-1715; Ennahar S. et al., Applied and Environmental Microbiology (2003) Vol.69, 444-451; Stevenson DM, et al., Applied microbiology 29 In either case, it is easily and frequently separated from each feed suspension by a medium for lactic acid bacteria. Although it is a bacterial species that is easily separated, it is not a known fact about the recognizability of guaiacol skeletal compounds. It is thought that the 4-methoxyphenol skeleton-containing compound recognition property of lactic acid strains such as plantarum can be evaluated.

(Example 6: Experiments with lactic acid bacteria other than L. plantarum)
In this embodiment, L.P. Experiments similar to those in Examples 1 to 4 were performed using lactic acid bacteria other than plantarum. In this example, Lactobacillus paraplantum, Lactobacillus sakei, and Lactobacillus casei were used.

  As representative examples thereof, Lactobacillus paraplanum LOOC 2020 (reception number NITE AP-1308), Lactobacillus sake SG SG171 (reception number NITE AP-1309), Lactobacillus case ILO CTE APN 310CLO AP Using.

  Using the above Lactobacillus paraplantarum LOOC2020 strain, Lactobacillus sakei SG171 strain, and Lactobacillus casei LOOC82 strain, capsaicin, vanillin, ferulic acid, silybin, guaiacol-promoted and activated activity were investigated. In addition, it isolate | separated from silage, such as each strain grass and rice for feed.

The method used is the same as the culture method, culture turbidity (OD600) measurement or culture solution pH measurement described in Example 1. Specifically, each strain is classified into No. 1 containing 500 (μM) capsaicin, vanillin, ferulic acid, silybin, guaiacol or eugenol. 189-091201 medium <1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum ), PH 6.5)> for 36 hours at 37 ° C. under anaerobic conditions, and the turbidity and pH of the culture solution after the culture were measured.

  The turbidity results are shown on the top of FIG. Moreover, culture solution pH is shown under FIG. In FIG. 11, the significant difference when compared with the non-added medium is indicated by *** (P <0.001), ** (P <0.01), * (P <0.05). Lactobacillus paraplantarum LOOC2020 strain, Lactobacillus sakei SG171 strain, and Lactobacillus casei LOOC82 strain showed growth activity (both turbidity and pH points) for all the compounds tested. In Lactobacillus paraplantarum LOOC2020, about 500 (μM) capsaicin, vanillin, ferulic acid, silybin, guaiacol and eugenol, the culture turbidity (OD600) was about 4.4 times, about 3.9 times and about 3.9 times, respectively. They were 5.9 times, about 2.7 times, about 5.8 times, and about 6.9 times. For the Lactobacillus sakei SG171 strain, the culture turbidity (OD600) of 500 (μM) capsaicin, vanillin, ferulic acid, silybin, guaiacol, and eugenol was about 3.8 times, about 2.7 times that of the control, respectively. They were about 6.8 times, about 3.0 times, about 6.4 times and about 7.3 times. For Lactobacillus casei LOOC82 strain, the culture turbidity (OD600) of about 500 (μM) capsaicin, vanillin, ferulic acid, silybin, guaiacol and eugenol was about 7.3 times, about 3.2 times and about 4 times, respectively. 0.9 times, about 9.2 times, about 9.2 times and about 9.8 times.

  In addition to the above strains, additional strains L. cerevisiae selected according to the method of Example 1. casei PR143 strain (reception number NITE AP-1311) and L. When experiments were conducted using casei PR150 strain (reception number NITE AP-1312), it showed a proliferative activity similarly to ferulic acid, silybin, guaiacol and eugenol.

  The strains obtained in this example are as follows: Lactobacillus paraplantarum LOOC 2020, receiving number: NITE AP-1308, Lactobacillus sakei SG171, receiving number: NITE AP-1309, Lactobacillus casei LOO82 Lactobacillus casei PR143 receipt number: NITE AP-1311, and Lactobacillus casei PR150 receipt number: NITE AP-1312, National Institute for Product Evaluation and Technology Patent Microorganism Depositary Center (2-5 Kazusa Kamashika, Kisarazu City, Chiba Prefecture) 8).

  Examples of other test bacteria include L. pentosus can also be used.

(Example 7: Calculation of multiplication activity multiplication factor)
In this example, to better characterize each strain, the growth activity was 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH. Changes in growth activity in media containing ₂ PO ₄ and 30% bovine serum (pH 6.5) were calculated. The same experiment as in Examples 1 to 5 was performed, and the results were calculated as follows. Below, the result put together by the strain of Lactobacillus plantarum is shown first.

  The proliferation activity of each of the non-added group and the compound having a guaiacol skeleton and / or the compound having a 4-methoxyphenol skeleton was analyzed based on the turbidity (optical density, OD 600 nm) of the culture solution after the culture. By calculating the average value of each group and calculating the relative value with the non-addition group (each test group / no-addition group), depending on the compound having each guaiacol skeleton and / or the compound having a 4-methoxyphenol skeleton The multiplication activity was taken as the multiplication factor.

  The following results were obtained in the above-mentioned medium for all strains of Lactobacillus plantarum tested in this specification including the JCM1149T strain.

An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4 times in the presence of 500 μM vanillin compared to the absence of vanillin,
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
An increase of at least about 5.1 fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to the absence of DL-4-hydroxy-3-methoxymandelic acid (VMA). ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride;
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride.
The tested Lactobacillus plantarum also has the following activities:
An increase of at least about 7.9 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 3.4-fold in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 7 in the presence of 500 μM eugenol compared to the absence of eugenol. It rises 8 times.

  The following results were obtained for the deposited strain.

An increase of at least about 5.5 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4 times in the presence of 500 μM vanillin compared to the absence of vanillin,
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
An increase of at least about 5.1 fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to the absence of DL-4-hydroxy-3-methoxymandelic acid (VMA). ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride;
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8 times in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride;
An increase of at least about 7.9 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 3.4-fold in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 7 in the presence of 500 μM eugenol compared to the absence of eugenol. It rises 8 times (however, only TO0002 strain is measured below Guaiacol).

  And about all the strains tested in this specification including Example 6, the following results were obtained in the above-mentioned culture medium. Hereinafter, the results including those shown in FIG. 11 are shown.

An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 2.7 times in the presence of 500 μM vanillin compared to the absence of vanillin,
An increase of at least about 3.8 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
An increase of at least about 5.8 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 2.7-fold in the presence of 500 μM silybin compared to the absence of silybin, and an increase of at least about 6.9-fold in the presence of 500 μM eugenol compared to the absence of eugenol .

Next, L. casei PR143 strain and L. A summary of the results obtained with the casei PR150 strain is shown below.
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.2 times in the presence of 500 mM guaiacol compared to the absence of guaiacol,
An increase of at least about 2.7-fold in the presence of 500 μM silybin compared to the absence of silybin, and an increase of at least about 4.3-fold in the presence of 500 μM eugenol compared to the absence of eugenol .

(Example 8: Example of prebiotic tablet)
For the compound identified by the present invention, a tablet having the following composition is produced by a conventional method. In the case of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, the component of the present invention is prepared in the same manner as a general low molecular weight compound. When the strain is used in combination, it is dried to make the strain powder. These tablets can be formulated together with, for example, fermented foods using the strain of the present invention.
Ingredient 100 mg of the present invention
Lactose 60mg
Potato starch 30mg
Polyvinyl alcohol 2mg
Magnesium stearate 1mg
Tar pigment Trace amount.

(Example 9: Example of prebiotic powder)
About the compound identified by this invention, the powder which consists of the following composition is manufactured by a conventional method. In the case of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, the component of the present invention is prepared in the same manner as a general low molecular weight compound. When the strain is used in combination, it is dried to make the strain powder. These powders can be formulated together with, for example, fermented foods using the strain of the present invention.
Ingredient 150 mg of the present invention
Lactose 280 mg.

(Example 10: Example of prebiotic syrup)
About the compound and strain identified by this invention, the syrup agent which consists of the following composition is manufactured by a conventional method. In the case of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, the component of the present invention is prepared in the same manner as a general low molecular weight compound. When the strain is used in combination, it is dried to make the strain powder. These syrups can be used in combination with, for example, fermented foods using the strain of the present invention.
Ingredient 100 mg of the present invention
Purified white sugar 40 g
40 mg ethyl p-hydroxybenzoate
Propyl p-hydroxybenzoate 10mg
Chocolate flavor 0.1cc
Water is added to make a total amount of 100 cc.

(Example 11: Example of probiotics)
About the probiotic lactic acid bacteria identified by this invention, fermented milk which consists of the following composition is manufactured by a conventional method. The strain of the present invention is co-fermented by adding it simultaneously with the yogurt fermentation starter. The lactic acid bacteria starter is not particularly limited, and those usually used for fermentation of yogurt can be used.

Of the raw materials described in the following recipe, all except the strain of the present invention and the starter are mixed together, heated, filtered, and homogenized. Next, after sterilization, cooling, addition of the strain of the present invention and a starter according to a conventional method, the container is filled, fermented in a stationary state, and then cooled to obtain plain yogurt or hard yogurt. Moreover, after adding the strain of the present invention and a starter, soft yogurt and drink yogurt can be produced by carrying out a conventional tank fermentation system.
10 kg of the strain of the present invention
Lactic acid bacteria starter 20kg
Reduced milk 800kg
Sugar 10kg
Stabilizer 1kg.

(Example 12: Example of probiotics)
About the probiotic lactic acid bacteria identified by this invention, the livestock storage fermented feed which consists of the following composition is manufactured by a conventional method. The strain of the present invention is added to, for example, feed crops, pastures, and food by-products, and preservation processing such as a conventional roll veil method is performed. Forage crops, pasture, and food by-products are not particularly limited, and those usually used for livestock feed can be used. In the case of the strain of the present invention, it is dried to obtain a strain powder, and when used, 50 g is dissolved in 10 L of water and sprayed on forage crops, pasture and food by-products.
50 g of the strain of the present invention
10kg of mixed water
Forage crops, pasture, food by-products 10 t.

(Example 13: Example of synbiotics)
With respect to the probiotic lactic acid bacteria identified by the present invention, fermented milk having the following composition is produced by a conventional method. The strain of the present invention is co-fermented by adding it simultaneously with the yogurt fermentation starter. The lactic acid bacteria starter is not particularly limited, and those usually used for fermentation of yogurt can be used. In the case of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, it is prepared in the same manner as a general low molecular weight compound.

Of the raw materials described in the following recipe, all except the strain of the present invention and the starter are mixed together, heated, filtered, and homogenized. Then, sterilization, cooling, adding the strain and starter of the present invention and the compound of the present invention according to a conventional method, filling into a container, fermenting in a stationary state, and then cooling to obtain plain yogurt or hard yogurt . Moreover, after adding the strain and starter of the present invention, soft yogurt and drink yogurt can be produced by carrying out a conventional tank fermentation system.
10 kg of the strain of the present invention
10 kg of the compound of the present invention
Lactic acid bacteria starter 20kg
Reduced milk 800kg
Sugar 10kg
Stabilizer 1kg.

(Example 14: Example of synbiotics)
About the probiotic lactic acid bacteria identified by this invention, the livestock storage fermented feed which consists of the following composition is manufactured by a conventional method. The strain of the present invention is added to, for example, feed crops, pastures, and food by-products, and preservation processing such as a conventional roll veil method is performed. Forage crops, pasture, and food by-products are not particularly limited, and those usually used for livestock feed can be used. In the case of the strain of the present invention, it is dried to obtain a strain powder. When used, 50 g of the compound of the present invention is dissolved in 10 L of water simultaneously with 50 g of the compound of the present invention and sprayed on feed crops, pastures and food by-products. In the case of a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, it is prepared in the same manner as a general low molecular weight compound.
50 g of the strain of the present invention
50 g of the compound of the present invention
10kg of mixed water
Forage crops, pasture, food by-products 10 t.

  As mentioned above, although this invention has been illustrated using preferable embodiment of this invention, this invention should not be limited and limited to this embodiment. It is understood that the scope of the present invention should be construed only by the claims. It is understood that those skilled in the art can implement an equivalent range based on the description of the present invention and the common general technical knowledge from the description of specific preferred embodiments of the present invention. Patents, patent applications, and documents cited herein should be incorporated by reference in their entirety, as if the contents themselves were specifically described herein. Understood.

NITE P-958
NITE P-959
NITE P-960
NITE P-961
NITE AP-1308
NITE AP-1309
NITE AP-1310
NITE AP-1311
NITE AP-1312

In one embodiment, in the present invention, the strain may be Lactobacillus plantarum TO1000, Lactobacillus plantarum TO1001, Lactobacillus plantarum TO1002 or Lactobacillus plantarum TE100I (Accession No. . 961 is Alternatively, the strain, Lactobacillus paraplantarum LOOC 2020 (consignment number: NITE P -1308), Lactobacillus sakei SG171 ( consignment number: NITE P -1309), Lactobacillus casei LOOC82 ( commissioned number: May be NITE P -1312): ITE P -1310 ), Lactobacillus casei PR143 ( consignment number: NITE P -1311) or Lactobacillus casei PR150 (entrusted number.

In one embodiment, in the present invention, the strain is Lactobacillus plantarum TO1000 (Accession No. NITE P-958), Lactobacillus plantarum TO1001 (Accession No. NITE P-959), Lactobacillus planta No. Lactobacillus plantarum TO1003 (accession number NITE P-961), Lactobacillus plantarum JCM1149, Lactobacillus paraplantarum LOOC 2020 ( consignment number: NITE P -1308), Lactobacillus sakei SG171 ( consignment number: NITE P -1309), La tobacillus casei LOOC82 (consignment number: NITE P -1310), Lactobacillus casei PR143 ( consignment number: NITE P -1311) or Lactobacillus casei PR150 (consignment number: NITE P -1312) is.

For example, L. plantarum TO1000, TO1001, TO1002 or TO1003 (accession number NITE P-958, NITE P-959, NITE P-960 or NITE P-961 (reception number NITE AP-958, NITE AP-959, NITE AP-960 or NITE respectively) corresponding to the AP-961)), corresponding to Lactobacillus paraplantarum LOOC 2020 (accession number NITE P-1308 (receipt number NITE AP-1308))), Lactobacillus sakei SG171 ( accession number NITE P-1309 (receipt number NITE AP-1309) the corresponding)), Lactobacillus casei LOOC82 (accession No. NITE P-1310 (receipt number NITE Corresponding to AP-1310 )) ), Lactobacillus casei PR143 ( corresponding to accession number NITE P-1311 (reception number NITE AP-1311)) ) or Lactobacillus casei PR150 ( accession number NITE P-1312 ( reception number NITE AP-1312 ) )) ). These strains are about 2.3 to 6.3 times or more than the control group by adding a compound having a guaiacol skeleton such as ferulic acid and / or a 4-methoxyphenol skeleton. It was found in the present invention that the proliferation activity was significantly increased. As another strain of Lactobacillus genus lactic acid bacteria, the JCM1149T strain can be mentioned. plantarum reference strain, which can be obtained from the same site. It should be noted that the L.A. The plantarum reference strain was also found to be reactive with ferulic acid according to the present invention (addition of 500 μM of the test compound was about 3.0 times that of the control group). Proliferative activity is increased.)

In one embodiment, the strain of the present invention is a strain belonging to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei and Lactobacillus casei, for example, in the exemplary embodiment L. plantarum TO1000, L. plantarum TO1001, L. plantarum TO1002 or L. plantarum TO1003 (Accession Number NITE P-958, NITE P-959, NITE P-960 or NITE P-961). Alternatively, Lactobacillus plantarum JCM1149, Lactobacillus paraplantarum LOOC 2020 ( consignment number NITE P -1308), Lactobacillus sakei SG171 ( consignment number NITE P -1309), Lactobacillus casei LOOC82 ( consignment number NITE P -1310), Lactobacillus casei PR143 ( consignment number NITE P -1311) or Lactobacillus casei PR150 (consignment number NITE P -1,312) may also be used.

In one embodiment, the strain of interest of the composition of the present invention is L. plantarum TO1000, TO1001, TO1002 or TO1003 (accession number NITE P-958, NITE P- 959, NITE P-960 or NITE P-961), JCM1149, Lactobacillus paraplantarum LOOC 2020 ( consignment number NITE P -1308), Lactobacillus sakei SG171 (consignment number NITE P -1309), Lactobacillus casei LOOC82 ( consignment number NITE P -1310), Lactobacillus casei PR143 ( consignment number NITE P -1311) or Lactobacillus casei PR150 (consignment number NITE P -1312) or These are multiple strains.

As a typical example, Lactobacillus paraplantarum LOOC 2020 which were selected according to the method of Example 1 (consignment number NITE P -1308), Lactobacillus sakei SG171 ( consignment number NITE P -1309), Lactobacillus casei LOOC82 ( consignment number NITE P - 1310) was used.

In addition to the above strains, additional strains L. cerevisiae selected according to the method of Example 1. casei PR143 shares (consignment number NITE P -1311) and L. casei PR150 strain using (entrusted number NITE P -1 312) was the experiment showed similar proliferation activity against ferulic acid, silybin, guaiacol and eugenol.

The resulting strain in the present embodiment, Lactobacillus paraplantarum LOOC 2020 for consignment numbers: as NITE P -1308, Lactobacillus sakei SG171 for consignment numbers: as NITE P -1309, Lactobacillus casei LOOC82 for consignment ID: NITE P - as 1310, Lactobacillus casei PR143 for the consignment number: as NITE P -1311, and Lactobacillus casei PR150 for the consignment number: NITE P -1312 as the National Institute of technology and evaluation Patent microorganisms Depositary Center (Kisarazu, Chiba Kazusa sickle Deposited at foot 2-5-8).

NITE P-958
NITE P-959
NITE P-960
NITE P-961
NITE P -1308
NITE P- 1309
NITE P- 1310
NITE P- 1311
NITE P- 1312

Claims (25)

Lactobacillus lactic acid strains whose growth activity is increased by a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton. The proliferation activity is a medium containing 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum. (PH 6.5)
An increase of at least about 2.3 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4-fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
Elevated at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to absence of DL-4-hydroxy-3-methoxymandelic acid (VMA) ,
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride,
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride,
An increase of at least about 4.2 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 2.7 times in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 4 in the presence of 500 μM eugenol compared to the absence of eugenol. 2. The strain of claim 1 having at least one characteristic selected from the group consisting of a threefold increase. The proliferation activity is a medium containing 1.45 mM MgSO ₄ , 8.92 mM NH ₄ NO ₃ , 4.79 mM KCl, 3.96 mM C ₆ H ₁₂ O ₆ , 2.62 mM KH ₂ PO ₄ and 30% bovine serum. (PH 6.5)
An increase of at least about 5.5 times in the presence of 500 μM ferulic acid compared to the absence of ferulic acid,
An increase of at least about 4.4-fold in the presence of 500 μM vanillin compared to the absence of vanillin;
An increase of at least about 8.2 times in the presence of 500 μM hydroxyferulic acid compared to the absence of hydroxyferulic acid,
An increase of at least about 5.0 times in the presence of 500 μM capsaicin compared to the absence of capsaicin;
In the presence of 500 μM 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG) as compared to in the absence of 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), at least about 2.8 times increase,
An increase of at least about 5.4-fold in the presence of 500 μM DL-normetanephrine hydrochloride compared to the absence of DL-normetanephrine hydrochloride,
An increase of at least about 5.1-fold in the presence of 500 μM DL-4-hydroxy-3-methoxymandelic acid (VMA) compared to the absence of DL-4-hydroxy-3methoxymandelic acid (VMA);
An increase of at least about 5.8 fold in the presence of 500 μM 3-methoxytyramine (3MT) hydrochloride compared to the absence of 3-methoxytyramine (3MT) hydrochloride,
An increase of at least about 5.5-fold in the presence of 500 μM homovanillic acid (HVA) compared to the absence of homovanillic acid (HVA),
An increase of at least about 6.3 times in the presence of 500 μM vanillic acid (VA) compared to the absence of vanillic acid (VA);
An increase of at least about 3.9 times in the presence of 500 μM homovanillyl alcohol (MOPET) compared to the absence of homovanillyl alcohol (MOPET);
An increase of at least about 4.8-fold in the presence of 500 μM Metanephrine hydrochloride compared to the absence of Metanephrine hydrochloride,
An increase of at least about 7.9 times in the presence of 500 μM guaiacol compared to the absence of guaiacol,
An increase of at least about 8.5 times in the presence of 500 μM 4-methoxyphenol compared to the absence of 4-methoxyphenol,
An increase of at least about 6.1 times in the presence of 1.5 mM Gingerol compared to the absence of Gingerol;
An increase of at least about 2.3 fold in the presence of 0.25 mM shogaol compared to the absence of shogaol,
An increase of at least about 3.7-fold in the presence of 500 μM vanillic acid diethylamide compared to the absence of vanillic acid diethylamide;
An increase of at least about 4.6 fold in the presence of 500 μM isoeugenol compared to the absence of isoeugenol,
An increase of at least about 5.7-fold in the presence of 500 μM isovanillic acid compared to the absence of isovanillic acid,
an increase of at least about 6.1-fold in the presence of 500 μM o-vanillin as compared to the absence of o-vanillin;
An increase of at least about 4.3 times in the presence of 500 μM of transferuric acid compared to the absence of transferuric acid,
An increase of at least about 4.4 times in the presence of 500 μM isoferulic acid compared to the absence of isoferulic acid;
An increase of at least about 12.6 times in the presence of 500 μM of isocresol compared to the absence of isoceosol;
Of 500 μM 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one compared to the absence of 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one In the presence, rise at least about 9.8 times,
An increase of at least about 6.6-fold in the presence of 500 μM hesperetin compared to the absence of hesperetin,
Elevated at least about 5.9 times in the presence of 500 μM 3- (4-hydroxy-3-methoxyphenyl) propionic acid compared to the absence of 3- (4-hydroxy-3-methoxyphenyl) propionic acid. ,
An increase of at least about 4.4 times in the presence of 500 μM isovanillyl alcohol as compared to the absence of isovanillyl alcohol;
An increase of at least about 9.7-fold in the presence of 500 μM cleosol compared to the absence of cleosol;
an increase of at least about 7.2-fold in the presence of 500 μM o-vanillic acid compared to the absence of o-vanillic acid;
An increase of at least about 7.4 times in the presence of 500 μM methyl vanillate compared to the absence of methyl vanillate;
In the presence of 500 μM methyl-3- (4-hydroxy-3-methoxyphenyl) propionate as compared to the absence of methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, at least about 6.6 times higher,
An increase of at least about 3.6-fold in the presence of 500 μM 5-nitroguaiacol compared to the absence of 5-nitroguaiacol;
An increase of at least about 8.8 times in the presence of 500 μM scopoletin compared to the absence of scopoletin,
An increase of at least about 3.4-fold in the presence of 500 μM silybin compared to the absence of silybin,
An increase of at least about 6.5 times in the presence of 500 μM vanillyl alcohol compared to the absence of vanillyl alcohol;
An increase of at least about 1.5 times in the presence of 500 μM ethyl vanillate compared to the absence of ethyl vanillate;
An increase of at least about 1.8 fold in the presence of 500 μM coniferyl alcohol compared to the absence of coniferyl alcohol, and at least about 7 in the presence of 500 μM eugenol compared to the absence of eugenol. 2. The strain of claim 1 having at least one characteristic selected from the group consisting of an 8-fold increase. The compound is ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride, DL-4-hydroxy-3 -Methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine hydrochloride, guaiacol (2- Methoxyphenol), 4-methoxyphenol, gingerol, gingerol, vanillic acid diethylamide, isoeugenol, eugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid , Isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, cresol O-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol and the like The strain according to claim 1, comprising at least one selected from the group consisting of a salt and a solvate. The strain according to claim 1, wherein the compound is selected from the group consisting of ferulic acid, hydroxyferulic acid, vanillic acid and capsaicin. The strain according to claim 1, wherein the compound comprises ferulic acid or a derivative thereof. The strain according to claim 1, wherein the strain belongs to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei, and Lactobacillus casei. The strains are Lactobacillus plantarum TO1000 (Accession No. NITE P-958), Lactobacillus plantarum TO1001 (Accession No. NITE P-959), Lactobacillus plantarum TO1002 (Accession No. ), Lactobacillus paraplanum LOOC 2020 (reception number: NITE AP-1308), Lactobacillus sakei SG171 (reception number: NITE AP-1309), Lactobacillus casei LOOCTE 82 (reception number-1 NITE AP-1308 , Lactobacillus casei PR143 (receipt number: NITE AP-1311) or Lactobacillus casei PR150 (receipt number: NITE AP-1312) strain of claim 1 in which. A composition for growth of Lactobacillus lactic acid strains, comprising a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton. The compound is ferulic acid, hydroxyferulic acid, vanillic acid, capsaicin, 4-hydroxy-3-methoxyphenylglycol hemipiperazinium salt (MHPG), DL-normetanephrine hydrochloride, DL-4-hydroxy-3 -Methoxymandelic acid (VMA), 3-methoxytyramine (3MT) hydrochloride, homovanillic acid (HVA), vanillic acid (VA), homovanillyl alcohol (MOPET), and metanephrine hydrochloride, guaiacol (2- Methoxyphenol), 4-methoxyphenol, gingerol, gingerol, vanillic acid diethylamide, isoeugenol, eugenol, isovanillic acid, o-vanillin, transferuric acid, isoferulic acid , Isocresol, 4- (4-hydroxy-3-methoxyphenyl) -3-buten-2-one, hesperetin, 3- (4-hydroxy-3-methoxyphenyl) propionic acid, isovanillyl alcohol, cresol O-vanillic acid, methyl vanillate, methyl-3- (4-hydroxy-3-methoxyphenyl) propionate, 5-nitroguaiacol, scopoletin, silybin, vanillyl alcohol, ethyl vanillate, coniferyl alcohol and the like The composition according to claim 9, comprising at least one selected from the group consisting of a salt and a solvate. 10. The composition of claim 9, wherein the compound is selected from the group consisting of ferulic acid, hydroxyferulic acid, vanillic acid, and capsaicin. The composition according to claim 9, wherein the compound is ferulic acid or a derivative thereof. The composition according to claim 9, wherein the strain belongs to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei, and Lactobacillus casei. The strains are Lactobacillus plantarum TO1000 (Accession No. NITE P-958), Lactobacillus plantarum TO1001 (Accession No. NITE P-959), Lactobacillus plantarum TO1002 (Accession No. ), Lactobacillus plantarum JCM1149, Lactobacillus paraplanum LOOC 2020 (reception number: NITE AP-1308), Lactobacillus sakei SG171 (reception number: NITE AP-1309il), Lactobac casei LOOC82 (receipt number: NITE AP-1310), Lactobacillus casei PR143 (receipt number: NITE AP-1311) or Lactobacillus casei PR150 (receipt number: NITE AP-1312) is a composition of claim 9. A probiotic composition comprising the strain according to any one of claims 1 to 8. A synbiotic composition comprising a combination of the strain according to any one of claims 1 to 8, a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton. 17. The composition of claim 16, wherein said compound is present at least 10 [mu] M per 1 x 10 < ⁶ > colony forming units (cfu) of said strain when present in the body. A prebiotic composition comprising a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton for use in combination with the strain according to any one of claims 1 to 8. A method for selecting lactic acid bacteria whose proliferation activity is increased by a compound having a guaiacol skeleton and / or a compound having a 4-methoxyphenol skeleton, the method comprising:
A) a providing step of providing a sample containing lactic acid bacteria;
B) magnesium sulfate (MgSO _4), ammonium nitrate _(NH 4 _NO 3), potassium chloride (KCl), D - (+ ) - glucose, potassium dihydrogen phosphate _(KH 2 PO ₄₎ and in a medium containing calf serum Culturing the sample in the presence or absence of a compound having a guaiacol skeleton; and C) not growing in the absence of the compound and growing in the presence or the presence of the compound A method comprising a separation step of separating a strain whose growth is promoted more than in the absence thereof. The magnesium sulfate is 1.16 to 1.74 mM, the ammonium nitrate is 8.02 to 9.81 mM, the potassium chloride is 3.83 to 5.74 mM, and the D-(+)-glucose is 3.96. 39.64 mM, the potassium dihydrogen phosphate is 2.09-3.14 mM, and the bovine serum is 15-30% in the medium, and the medium has a pH of 6.0-7.0. 20. The method of claim 19, wherein The method according to claim 19, wherein the culturing step is performed for 32 to 48 hours. The method according to claim 19, wherein the culturing step is performed at 25 to 40 ° C. The method according to claim 19, wherein the culturing step is performed under conditions of 0 to 5% CO ₂ and 1 to 20% O ₂ . 20. The method according to claim 19, wherein the lactic acid bacteria belong to a species selected from the group consisting of Lactobacillus plantarum, Lactobacillus paraplanum, Lactobacillus sakei, and Lactobacillus casei. 25. The method of claim 24, wherein the sample comprises the lactic acid bacteria between 5 x 10 < ⁵ > to 5 x 10 < ⁶ > colony forming units / well.