WO2006067940A1

WO2006067940A1 - Lactic acid bacteria binding to human abo blood types

Info

Publication number: WO2006067940A1
Application number: PCT/JP2005/022096
Authority: WO
Inventors: Tadao Saito; Yasushi Kawai; Hideaki Uchida; Katsunori Kimura; Kakuhei Isawa; Keisuke Furuichi
Original assignee: Meiji Dairies Corporation
Priority date: 2004-12-01
Filing date: 2005-12-01
Publication date: 2006-06-29
Also published as: CA2588940A1; CA2588940C

Abstract

Lactic acid bacteria are screened by the surface plasmon resonance spectroscopy with the use of human intestinal mucin and blood type antigens as probes. To adapt to the mass-scaled screening, it is attempted to set selection standards for the screening of lactic acid bacteria as described above. As a result, it is found out that lactic acid bacteria respectively matching ABO blood types can be screened by using 100RU under definite conditions as the standard of binding of bacteria. 238 strains of lactic acid bacteria are subjected to this screening. Further, a test for examining the suitability for yogurt production is performed. Finally, strains matching individual blood types of A, B and O are clarified in practice.

Description

Human ABO blood group-binding lactic acid bacteria

[0001] The present invention relates to lactic acid bacteria and screening of lactic acid bacteria.

Background art

[0002] The human intestinal tract is inhabited by approximately 200 types of intestinal bacteria (intestinal resident microorganisms). Microorganisms called probiotics improve the balance between useful and harmful bacteria in the human intestine and contribute to the health of the host. Recently, there has been a trend toward applying this probiotic microorganism to food. For example, several functional yogurts produced using lactobacilli having a probiotic function have been commercialized. Therefore, there is a need for mass screening technology that selects better probiotics.

[0003] Intestinal colonized lactic acid bacteria adhere to and proliferate in the human intestinal tract. For this reason, intestinal binding of lactic acid bacteria is very important for the probiotic function to be demonstrated. The binding mechanism of lactic acid bacteria in the human intestine has not yet been elucidated. From previous studies on intestinal lactic acid bacteria, it has been confirmed that L. casei has binding properties to glycolipid sugar chains, and L. reuteri and L. cri spatus have binding properties to collagen. Furthermore, a lectin-like protein that binds to the intestinal lactobacilli has been identified. However, in the intestinal epithelium of many healthy subjects, there are very few exposed sites of cytoskeletal protein (collagen), and colonization of lactic acid bacteria by lectin-like protein in the intestinal epidermis is unlikely. Therefore, it is thought that sugar chains that bind to intestinal mucin play an important role in the intestinal binding of intestinal settled lactic acid bacteria. A lectin-like protein, a protein that recognizes carbohydrates, exists in the surface layer protein (SLP) of many intestinal settled lactic acid bacteria. Intestinal mucin exists on the surface of the intestinal tract.

Intestinal mucin is a mucous high molecular weight glycoprotein having an infinite number of mucin-type sugar chains linked to a polypeptide (core protein, apomucin) through a 0-glycoside bond. In other words, intestinal settled lactic acid bacteria acquire intestinal connectivity by binding to the sugar chains of intestinal mucin through lectin-like proteins on the surface of the bacteria, and are considered to achieve stable growth. available.

[0004] On the other hand, recently, an interesting fact has been reported that the chemical structure of the sugar chain constituting human colonic mucin (HCM) varies depending on the AB0 blood type (Non-Patent Document 1-4). ).

[0005] Human ABO blood groups are distinguished by the type of antigenic substance expressed on the surface of erythrocytes. The antigenic site of this ABO blood group substance is a sugar chain having a specific chemical structure (ABO blood group antigen). Both type A and type B antigens are molecules composed of three sugars, and a -N-acetylethyl latatosamine is bound to a basic structure called a type H antigen composed of two sugars in a specific binding mode. Is type A antigen and α-galactose is bound to type B antigen. Humans of type IV blood express the type A antigen, humans of type B blood express the type B antigen, and humans of type AB blood express both the type A antigen and the type B antigen on the erythrocyte surface. In contrast, type 0 blood humans express H-type antigens of the basic structure.

[0006] The above-mentioned scientific fact that the sugar chain structure of gastrointestinal mucin varies depending on the blood type suggests that the type of probiotic lactic acid bacteria that bind and proliferate in the gastrointestinal tract varies depending on the blood type. If a lactic acid bacterium suitable for each blood type is found, it will be possible to develop functional yogurt corresponding to the individual level. So far, the present inventors have focused on this point and developed a method for screening human intestinal-binding lactic acid bacteria by the adsorptive power with ABO blood group antigen (Patent Document 1). This method is an epoch-making method in which the adsorptive power between lactic acid bacteria and ABO blood group antigens is detected using surface plasmon resonance spectrum (SPR), and lactic acid bacteria that are compatible with the blood type are selected. Specifically, when ABO blood group antigen or intestinal mucin is used as a ligand and lactic acid bacteria are brought into contact with the ligand immobilized on the sensor chip, the binding between the lactic acid bacteria and the ligand is accompanied by the binding. This is a method to detect the mass change that occurs on the sensor chip as a surface plasmon resonance (SPR) signal. The mass change is expressed in Resonance Unit (= RU resonance unit), where lRU = lpg / mm ² , and 1RU indicates that the substance lpg is bound to lmm ² . The present inventors have carried out the above method and confirmed that the Lactobacillus crispatus JCM8778 strain and the Lact obacillus acidophilus OLL2769 strain are A-type antigen recognizable (Patent Document Non-Patent Document 5). However, probiotic lactic acid bacteria including yogurt Because of the expected increase in demand for foods that use blood, we have been waiting for the acquisition of blood group-binding lactic acid bacteria with even better binding properties!

[0007] Patent Document 1: JP 2004-101249 A

Non-Patent Document 1: Junko Amano, Biochemistry, Japan Biochemical Society, 1999, No. 71, p.274 -277

Special Reference 2: Holgersson, J., Stromberg, N., and Breimer, ME, ulyconpids of hu man large intenstine: glycolopid expression related to anatomical localization, epithe lial I non-epithelial tissue and the ABO, Le and Se phenotypes of the donors. Bioch imie, 70, 1565—1574 (1988).

Patent Document 3: Holgersson, J., Jovall, PA, and Breimer, ME, Glycosphingolipids of human large intenstine: detailed structural characterization with special reference to blood group compounds and bacterial receptor structures.J. Biochem, (Tokyo), 110, 120 -131 (1991).

Non-Patent Document 4: Vanak, J., Ehrmann, J., Drimalova, D., Nemec, M., monoclonal antibo dies in the detection of blood group antigens A and B in the mucosa of the large inte stine.Cas Lek Cesk , 18, 364-367 (1988).

Non-Patent Document 5: Uchida, H. et al, Biosci. Biotechnol. Biochem., 68 (5), 1004-1010 (20 04).

Patent Document 6: Holmes, S. D. et al, Studies on the interaction of Staphylococcus aure us and Staphylococcus epidermidis with fibronectin using surface plasmon resonance (BIACORE). J. Microbiological Methods, 28, 77-84 (1997).

Non-Patent Document 7: Fratamico, P. M. et al, Detection of Escherichia coli 0157: H7 using a surface plasmon resonance biosensor. Biotechnol. Techniques., 7, 571-576 (1998). Disclosure of the Invention

Problems to be solved by the invention

[0008] The present invention has been made in view of such circumstances, and the problem to be solved by the present invention is to find a novel intestinal-binding lactic acid bacterium suitable for human ABO blood group. Means for solving the problem [0009] The inventors of the present invention who have solved the above-mentioned problems have implemented the lactic acid bacteria screening method based on the surface plasmon resonance spectrum described above. Although the above method has already been established as a screening method for lactic acid bacteria, an attempt has been made by the present inventors to set a “selection reference value” in the screening method for lactic acid bacteria in order to make it suitable for large-scale screening. As an example of the investigation of bacteria by surface plasmon resonance spectrum, Escherichia coli 0157, an example in which the binding of Staphylococcus aureus to fibronectin was measured by surface plasmon resonance spectrum, Non-patent document 6 Examples of detection using protein A or protein G immobilized on a chip via an antibody (Non-Patent Document 7) are known. In these papers, the RU value indicating the binding of viable cells is about 100 It is 1000R U. However, examples of lactic acid bacteria studied by surface plasmon resonance spectrum are not known except for reports by the present inventors (Patent Document 1, Non-Patent Document 5). In addition, even if the specimen is the same, the RU value can change depending on various measurement conditions. Therefore, as a result of diligent efforts, the present inventors established a screening method for lactic acid bacteria compatible with the ABO blood group by using 100 RU as a criterion for bacterial binding under certain conditions.

[0010] Furthermore, the present inventors conducted the screening method described above for 238 strains of lactic acid bacteria of human intestinal origin isolated from the human intestinal tract, and further conducted a test for judging suitability for yogurt production. Finally, We specifically found strains compatible with blood group A and type 0. That is, the present invention relates to a lactic acid bacterium suitable for blood group compatible yogurt and a method for screening the lactic acid bacterium, and specifically provides the following.

(1) Lactobacillus gasseri lactic acid bacteria capable of binding to the human ABO blood group antigen represented by any one of the following formulas (a) to (c)

(a) [GalNAc a 1-3 (Fuc a 1-2) Gal-]

(b) [Gal a 1-3 (Fuc a 1-2) Gal-]

(c) [Fuc a l-2Ga 卜],

(2) Accession number: NITE BP-25, Accession number: NITE BP-26, Accession number: NITE BP-27, Accession number: NITE BP-28, Accession number: NITE BP-145, Accession number: NITE BP-146 Lactobacillus gasseri lactic acid bacteria described in (1) above,

(3) Human ABO formula containing the Lactobacillus gasseri lactic acid bacterium according to (1) or (2) above Blood type compatible fermented milk and starter for dairy production,

(4) A food or drink containing the Lactobacillus gasseri lactic acid bacterium according to (1) or (2) above

P

TO,

(5) Fermented milk and lactic acid bacteria beverages containing the Lactobacillus gasseri lactic acid bacteria described in (1) or (2) above,

(6) A method for screening lactic acid bacteria using a surface plasmon resonance spectrum, using as an index the following condition (i) Kaza et al. (Iii):

(i) RU value that indicates the ability to bind human ABO blood group antigen of type A is 100RU or more

(ii) The RU value indicating the ability to bind to human ABO blood group antigens of type B and Z or H is 100RU or less

(iii) The RU value indicating the ability to bind to human type A intestinal mucin is 100 RU or more,

(7) A method for screening lactic acid bacteria using a surface plasmon resonance spectrum, using as an index the following condition (i) Kazu et al. (Iii):

(i) The RU value that indicates the ability to bind B-type human ABO blood group antigen is 100RU or more

(ii) The RU value indicating the ability to bind to human ABO blood group antigens of type A and Z or H is 100RU or less

(iii) The RU value that indicates the ability to bind human B-type intestinal mucin is 100 RU or more

(8) A method for screening lactic acid bacteria using a surface plasmon resonance spectrum, using as an index the following condition (i) Kazu et al. (Iii):

(i) The RU value that indicates the ability to bind H-type human ABO blood group antigen is 100RU or more

(ii) The RU value that indicates the ability to bind human ABO blood group antigens of type A and Z or B is 100RU or less

(iii) The RU value indicating the ability to bind human type 0 intestinal mucin must be 100 RU or more.

(9) A screening method for lactic acid bacteria using a surface plasmon resonance spectrum, using as an index the following condition (i) Kazu et al. (Iii):

(ii) The RU value, which indicates the ability to bind human ABO blood group antigens of type A and H, is 100RU or less (iii) The RU value indicating the ability to bind to human type B intestinal mucin must be greater than the RU value indicating the ability to bind to human type A intestinal mucin and the RU value indicating the ability to bind to human type 0 intestinal mucin.

Brief Description of Drawings

[0011] Fig. 1 is a diagram showing the chemical structure of a Pioti-Louis polymer (BP) probe having a sugar chain antigen site of human ABO blood group.

[Fig. 2] Diagram showing the sugar chain part of the A-type antigen BP-probe (top), the B-type antigen BP-probe sugar chain part (middle), and the type 0 antigen BP-probe sugar chain part (bottom) is there.

BEST MODE FOR CARRYING OUT THE INVENTION

[0012] The present invention comprises (a) [GalNAc a 1-3 (Fuc a 1-2) Gal-], (b) [Gal a 1-3 (Fuc a 1-2) Gal-], (c) An intestinal-binding Lactobacillus gasseri lactic acid bacterium having binding ability to a human ABO blood group antigen represented by any formula of [Fucal-2Ga a] is provided.

[0013] In general, lactic acid bacteria are a general term for a group of bacteria that produce glucose strength lactic acid in a molar ratio of 50% or more. O Lactobacillus genus, Lactococcus rot, Streptococcus moth, Leuconostoc moth are representative genus of lactic acid bacteria. is there. The genus Bifidobacterium is also included in the lactic acid bacteria in this specification. The genus Lactobacillus is further classified into species. As a typical example of the species of the genus Lactobacillus, Lactobacillus delbruekn suosp. Bulgancus (and bulgaricus), Lactobacillus de lbruekn subsp. Delbruekii (L. Delbruekii), Lactobacillus acidophilus Group ~ ~ flop lactic acid bacteria (L. acidophilus Gunore ¹ ~ ~-flops), Lactobacillus casei (L. casei), Lactobacillus plantarum (L. plantarum.), Lactobacillus brevis (L. brevis), Lactobacillus buchneri (L. buchneri), Lactobacillus fermentum (L. fermentum), Lactobacillus helveticus (L. helveticus) Can be mentioned. L. acidophilus group lactic acid bacteria are classified into Lactobacillus acidophilus (A-1), Lactobacillus cnspatus (A-2), Lactobacillus amylovorus (Λ-3), Lactobacillus gallinarum (A-4) according to the results of DNA-DNA homology and cell wall composition analysis. ), L actobacillus gasseri (B-1), and Lactobacillus johnsonii (B-2).

[0014] The Lactobacillus gasseri (hereinafter abbreviated as "L. gasserij") lactic acid bacteria of the present invention is an intestinal tract Lactobacillus gasseri characterized by having a binding ability to a human ABO blood group antigen. ABO human blood group antigen is a sugar chain that determines blood group Specifically, type A sugar chain (type A antigen): [GalNAc a 1-3 (Fuc a 1-2) Gal-], type B sugar chain (type B antigen): [Gal a 1-3 ( Fuc a 1-2) Gal-], O-type sugar chain (hereinafter referred to as a sugar chain that determines blood group 0, also referred to as H-type antigen, 0-type antigen): [Fuc a to 2Ga]. As described above, intestinal mucins present on the surface of the intestinal tract have different sugar chains depending on the ABO blood group. The present inventors confirmed that there is L. gasseri lactic acid bacteria that bind to type A sugar chains, which are antigens that determine human blood group A, among lactic acid bacteria that bind to mucin prepared for human intestinal force of type A did. It was also confirmed that type A sugar chains were expressed in the mucin. Therefore, in the L. gasseri lactic acid bacterium of the present invention, the lactic acid bacterium that binds to the A-type sugar chain and does not adsorb to other types of sugar chains binds to the A-type sugar chain on the intestinal mucin, whereby the ABO blood group It has acquired intestinal connectivity for type A humans. The L. gasseri lactic acid bacterium is considered to stably bind to and proliferate in the intestinal tract of blood group A humans and exert a probiotic function. That is, it can be said that the L. gasseri lactic acid bacteria can contribute to the health of blood group A humans and is suitable for blood group A humans. Similarly, the L. gasseri lactic acid bacterium of the present invention that binds to the B-type sugar chain and does not adsorb to other types of sugar chains is compatible with ABO blood group B humans. The L. gasseri lactic acid bacterium of the present invention that binds to the type 0 sugar chain and does not adsorb to other types of sugar chains is suitable for ABO blood group 0 type humans.

[0015] The L. gasseri lactic acid bacteria of the present invention (hereinafter sometimes referred to as "blood group-specific L. gasseri lactic acid bacteria") can be separated from human feces. If it is L. gasseri lactic acid bacteria (hereinafter abbreviated as "A-type compatible lactic acid bacteria") compatible with blood group A humans, it is compatible with humans of blood group B from ABO blood group A human feces L. gasseri lactic acid bacteria (hereinafter abbreviated as “B-type compatible lactic acid bacteria”) from ABO blood type B human feces to L. gasseri lactobacilli compatible with blood type 0 humans ( (Hereinafter abbreviated as “type 0 compatible lactic acid bacteria”), there is a high possibility of more efficient separation from human feces of ABO blood type 0. For the separation, the properties of L. gasseri lactic acid bacteria well known to those skilled in the art can be used as a guide. For example, gonococcus, homo-fermentation, aerobic growth, no gas generation, etc. can be used as a guide.

[0016] In order to culture L. gasseri lactic acid bacteria according to the present invention, any medium suitable for culturing Lactobacillus is generally used. Glucose, latatoses, galactose, fructose, trenorose, sucrose , Carbon sources such as mannose and cellobiose, meat extract, peptone, A medium containing a nitrogen source such as yeast extratate, casein, or whey protein, and inorganic nutrients such as magnesium sulfate, iron sulfate, or manganese sulfate can be used. One suitable example is Lactobacilli MRS broth (Difco, Ref. No. 288130). The culture conditions are not particularly limited as long as the intestinal lactic acid bacteria can grow. Preferred conditions are, for example, pH 5.0-pH 8.0, temperature 20 ° C-45 ° C, and more preferred conditions are anaerobic, pH 5.0-pH 7.0, temperature 30 ° C-40. ° C.

[0017] Whether or not L. gasseri separated and cultured as described above has intestinal tract-binding properties and blood group antigens by blood group is determined by binding to human intestinal mucin or blood group antigens. It can be known by judging the presence or absence of sex. For example, a cell surface protein (SLP) may be prepared from the surface layer of the test bacterium, and the binding between SLP labeled with piotin or the like and intestinal mucin or blood group antigen may be detected. Alternatively, detection may be performed by a hybridization method after electrophoresis of SLP of a test bacterium using labeled intestinal mucin or labeled blood group antigen. Further, as in the examples described later, if a surface plasmon analyzer (for example, BIACORE) is used, it is possible to detect viable bacteria as they are.

[0018] In order to prepare human intestinal mucin by blood type, the surface part of the human intestinal force of a specific blood type is also collected and subjected to gel filtration using a solubilizing agent such as guanidine hydrochloride to reduce protein absorption. Purification can be carried out based on the high neutral sugar content. For example, the method described in Purushothaman, b. B. Et al, Adherence of Shigella dysentenae 1 to Human colonic Mucin. Curr. Ic robiol, 42 (6), 381-387 (2001). It is more preferable to confirm that the blood group antigen is expressed in the prepared human intestinal mucin using an anti-blood group antigen antibody. Specific examples include the methods described in the examples. On the other hand, blood group antigens can be synthesized on the basis of the sugar chain sequence shown in the formula below or in FIG. 2 (eg, commercially available sugar chain probes (eg, manufactured by Seikagaku Corporation) or neoglycoprotein 'Blood Group A Trisaccaride- BSA (eg Calbiochem) or neoglycoprotein 'Blood Group B Trisaccaride- BSA (eg Calbiochem) may be used.

[0019] As representative examples of L. gasseri lactic acid bacteria according to blood type of the present invention, accession number: NITE BP-25, accession number: NITE BP-26, accession number: NITE BP-27, accession number: NITE BP -Lactobacillus gasseri specified by -28. Of these, accession number: NITE BP- Lactic acid bacteria identified by 26 and accession number: NITE BP-27 are type A compatible lactic acid bacteria, and lactic acid bacteria specified by accession number: NITE BP-25 and accession number: NITE BP-28 are type 0 compatible lactic acid bacteria. These strains are lactic acid bacteria that have been confirmed by the present inventors to be compatible with the above-mentioned blood types. Further, representative examples of L. gasseri lactic acid bacteria according to blood group of the present invention include Lactobacillus gasser i specified by accession number: NITE BP-145 and accession number: NITE BP-146. The lactic acid bacteria specified by the accession number: NITE BP-145 and the accession number: NITE BP-146 are B-type compatible lactic acid bacteria. These strains were confirmed by the present inventors to be compatible with blood group B.

The present inventors deposited these strains with the Patent Microorganism Depositary Center, National Institute of Product Evaluation Technology, based on the Budapest Treaty concerning the international recognition of the deposit of microorganisms in the patent procedure. The contents specifying the deposit are described below.

Accession numbers NITE BP-25, 26, 27, 28!

(I) Depositary Institution: National Institute of Technology and Evaluation Patent Microorganisms Deposit Center (Location: Kisarazu Kazusa Kamashita 2-5-8 Postcode 292-0818, Chiba Prefecture, Japan) (Kuchi) Original deposit date: Heisei 16 Year (2004) October 8

Date of request for transfer to deposit under the Budapest Treaty (transfer date): May 6 2005

(C) Accession number:

Lactobacillus gasseri OLL2915 strain (Accession number NITE BP-25)

Lactobacillus gasseri OLL2804 strain (Accession number NITE BP-26)

Lactobacillus gasseri OLL2818 strain (Accession number NITE BP-27)

Lactobacillus gasseri OLL2827 strain (Accession number NITE BP-28)

Accession numbers NITE BP-145 and 146 are as follows.

(I) Depositary Institution: National Institute of Technology and Evaluation Patent Microorganism Depositary Center (Location: Kisarazu Kazusa Kamashika 2-5-8 Postal Code 292-0818, Chiba Prefecture, Japan) (Date) Hara Deposit Date: Heisei 17 Year (2005) October 4

Date of receipt of request for transfer to deposit under the Budapest Treaty (transfer date): November 25, 2005 (C) Contract number:

Lactobacillus gasseri OLL2877 strain (Accession number MTE BP-145)

Lactobacillus gasseri OL 2901 strain (Accession number MTE BP-146)

[0021] The gasseri lactic acid bacterium according to the blood type of the present invention can be used for the production of foods and drinks suitable for each blood type. The foods and beverages produced by using gasseri lactic acid bacteria according to the present invention may be functional foods, specific health foods, health foods, and nursing foods that are not restricted in categories or types. It may be a dairy product such as cheese or yogurt, a seasoning or the like. There are no restrictions on the shape of the food or drink, and any form of food or drink that can normally be distributed can be used, such as solid, liquid, liquid food, jelly, tablet, granule, and capsule. Manufacture of the said food-drinks can be performed by those skilled in the art. In the production of the above food and drink, as long as it does not interfere with the growth of lactic acid bacteria, carbohydrates, proteins, fats, dietary fibers, vitamins, biologically essential trace metals (manganese sulfate, zinc sulfate, magnesium chloride, potassium carbonate, etc.), Perfumes and other blends can also be added.

[0022] The gasseri lactic acid bacterium according to the blood type of the present invention can be mixed with general foods and drinks, and can be used as a starter for producing dairy products and fermented milk such as yogurt and cheese. . In the case of a starter, as long as there is no hindrance to growth and dairy product production, the presence of L. gasseri? Good. For example, it may be mixed with Lacto bacillus delbruek subsp.bulgancus, Streptococcus thermophilus, Lactobacillus aci dophilus, etc. It can be mixed to make a starter. Yogurt production using the above starter can be carried out according to a conventional method. For example, plain milk dart can be produced by mixing the above starter with milk or dairy products cooled after warming 'mixing' and 'homogenizing' and sterilizing, followed by fermentation and cooling.

[0023] The food and drink produced by using the gasseri lactic acid bacterium according to the present invention according to blood type contains the same bacterium in the food and drink. When this food or drink is ingested by a human who is compatible with the AB0 blood group, the gasseri lactic acid bacteria bind and grow in the intestinal tract according to the blood type of the present invention, and the intestinal balance is adjusted and maintained. Early take-out function is expected. Therefore, the

Corrected ^ (Rule 91) The foods and drinks are useful not only as general foods and drinks but also as functional foods for the purpose of promoting health, such as “health and nutrition foods”, “care foods” and “health foods”.

[0024] The present invention also provides a method for screening lactic acid bacteria by surface plasmon resonance spectrum. The screening method of the present invention is a method for selecting lactic acid bacteria suitable for each blood group. The screening method of the present invention comprises screening by measuring the binding of lactic acid bacteria to blood type human intestinal mucin and ABO blood group antigens by surface plasmon resonance spectrum, and (i) specific blood (Ii) ABO blood group antigens other than the specific blood group in (i) above and a certain amount or more of adsorbing ability. (Iii) Selection is made using as an index the ability to bind to a specific blood group of human intestinal mucin in (i) above. By measuring the ability to bind to ABO blood group antigens, parts other than antigens that mainly determine blood groups (for example, the partial structure of the non-terminal part of sugar chains, the expression sites of sialic acid and sulfate groups, mucins) Lactic acid bacteria that bind to the protein site) can be eliminated, and screening for lactic acid bacteria is possible with high blood group specificity. In the present invention, human type A intestinal mucin refers to type A human intestinal mucin of the ABO blood group. Similarly, human type B intestinal mucin is a blood group B human-derived intestinal mucin, and human type 0 intestinal mucin is a blood group 0 human intestinal mucin.

[0025] In the screening method of the present invention, surface plasmon resonance spectrum analysis is performed using ABO blood group antigen and human intestinal mucin as a probe. For example, BIACORE (Biacore Co., Ltd.) can be used as a biomolecular interanalyzer utilizing the surface plasmon resonance spectrum. The preparation method of ABO blood group antigen and human intestinal mucin is as described above. In the case of immobilizing the probe, it can be performed by a known immobilization method. The immobilization method may be a physical adsorption method or a covalent bond adsorption. For example, if the chip is coated with streptavidin and the probe is piotinated, it can be easily fixed by a piotine-avidin bond. You can also use a commercially available streptavidinized chip (BIACORE)! /.

Since the method of the present invention is based on a surface plasmon resonance spectrum, the “binding ability” is represented by a resonance unit (RU). 1RU is, to 1mm ² of lpg substance binding child Represents. In the method of the present invention, the “binding ability of a certain amount or more” is determined based on “100RU”. That is, it is determined that a certain amount or more of lactic acid bacteria binds to the probe based on a measured value of “100RU” or more. For example, if the measurement result using a blood group A human-derived intestinal mucin as a probe is 100 RU or more, the lactic acid bacterium is a lactobacillus that binds to the intestinal mucin. The RU value can vary depending on the measurement conditions. The temperature condition in this method is, for example, 20-40 ° C, preferably 20-30 ° C, more preferably 23-28 ° C. The preferred sample concentration in this method is 0.1-0.5 mg / mL, and the preferred flow rate in this method is 3-1 で / zl / min. If it is within the above range, the RU value will not change even if the above conditions are changed. Furthermore, even when various conditions such as sample concentration are changed outside the above range, they are included in “100RU” in this method if they are substantially equivalent to 100RU under the above conditions. In addition, regarding the ability to bind to the ABO blood group antigen probe, selection may be made by a value other than “100RU” for the purpose of making the selection criteria stricter. In the method of the present invention, a bacterium having a higher RU value means a bacterium having a stronger ability to bind to a probe (ligand). Therefore, for the purpose of finding bacteria with stronger binding power, it is possible to screen for bacteria that bind to ABO blood group antigen probes with RU values higher than 100! For example, screening may be performed based on 2000 RU as in the examples described later, which may be 150, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 RU. Conversely, for recognition of blood types that are not intended, screening may be based on numerical values that are stricter than "100RU or less". For example, 90,80,70,60,50RU may be used as a standard.

The blood group-binding lactic acid bacteria were measured when the ABO blood group probe was used as the measured value (RU) when using the blood type human intestinal mucin as a probe according to the screening method of the present invention. Expected to be lower than the value (RU). This is because blood group antigens (sugar chains) bound to human intestinal mucins when human intestinal mucins are immobilized on a chip are compared only with blood group antigens (sugar chains) on a chip unit area. This is considered to be the force that is reduced when fixed. The measured value when using a specific ABO blood group probe is sufficiently high compared to the measured value when using another ABO blood group probe, for lactic acid bacteria! Measured value when using human intestinal mucin by type as a probe However, whether or not the measured value when using the specific blood type human intestinal mucin as a probe is sufficiently higher than the measured value when using other blood type human intestinal mucin probes, When it can be judged that the usefulness for the purpose can be obtained, the blood group-specific binding lactic acid bacteria of the present invention can be obtained. For example, in type A compatible lactic acid bacteria, (i) the RU value indicating the ability to bind to human ABO blood type antigen of type A is 100RU or more, (ii) human ABO blood of type B and H RU value indicating ability to bind to type A antigen is 100 RU or less, (iii) RU value indicating ability to bind to human type A intestinal mucin RU value and human ability to bind to human type B intestinal mucin It can be screened using as an indicator that it is larger than the RU value indicating the ability to bind to type 0 intestinal mucin. In B-type compatible lactic acid bacteria, (i) the RU value indicating the binding ability with human ABO blood group antigen of type B is 100RU or more, (ii) human ABO blood group antigens of type A and H (Iii) RU value indicating the ability to bind to human type B intestinal mucin is the RU value indicating the ability to bind to human type A intestinal mucin and human type 0 It is possible to screen for an index that is greater than the RU value indicating the ability to bind to intestinal mucin. In type 0 compatible lactic acid bacteria, (i) the RU value that indicates the ability to bind to H-type human ABO blood group antigens is 100 RU or more, and (ii) the human ABO blood group antigens of type A and B RU value showing binding ability is 100RU or less, (iii) RU value showing binding ability with human type 0 intestinal mucin Force RU value showing binding ability with human type A intestinal mucin and human type B intestinal mucin It is possible to screen using an index that is larger than the RU value indicating the binding ability of.

[0028] During surface plasmon resonance spectrum measurement, the test bacterium may cause bacterial aggregation due to non-specific adsorption, and the RU value may become abnormally high. Therefore, adsorption with human-derived intestinal mucin and ABO blood group antigen is not correctly reflected in the measurement results. Therefore, if the RU value is abnormally high, it is suspected that nonspecific adsorption (bacterial aggregation) has occurred, and the lactic acid bacteria can be excluded from the screening.

[0029] The subject of the screening method of the present invention is not particularly limited as long as it is a lactic acid bacterium. Despite being listed as an object, Lactobacillus acidopnilus, Lactobacillus crispatus, Lactooaci llus amylovorus, Lactobacillus gallinarum, Lactobacillus gasseri, Lactobacillus johns onii, Lactobacillus caselus, Lactobacillus caselus Lactobacillus delbruekn subsp. 1 Suitable for use with actis, Lactobacillus fermentum, Lactobacillus murinus, Bifidobacterium animalis, Bifi dobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacteriu m longum, Bifidobacterium pseudolongum, Enterococcus faecium, Enterococcus feca lis, Streptococcus thermophilus. A more suitable subject example is the genus Lactobacillus, and the most preferred subject example is: Lactobacillus acidophilus group lactic acid bacteria containing L. gasseri.

[0030] The lactic acid bacteria screened by the method of the present invention is a lactic acid bacterium that easily binds to the human intestinal tract of a specific ABO blood group. By taking the lactic acid bacteria screened by the method of the present invention, the human of the specific blood group can improve health due to the effect of improving the intestinal balance in a short period of time. Therefore, the lactic acid bacteria screened by the method of the present invention can be applied to foods and drinks for specific blood groups, functional foods, foods for specific health use, dairy products, lactic acid bacteria drinks, and the like.

It should be noted that all prior art documents cited in this specification are incorporated herein by reference.

Example

[0031] Hereinafter, the present invention will be described more specifically based on examples. However, the present invention is not limited to the following examples.

[0032] [Example 1] Gas production test

It is desirable that the lactic acid bacteria used for yogurt production do not produce gas. This is because Japanese laws require that the product be marketed in a sealed container, and also to prevent product defects and rupture due to container expansion associated with gas production. Therefore, a gas production test was conducted for 238 strains.

[0033] Lactic acid bacteria were activated and cultured twice (37 ° C, 18h) with MRS Broth (DIFCO). A Durham tube and MRS Broth (5 ml) were put together in a test tube with an aluminum cap and sterilized (121 ° C, 15 minutes). The Durham tube has an opening facing down, and bubbles are removed from the Durham tube during sterilization. The sterilized test tube with an aluminum cap was inoculated with 10 μL of about 10 ⁹ cfo / mL lactic acid bacteria solution, and anaerobically cultured at 37 ° C. for 24 hours. After completion of the culture, the presence or absence of gas accumulated in the Durham tube was visually observed. Gas production occurs when clear bubbles are observed (+ (Table 1).

[0034] [Example 2] Analysis of blood group antigen recognition

In order to obtain lactic acid bacteria for blood group compatible yogurt, lactic acid bacteria that recognize and bind to blood group antigens were selected by surface plasmon resonance spectrum measurement. Prepare 238 strains belonging to Lactobacillus acidophilus group (L. gasseri, L. plant arum, L. crispatus, L. amylovorus, L. casei, L. salivarius, L. brevis, L. fermentum, etc.) as test bacteria did. BIACORE1000 was used as the surface plasmon resonance spectrum analyzer.

[0035] 2- (1) Preparation of analyte

Each strain was subcultured 3 times in MRS medium after inoculation, and after culturing for 12 hours, 500 μl was poured into a 1.5 ml tube. The cells obtained by centrifuging this solution (6,000 rpm, 4 ° C, lOmin) were washed twice in PBS (pH 7.2) and then lyophilized. This bacterial cell was prepared to an O.lmg / ml concentration using HBS-EP buffer (0.01M HEPES pH7.4, 0.15M NaCl, 3mM EDTA, 0.005% Surfactant P20) to prepare an analyte solution.

[0036] 2- (2) Blood antigen probe and immobilization to chip

The antigen structure of human ABO blood group substance was used as a probe for selection of bacteria. Specifically, the following trisaccharide sugar chain polymer probe (manufactured by Seikagaku Corporation) (hereinafter also referred to as “BP-probe”) was used.

A antigen probe: [GalNAc a 1-3 (Fuc a 1-2) Gal-]

B antigen probe: [Gal a 1-3 (Fuc a 1-2) Gal-]

H antigen probe: [Fuc a l-2Gal-]

The chemical structure of the BP-probe is shown in FIG. 1 and the sugar chain structure is shown in FIG.

[0037] The blood antigen BP-probe was prepared to O.lmg / lOml with HBS-EP buffer (pH 7.4).

This solution was added to the sensor chip SA, and a commercially available probe was fixed to the surface of the sensor chip by the piotin-avidin reaction. Sensor chip SA is a BIACORE dedicated chip on which streptavidin is immobilized. The immobilized amount of the A antigen probe was 750 RU, and the immobilized amount of the B antigen probe was 850 RU. The 0 antigen probe was immobilized as follows. l-mg A antigen sugar chain probe (Seikagaku Corporation) α- 酵素 -Acetylgalactosaminidase (EC3.2.1) .97 Streptococcus pneumoniae derived, SIGMA, JAPAN) 0.5U was used to react in acetate buffer pH 4.5 at 55 ° C for 20 hours to remove terminal GalNAc. (It was confirmed that 80% or more of GalNAc was removed by measuring the removal rate with the A antigen antibody!) This biotinylated H-type antigen sugar chain probe was treated in the same manner as the A-type antigen sugar chain probe. Then, it was immobilized on a sensor chip to which avidin was bound. The amount of binding at this time was about 1000 RU.

[0038] 2- (3) Preparation of human intestinal mucin and fixation to chip

Human large intestine mucin was prepared as another ligand used for selection of bacteria. Human type A intestinal tract (large intestine) Human type B intestinal tract (large intestine) and human type 0 intestinal tract (large intestine) were distributed by the Graduate School of Medicine, Tohoku University as sampling samples. The mucous mucin layer was collected from the normal colon by the surface layer removal method. The mucin layer was degreased with Folch's solvent and jetyl ether, dried, and extracted with 4M guanidine hydrochloride solution at 37 ° C for 2 hours. Purified human A type, human B type, or human type 0 intestinal mucin (human colon mucin HCM, sometimes referred to as A-HCM type B intestinal mucin, type B-HCM type 0) Intestinal mucin was abbreviated as 0-HCM) and used for the test. Gel filtration purification is a purification method for human colon mucin described in Purushothaman, SS et al, Adherence of Shigella dysentenae 1 to Human colonic Mucin. Curr.icrobio 1., 42 (6), 381-387 (2001). Based on. The moving bed was 4M guanidine hydrochloride solution, and the column was Toyopearl HW-65F (90 cm X 2.6 cm, To soh. Tokyo. Japan). For detection, neutral sugars were measured by the phenol sulfate method (490 nm) and proteins were measured at 280. The peak with the highest protein absorption and neutral sugar content was selected, and a fraction with a molecular weight of about 2 million or more was collected as a standard to obtain human large intestine mucin (HCM). About the obtained HCM, the human blood group substrate antigenicity of each blood group was confirmed with an antibody. The above sample collection was conducted through the ethics committee of the Graduate School of Medicine, Tohoku University, and patient consent was obtained.

[0039] Immobilization of the HCM to the BIACORE chip was performed by the amine coupling method. First, 75.0 mg / ml N-ethyto-N-dimethyl aminopropyto carbodiimide hydrochloride (EDC) 50 μ 1 and 11.5 mg / ml Ν-hydroxysuccinimide were applied to sensor chip CM5 into which carboxymethyl dextran group had been previously introduced. A mixed reagent in which 50 μl of (NHS) was mixed was flowed to activate the carboxyl group introduced at the dextran end. There 120 A mixed solution containing HCM-A 30 1 was poured into an acetic acid buffer (ρΗ4.0) for immobilizing μ 1 and covalently bound by amin coupling reaction. Subsequently, 1M ethanolamine hydrochloride-NaOH pH 8.5 was used to block the remaining active groups in the places where the ligand was not bound. HBS-EP buffer was used as Running Buffer. The amount of HCM immobilized was set to 1000 to 2000 RU, considering that the screening method of the present invention can be performed with good reproducibility.

[0040] 2- (4) Measurement with BIACORE1000

Using the aforementioned blood group antigen probe-immobilized chip or HCM-immobilized chip, the interaction of the above-mentioned analyte with the blood group antigen was analyzed by BIACORE. The measurement conditions for BIACORE are shown below.

Running buffer: HBS-EP buffer (pH7.4)

Sample amount: 20 1

Flow rate: 3 μ 1 / min

Temperature: 25 ° C

Regeneration solution: 1M guazine hydrochloride solution 5 μ 1

[0041] In the analysis by BIACORE, the interaction between the analyte and the blood group antigen is expressed by a resonance unit (RU). 1 Resonance unit (RU) means that the substance lpg is bound to 1 mm ² . The results are shown in Table 1.

[0042] [Table 1]

Selection of type A compatible lactic acid bacteria

Selection of type 0 compatible lactic acid

Selection of B-type compatible lactic acid bacteria

B-HCM A-HC 0-HCM

Strain name B antigen recognition Gas production A antigen recognition B antigen recognition H antigen recognition B antigen recognition Bile acid resistance Gastric acid resistance Recognizability Recognizability Recognizability

/ A antigen recognition / H antigen recognition

(RU) (RU) (RU) (RU) (RU) (RU) (0Ό _65ΰ ) (survival rate%) EP 1 65507 0-224.8 0.0 0 0.0 0.0 0.0 2.4 0.339 0.01 1

MEP 1 65 508 0 + 0 0.0 96.1 0.0 18.7 0.0 531 .4 2.020 0.010

MEP 165 509 0-0 0.0 0 0.0 0.0 0.0 154.0 0.000 1.499

L gasseri 0 2877 4220.9-66.4 63.6 0 ∞ 1 406.3 218.2 0.0 0.651 10.81 2

L gasseri OLL 2901 2000.9-0 0 1 7.1 0.0 7.6 0.031 8.923

[0043] 2- (5) Selection of bacteria (part 1)

Lactic acid bacteria were selected from the results so far. First, selection was performed from the BIACORE measurement result and the gas production test result of Example 1.

[0044] Blood type A yogurt lactic acid bacteria were selected from the viewpoints of (i) A antigen recognition and (ii) gas production. The selection criteria are

(i) A antigen recognizability: As measured by BIACORE, the result of A antigen probe was 2000 RU or more (upper 35%)

(ii) Gas production: Gas production is not recognized in the gas production test (-)

It was. As a result, Lactobacillus 238 was selected to 43 strains.

[0045] Similarly, yogurt lactic acid bacteria for blood group 0 were selected by (i) H antigen recognition and (ii) gas production. The selection criteria are

(i) H antigen recognizability: As measured by BIACORE, the result of H antigen probe is 700RU or higher (upper 13%)

It was. As a result, Lactobacillus 238 strain was narrowed down to 14 strains.

[0046] Similarly, yogurt lactic acid bacteria for blood group B were selected by (i) B antigen recognition and (ii) gas production. The selection criteria are

(i) B antigen recognizability: BIACORE measurement results showed a B antigen probe of 2000 RU or more (upper 14%)

It was. As a result, Lactobacillus 238 strain was narrowed down to 25 strains.

[0047] 2- (6) Selection of bacteria (part 2)

As described above, 43 strains that recognized the A antigen and showed no gas production were selected as blood group A yogurt lactic acid bacteria candidates. Furthermore, they were selected from the viewpoint of (i) that other blood group antigens (B antigen and H antigen) were not recognizable (ii) that A antigen was specifically recognized. The selection criteria are

(i) Recognition of other blood group antigens (B antigen and H antigen)

B antigen recognition: When measured with BIACORE, the result of B antigen probe is 100RU or more. under,

H antigen recognition: 100 RU or less with H antigen probe as measured by BIACORE (ii) Specific recognition of A antigen

A antigen recognition / B antigen recognition ratio is 70 or more, A antigen recognition / H antigen recognition ratio is 100 or more

It was. From these, 13 strains were selected from 43 strains.

[0048] Similarly, yogurt lactic acid bacteria candidates for blood group 0 were also selected. The selection criteria are

(i) Recognition of other blood group antigens (A antigen and B antigen)

A antigen recognition: In BIACORE measurement, the result of A antigen probe is 100RU or less.

B antigen recognition: In BIACORE measurement, B antigen probe results are less than 100RU,

(ii) Specific recognition of H antigen

H antigen recognition / A antigen recognition ratio is 800 or more, H antigen recognition / B antigen recognition ratio is 20 or more

It was. From these, 5 strains were selected from 14 strains.

[0049] Similarly, a yogurt candidate strain for blood group B was also selected. The selection criteria are

(i) Recognition of other blood group antigens (A antigen and H antigen)

H antigen recognition: In BIACORE measurement, the result with H antigen probe is 100RU or less.

(ii) Specific recognition of B antigen

The ratio of B antigen recognition / A antigen recognition was 50 or more, and the ratio of B antigen recognition / H antigen recognition was 50 or more. Based on these, 6 strains were selected from 25 strains.

[0050] 2- (7) Selection of bacteria (part 3)

The strains selected above were further selected based on blood type human colon mucin (HCM) recognition. [0051] Regarding blood group A-type yogurt lactic acid bacteria candidates, (i) those that recognize A-HCM were selected, and (ii) those that recognize O-HCM were excluded. In other words, the selection criteria were (i) A-HCM recognizability of 100RU or higher and (ii) O-HCM recognizability of 100RU or lower in BIACORE measurement results by human HCM. MEP165511 strain and MEP165530 strain were excluded because A-HCM recognizability was 1000 RU or more and bacterial aggregation was suspected. From the above selection, 2 strains (L. gasseri OLL 2804, L. gasseri OLL 2818) were selected from 13 strains.

[0052] Similarly, yogurt lactic acid bacteria candidates for blood group 0 are selected from (i) those that recognize 0-HCM, and (ii) those with relatively high recognition of A-HCM. Was excluded. In other words, the selection criteria were (i) O-HCM recognizability of 100RU or higher and (ii) A-HCM recognizability of 1000RU or lower in the BIACORE measurement results by human HCM. From the above selection, 2 strains (L. gasseri OLL 2827, L. gasseri OLL 2915) were selected from 5 strains.

Similarly, the selection criteria for blood type B yogurt candidate strains were that B-HCM recognition was higher than A-HCM and H-HCM recognition. From the above selection, 2 strains (L. gasseri OLL2877, L. gasseri OLL2901) were selected from 6 strains.

[Example 3] Gastric acid tolerance test and bile acid tolerance test

In order for the lactic acid bacteria taken into the body as yogurt to fully perform its function, it is desirable to stay alive in the intestine. Therefore, a gastric acid tolerance test and a bile acid tolerance test were conducted.

[0055] 3- (1) Gastric acid tolerance test

Artificial gastric fluid PH2 sterilized by filtration [NaCl (0.2%), Pepsin (l: 5000, Tokyo Chemical Industry) (0.35%): adjusted to pH 2 with 1N hydrochloric acid] 9 ml in Lactobacilli MRS broth (DIFCO) 37. C, 18 hours), add lml of lactic acid bacteria suspension washed twice with physiological saline, contact aerobically at 37 ° C for 2 hours, remove lml, 67mM phosphate buffer The reaction was stopped by adding 9 ml of the solution (pH 6.5). The number of viable bacteria before and after contact with the artificial gastric juice was measured using Lactobacilli MRS Agar (DIFCO), and the survival rate (%) was calculated.

[0056] Selection criteria vary depending on the type of bacteria Since all of the bacterial species selected this time are Lactobacillus gas seri, they were set to 0.50 or more. Blood group type A yogurt lactic acid bacteria (L.gasseri OLL 280 4, L.gasseri OLL 2818), blood group type yoghurt (L.gasseri OLL 2827, L.gasseri O LL 2915) and blood group B yogurt lactic acid bacteria (L.gasseri OLL2877, L.gasseri OLL2901) both met the above criteria (Table 1).

[0057] 3- (2) Bile acid tolerance test

Lactobacilli MRS broth (DIFCO) 2 times of activation culture (37 ° C, 18 hours) Lactobacilli MRS broth (DIFCO) containing 5% of Lactobacilli MRS broth (DIFCO) Cultured at 37 ° C. After 18 hours of culture, the turbidity (OD) of the medium was measured.

650

[0058] The selection criteria were 0.08 or more for blood type A yogurt lactic acid bacteria and blood group 0 yogurt lactic acid bacteria, which differ depending on the type of bacteria. Both blood type A yogurt lactic acid bacteria (L.gasseri OLL 2804, L.gasseri OLL 2818) and blood type O yogurt lactic acid bacteria (Lg asseri OLL 2827, L.gasseri OLL 2915) met the above criteria. (table 1). Among blood type B yogurt lactic acid bacteria, L. gasseri OLL2901 had a bile acid tolerance of 0.031, but it is considered that there is no particular problem with intestinal retention. Table 2 shows the scientific characteristics of the above six strains.

[0059] [Table 2]

[0060] [Example 4] Production of yogurt by blood group-binding lactic acid bacteria

Using the lactic acid bacteria selected above (yogurt lactic acid bacteria for blood group A: L.gasseri OLL 2804, L.ga sseri OLL 2818, yogurt lactic acid bacteria for blood group O: L.gasseri OLL 2827, L.gasseri OL L 2915) Yogurt was manufactured.

[0061] 4 (1) Example of yogurt production using L. gasseri OLL 2804

Plain yogurt was prepared using L. gasseri OLL 2804 strain. First, inoculate 1% each of L. gasseri OLL2804 strain, L. bulgaricus JCM 1002 ^T and S. thermophilus ATCC19258 in 10% skimmed milk powder culture medium, and incubate at 37 ° C for 15 hours to prepare a Balta starter. [0062] Yogurt mix (SNF: 9.5%, FAT: 3.0%) heated at 95 ° C for 5 minutes, 1% each of starter of L. bulgar icus JCM 1002 ^τ and S. thermophilus ATCC 19258, L. gasseri OLL 2804 strain starter was inoculated 5% and fermented at 43 ° C for 4 hours.

[0063] The viable cell counts of L. gasseri OLL 2804, L. bulgaricus JCM 1002 ^T , and S. thermophilus ATCC 19258 were 12.5 X 10 ⁷ CFU / mL, 14.0 X 10 ⁷ CFU / mL, 11.8 X 10 ⁸ CFU / mL. The flavor and physical properties were both good. In addition, when the above yogurt was stored at 10 ° C, the viable count of gasseri OLL 2804, L. bulgaricus JCM1002 ^T , S. thermophilus ATCC 19258 was 9.60 X 10 ⁷ CFU / mL, 7.50 X 10 ⁷ CFU / mL, 11.0 X 10 ⁷ CFU / mL, and the survival rate of the L. gasseri OLL 2804 strain was 77% on the first day of storage, with a slight decrease in the number of viable bacteria. The preserved product has good flavor and physical properties.

[0064] 4- (2) Example of yogurt production using L. gasseri OLL 2818

Plain yogurt was prepared using L. gasseri OLL 2818 strain. First, 1% each of L. gasseri OLL2818 strain, L. bulgaricus JCM 1002 ^T and S. thermophilus ATCC19258 was inoculated into 10% nonfat dry milk medium, and cultured at 37 ° C for 15 hours to prepare a Balta starter .

[0065] Yogurt mix (SNF: 9.5%, FAT: 3.0%) heated at 95 ° C for 5 minutes, 1% each of L. bulga ricus JCM 1002T and S. thermophilus ATCC 19258 starter, L. gasser i OLL 2818 starters were inoculated with 5% and fermented at 43 ° C for 4 hours.

[0066] The viable cell counts of L. gasseri OLL 2818, L. bulgaricus JCM 1002 ^T and S. thermophilus ATCC 19258 were 14.0 X 10 ⁷ CFU / mL, 20.0 X 10 ⁷ CFU / mL, 11.4 X 10 ⁸ CFU / mL. The flavor and physical properties were both good. In addition, when this yogurt was stored at 10 ° C, the viable count of gasseri OLL 2818, L. bulgaricus JCM1002T and S. thermophilus ATCC 19258 was 11.4 X 10 ⁷ CFU / mL, 7.00 X The survival rate of L. gasseri OLL 2818 was 82% on the first day of storage, and the viable cell count decreased only slightly, 10 ⁷ CFU / mL, 10.0 X 10 ⁷ CFU / mL. The preserved product has good flavor and physical properties.

Industrial applicability According to the present invention, a lactic acid bacterium suitable for human ABO blood group and a screening method thereof are provided. The lactic acid bacterium of the present invention or the lactate strain obtained by the screening method of the present invention is a lactic acid bacterium having a high intestinal binding property for each ABO blood type, and functional yogurt classified by blood type is obtained by applying the lactic acid bacterium to food and beverage production. New probiotic foods and drinks can be provided.

Claims

The scope of the claims

[1] An intestinal-binding Lactobacillus gasseri lactic acid bacterium having a binding ability to a human ABO blood group antigen represented by any one of the following formulas (a) to (c):

(a) [GalNAc a 1-3 (Fuc a 1-2) Gal-]

(b) [Gal a 1-3 (Fuc a 1-2) Gal-]

(c) [Fuc a l-2Gal-]

[2] Accession number: NITE BP-25, Accession number: NITE BP-26, Accession number: NITE BP-27, Accession number: NITE BP-28, Accession number: NITE BP-145, Accession number: NITE BP- The Lactobacillus gasseri lactic acid bacterium according to claim 1, which is specified by any one of 146.

[3] A human ABO blood group-compatible fermented milk and a starter for producing a dairy product, comprising the Lactobacillus gasseri lactic acid bacterium according to claim 1 or 2.

[4] A food or drink containing the Lactobacillus gasseri lactic acid bacterium according to claim 1 or 2.

[5] Fermented milk and lactic acid bacteria beverages containing the Lactobacillus gasseri lactic acid bacteria according to claim 1 or claim 2.

[6] A screening method for lactic acid bacteria by surface plasmon resonance spectrum using the following condition (i) force (iii) as an index.

(iii) The RU value indicating the ability to bind to human type A intestinal mucin must be 100 RU or more

[7] A screening method for lactic acid bacteria by surface plasmon resonance spectrum using the following condition ω force (iii) as an index.

[8] A screening method for lactic acid bacteria by surface plasmon resonance spectrum, using the following condition (i) force (iii) as an index. (i) The RU value that indicates the ability to bind H-type human ABO blood group antigen is 100RU or more

(iii) The RU value that indicates the ability to bind human type 0 intestinal mucin is 100 RU or more

A screening method for lactic acid bacteria by surface plasmon resonance spectrum, using the following condition (i) force (iii) as an index.

(ii) The RU value, which indicates the ability to bind human ABO blood group antigens of type A and H, is 100RU or less

(iii) The RU value indicating the ability to bind to human type B intestinal mucin must be greater than the RU value indicating the ability to bind to human type A intestinal mucin and the RU value indicating the ability to bind to human type 0 intestinal mucin.