KR102114046B1 - Lactobacillus brevis BHN-LAB 127 having acetylcholinesterase inhibitory activity - Google Patents

Abstract

The present invention provides a Lactobacillus brevis BHN-LAB 127 strain (Accession No: KCTC13441BP) having an acetylcholinesterase inhibitory activity, and a composition comprising the same. The Lactobacillus brevis BHN-LAB 127 strain of the present invention significantly inhibits acetylcholinesterase activity, and thus, as a material for a pharmaceutical composition for treating Alzheimer′s disease or a food composition for ameliorating Alzheimer′s symptoms, is useful in the field of food processing, functional health food and pharmaceutical industries.

Description

Lactobacillus brevis BHN-LAB 127 having acetylcholinesterase inhibitory activity}

The present invention relates to a strain having an acetylcholinesterase (AChE) inhibitory activity and a composition comprising the same, and more specifically, Lactobacillus brevis BHN- having an acetylcholinesterase inhibitory activity related to anti-dementia efficacy LAB 127 and relates to a composition comprising the same.

Lactobacillus brevis ( Lactobacillus brevis) is a lactobacillus that is abundant in traditional fermented foods such as kimchi, miso, and salted fish, and is also called probiotics. Lactobacillus is a representative intestinal beneficial flora, and has various physiological activities such as membranous immune activity, antimutagenic activity, intestinal action, antibacterial activity, anti-cholesterol action, antioxidant activity, and so on. It is widely used as a material for medicines, cosmetics, and probiotics, and is one of the fastest growing health food materials in recent years.

In order to be effective as a lactic acid bacterium, it is required to reach the small intestine after ingestion orally and adhere to the surface of the intestine, so it is necessary to overcome the strong acidic environment up to pH 2.5 when fasting. As a lactic acid bacterium having excellent resistance to pancreatic enzyme, it must reach the small intestine without dying.

Alzhimer's disease (AD) is a chronic and progressive neurodegenerative disease that causes memory loss, disorientation, cognitive dysfunction and behavioral disorders. Alzheimer's disease is associated with the loss of choline at the base of the whole brain and cortex. It has been reported that activating choline function by inhibiting cholinesterase may be effective in the treatment of Alzheimer's disease, because lack of choline neurotransmission plays a critical role in declining learning and memory of Alzheimer's disease patients. Synthetic acetylcholinesterase inhibitors, including Tacrine, Donepezil, Rivastigmine, and Galanthamine, have been used to treat Alzheimer's disease. However, these drugs are known to induce side effects such as insufficient activity, hepatotoxicity, and gastrointestinal disorders, and thus have a great interest in the development of new acetylcholinesterase inhibitors that are harmless to the human body as substitutes for synthetic drugs. Particularly, some studies have been conducted to search for acetylcholinesterase inhibitors from natural products, but studies on acetylcholinesterase inhibitory activity using lactic acid bacteria have been insufficient. Therefore, development of lactic acid bacteria having acetylcholinesterase inhibitory activity is required.

Korean Patent Publication No. 10-2014-0064580 (2014.05.28.) Korean Patent Publication No. 10-2016-0065280 (2016.06.09.)

Juan Xiao, et al., PLoS One. 2014 Nov 14; 9 (11): e112773

The inventors of the present invention, while studying lactic acid bacteria having an acetylcholinesterase (AChE) inhibitory activity related to anti-dementia effects, a new Lactobacillus brevis strain isolated from kimchi, miso, salted fish, etc. It was found that the survival rate was high and exhibited acetylcholinesterase inhibitory activity.

Accordingly, an object of the present invention is to provide a novel Lactobacillus brevis strain having an acetylcholinesterase inhibitory activity and a composition comprising the same.

According to an aspect of the present invention, a Lactobacillus brevis BHN-LAB 127 (Accession No .: KCTC13441BP) strain having acetylcholinesterase inhibitory activity is provided.

In one embodiment, the strain may include a 16S rRNA of SEQ ID NO: 3, acid-resistant, bile-resistant, or a strain having resistance to pancreatic enzymes.

According to another aspect of the invention, there is provided a culture medium cultured by inoculating the strain.

According to another aspect of the present invention, a method of manufacturing a fermentation product comprising the step of fermenting using the strain is provided.

According to another aspect of the present invention, a fermented product prepared by the above manufacturing method is provided.

According to another aspect of the present invention, there is provided a pharmaceutical composition for treating Alzheimer's, comprising the strain or a culture medium thereof as an active ingredient.

According to another aspect of the present invention, there is provided a food composition for improving symptoms of Alzheimer's disease, comprising the strain or a culture medium thereof.

According to another aspect of the present invention, there is provided a food additive for improving Alzheimer's symptoms including the strain or a culture medium thereof.

According to the present invention, the Lactobacillus brevis BHN-LAB 127 strain (KCTC13441BP) isolated from traditional foods exhibits acid resistance, bile acid resistance, and resistance to pancreatic enzymes suitable for Koreans' diets, resulting in high intestinal survival and anti-dementia efficacy It was found to exhibit about 40% inhibitory activity against acetylcholinesterase related to.

Therefore, Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) of the present invention has anti-dementia efficacy that significantly inhibits acetylcholinesterase activity, so food processing, health as a material for a pharmaceutical composition for treating Alzheimer's disease and a food composition for improving Alzheimer's symptoms It can be usefully used in the functional food and pharmaceutical industries.

1 is a 16S rRNA gene sequence of Lactobacillus brevis BHN-LAB 127 strain.
2 is a phylogenetic tree of Lactobacillus brevis BHN-LAB 127 strain.
Figure 3 is a result showing the acetylcholinesterase inhibitory activity of Lactobacillus Brevis BHN-LAB 127 strain (Donepezil as a positive control).

The present invention provides a Lactobacillus brevis BHN-LAB 127 (Accession No .: KCTC13441BP) strain having acetylcholinesterase inhibitory activity.

The Lactobacillus Brevis BHN-LAB 127 (Accession No .: KCTC13441BP) strain is diluted in distilled water with traditional foods such as kimchi, miso, and salted fish, and then plated on a selective medium (bromine cresol purple flat plate medium) to be separated from the yellow colonies. Can be. After extracting the DNA of the isolated strain, PCR was performed using primers of SEQ ID NO: 1 (forward) and primers of SEQ ID NO: 2 (reverse), PCR products, all-in-one (Biofact) vectors, and E. coli DH5α. Gene transformation may be performed using a back. A 16S rRNA nucleotide sequence can be obtained by requesting a plasmid of a selected colony for which genetic transformation has been confirmed to a sequencing organization. As a result of nucleotide sequence analysis of the obtained 16S rRNA, a nucleotide sequence (SEQ ID NO: 3) of about 1,438 bp was obtained, showing 100% homology with the Lactobacillus brevis ATCC 14869 strain, and Lactobacillus brevis BHN-LAB 127 (Accession No .: KCTC13441BP) strain It was identified as.

The strain obtained by the above process has acid resistance, bile resistance and resistance to pancreatic enzyme. The strain may be inoculated into a liquid medium (artificial gastric juice environment) that maintains a pH of 2.5 by adding pepsin enzyme, followed by culturing for a period of time, followed by plating on a normal solid medium to further culture. As a result, it was confirmed that the number of viable cells according to the culture time in the artificial gastric juice environment was generally maintained. In addition, the strain can be cultured by plating on a solid medium containing bile acid (0.3% oxgal). As a result, it was confirmed that the strain maintains the viable cell count even in an environment containing bile acids. In addition, the strain may be inoculated into a liquid medium containing pancreatin (pancreatic enzyme) present in the small intestine, cultured for a period of time, and then further cultured by plating on a general solid medium. As a result, it was confirmed that the number of viable cells was maintained or increased according to the incubation time in the conditions containing the pancreatic enzyme.

In addition, the present invention provides a culture medium cultured by inoculating the strain. The acetylcholinesterase activity experiment can be performed using the culture solution obtained by inoculating the strain and culturing. After adding the culture solution and acetylcholinesterase to the phosphate buffer at pH 8 and reacting at room temperature, the enzyme reaction is initiated by adding 5,5'-dithio-bis- (2-nitrobenzoic acid) and acetylcholine iodide. Can be. Absorbance was measured at 412 nm using a spectrophotometer to confirm the degree of decomposition of acetylcholine. The negative control group was set as the untreated group, and the positive control group was set as the group treated with donepezil, a dementia treatment. As a result, the culture medium exhibited an inhibitory activity of about 40% against acetylcholinesterase related to anti-dementia efficacy.

In addition, the present invention provides a method of manufacturing a fermentation product comprising the step of fermentation using the strain. Food to be fermented using the strain may include fruits, vegetables, herbal medicines, mushrooms, seaweeds, and surplus by-products. In addition, the present invention provides a fermentation product fermented by the fermentation method.

In addition, the present invention provides a pharmaceutical composition for treating Alzheimer's, comprising the strain or a culture medium thereof as an active ingredient.

In addition, the present invention provides a food composition and food additives for improving Alzheimer's symptoms, including the strain or a culture medium thereof. The food composition comprising the strain may be utilized in the manufacture of foods having anti-dementia effects, such as powders, granules, tablets, capsules or beverages. In addition, food additives containing the strain can be easily utilized as additives in the production of functional foods or beverages for improving Alzheimer's symptoms.

The Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain of the present invention exhibits acid resistance, bile acid resistance, and resistance to pancreatic enzymes suitable for the Korean diet, so that it has a high intestinal survival rate and has anti-dementia effects that significantly inhibit acetylcholinesterase activity. It was shown. Therefore, the strain and its culture medium can be usefully used in the food processing, health functional food and pharmaceutical industries as a material for pharmaceutical compositions for treating Alzheimer's disease, food compositions for improving Alzheimer's symptoms, and food additives, as well as cosmetic compositions, immunity It can also be usefully used in various fields as a material for the enhancement composition, bowel disease composition, feed and feed additives.

Hereinafter, the present invention will be described in more detail through examples. However, the following examples are intended to illustrate the present invention, and the scope of the present invention is not limited thereto.

<Example>

1. Isolation of strains

After diluting 1 g of traditional fermented foods such as kimchi, miso, and salted fish in 10 ml of distilled water, yellow colonies were separated by plating on lactic acid bacteria selective medium (Brom Cresol Purple) plate medium (Difco, USA) through a multi-step dilution method. . The separated colonies were selected for excellent strains having acid resistance, bile resistance, and resistance to pancreatic enzyme through an experiment to confirm resistance to acid resistance, bile resistance, and pancreatic enzyme, and named BHN-LAB 127.

2. Identification of strains

The BHN-LAB 127 strain was extracted with DNA using the FastDNA SPIN for soil kit (MP biomedicals), and PCR was performed using the primers in Table 1. In addition, the composition of the PCR reaction solution is shown in Table 2 below.

  27 forward primer   5'-AGA GTT TGA TCM TGG CTC AG-3 '(SEQ ID NO: 1)   1492 reverse primer   5'-GGY TAC CTT GTT ACG ACT T-3 '(SEQ ID NO: 2)

  Components required for PCR reaction usage
(Μl)   DNA 1.0   forward PCR primer (12.5 pmol) 2.5   reverse PCR primer (12.5 pmol) 2.5   dNTP mixture (10mM) 2.0   10 × PCR buffer 5.0   Bovine serum albumin (BSA) 0.5   Taq DNA polymerase 0.5   Distilled water 36.0   Total volume 50.0

PCR reaction conditions were 95 ° C., 5 minutes, then 94 ° C. (1 minute) [denaturation step] → 58 ° C. (1 minute) [annealing step] → 72 ° C. (1 minute 30 seconds) [synthesis ( extension) After repeating the process of 30 cycles, an additional extension process of 72 ° C and 10 minutes is included, and the results of the PCR reactions are confirmed in agarose gel.

3. Gene transformation for strain identification

PCR was performed using DNA extracted from the BHN-LAB 127 strain, and gene transformation was performed to obtain a 16S rRNA sequence. For gene transformation, the all-in-one (Biofact) vector shown in Table 3 was used, and E. coli DH5α was used for transformation. After transformation, white colonies were selected by plating on LB solid medium containing kanamycin, X-gal and IPTG.

  Components required for colonization reaction usage
(Μl)   Vector (all-in-one) One   T4 ligase One   PCR products 3   2 × buffer 5   Total volume 10

The specific transformation method in the gene transformation is as follows. (a) The mixture containing the components of Table 3 was first reacted at room temperature for 2 hours. (b) E. coli DH5α is thawed on ice. (c) The reactant obtained in step (a) was added to E. coli DH5α and reacted at 42 ° C. for 90 seconds, followed by additional reaction for 30 minutes on ice. (d) The reactant obtained in step (c) is plated on an LB solid medium containing kanamycin, X-gal and IPTG, and then incubated at 37 ° C overnight.

4. 16S rRNA sequence analysis and identification of strain

The selected white colonies were extracted using a Qiagen spin miniprep kit (Qiagen), and the presence or absence of inserts was determined using the restriction enzyme EcoRI. A plasmid having an insert size of about 1,400 bp was selected and commissioned to Solgent (Daejeon) to identify it through 16S rRNA sequencing (see FIG. 1, SEQ ID NO: 3). As a result of identification of the BHN-LAB 127 strain, it was confirmed as Lactobacillus brevis It was named Lactobacillus brevis BHN-LAB 127 ( Lactobacillus brevis BHN-LAB 127, KCTC13441BP) (see FIG. 2).

5. Acid resistance experiment

The isolated strain was pre-incubated for 18 hours at 37 ° C. in MRS liquid medium (Difco, USA), and the culture medium passaged three times was used for acid resistance experiment. In order to prepare artificial gastric juice, after adding pepsin enzyme 1% to the MRS liquid medium, the pH was adjusted with hydrochloric acid (HCl) to prepare a MRS liquid medium with a pH of 2.5. The isolates were inoculated into the MRS liquid medium at pH 2.5, incubated at 37 ° C for 1 hour, 2 hours, and 3 hours, respectively, and then spread on MRS agar for 48 hours in a 37 ° C incubator. Colonies were counted and compared.

The initial number of individuals inoculated with the isolated strain was 5.1 × 10 ⁹ ± 0.6 cfu / ml, the number of live bacteria 1 hour after inoculation was 6.6 × 10 ⁹ ± 0.5 cfu / ml, and the number of live bacteria 2 hours after inoculation was 4.8 × 10 ⁹ ± 0.4 cfu / ml and the number of viable cells 3 hours after inoculation was 4.2 × 10 ⁹ ± 0.7 cfu / ml, confirming that the population was generally maintained (see Table 4). This shows that the number of individuals increases rather than 129% after 1 hour of inoculation compared to the initial number of inoculations of the isolated strains, and it is confirmed that it shows a high survival rate of 94% after 2 hours and 82% after 3 hours. Despite the artificial gastric juice environment, which is a very strong acidic condition of pH 2.5, Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) was confirmed to be an excellent acid resistance strain.

Strain name 0 hours 1 hours 2 hours 3 hours  Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) 5.1 × 10 ⁹ ± 0.6 cfu / ml 6.6 × 10 ⁹ ± 0.5 cfu / ml 4.8 × 10 ⁹ ± 0.4 cfu / ml 4.2 × 10 ⁹ ± 0.7 cfu / ml Survival rate 100% 129% 94% 82%

6. Bile resistance test

The isolated strains were pre-incubated for 18 hours at 37 ° C. in a MRS liquid medium, and then the culture medium of the three-passaged strains was smeared on MRS agar containing 0.3% oxgall and MRS agar not containing oxgal, respectively. After incubation for 48 hours in a 37 ℃ incubator, the colonies produced were counted and compared.

As shown in Table 5, under the condition not containing bile acid, the number of live cells of 5.1 × 10 ⁹ ± 0.5 cfu / ml was confirmed, and under the condition including bile acid, the number of live bacteria of 4.8 × 10 ⁹ ± 0.8 cfu / ml was confirmed. . The Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain showed a very high survival rate of 94% in media containing bile acids compared to the control group without bile acids. Therefore, it was confirmed that the Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain is an excellent bile acid-resistant strain.

Strain name Without bile acid Bile acid included Lactobacillus brevis
BHN-LAB 127
(KCTC13441BP) 5.1 × 10 ⁹ ± 0.5 cfu / ml 4.8 × 10 ⁹ ± 0.8 cfu / ml Survival rate 100% 94%

7. Pancreatic Resistance Test

The isolated strain was pre-incubated for 18 hours at 37 ° C. in MRS liquid medium, and the culture medium passaged three times was used for resistance test to pancreatic enzyme. In order to investigate the resistance to pancreatin, a pancreatic enzyme present in the small intestine, 0.1% of pancreatin was added to the MRS liquid medium to prepare a culture solution, and then inoculated isolate strains were inoculated at 37 ° C for 1 hour, 2 hours, 3 hours, After culturing, the cells were spread on MRS agar, and cultured in a 37 ° C. incubator for 48 hours, and the resulting colonies were counted and compared.

As shown in Table 6, the initial viable count of the isolated strain was 5.1 × 10 ⁹ ± 0.4 cfu / mL, the viable count after 1 hour of inoculation was 5.0 × 10 ⁹ ± 0.7 cfu / mL, and the viable count after 2 hours of inoculation was 8.3. × 10 ⁹ ± 0.6 cfu / ml, the number of viable cells after 3 hours of inoculation was confirmed to be 5.0 × 10 ⁹ ± 0.7 cfu / ml. The Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain was found to have a survival rate of 98% after 1 hour of inoculation, a survival rate of 162% after 2 hours of inoculation, and a survival rate of 98% after 3 hours of inoculation compared to the number of live cells inoculated, and 2 hours of inoculation. In the survival rate afterwards, it was confirmed that the number of bacteria increased more than 60%. As a result of confirming resistance to pancreatic enzymes, the Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain showed a high survival rate in an environment containing pancreatic enzymes, confirming that it is an excellent strain for pancreatic enzyme resistance.

Strain name 0 hours 1 hours 2 hours 3 hours Lactobacillus brevis
BHN-LAB 127
(KCTC13441BP) 5.1 × 10 ⁹ ± 0.4 cfu / ml 5.0 × 10 ⁹ ± 0.7 cfu / ml 8.3 × 10 ⁹ ± 0.6 cfu / ml 5.0 × 10 ⁹ ± 0.7 cfu / ml Survival rate 100% 98% 162% 98%

8. Acetylcholinesterase (AChE) activity experiment

100 μl of 100 mM phosphate buffer (pH 8.0) was mixed well with 10 μl of the isolated strain culture supernatant and 10 μl of AChE (0.5 U / ml) and reacted at room temperature (25 ° C.) for 15 minutes. Thereafter, 10 μl of 5,5'-dithio-bis- (2-nitrobenzoic acid) [5,5'-dithio-bis- (2-nitrobenzoic acid), DTNB, 10 mM] and acetylcholine iodide , ATCI, 14 mM) 10 μl was added to initiate the enzyme reaction. After the reaction, the absorbance was measured at 412 nm using a spectrophotometer, and the degree of decomposition of acetylcholine was expressed as a percentage change in absorbance for the untreated group (negative control). As a positive control, Donepezil (10 μl), a dementia treatment agent of the 'acetylcholinesterase inhibitor' family, was used (see FIG. 3).

It was confirmed that the acetylcholinesterase inhibitory activity of Donepezil, a positive control group, was 80.2 ± 4.5%, and the acetylcholinesterase inhibitory activity of the culture supernatant of the isolate strain was 40.1 ± 5.2%. Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) culture supernatant showed that acetylcholinesterase inhibitory activity was lower than that of donepezil, a drug used to treat dementia, but donepezil was a commercially available dementia drug. Considering that, the supernatant of Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain culture is also considered to be excellent because it exhibits an inhibitory activity of acetylcholinesterase of 40% or more.

Therefore, the Lactobacillus brevis BHN-LAB 127 (KCTC13441BP) strain exhibits acid resistance, bile resistance and resistance to pancreatic enzymes, and thus has a high intestinal survival rate, and has anti-dementia effects that inhibit acetylcholinesterase activity, thereby preventing Alzheimer's As a material, it can be usefully used in food processing, dietary supplement and pharmaceutical industries.

Korea Research Institute of Bioscience and Biotechnology KCTC13441BP 20171220

<110> BHNBIO CO., LTD <120> Lactobacillus brevis BHN-LAB 127 having acetylcholinesterase          inhibitory activity <130> P181021 <160> 3 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> 27 forward primer <400> 1 agagtttgat cmtggctcag 20 <210> 2 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> 1492 reverse primer <400> 2 ggytaccttg ttacgactt 19 <210> 3 <211> 1438 <212> RNA <213> Lactobacillus brevis <400> 3 gcaagtcgaa cgagcttccg ttgaatgacg tgcttgcact gatttcaaca atgaagcgag 60 tggcgaactg gtgagtaaca cgtggggaat ctgcccagaa gcaggggata acacttggaa 120 acaggtgcta ataccgtata acaacaaaat ccgcatggat tttgtttgaa aggtggcttc 180 ggctatcact tctggatgat cccgcggcgt attagttagt tggtgaggta aaggcccacc 240 aagacgatga tacgtagccg acctgagagg gtaatcggcc acattgggac tgagacacgg 300 cccaaactcc tacgggaggc agcagtaggg aatcttccac aatggacgaa agtctgatgg 360 agcaatgccg cgtgagtgaa gaagggtttc ggctcgtaaa actctgttgt taaagaagaa 420 cacctttgag agtaactgtt caagggttga cggtatttaa ccagaaagcc acggctaact 480 acgtgccagc agccgcggta atacgtaggt ggcaagcgtt gtccggattt attgggcgta 540 aagcgagcgc aggcggtttt ttaagtctga tgtgaaagcc ttcggcttaa ccggagaagt 600 gcatcggaaa ctgggagact tgagtgcaga agaggacagt ggaactccat gtgtagcggt 660 ggaatgcgta gatatatgga agaacaccag tggcgaaggc ggctgtctag tctgtaactg 720 acgctgaggc tcgaaagcat gggtagcgaa caggattaga taccctggta gtccatgccg 780 taaacgatga gtgctaagtg ttggagggtt tccgcccttc agtgctgcag ctaacgcatt 840 aagcactccg cctggggagt acgaccgcaa ggttgaaact caaaggaatt gacgggggcc 900 cgcacaagcg gtggagcatg tggtttaatt cgaagctacg cgaagaacct taccaggtct 960 tgacatcttc tgccaatctt agagataaga cgttcccttc ggggacagaa tgacaggtgg 1020 tgcatggttg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa 1080 cccttattat cagttgccag cattcagttg ggcactctgg tgagactgcc ggtgacaaac 1140 cggaggaagg tggggatgac gtcaaatcat catgcccctt atgacctggg ctacacacgt 1200 gctacaatgg acggtacaac gagttgcgaa gtcgtgaggc taagctaatc tcttaaagcc 1260 gttctcagtt cggattgtag gctgcaactc gcctacatga agttggaatc gctagtaatc 1320 gcggatcagc atgccgcggt gaatacgttc ccgggccttg tacacaccgc ccgtcacacc 1380 atgagagttt gtaacaccca aagccggtga gataaccttc gggagtcagc cgtctaag 1438

Claims (12)

Lactobacillus brevis BHN-LAB 127 (Accession No .: KCTC13441BP) strain comprising the 16S rRNA of SEQ ID NO: 3 and having acetylcholinesterase inhibitory activity. delete The strain according to claim 1, wherein the strain has acid resistance, bile resistance and resistance to pancreatic enzyme. A culture medium cultured by inoculating the strain of any one of claims 1 and 3. Method of producing a fermented product comprising the step of fermenting using the strain of any one of claims 1 and 3. Fermented product prepared according to claim 5. A pharmaceutical composition for inhibiting acetylcholinesterase activity, comprising the strain of any one of claims 1 and 3 as an active ingredient. A pharmaceutical composition for inhibiting acetylcholinesterase activity, comprising the culture medium of claim 4 as an active ingredient. A food composition for inhibiting acetylcholinesterase activity, comprising the strain of any one of claims 1 and 3. A food composition for inhibiting acetylcholinesterase activity, comprising the culture medium of claim 4. A food additive for inhibiting acetylcholinesterase activity, comprising the strain of any one of claims 1 and 3. A food additive for inhibiting acetylcholinesterase activity, comprising the culture medium of claim 4.

Images