(b) Description of the Related Art Helicobacter pylori is a gram-negative microorganism. In 1983, Warren and Maeshall first isolated and characterized Helicobacter in the human gastric mucous membrane. Helicobacter initially was called Campylobacter pyloridis, and was later called Helicobacter pylori after morphology in the body and name of the habitat, pylorus.

Helicobacter pylori cause gastritis and gastric carcinoma (Dubois, A.

1995. Spiral bacteria in the human stomach: the gastric Helicobacters.

Emerg. Infect. Dis. 1: 79-85; Slomiany, B. L. and A. Slomiany. 1992.

Mechanism of Helicobacter pylori pathogenesis: focus on mucus. J. Clin.

Gastroenterol. 14 Suppl 1 S114-21). Once Helicobacter pylori infects, it remains for several decades and is not eliminated naturally and thus, Helicobacter pylori is a major cause of chronic gastritis.

Helicobacter pylori infect through the intake of food and Helicobacter pylori attach to the gastric mucous membrane and the duodenal mucous membrane. A disease-causing factor of Helicobacter pylori is a urease secreted for surviving in highly acidic condition of stomach, a flagellum for maintaining mobility and the outer membrane protein having adherence to the gastric mucous membrane.

In the attachment of Helicobacter pylori to the human gastric epithelium, Helicobacter pylori binds the same antigens identified by antigens of red blood cells (Alkout, A. M., C. C. Blackwell, D. M. Weir, 1. R.

Poxton, R. A. Elton, W. Luman, and K. Palmer. 1997. Gastroenterol. 112: 1179-1187; Boren, T., P. Flak, K. A. Roth, G. Larson, and S. Normark. 1993.

Science 262: 1892-1895; Clyne, M. and B. Drumm. 1997. Gastroenterol.

113: 72-80). Antigens like the Lewis antigen isolated in human blood type O, are identified in the gastric mucous membrane. Therefore, gastritis breaks out mostly in blood type O cases (Heneghan, M. A., A. P. Moran, K.

M. Feeley, E. L. Egan, J. Goulding J, C. E. Connolly, and C. F. McCarthy.

1998. FEMS Immunol. Med. Microbiol. 20: 257-266; Kobayashi, K., J.

Sakamoto, Y. Yamamura, T. Kito, H. Inagaki, T. Watanabe, and H. Nakazato.

1991. Nippon Geka Gakkai Zasshi 92 : 813-819; Kobayashi, K., J.' Sakamoto, Y. Kito, Y. Yamamura, T. Koshikawa, M. Fugita, T. Watanabe, and H. Nakazato. 1993. Am. J. Gastroenterol. 88: 919-924).

For the removal of Helicobacter pylori, antibiotic drugs, restrainers on proton pumping, and gastric acid removers have been used. It is difficult

that Helicobacter was cultured with a large scale, thereby development of a vaccine using the entire germ remains unsuccessful. The method using antibiotic drugs has a side effect in that the Helicobacter pylori become resistant to the antibiotic and the possibility of re-infection is not prevented.

The method using stomach acid remover that suppresses stomach acid secretion is a not basic solution. In addition, although the vaccine using urease has been developed, it is not effective. In the future, the development of a vaccine against Helicobacter pylori will be difficult, due to its complex culture conditions, which make it difficult to determine an active area for a vaccine.

Lactobacillus produce lactate and an unknown material, and it inhibit Helicobacter pylori growth. In the case of Lactobacillus salivarius, the culture solution is able to inhibit growth of Helicobacter pylori (Aiba, Y., N.

Suzuki, A. M. Kabir, A. Takagi, and Y. Koga. 1998 Am. J. Gastroenterol. 93: 2097-2101). Also, Lactobacillus inhibits the binding between the Lewis antigen and Helicobacter pylori (Lee, Y., E. Shin, J. Lee, and J. H. Park. 1999.

Lactobacillus acidophilus inhibits the Helicobacter pylori adherence. J.

Microb. Biotech. 9: 794-797).

SUMMARY OF THE INVENTION Accordingly, the present invention is designed by the necessities of the prior art, and it is an object of the present invention to provide bacteria that suppress stomach ulcers.

Also, it is an object to provide bacteria that inhibits the adherence of Helicobacter pylori..

Also, it is an object to provide bacteria that inhibits the activity of Helicobacter pylori.

Also, it is an object to provide bacteria that inhibits the growth of Helicobacter pylori.

Also, it is an object to provide bacteria that inhibits the growth of bacillus causing food poisoning.

Also, it is an object to provide bacteria that inhibits the growth of anaerobic bacteria or bacillus causing acne.

In order to achieve these objects, the present invention also provides a lactic acid bacteria containing inhibitory activity on Helicobacter pylori adherence of the gastric mucous membrane. More preferably, the lactic acid bacteria is at least one selected from the group consisting of Lactobacillus coprophilus PL 9001 (KCCM-10245), Enterococcus durans PL 9002 (KCCM-10246), Streptococcus faecalis PL 9003 (KCCM-10247), Lactobacillus coprophilus PL 9004 (KCCM-10248). Lactobacillus fermentum PL 9005 (KCCM-10250), and Lactobacillus fermentum PL 9006 (KCCM- 10251).

The present invention also provides a medicine for the prevention or treatment of infectious bacillus by the use of lactic acid bacteria. The bacillus is Helicobacter pylori, bacillus causing food poisoning, anaerobic bacteria or bacillus causing acne.

The present invention also provides a cosmetic composition containing lactic acid bacteria or spent culture of lactic acid bacteria.

The present invention also provides a food additive containing lactic acid or spent culture of lactic acid bacteria.

The present invention also provides a food prepared by fermentation with lactic acid bacteria.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a slide showing Helicobacter pylori attachment to glycolipid isolated from red blood cells, wherein the greater the level of glycolipid (0 ug, 0.14 ug, 1.4 ug, 14 ug, 70 ug, 140 ug, 280 ug), the greater the degree of attachment.

FIG. 2 is TLC plate showing that lactobacillus inhibit Helicobacter pylori adherence on glycolipid.

FIG. 3 shows that materials produced from PL bacteria, suppress the growth of Helicobacter pylori.

FIG. 4 is a schematic diagram showing the diameter of the growth- inhibited-zone for Helicobacter pylori.

FIG. 5 is a graph showing the growth-inhibited degree of Helicobacter pylori by spent culture of PL bacteria.

FIG. 6 is a graph showing growth inhibition of bacillus causing food

poisoning by Lactobacillus coprophilus PL 9001.

FIG. 7 is a graph showing growth inhibition of bacillus causing food poisoning by Enterococcus durans PL 9002.

FIG. 8 is a graph showing growth inhibition of bacillus causing food poisoning by Streptococcus faecalis PL 9003.

FIG. 9 is a graph showing growth inhibition of bacillus causing food poisoning by Lactobacillus coprophilus PL 9004.

FIG. 10 is a graph showing growth inhibition of bacillus causing food poisoning by Lactobacillus ferment PL 9005.

FIG. 11 is a graph showing growth inhibition of bacillus causing food poisoning by Lactobacillus fermentum PL 9006.

FIG. 12 is a graph showing growth inhibition of bacillus causing acne by PL bacteria.

FIG. 13 is a graph showing effect of immune improvement by PL bacteria.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be explained in more detail.

The present invention provides bacteria, inhibiting the adherence of Helicobacter pylori to the gastric mucous membrane. The bacteria are lactic acid bacteria and preferably Lactobacillus sp. or Enterococcus sp.

More preferably, the bacteria are Lactobacillus coprophilus, Enterococcus durans, Streptococcus faecalis and Lactobacillus fermentum, and most preferably, are Lactobacillus coprophilus PL 9001, Enterococcus durans PL

9002, Streptococcus faecalis PL. 9003, Lactobacillus coprophilus PL 9004, Lactobacillus ferment PL 9005 and Lactobacillus fermentum PL 9006.

The Streptococcus faecalis is identical to Enterococcus faecalis and then it was termed as Streptococcus faecalis PL 9003.

The PL bacteria (Lactobacillus coprophilus PL 9001, Enterococcus durans PL 9002, Streptococcus faecalis PL 9003, Lactobacillus coprophilus PL 9004, Lactobacillus fermentum PL 9005 and Lactobacillus ferment PL 9006) are internationally deposited in the Korean culture center of microorganisms and given an accession number. The accession number given by the international depositary authority are KCCM-10245 for Lactobacillus coprophilus PL 9001, KCCM-10246 for Enterococcus durans PL 9002, KCCM-10247 for Streptococcus faecalis PL 9003, KCCM-10248 for Lactobacillus coprophilus PL 9004, KCCM-10250 for Lactobacillus ferment PL 9005 and KCCM-10251 for, Lactobacillus ferment PL 6.

Also, the present invention provides a gastric ulcer restrainer. The gastric ulcer restrainer contains Lactobacillus sp. or Enterococcus sp. and more preferably, contains the PL bacteria. The Lactobacillus sp. or Enterococcus sp. have inhibitory activity on Helicobacter pylori adherence to the gastric mucous membrane as well as for Helicobacter pylori growth.

These effects are confirmed with Lactobacillus coprophilus PL 9001 (KCCM- 10245), Enterococcus durans PL 9002 (KCCM-10246), Streptococcus faecalis PL 9003 (KCCM-10247), Lactobacillus coprophilus PL 9004 (KCCM- 10248). Lactobacillus fermentum PL 9005 (KCCM-10250), and Lactobacillus

ferment PL 9006 (KCCM-10251).

Also, the PL bacteria produce materials that inhibit the growth of Helicobacter pylori and adherence to the gastric mucous membrane, and thus, spent culture of the PL bacteria can be used for the gastric ulcer restrainer. The PL bacteria have antibiotic-resistant activity, acid-resistant activity, and bile-resistant activity, and are stable in vivo, whereby the PL bacteria can be provided as raw bacteria, dehydrated bacteria and dead bacteria.

The gastric ulcer restrainer of the present invention can be adjusted for use. The drug is administered orally, by injection, and ointment.

Preferably, the drug would be administered orally or by injection; but most preferably, the drug would be administered orally. The gastric ulcer restrainer can be prepared as a single compound drug or complex drug and the complex drug can contain more pharmaceutically acceptable material.

It is also reasonable that the blending ratio of the PL bacteria is controlled by using methods applicable to the type of drug.

The spent culture of PL bacteria is preferably solution without bacteria that is prepared by filtering or centrifuging culture fluid, and more preferably, the spent culture is a supernatant solution prepared by inoculating the PL bacteria in MRS liquid broth, culturing at 37 °C, for 16 hr- 48hr, and centrifuging.

Also, Lactobacillus sp. and Enterococcus sp. can be applied to inhibit the activity of other bacilli. More preferably, the PL bacteria (PL9001,

PL9002, PL9003, PL9004, PL9005, and PL9006), fragmented cell wall of the PL bacteria or spent culture of the PL bacteria can be used to inhibit the growth of bacillus causing food poisoning, anaerobic bacteria, bacillus causing acne and so on.

The anaerobic bacteria are general disease-causing germs. The anaerobic bacteria prefer tetanus bacteria, gas gangrene bacteria and Clostridium sp., and more preferably, Clostridium sp.

The bacillus causing food poisoning is a listeria, a dysentery bacillus, diarrhea bacillus, 0157, and bacillus mediated food poisoning.

Also, Lactobacillus sp. and Enterococcus sp. have activity for improving immunity and can be applied to improve health.

Also, the Lactobacillus sp. and the Enterococcus sp. of the present invention is preferably used for feed, feed additives, food, food additives, medicine, or cosmetic composition. Moreover, the cell wall prepared by grinding the PL bacteria, raw, dead and dehydrated bacteria, and spent culture, are contained. The Lactobacillus sp. and the Enterococcus sp. of the present invention underwent a toxic test, which confirmed that the lactic acid bacteria were safe in vivo.

The food or food additives prefer yogurt, baby food, dairy goods, cheese, Kimchi, drinks, or food and is prepared by fermenting with the Lactobacillus sp. or the Enterococcus sp. of the present invention.

The medicine contained the Lactobacillus sp. or the Enterococcus sp. of the present invention is preferably formulated as a solution, powder, solid,

or capsule and most preferably, is a capsule type prepared by dehydrating.

The cosmetic composition can be used for massage, pack, solution type ointment, or as an additive in order to prevent and treat bacillus, such as acne. The cosmetic composition is preferably for external application.

Also, the present invention provides a material for inhibiting the growth of harmful bacteria. The material for inhibiting the growth of harmful bacteria is able to suppress the growth of Helicobacter pylori, anaerobic bacteria, bacillus causing acne or bacillus causing food poisoning and is prepared as spent culture in bacillus culture. Thus, the material for inhibiting the growth of harmful bacteria produced by Lactobacillus sp. or Enterococcus sp., can also. be used for the treatment and prevention of bacteria.

As mentioned above, Lactobacillus coprophilus PL 9001, Enterococcus durans PL 9002, Streptococcus faecalis PL 9003, Lactobacillus coprophilus PL 9004. Lactobacillus ferment PL 9005, and Lactobacillus fermentum PL 9006 of the present invention suppress the growth and adhesion of Helicobacter pylori. Also, PL bacteria, fragment of PL bacteria, or spent culture of PL bacteria is able to be used for the treatment and prevention of bacteria. PL bacteria, and spent culture of PL bacteria have activity of inhibiting the growth of bacillus causing food poisoning, bacillus causing acne and anaerobic bacteria, and improving effect of immunity, thereby PL bacteria, and spent culture of PL bacteria can be used for treatment and prevention.

The present invention will be explained in more detail with reference to the following Examples. However, the following Examples are to illustrate the present invention and the present invention is not limited to them.

EXAMPLE MRS+BPB are prepared with MRS broth (Difco, bacto proteose peptone No. 3 10 g, bacto beef extract 10 g, bacto yeast extract 5 g, bacto dextrose 20 g, polysorbate 80 g, ammonium citrate 2 g, sodium acetate 5 g, magnesium sulfate 0.1 g, manganese sulfate 0.05 g, dipotassium phosphate 2 g/L) containing 0.002 % of bromophenol blue. Kimchi samples are diluted with 10 X pepton solutions. 0.1 mL of the diluted solution are inoculated in MRS+BPB medium and spread. The feces from an. infant were picked by a cotton swab and inoculated in MRS+BPB medium. After incubation for 3-4 days in an incubator (25 °C), Lactobacillus sp. were observed and isolated according to the formed colonies.

In order to distinguish a group of Lactobacillus sp., MRS solid media was added with BPB which displays yellow color in pH 3.0 and purple color in pH 4.6, and Lactobacillus sp. was classified according to a coloration by BPB. The coloration was determined by producing degrees of lactic acid, pH-resistant, and the length of life. P. acidolactic and S. faecalis are normal lactic acid bacterium, L. mesenteroides and L. brevis are abnormal lactic acid bacterium, and L. plantarum are random lactic acid bacterium. Due to ferment lactic acid, the production of a lot of lactic acid, S faecalis has

formed a white colony where the medium of it has changed to light yellow.

L. mesenteroides ferment abnormally lactic acid and produce a low level of lactic acid, the colony is deep blue as a whole and has a small size without a ring. Lactobacillus sp. have a light blue ring, being deep blue in center, or generally white color, and the size of the colony is large. P. acidolactic and L. mesenteroides express a deep-blue for a short life time and low pH.

For example, L. mesenteroides cannot growth below pH 4.8.

All bacteria of the present invention formed a white colony of over 0.3 mm (diameter) and are classified into Lactobacillus sp.

Bacteria were isolated from single colony and, according to Bergy's manual of systematic bacteriology, exhibited morphological and biochemical properties. After performed by gram stain and catalase-reaction, the bacteria were observed in an API system (La Balme-les-Grottes, France).

After the colony was picked with the cotton wrap, it was floated with 2 ml of distilled water. The floating solution was added to the API 50 CHL media and mixed. Each 200 ul of the mixture was inoculated into 50 tubes, was covered with mineral oil, and then was incubated at 37°C, for 48 h. The fermentation pattern of the 49 kinds of saccharide were analyzed with API 50 CH and API 50 CHL, the resulting data was fed into the ATB identification computer system (bio Merieux France), and then bacterium was identified.

The result of the first bacterium among the isolated microorganisms is presented in Table 1.

[Table 1]

Strip No. 1 Strip No. 2 Strip No. 3 Strip No. 4 Strip No. 5 tube/substrate tube/substrate tube/substrate tube/substrate tube/substrate -Control-Galactose-D-Mannoside-Melibiose-D-Turanose -Giycerol + D-Glucose-D-Glucoside + Saccharose-D-Lyxose -Erythritol + D-Fructose + Glucosamine-Trehalose-D-Tagatose -D-Arabinose + D-Mannose + Amygdaline-Inuline-D-Fucose -L-Arabinose-L-Sorbose + Arbutine-Melezitose-L-Fucose -Ribose-Rhamnose + Esculine-D-Raffinose-D-Arabitol +D-Xylose-Dulcitol + Salicine-Amidon-L-Arabitol -L-Xylose-Inositol + Cellobiose-Glycogene + Gluconate -Adonitol-Mannitol + Maltose-Xylitol-2-Gluconate -Xyloside-Sorbitol-Lactose + Gentiobiose-5-Gluconate The first bacterium was identified 99.9 % to Lactobacillus coprophilus and 0.1 % to Lacto. brevis. 16S rRNA of the first bacterium was sequenced with 16S rRNA gene kit (Perkin Elmer Applied Biosystem) and sequence was shown in Sequence No. 1. The base sequence of Sequence No. 1 is identical to 16S rRNA base sequence of Lactobacillus coprophilus as the result of BLAST search (http ://www. ncbi. nlm. nih. gov/blast) other name of the Lactobacillus coprophilus is Weissella confusa and Lactobacillus confuses.

The bacterium was deposited under Lactobacillus 9001 (KFCC-11240) in the Korean federation of culture collections, and internationally deposited under Lactobacillus coprophilus PL 9001 (KCCM-10245) in the Korean culture center of microorganisms.

The second bacterium was characterized with API STREP Kit and the result was presented in Table 2.

[Table 2] VP + ARA + HIP MAN + ESC + SOR PYRA + LAC AGAL TRE GUR - INU GAL - RAF PAL AMD LAP GLYG ADH +-HEM RIB + For the second bacterium was not classified, 16S rRNA base

sequence was analyzed. 16S rRNA base sequence of the second bacterium is Sequence No. 2, and identical to that of Enterococcus durans. According to the sequencing result, the second bacterium was called Enterococcus durans PL 9002, was deposited under Enterococcus durans PL9002 (KFCC- 11241) in the Korean federation of culture collections, and internationally deposited under Enterococcus durans PL 9002 (KCCM-10246) in the Korean culture center of microorganisms.

The result of the third bacterium is presented in Table 3.

[Table 3] VP + ARA HIP-MAN + ESC + SOR + PYRA + LAC + AGAL - TRE + GUR - INU - GAL - RAF - PAL - AMD + LAP + GLYG ADH +-HEM RIB The third bacterium was identified 99.1 % to Streptococcus faecalis

(Enterococcus faecalis). The bacterium was deposited under Streptococcus faecalis 9003 (KFCC-11242) in the Korea federation of culture collections, and internationally deposited under Streptococcus faecalis PL 9003 (KCCM-10247) in the Korea culture center of microorganisms.

16S rRNA base sequence of the third bacterium was analyzed as Sequence No. 3, and identical to that of Streptococcus faecalis (Enterococcus faecalis).

The result of the fourth bacterium is presented in Table 4.

[Table 4] Strip No. 1 Strip No. 2 Strip No. 3 Strip No. 4 Strip No. 5 tube/substrate tube/substrate tube/substrate tube/substrate tube/substrate -Control-Galactose-D-Mannoside-Melibiose-D-Turanose -Glycerol + D-Glucose-D-Glucoside + Saccharose-D-Lyxose -Erythritol + D-Fructose + Glucosamine-Trehalose-D-Tagatose -D-Arabinose + D-Mannose + Amygdaline-inuline-D-Fucose +L-Arabinose-L-Sorbose-Arbutine-Melezitose-L-Fucose -Ribose-Rhamnose + Esculine D-Raffinose-D-Arabitol + D-Xylose-Dulcitol + Salicine-Amidon-L-Arabitol -L-Xylose-Inositol + Cellobiose-Glycogene + Gluconate -Adonitol-Mannitol + Maltose-Xylitol-2-Gluconate -Xyloside-Sorbitol-Lactose + Gentiobiose-5-Gluconate The fourth bacterium was identified 98.2 % to Lactobacillus. coprophilus and 1.3 % to Lacto, brevis. Also, 16S rRNA base sequence of the bacterium was analyzed as Sequence No. 4, and identical to that of Lactobacillus coprophilus as the result of BLAST search (http://www. ncbi. nlm. nih. gov/blast). The bacterium was deposited under Lactobacillus coprophilus PL 9004 (KFCC-11243) in the Korea federation of culture collections, and internationally deposited under Lactobacillus coprophilus PL 9-4 (KCCM-10248) in the Korean culture center of microorganisms The result of the fifth bacterium is presented in Table 5.

[Table 5] Strip No. 1 Strip No. 2 Strip No. 3 Strip No. 4 Strip No. 5 tube/substrate tube/substrate tube/substrate tube/substrate tube/substrate -Control + Galactose-D-Mannoside + Melibiose-D-Turanose

-Glycerol + D-Glucose-D-Glucoside + Saccharose-D-Lyxose -Erythritol + D-Fructose + Glucosamine-Trehalose-D-Tagatose -D-Arabinose + D-Mannose-Amygdaline-Inuline-D-Fucose -L-Arabinose-L-Sorbose-Arbutine-Melezitose-L-Fucose +Ribose-Rhamnose-Esculine + D-Raffinose-D-Arabitol -D-Xylose-Dulcitol-Salicine-Amidon-L-Arabitol -L-Xylose-Inositol-Cellobiose-Glycogene + Gluconate -Adonitol-Mannitol + Maltose-Xylitol-2-Gluconate -Xyloside-Sorbitol-Lactose-Gentiobiose-5-Gluconate The fifth bacterium was identified 93.2 % to Lactobacillus fermentum and 6.7 % to Leuoconostoc lactis. Also, 16S rRNA base sequence of the bacterium was analyzed as Sequence No. 5, and identical to that of Lactobacillus fermentum as the result of BLAST search (http://www. ncbi. nlm. nih. gov/blast). The bacterium was deposited under Lactobacillus fermentum PL 9005 (KCCM-10250) in the Korean culture center of microorganisms The result of the sixth bacterium is presented in Table 6.

[Table 6] Strip No. 1 Strip No. 2 Strip No. 3 Strip No. 4 Strip No. 5 tube/substrate tube/substrate tube/substrate tube/substrate tube/substrate -Control + Galactose + D-Mannoside + Melibiose-D-Turanose -Glycerol + D-Glucose-D-Glucoside + Saccharose-D-Lyxose -Erythritol + D-Fructose-Glucosamine + Trehalose-D-Tagatose -D-Arabinose + D-Mannose-Amygdaline-Inuline-D-Fucose -L-Arabinose-L-Sorbose-Arbutine-Melezitose-L-Fucose + Ribose-Rhamnose-Esculine + D-Raffinose-D-Arabitol -D-Xylose-Dulcitol-Salicine-Amidon-L-Arabitol -L-Xylose-Inositol-Cellobiose-Glycogene-Gluconate -Adonitol-Mannitol + Maltose-Xylitol-2-Gluconate -Xyloside-Sorbitol + Lactose + Gentiobiose-5-Gluconate The sixth bacterium was identified 94.4 % to Lactobacillus fermentum and 5.4 % to Lactobacillus lactis. Also, 16S rRNA base sequence of the bacterium was analyzed as Sequence No. 6, and identical to that of Lactobacillus ferment as the result of BLAST search

(http://www. ncbi. nlm. nih. gov/blast). The bacterium was deposited under Lactobacillus fermentum PL 9006 (KCCM-10251) in the Korean culture center of microorganisms TEST 1. Test for inhibiting adherence of Helicobacter pylori on the gastric mucous membrane.

(1) Preparation of bacteria Helicobacter pylori (ATCC 43504) cultured for 48 hr on a Brucella solid medium [Brucella broth, fungizone (2.5 g/ml amphotericin B), and Skirrow's supplement (0.16 mg/ml polymyxin B, 5 mg/ml vancomycin, 2.5 mg/ml trimethoprim)] supplemented with 10% horse serum under 5-10% COg incubator. Cultured cells were collected by scraping, washed twice with phosphate-buffered saline (PBS, pH 7.4), and then kept in 10 mM Tris-CI buffer at-20 C until used.

The bacteria prepared by EXAMPLE 1 referred as PL bacteria. The PL bacteria (Lactobacillus coprophilus PL 9001 (KCCM-10245), Enterococcus durans PL 9002 (KCCM-10246), Streptococcus faecalis PL 9003 (KCCM-10247), Lactobacillus coprophilus PL 9004 (KCCM-10248).

Lactobacillus ferment PL 9005 (KCCM-10250), and Lactobacillus ferment PL 9006 (KCCM-10251)) were grown for 24 h, at 37°C in MRS broth, collected by centrifugation, pellet washed 2-3 times with PBS (pH 7.4), and then kept in 10 mM Tris-cl at-20 C until used.

(2) Extraction and purification of glycolipid Glycolipid was isolated from human O-type red blood cells (RBCs),

as described in the report with a slight modification (Lee, Y., E. Shin, J. Lee, and J. H. Park. 1999. J. Microb. Biotech. 9: 794-797). The human 0-type RBCs were dispersed in a minimum volume of water, repeated process of freezing at-70 C and melting, and then destructed. After the lysate was fixed, the supernatant was removed. The remains collected and extracted as mentioned below. The remains were mixed with a chloroform-methanol mixture (2: 1, v/v) and a lower phase (containing lipid) collected. The material of lipid layer was dried in a rotary vacuum evaporator. The dried solid was dissolved in chloroform containing 2 % of methanol and loaded on a column (bed volume = 20 ml) that was equilibrated with silicic acid (H2Si03).

The column fraction was collected by sequentially eluting with chloroform, acetone-methanol (3: 1, v/v), and methanol. The methanol fraction was then dried in a rotary vacuum evaporator, dissolved in a minimum volume of methanol, and stored in microcentrifuge tube at-70 C.

(3) Competitive lipid binding assay Competitive lipid binding assay was performed with a slight. modification (Lee, Y., E. Shin, J. Lee, and J. H. Park. 1999. J. Microb. Biotech.

9: 794-797). The extracted glycolipid (14 ug/5 ul) was spotted on a thin layer chromatography plate. (TLC, Merck, Kreselge160, EM Separations, Gibbstown, NJ, U. S. A) The plate was soaked in 10 mM Tris buffer (pH 7.6) containing 3 % gelatin for 2 h at 37°C to prevent non-specific binding. The TLC plate was rinsed two times with the same buffer and was added culture solution of PL bacteria (OD6oo=1, CFU 2.4 X 10"). After 5 to 30 min, H.

pylori (2.0 X 1 o8 CFU) was added and mixture was gently agitated for 2 hr at 37°C. In addition, except PL bacteria, the control plate was reacted with only Helicobacter pylori as described above, and agitated for 2 hr. The plate was rinsed 3 times with the same buffer for 10 min/time. The first antibody, rabbit antiserum raised against H. pylori was added to the buffer (1: 600) and further incubated for 2 hr at room temperature with gentle shaking. After the plate was washed to remove the first antibody (1: 1000), IgG, the second antibody conjugated with alkaline phosphatase, was added to the plate and the plate was incubated for 1 hr at room temperature. A chromogenic reaction was observed by adding BCIP/NBT (5-bromo-4-chloro- 3-indolyl phosphate disodium salt/nitro blue tetrazolium chloride). Therefore, the chromogenic level shows the adherence degree of Helicobacter pylori on glycolipid derived from red blood cells.

FIG. 1 is TCL plate showing that bound Helicobacter pylori to glycolipid from red blood cells, the amounts of bound H. pylori increased linearly as the amount of spotted glycolipid (0 ug, 0.14 ug, 1.4 ug, 14 ug, 70 ug, 140 ug, 280 ug) increased.

FIG. 2 is TLC plate observed whether or not Lactobacillus inhibit adherence of Helicobacter pylori on glycolipid. PL bacteria (Lactobacillus coprophilus PL 9001 (KCCM-10245), Enterococcus durans PL 9002 (KCCM- 10246), Streptococcus faecalis PL 9003 (KCCM-10247), Lactobacillus coprophilus PL 9004 (KCCM-10248). Lactobacillus fermentum PL 9005 (KCCM-10250), and Lactobacillus fermentum PL 9006 (KCCM-10251)) was

reacted with Helicobacter pylori. As shown in Fig. 2, PL bacteria of the EXAMPLE inhibited the adherence of Helicobacter pylori.

TEST 2. Test of inhibitina Helicobacter pviori growth.

(1) Test of inhibiting Helicobacter pylori growth using spent culture of PL bacteria.

The PL bacteria (Lactobacillus coprophilus PL 9001 (KCCM-10245), Enterococcus durans PL 9002 (KCCM-10246), Streptococcus faecalis PL 9003 (KCCM-10247), Lactobacillus coprophilus PL 9004 (KCCM-10248).

Lactobacillus fermentum PL 9005 (KCCM-10250), and Lactobacillus ferment PL 9006 (KCCM-10251)) cultured for 24 h, at 37 °C in MRS broth, and the supernatant was prepared spent culture by centrifugation.

Inhibition test of Helicobacter pylori growth using spent culture of PL bacteria was performed.

Wells were formed on Brucella solid media using a sterilized Pasteur pipet. Helicobacter pylori spread on the media and the well was added with the spent culture of PL bacteria. After incubating in a COg incubator (5 % to 10 % CO2) for 2 days, the diameter of the zone that Helicobacter pylori growth inhibited due to the inhibitory material of the spent culture, was measured.

FIG. 3 is a picture showing that the spent culture has growth inhibitory activity against Helicobacter pylori and FIG. 4 shows a diameter of the growth-inhibited-zone of Helicobacter pylori by the spent culture.

Therefore, it was confirmed that PL bacteria prepared by the EXAMPLE,

secreted material that inhibited Helicobacter pylori growth.

(2) Test of inhibiting Helicobacter pylori growth by lyophilized spent culture of PL bacteria.

4 ml of MRS spent culture of PL bacteria was mixed with 2 mi of distilled MQ and the mixture was lyophilized. The lyophilized spent culture was suspended on 6 ml of skim milk (10 mg/ml) and after 1 ml of the solution was inoculated with 10 ul of Helicobacter pylori (OD625=1. 0), it was incubated for 1hr, at 37 °C CO2 incubator. The incubated solution was centrifuged (5000 rpm, 10min) and supernatant was diluted half with distilled MQ. The quantity of urease of diluted solution was measured by indolphenol method.

The quantity of urease is proportional to urease activity, and amount of Helicobacter pylori in the incubated solution was analyzed. Through these methods, growth inhibition of Helicobacter pylori by spent culture of PL bacteria was analyzed.

FIG. 5 is a graph showing the growth-inhibition degree of Helicobacter pylori by spent culture of PL bacteria, it was known that lyophilized spent culture of PL bacteria maintains the growth inhibition activity for Helicobacter pylori.

TEST 3. Toxicity test The oral toxic test for the PL bacteria of EXAMPLE was based on the'toxic test standard for drug (1999.12.22)'referred as Notification No.

1999-61 of the Korean Food and Drug Administration.

Tests were conducted on Sprague-Dawley rats, which were 5 weeks old (female : 100-120g/male : 110-130g). The rats were reared in 260 x 420 x 180 mm (W x L x H) cages at 23 2 C and 50 10 an.

Referring to established rule No. 10 of the Korean Food and Drug Administration, when the value of LD50 is up to 5,000 mg/kg in an oral test, the material is low toxicity material in body. Therefore, in the experiment, to calculate the value of LDgo of PL bacteria, the dosage was set to 5,000 mg/kg (20 ml/kg B. W.) 5,000 mg/kg was the maximum dosage given to the rat to determine the toxicity of the material.

[Table 7] Bacteria Sex number Dosage(mg/kg B.W) Dosage (ml/kg B.W) Male 5 I(PL 9001) 5,000 20 Female 5 Male 5 II(PL 9002) 5,000 20 Female 5 Male 5 III(PL 9003) 5,000 20 Female 5 Male 5 IV(PL 9004) 2,500 20 Female 5 Male 5 V(PL 9004) 5,000 20 Female 5 Male 5 VI(control) 0 20 Female 5 After 14 days, no deaths were observed. Thus the value of LD50

could not be estimated.

Table 8 shows the weight change of the test group and there were no significant changes.

[Table 8] Sex Days I II III IV V VI 0 120.1~ 120.2~ 120.1~ 120.4~ 120.2~ 120.1~ 3.10 3.08 3.08 2.86 3.25 3. 31 3 142. 6 146. 9 140. 5 140. 7 145. 4 143. 0 6.81 4. 15 5. 46 8. 85 7. 73 3.68 7 162.7~ 161.0~ 155.5~ 156.6~ 162.1~ 159.8~ 7.96 5. 87 2. 92 11. 59 5. 64 3.06 14 212. 3 193. 1 194. 9 201. 1 195. 5 193. 6 8. 40 1.03 9.1 22.72 11.14 16. 24 0 114. 0 113. 9 113. 9 113. 5 113. 5 113. 6 4.34 4. 11 4. 17 4. 41 4. 45 4.50 3 134. 3 132. 6 131. 0 132. 5 129. 8 129. 2 6.81 4.15 5.46 8.85 7.73 3.68 F 7 155.4~ 151.5~ 149.5~ 152.2~ 148.6~ 148.8~ 8. 01 5.69 5.86 7.55 5.81 6. 57 14 192. 7 192. 5 193. 8 194. 7 192. 6 8. 16 10. 87 10. 0 12. 06 7. 55 9. 35 In addition, all the rats were examined to determine the cause of death, but there was no symptom visible to ordinary sight (Table 9).

[Table 9] Organ I m V Vl Brain No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Kidney-Left No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Kidney-Right No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Heart No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Lung No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Spleen No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Liver No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Stomach No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 No. of observations 5 5 5 5 5 5 Intestine No gross findings 5 5 5 5 5 5 Pancreas No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Adrenal gland (left) No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Adrenal gland (right) No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Pituitary gland No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5

Ovary-L No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 Ovary-R No. of observations 5 5 5 5 5 5 No gross findings 5 5 5 5 5 5 No. of observations 5 5 5 5 5 5 Other organs No gross findings 5 5 5 5 5 5 As shown above, PL bacteria of the EXAMPLE is safe without oral toxicity and side effects.

TEST 4. Growth inhibition test for bacillus causing food poisoning MRS spent culture of each PL bacterium was mixed with 2 X concentrated BHI media. Each of Salmonella typhimurum, Salmonella Enteriditis, E. coli O157:HY7, Aeromonas hydrophila, and Listeria monocytogenes cultured in BHI (Brain Heart Infusion : Brain Hear, Infusion form, 6.0 g, Peptic digest of animal tissue 6.0 g, sodium chloride 5.0 g, dextrose 3.0 g, pancreatic digest of gelatin 14.5 g, disodium phosphate 2.5 g) inoculated with 1 % final concentration on the mixture and incubated at 37°C. After 5 hr and 12 hr, the number of survived bacteria was measured.

For measurement, Listeria monocytogenes (L. M) incubated in blood-agar plate. Salmonella typhimurium (S. T), Salmonella Enteriditis (S. E), E. coli 0157 : H7 (0157) and Aeromonas hydrophila (A. H) incubated in MacConkey media and number of survived bacteria was measured at O. D600.

Table 10 shows a O. D600 of bacillus causing food poison by PL 9001 spent culture.

[Table 10] Condition 0 hour 5 hour 24 hour L. M 0. 055 0. 487 0. 335 L. M + spent culture (PL9001) 0.059 0. 069 0.068 S.T. 0.053 0.538 0. 748

S. T+ spent culture (PL9001) 0.063 0. 073 0.072 S. E 0. 053 0. 572 0. 966 S. E+ spent culture (PL9001) 0.059 0. 073 0. 07 0157 0. 051 0. 408 0. 468 0157+ spent culture (PL9001) 0. 06 0. 074 0. 065 A. H 0. 077 0. 418 0. 432 A. H+ spent culture (PL9001) 0.069 0. 093 0.083 FIG. 6 is a graph showing growth inhibition of bacillus causing food poisoning by Lactobacillus coprophilus PL 9001 and shows the result of table 10 (absorbance of O. D600 at 0 hr calculate 100). As mentioned above in table 10 and Fig. 6, Lactobacillus coprophilus PL 9001 produced materials that inhibit the growth of bacillus causing food poisoning.

Also, Enterococcus durans PL 9002 inhibited the growth of bacillus causing food poisoning. Inhibition result shows in table 11 and Fig. 7.

[Table 11] Condition 0 hour 5 hour 24 hour L. M 0. 055 0. 487 0. 335 L. M+ spent culture (PL 9002) 0.059 0. 071 0. 17 S. T 0. 053 0. 538 0. 748 S. T+ spent culture (PL 9002) 0.064 0. 073 0.196 S. E 0. 053 0. 572 0. 966 S. E+ spent culture (PL 9002) 0.061 0. 074 0.167 0157 0. 051 0. 408 0. 468 0157+ spent culture (PL 9002) 0.59 0. 073 0.17 A. H 0. 077 0. 418 0. 432 A. H+ spent culture (PL 9002) 0.065 0. 086 0.235 The inhibition effect of Streptococcus faecalis PL 9003 for growth of bacillus causing food poisoning was measured. The above table 12 shows O. D600 of bacillus causing food poisoning in culture solution containing Streptococcus faecalis PL 9003, and it makes a graph (Fig 8) of data of table

12. Streptococcus faecalis PL 9003 produced materials that inhibited the growth of bacillus causing food.

[Table12] Condition 0 hour 5 hour 24 hour L. M 0. 055 0. 487 0. 335 L. M+ spent culture (PL 9003) 0.058 0. 007 0.075 S. T 0. 053 0. 538 0. 748 S. T+ spent culture (PL 9003) 0.063 0. 074 0.084 S. E 0. 053 0. 572 0. 966 S. E+ spent culture (PL 9003) 0. 062 0. 074 0. 082 0157 0. 051 0. 408 0. 468 0157+ spent culture (PL 9003) 0.058 0. 07 0.071 A. H 0. 077 0. 418 0. 432 A.H+ spent culture (PL 9003) 0.066 0.085 0.094 The inhibition effect of Lactobacillus coprophilus PL 9004 for the growth of bacillus causing food poisoning was measured. The above table 13 shows O. D600 of bacillus causing food poisoning in culture solution containing Lactobacillus coprophilus PL 9004, and it makes a graph (Fig. 9) of data of table 13. Lactobacillus coprophilus PL 9004 produced materials that greatly inhibited the growth of bacillus causing food, but the inhibition effect was decreased.

[Table 13] Condition 0 hour 5 hour 24 hour L. M 0.055 0. 487 0.335 L. M+ spent culture (PL 9004) 0. 06 0. 075 0. 287 S. T 0. 053 0. 538 0. 748 S. T+ spent culture (PL 9004) 0.063 0. 071 0.266 S. E 0. 053 0. 572 0. 966 S. E+ spent culture (PL 9004) 0.061 0. 073 0.227 0157 0. 051 0. 408 0. 468 0157+ spent culture (PL 9004) 0.062 0. 073 0.267 A. H 0. 077 0. 418 0. 432

A.H+ spent culture (PL 9004) 0.065 0.087 0.251 The inhibition effect of Lactobacillus fermentum PL 9005 for the growth of bacillus causing food poisoning was measured. The above table 14 shows O. D600 of bacillus causing food poisoning in culture solution containing Lactobacillus fermentum PL 9005, and Fig 10 is a graph showing the growth inhibition for Listeria. Lactobacillus fermentum PL 9005 produced materials that greatly inhibited the growth of bacillus causing food.

[Table 14] Condition 0 hr 5 hr 24 hr L. M 0. 055 0. 487 0. 335 L. M+ spent culture (PL 9005) 0.06 0. 068 0.413 S. T 0. 053 0. 538 0. 748 S. T+ spent culture (PL 9005) 0.065 0. 074 0.434 S. E 0.053 0. 572 0.966 S. E+ spent culture (PL 9005) 0.062 0. 075 0.448 0157 0.051 0. 408 0.468 0157+ spent culture (PL 9005) 0.06 0. 077 0.389 A. H 0.077 0. 418 0. 432 A. H+ spent culture (PL 9005) 0.067 0. 087 0.245 The inhibition effect of Lactobacillus fermentum PL 9006 for the growth of bacillus causing food poisoning was measured. The above table 15 shows a O. D600 of bacillus causing food poisoning in culture solution containing Lactobacillus ferment PL 9006, and Fig 11 is a graph showing the growth inhibition for Listeria. Lactobacillus fermentum PL 9006 produced materials that greatly inhibited the growth of bacillus causing food.

[Table 15] Condition 0 hr 5 hr 24 hr L. M 0. 055 0. 487 0. 335 L. M+ spent culture (PL 9006) 0.058 0.068 0. 283 S. T 0.053 0. 538 0.748 S. T+ spent culture (PL 9006) 0. 061 0.073 0. 303 S. E 0.053 0.572 0. 966

S. E+ spent culture (PL 9006) 0.065 0. 069 0.428 0157 0. 051 0. 408 0. 468 0157+ spent culture (PL 9006) 0.063 0. 077 0.435 A. H 0. 077 0. 418 0. 432 A. H+ spent culture (PL 9006) 0. 067 0. 087 0. 112 As mentioned above, PL bacteria (Lactobacillus coprophilus PL 9001, Enterococcus durans PL 9002, Streptococcus faecalis PL 9003, Lactobacillus coprophilus PL 9004, Lactobacillus fermentum PL 9005 and Lactobacillus fermentum PL 9006) of the present invention produced materials that inhibited the growth of bacillus causing food poisoning. Also, the decreased inhibition effect is caused by depleting material derived from spent culture, and then it can maintain the growth inhibition effect for bacillus causing food poisoning by using PL bacteria.

TEST 5. Growth inhibition effect for bacillus causing acne Propionibacterium acne is a normal bacilli existing on skin and causing acne. Propionibacterium acne (KCTC 3314) was inoculated in 5 mi of actinomyces broth, and after covering by parapin-oil, Propionibacterium acne was anaerobically incubated for 2 days (BR Vowels, S Yang, JJ Leyden.

1995. Induction of proinflammatory cytokines by a soluble factor of Profionibacterium acnes: Implications for chronic inflammatory acne.

Infection and Immunity 63: 3158-3165). 45 mi of PL spent culture breed on MRS liquid media was mixed with 5 mi of Profionibacterium acnes culture solution. The mixture 50 ul was inoculated in 5 mi of fresh media, actinomyces broth and cultured anaerobically for 2 days without shaking.

For measurement of survival Profionibacterium acnes, culture solution was

diluted (When OD6oo=2. 4,9.6 X 108 CFU/ml) with diluent for anaerobic bacteria containing 0.05 % L-cysteine) and 100 ul of diluted solution was inoculated on actinomyces agar plate. After culture in anaerobic jar, at 37 °C, for 6-7 days, the number of colony on the plate was measured as the number of survival Profionibacterium acnes.

FIG. 12 is a graph showing the growth inhibition activity for bacillus causing acne by PL bacteria, Lactobacillus coprophilus PL 9001 did not inhibit nearly the growth of Profionibacterium acnes, Enterococcus durans PL 9002 and Streptococcus faecalis PL 9003 inhibited completely the growth of Profionibacterium acnes, and Lactobacillus coprophilus PL 9004, Lactobacillus ferment PL 9005 and Lactobacillus ferment PL 9006 inhibited the growth of Profionibacterium acnes.

TEST 6. Growth inhibition test for anaerobic bacillus.

Clostrodium perfringens was inoculated on BHI liquid broth containing Hemin (0.01 g/L) and L-cysteine (0.5 g/L) and anaerobically cultured at 37 °C, for 24 hours (Balows A Hausler WJ Herrmann KL Isenberg HD Shadomy HJ. Chapter 50. Clostridium. p505-521. Manual of Clinical Microbiology, 5th ed. ASM Washington D. C. U. S. A.). Spent culture of PL bacteria was mixed with the same volume of 2X concentrated BHI broth and 1 % final concentration of Clostrodium perfringens was inoculated on the mixture. After covering mixture with a parapin-oil, it was anaerobically cultured at 37 °C. After 24 hour, the culture solution was diluted and inoculated on blood agar plate. The plated was incubated for 37 days and 48 days, in anaerobic incubator. The survival Clostrodium perfringens was measured and present by log.

[Table 16] Number of survival bacteria (log cfu) 0 hrs 24 hrs Clostrodium perfringens 6. 60 8. 30 Lactobacillus coprophilus PL 9001 6. 60 7. 60 Enterococcus durans PL 9002 6.78 7.66 Streptococcus faecalis PL 9003 6. 60 7. 48 Lactobacillus coprophilus PL 9004 6. 90 8. 00 Lactobacillus ferment PL 9005 6.60. 7. 78 Lactobacillus fermentum PL 9006 6.60 7.41

It was confirmed that all of PL bacteria produced materials that inhibited growth of anaerobic bacillus.

TEST 7. improving effect of immunity 50 ul of coating buffer containing purified rabbit anti-TNF or rabbit anti-IL-6 was coated within a microplate well and the microplate was stayed at-4 °C for 12 hr. The microplate well was washed three times with PBST (tween80+phosphate buffer) solution and reacted with 3 % of BSA-PBST, for 30 min. The microplate well was washed four times with PBST and reacted with 50 ul of TNF-standard/IL-6 standard (recombinant mouse TNF-alpha/IL- 6, SEROTEC. UK) or supernatant of macrophage cultured in media containing PL bacteria, 37 oC, for 1 hr. The microplate well was washed four times with PBST, one time with distilled water, and was reacted with 50 ul of mixture prepared by adding 3 % BSA-PBST to biotinylated rat anti- mouse TNF-a or biotinylated rat anti-mouse TNF-alpha/IL-6 (1ug/ml), at room temperature for 1 hr. The microplate well was washed six times with

PBST, one time with distilled water, and reacted with streptavidin peroxidase, for one hour. The microplate well was washed eight times with PBST, two times with distilled water, and reacted with TMB substrate (25 mi citric- phosphate buffer+400 ul TMB stock+100 ul of 1 % H202) at room temperature till showing color. The reaction was stopped to add 6 N of H2SO4 and for measurement of immune reactivity, adsorption at OD450 was observed.

Therefore, PL bacteria of the present invention can be applied to promote immunity and more particularly, can be used for health food and as a treatment drug that promotes the health for aged persons and children.

TEST 8. Test for acid-resistance and bile-resistance MRS medium contained cysteine was optimized pH (7,4.5 or 4.0) with 4 N HCI and 0.1 N NaOH and sterilized. To observe an effect of bile, oxgall powder was added to the medium in 0 %, 0.25 %, 0.50 % conc and sterilized. Each PL bacteria was inoculated medium with total 1 % conc and optical density was determined in 620 nm at 24-hours intervals. Table 17 shows the acid-resistance and bile-resistance. It was shown that PL 9003 was weak in acid, but PL 9001, PL 9002, PL 9004, PL 9005 and PL 9006 had the strong acid-resistant activity and all of PL bacteria had the strong ) bile-resistant activity. Therefore, all PL bacteria are safe in the stomach and intestine (Conway PL Gorback SL goldin BR. 1987. Survival of lactic acid bacteria in the human stomach and adhesion to intestinal cells. J. Dairy Sci.

70: 1-12.: Ibrahim SA Bezkorovainy A. 1993. Survival of bifidobacteria in the presence of bile salt. J. Sci. Food Agric. 62: 351-354) [Table 17] Time PL 9001 PL 9002 PL 9003 PL 9004 PL 9005 PL 9006 24 hr 1.40 1.20 1.00 1.40 1.50 1.50 pH 7 48 hr 1.50 1.50 1.20 1.50 1.50 1.50 pH 5.0 24 hr 1. 00 0. 70 0. 31 0. 90 1. 50 1. 50 48 hr 1. 50 0. 80 0. 45 1. 10 1. 50 1. 50 H 4 5 24 hr 0. 18 0. 21 0. 06 0. 18 1. 50 1. 50 P 48 hr 0. 70 0. 28 0. 09 0. 15 1. 50 1. 50 Oxgall 24 hr 1. 40 1. 20 1. 00 1. 20 1. 10 1. 20 0.25% 48 hr 1. 50 1. 50 1. 40 1. 50 1. 40 1. 20 Oxgall 24 hr 1. 10 1. 20 1. 10 0. 85 0. 75. 70 0.50 % 48 hr 1.40 1.50 1.40 1.00 0.76 0. 70 TEST 9. Antibiotic-resistant

An antibiotic-resistant for PL bacteria prepared by the EXAMPLE was observed. PL bacteria was inoculated on MRS solid medium and a filter (diameter 6 mm) containing the antibiotic of Table 18 was put on the medium.

After incubation for 24-48 h, the diameter of the growth inhibited-zone formed by antibiotics was determined. The decreased diameter of the growth inhibited-zone means that PL bacteria have a resistant to the antibiotics. The result is presented in Table 18.

[Table 18] Antibiotics PL9001 PL9002 PL9003 PL9004 PL9005 PL9006 Penicillin (10 IU/E/UI) 20 mm 20 mm 24 mm 23 mm 30 mm 30 mm Ampicillin (10 ug) 30 mm 24 mm 26 mm 30 mm 30 mm 32 mm Cephalothin (30 ug) 30 mm 20 mm 20 mm 24 mm 30 mm 30 mm Gentamycin(10 ug) 13 mm 12 mm 15 mm 16 mm Vancomycin(30 ug) 20 mm 16 mm Erythromycin (15 ug) 21 mm - - 22 mm 26 mm 29 mm Tetracycline (30 ug) 26 mm 30 mm 10 mm 24 mm 27 mm 30 mm As mentioned above, PL bacteria have high antibiotics resistant. INDICATIONS RELATING TO DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIAL (PCT Rule 13bis) A. The indications made below relate to the deposited microorganism or other biological material refereed to in the description on page 4, line 15-15 B. IDENTIFICATION OF DEPOSIT Further deposits are identified on an additional sheet Name of depositary institution Korea Culture Center of Microorganisms Address of depositary institution (including postal code and counb v) 361-221, Yurim B/D Hongje-1-dong, Sedaemun-gu SEOUL 120-091 Republic of Korea Date of deposit Accession Number December 2.2000 KCCM-10246 C. ADDITIONAL INDICATIONS (leave blank if not applicable) This information is continued on an additional sheet D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE (if the indications are notfor all designated States) E. SEPARATE FURNISHING OF INDICATIONS (leave blank if not applicable) The indications listed below will be submitted to the International Bureau later (specific J tlze general natgae of tlle indicanons e. g Accession Number ofDeposit') For receiving Office use only For International Bureau use only This sheet was received with the international application This sheet was received by the International Bureau on: Authorized officer Authorized officer A @e BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITIONOF THE DEPOSIT OF MCIROORGANISMS FOR THE PURPOSES 01''PATENT PROCEDURE INTERNATIONAL FORM To, Yeon-hee L Department of Biology and Culture Collection of Antibiotic Resistant Microbes, College of Natural Science, Seoul Woman's University, Seoul 139-774, Korea RECEIPT IN THE CASE OF AN ORIGINAL issued pursuant to Rule 7.1 by the INTERNATIONAL DEPOSITARY AUTHORITY identified at. the boltom of this page I IDEtfTIFICATION OF TI-E MICROORGANISM ....,... .', Accession number given by the OITO"' INTERNATIONAL DEPOSITARY AUTHORITY = DEPOSITOR- Lactobacillus plarz KCCM-10246 n. SCIENTIFIC DESCRBPTION AND/OR PROPOSED TAXONOMIC DESIGNATION The microorganisnl iden. tifted ttnder I above was acconvatied by : 1 a scien¢ desfon a proposed txononiic designation (MarkwL a cross where applicalDle) In, RECEIPT AND ACCEPTANCE the tmcroorganisna identified under I above was received by this intematjoaal Depositary Autllority on 0æ a request X conerert the awmad dePosit ; to a deposit under the Budapest Treaty was received by it on Jan. 31. 2001. ! V. INTERNATIONAL DEPOSITARY AUTHORITY Name : Korean Culture Center of Micron Sat) of perscn (s) having the power to represent the International Depositary Hongie-l-dons, Authonty or of autbc'''" ! '= Scodaemuft-gu Seodaemun-gu g , ? 1t jJ t SEOUL120-091 DaX : Jan. 3 0 ftepiC of ea _ rv aEfi BUDAFEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FC) THE PURPOSE OF FATENT PROCEDURE ATTESTATION CONCERNING THE LATER LINDICATION OR AN AMENDMENT OF THE SIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESTICINATION pursuant to Rule 8. 2 TO Yeonhee lee Department of Biology, Seoul woman's University, Scout 139-774, Korea TheeraclDscd cDlalmullication has bcen recc ; ved lBy this laternalionai Dcpositaly ADtherily on Tan, 21 2001. INDUSTRIAL IzEPOf A. Ry AtiTHORITY 'Name : Iirean Culure Center of Microorganisms SiSnamreRs) r persunts) having the power to Addresst tPat Inl. ernViUnr thorip or autilorized l 120- () 93 Seodael. n. un gu, __ | , . z. r"'. L. oui, 12o-om D2te; Jaa, 25_ 2a0t. Rerivblic of Korea ffmufg Enolosure: Communication of the later indication or an amendment of the scientific description proposed taxounic designation puruant to Rule 8.1 BUDAFEST TREATY ON THE INTERNATIONAL RECONTION OF THE DEPOST OF MICROORGANTISMS FOR THE PURPOSE OF PATENT PROCEDURE COMMUNICATION OF THE LATER INOICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION pursuant to Rule 8.1 To. KCCM 361-221 Yurim B/D Hongie-1-dong Seodaemun-gu Seoul. 120-091 Republic of Korea I.) DBNTinCA'nON OF THE MCKOOMAMtSM Accessionaumbw givm by the RqTZIRNAIT. O. N. XL I) E ( : SITARY AUTRORITY- KCCM46 I [. SC. PP-$Ck ON OR EXROPOSE : IP T. <C} tI10St YNATION CS Q SciaitMn : ctesodtption : p'Fast pccedmg scinc rsc. ripticn (if any) : [S sed tanss desion : BrntetOcoccus PL 9002 edipg PXUI) 0. 5ed t= ; UoMiC degigMtioU (it aayy La : lS CW 9-t s Mark with a okose if additional informationa. is given on an atenthed sheat.

, Mark with a cross the upplicable box or boxes. lUi, REQUEST POP. ATOSTkrl ( :) N ! D. REQUEST FOR ATTESTATION Tle un8etic"rpcd 111 Ihe wld « Sd 111 requests dn vot. rc. quest (he affestation reftred to in Rule 8, 2 IV. DEPOSITOR : : y d. o.: 1 ind Name ; Yeonhee lee $Igoah>4} X ut . AdX4 : » XtQt of Biolog, Seout womm's Uinvetsi. ty, Seol 139. 774, Date: Jan. 21. 2001 , Korea 9. Mark with a cross the application boxs.

1. Where the signature is required an begolf of a legal entity, the typerwritten nature(s) of the notural person(s) signing on the lgal entity should accompany the signature(s). INDICATIONS RELATING TO DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIAL (PCT Rule 13bis) A. The indications made below relate to the deposited microorganism or other biological material referred to in the description on page 4, line 16 B. IDENTIFICATION OF DEPOSIT Further deposits are identified on an additional sheet Name of depositary institution Korea Culture Center of Microorganisms Address of depositary institution (including postal code and country) 361-221, Turim B/D Hongje-l-dong, Seodaemun-gu SEOUL 120-091 Republic of Korea Date of deposit Accession Number December 2.2000 KCCM-10247 C. ADDITIONAL INDICATIONS (leave blank if not applicable) This information is continued on an additional sheet D. 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IDENTIFICATtON OF THE MfCROORGAMSM Accssion n2nk giva w the Zdni. i : zc ; itcr rft'ce ivn'y i : e -TRATTC. . L D'tI'"l, t' ?" AUTTItIT. DEPOSlTOR' Lactobact'lids pamparacasei I PL 9-3 °: L727, m SCIENTIFtC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION n. SCENTFIC N RM TGON DRIGNS N The micraorgusm idenffted mder I above was accompanied by : a a scienda cles íot a a. t ; roerl i ; axrncmit rinatict (Mark i a oss whet ap le) lUECEIPT AND ACCEPTANCE the microorganism identified under I above was received by this inteftiaonal Deposttary An< : hodty on Dec. 2. 2000. and a request to convert She ongtnal deposit to a deposit t : der the Biapeæl ; Treat mras red bY it on San 3 : OP1. N. s7J. u . . ilV J, LllLYt7, A. l. IIYL' ? La 7L t ?. AJ 1 a. Jlli 1 S Name : frean C e Ceatezf d Mkcroott isms Signatxrels) of >rson (S) having t1. Ze Dawer N : . cm C-of Mm. Signa (a) of person (s) hnving the Dower to Hongje 1-do A by ç bj \361-221, Yua-i B/D Authority cr of autlio, =A, PM' Scodaemun-gu bEOUL 0-091 mte : Jan. 31. 208 ! . : : ', ; cr, zo-csr t: o, ' ;. Jn. R lìc of Soree s ! BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS I'-'O, R THE PURPOSE OF PATENT PROCEDURE ATTESTATION CONCERNING THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRPTION AND/OR PROPOSED TAXONOMIC DESIGNATION pursuant to Rule 8. 2 TO Yeonhee lee Department of Biology, Seoul <BR> <BR> Seoul 139-774,<BR> <BR> Korea. The enclosed commllaication has bscn rcceìved by thts llttcrnatìonal Dquosìtaly. Authorit. y on JiliL. 21. 2001. INDUSTRIALDEPOSfTARY AUTHONTY Nsmtc* Korean Clllulre Center of Microorganisms Addr: 361-22], Ym B/D'""" ' "" reprMent the McmatioHatJemMJ'tan'Auihb rir 'represent the Intcrnatioiia'sixa mhurhority Hngje-1-Glprlg, A. r tutihurized ufficial ( ; s) ( Seutlcc-' ; mun- u, SEOUL 120-091 , , Republic of Kai-e-a {jfn Enclosure : Communication of the later indiention or an amendment of the scientific description proposed taxonomic designation pursuant to Rule 8. 1 BUDAFEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSE OF PATENT PROCEDURE COMMUNICATION OF THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION<BR> pursuent to Rule 8.1 TO KCCM 361-221, Yurim B/D Hongie-1-dong Seodaemun-gu Seouo, 120-091 Republic of Korea t'Nt CAllON OF E M ; ; 4R M AcceioH, n-mnber given'by ttie BOIEKMA'nONAL DBOSITARY AUTHORrTY KCCM M247 It, SCIENITFIC DESCRIPTION AND/OR PROPOSED TAXONOMC DESGNA. T [ON D' cictiiia iienx, ritin : 2 Last prereding isciisntifi tion (if 2zy) : :pposed taxtionomie'Jesigaatt<Ht : hterococsíí. i PL W) 3 , 2 I ; ; gt pre » c oposesl laxonomio designati, On (if nny') : Uctob=ilTus CHARM M Mark with a erous if additional information is given on an ettabobed sheet. i4nrk tvith a cross the applicable box or boxes. , REQUEST FOR A'tTESTATEON The mi (tM!; ij! < <]. 1! The imdeisipcd. I I LE, teqlBsls dnes aa. l7t rutt 'fhe attiestaton refafed ? m Rule 8. IV. DEPOSMOR . .....''""""***' taf., Nam. e : a''tb. ee l. ee iotrue , n. Y'r a. yYfll, Address : Depattntsnt of Bido . Seoul womai's UmvasHy. Seol 139-774, Date : jTme. 21. 2<'C ! Korea . Mark with a cross the application box.

4. Where the signature is required on behalf of a legal entity, the typewritten name(s) of the natural person9s) signing on the legal entity should accompany tha signature(s). INDICATIONS RELATING TO DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIAL (PCT Rule 13bis) A. The indications made below relate to the deposited microorganism or other biological material referred to in the description on page , line'15 B. IDENTIFICATION OF DEPOSIT Further deposits are identified on an additional sheet Name of depositary institution KOREA CULTURE CENTER OF MICROORGANISMS Address of depositary institution (including postal code arzd countly) 361-221, Yurim B/D Hongje-l-dong, Seodaemun-gu SEOUL 120-091 Republic of Korea Date of deposit Accession Number . December 2ç 2000 KCCM-10245 C. ADDITIONAL INDICATIONS (leave blank if not applicable) This infomlation is continued on an additional sheet D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE (if the In iitior < E. SEPARATE FURNISHING OF INDICATIONS (leave bllk if not applicable) The indications listed below will be submitted to the International Bureau later (specs dze genal tat¢re ofthe indicatios e. g.,"Accessioa Number of Deposit'J For receiving Office use only For International Bureau use only This sheet was received with the international application)) This sheet was received by the International Bureau on: Authorized officer Authorized officer , > BUD. TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSES PATENT PROCEDURE INTERNATIONAL FORM To. Yeon-hee Lee Department of Biology and Culture Collection of Antibiotic Resistant Microbes, College of Natllral Science, Seoul Woman's University, Seoul 139-774, Korea RECEPT IN THE CASE OF AN ORIGINAL issued pursuant to Rule 7.1 by the INTERNATIONAL DEPOSITARY AUTHORITY identified at the bottom of this page I. IDENTIFICATION OF THE MICROORGANISM .,,. .. ., Accession number given by the "' INTERNATIONAL DEPOSITARY AUTHORITY.- DE. POSITOR Zc&o'c/s ee'prepMMS'PL 9-1 Tn\/t KCM- x45 lI. SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION 'i'y micorganism. identiftati under I above was accompanied by ; a a scientific descnption 12 a proXsed t nomic slesignation (Mr1< wth a rms wlze apabte) m. RECEIPT AND ACCEPTANCE the microorganism idendRed under I above was received by this mtertiattonal DepositaiT Authority on Dec. 2. 2000. and a request bo convert the ofigi-nal deposit to a deposit -under the Budapest Treaty was received by it on Jan. 31. 2001. I, Nh'IIJT. lL DFPDIT, A, Y' A, JTIIII°J''Y tan : iorean uttute Center of Mlcrorxnxstrta. . ignau (s) of so. (sl lvang tm ; tower Addr.-36121, Yul B/D ' Imational Ddtajy HonNe-ldong Authonty or of ).- Seoda=un-gu SEOUL0w091 Dai : Jan, 31. 2N t Repube of Hjoifea ss t'mtfM IlepubNc of lorea BUDAPEST TREATY ON THE TNER NATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSE OF PATENT PROCEDURE ATTESTATION CONCERNING THE LATER INDICATION OR AN AMEND. M. ENT OF THE SCIENTIFIC DESCRIPTION AND/OR.

PROPOSED IAXONOMIC DESIGNATION pursuant to Rule 3.2 TO Yeonhee lee Department of Biology, Seoul woman's University, Seoul 139-774, Korea The enced communication has bMO. fved by ihbi Intcrnation&. i DepositHry Authority on Jim. 21. 2001. AUTHORITY - Waale ; Kt ; lrean Culx e Cenl. e. of hficroorzflanisms rcpt'eetU ! h6 Jntefnatipnat ueposttat'y Authont ;' rcprcsent the InteratJonal'1D sposxt ? ry tutborry or authorized official (s) SL-'OT-IL 120-091 s; zn ao-cm D : it : Jun. 25. 2001. Ft. epululsc o. f. o res 1 ? aS 1f Enclosure Communication of the later indication or an amendment of the scientific description proposed taxonomic designation pursuant to Rule 8,1 BUDAFEST TREATY ON THE INTERNATIONAL RECOGNITION OF TH EDEPOSIT OF MICROORGANISMS : FOR THE PURPOSE OF RATENT PROCEDURE COM. MUNICATION OF THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AN'D/OR PROPOSED TAXONOMIC DESIGNATION pursuant to Rule 8. 1 TO. K KCCM) 361-221 Yurim B/D Hongje-1-dong Seodaemun-gu Seoul, 120-091 Republic of Korea I. IDENTIFICA'noN < ?' THE MCROORGANISM Accessionmuntx iven by tk MTBKNA'fIONAL DEOSITARY AtJTHOmTY : KCCM M245 E SC N g/0R ON. SS aANOtGC D NANON D' Scienti : ric descxiptiom Ey Laat ptcdrng SMentijfic deocnptton (if any) : 2 Proposad tarmonoaic desimate LttctobacNtus PL 900} .. nst rt : cin l7tuPnscrl ta. nnomic desisttutiu t cmyr Lactobacilluvs ee 1.Mark with a cross if additional information is given on as attached sheet, 2.Mark with a cross the applicable box or boxes. 01. REQUEST FOR ATTATION The mdersjped signed 111 2 Pw ur. a does does npt request thé aMestation reined to m Rtt 8. 2 R E. POS2QR Nlme : Teonhee lee r , ; s / Ad<MS : Department of Bicloy,''"''" Seoul wOme : ufs Univew, tys Seoul'139-774, 1) Ze. 21. SOQI Kom 2. Mark with a cross the application box.

4. where the signature is required on belef of a legal entity, the typewritten name(s) of the natural person(s) singing on the legal entity should anompany the signature(s). INDICATIONS RELATING TO DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIAL (PCT Rule 13bis) A. The indications made below relate to the deposited microorganism or other biological material referred to in the description on page 4, line B. IDENTIFICATION OF DEPOSIT Further deposits are identified on an additional sheet Name of depositary institution KOREA CULTURE CENTER OF MICROORGANISMS Address of depositary institution (il7cluding postal code and colnhy) 361-221, Yurim B/D Hongje-l-dong, Sedaemun-gu SEOUL 120-091 Republic of Korea Date of deposit Accession Number Decémber"-2. 2000 KCCM-10248 C. ADDITIONAL INDICATIONS (leave blank if not applicable) This information is continued on an additional sheet D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE (if the indicatiolls are notfol all designated Star. es) | E. SEPARATE FURNISHING OF INDICATIONS (leave blank if not applicable) The indications listed below will be submitted to the International Bureau later (specify the generrl uanne of tle irdicutioTZS e. g.,'tccession Number of Deposit') For receiving Office use only For International Bureau use only This sheet was received with the international application This sheet was received by the International Bureau on: Authorized officer Authorized officer BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSES OF PATENT PROCEDURE INTERNATIONAL FORM To. Yeon-hee Lee Department of Biology and Culture Collection of Antibiotic Resist-ant Microbes, College of Natural Science, Seoul Woman's University, Seoul 139-774, Korea RECEIPT IN THE CASE OF AN ORIGINAL issued pursuant to Rule 7. 1 by the INTERNATIONAL DEPOSITIARY AUTHORITY identified at the bottom of this page I. IDENTIFICATION OF THE MICROORGANISM Accession number given by the IdMon reduce given by the INTERNATIONAL DEPOSITARY AUTHORITY-. DEPOSITOR : Lactobacillus coprophilus Pi, 9-4 KCCM 10248 K-: Lp29g H. SCENTEIC RWION X/OR PROSED TgONOMC DBIGNAT, ION The micr44rganism identified Wder I above was accompanied by : 12 a scien « s desmpdr C) a proposed taxotio. nnc designadjon (Mark with a cross where çlicable) Ill.RECEIPT AMD ACCEPTANCE the microorganism identified under I above was received by this intEnational Depositary Authority on Dec. 2.2000. and a repeat to convert, the original deposit : to a deposit under the Budapest Treaty was received by it on Jan. 31. 2001. IV.INTERNATIONAL DEPOSITARY AUTHORITY Name : Korean Culture Center of Mlcroorganisms Sigriaia. ue) of xson. (s) lvxr l; le parwer . . . to represent the International Depositary Address : 361-221, fm BS Hoiigje-l-dong. Authotity or of auth Seodaemun- n SEOUL 120-091 Date : Jan. 31. 200. ) r Republic of Yw : rea p Pi P- ," BUDAFEST TREATY ON THE INTRRNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSE OF PATENT PROCEDURE ATTESTATION CONCERNING THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION pursuant to Rule 8.2 dz(conhee lee Dcpanment of Biology, Seoul woman's University, Seoul 139-774, Korea The MClt'Sed communication hi been receive by this Internatiottvl, Depositaiy luthority on. Jnn,21.2001. 1I. 1'DUSTRIrIL D'h'1'f) SITARSc' AUTTC (, 71I. T7. 'lNDUSTRAL D'EPOSITßY AHOiRlTY Darne: Korean Culture Cfmter of MicfoofR'aMsms Stgnat. urcfs) of pN'sn'n (s) ha'vmg the power to represent @. Zc Intcrnational'Oepositat xtrllority or authoriye (I ofiioiiil (s) Secdaenunru, i SE07JL. 120-U51., " Datc ; Juii. 25. 2001. fiepublic of Ioru. 4- jj---! Enelosure: Communication of the later indication or an amendment of the scientific description proposed taxonomic designation pursuant to Rule. 8,1 BUDAFEST IREATY ON THE INTERNATIONAL RBCOGNTTION OF THE DEPOSIT OF MICROORGANISMS FOR mE PURPOSE OF PATENT PROCEDURE COM,MUNICATION OF THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION pursunut to Rule 8. 1 TO. KCCM 361-221 Yurim B/D Hongje-1-dong Seodaemun-gu Seoul, 120-091 Republic of Korea I. IDENTIFICATION OF THE MICROORGANISM A@ossion nt7mbx give : lay dle INTERNATIONAL DEOStTARY UTHORirY : KCCM H) 24K n. SOP. N'IC DESCRIPTION AND/OR. PROPOSED TAXONOMC KESIGNATION 0' 2 Scientitt das~tios : 02 I :,. &st prezedin ientific descsiplion (H 6ey). FYOpo3G. tn, ntotlaic designation : JLaetbaciUus PL 9004 Ust pMcediag pn) pos<M) taxOMmic degnation (if any) : Laolullacillus C : CAZ qX s,, Mark with a cross if additional information is given on an aattached sheet.

2, mark with a cross the applichle box or boxes. DL REQUEST : FOR ATTESTATION Dietmdzasigned 111 a Kquefs {J docs not request the aM,-, Otion rerLm-A to : in Rule 3. 2 IV.DEPOSIT*, N4me : Yeol, hee Ace (; i ; al ; ut ; ''' '5 A. ddres.' ! : DepM'tmenI ofBioIcgy,'<'.' Seoul WODíbn1s Universily, gead lS, 9-774s Dalc Jme. 21. 20 {; 11 Korea Y. Mark with a cross the application box.

''. Where the signature is required on behalf of a logal entity. the lypewriten name(s) of the natural person(s) signing on the legal entity should accomapny the signature(s). INDICATIONS RELATING TO DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIAL PCT Rule 13bis) A. The indications made below relate to the deposited microorganism or other biological material referred to in the description on page 4 line 18 B. IDENTIFICATION OF DEPOSIT Further deposits are identified on an additional sheet Name of depositary institution KOREA CULTURE CENTER OF MICROORGANISMS Address of depositary institution (inclztding postal code and coarrzh) 361-221, Yurim B/D Hongje-1-dong,'seodaemun-gu SEOUL 120-091 Republic of Korea Date of deposit Accession Number Mar. 2.2001 KCCM-10250 C. ADDITIONAL INDICATIONS (leave blank if not applicable) This information is continued on an additional sheet D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE (if the ttidicatioras are notfor all designated States) E. SEPARATE FURNISHING OF INDICATIONS (leave blank if not applicable) The indications listed below will be submitted to the International Bureau later (spec the general riarerre of the irdications e. g.,'Accession Nannber of Deposit') For receiving Office use only For International Bureau use only Et This sheet was received with the international application This sheet was received by the International Bureau on: Authorized officer Authorized officer BUDAPEST TREATY ON THE INTERNATIONAL RECOGNITION OP THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSED OF PATENT PROCEDURE INTERNATIONALFORM "To. Yeonhee Lee Department of Biology, Seoul Woman's University, Seoul 139-774, Korea RECEIPT IN THE CASE OF OF AN ORIGINAL. issued pursuit to Rule 7. 1 by the INTERNATIONAL DEPOSITARY AUTHORITY identified at the bottom of this page J. I. IDENTIFICATION OF THE MICROORGANISM IdeniEcatioti re'feretice s'iveti by the Accession number given by the D E P O S a T O R : I t E t A n O N A L. D E P O S I T M Y A R 1 w : lwct, lweillt4s PL 9-5 SCCM-10250 Jeocfs PL 9-5 CCM-10250 ad SCIENTIFIC TE} DESCSOION AND/OR PROPOSED TAXONOSC DESIGNAT. TON The microorganism identified undef I above waa accomnied by : Cl a sr. ientXc descri. Ption Cl a proposed laonomic designadon (al wl : h a cross where applicable) DI. RECEIPT AND ACCEPTANCE Th : a InternaHoRal Depoaittaty Authotiity accepts the tntcroorBanisn) ldent ed under I above, which was received by it on Mar. 2, 2001. (date of the orIginal deposit) Y' IV. INTERNATIONAL DEPOSMARY AUTHORnY Name : Korean Culture Center of Microorganisms Signature (a) offeMOti (s) having the power t. o represent the International Depoaitry Address-a6l-221, Yurim B/D ., < Author). tyofofa. uthou ! zed<mt {a ; Jsg Honsae"l''dc) ng, y7' ! S=nt Seodaemu-gLf 'S§ S ! SEOUL 130-091 Date : Mar. 8. 2001. Republic of Korea 1 Where Rule 6. 4 (d) applies, such date is the date on which the status of international depositary authority was acquired : where a deposit made outside the Budapest : Treaty after the acquisition of the status of i ni : ernati. on. al depositary authority is converted into a deposit under the Budapest Treaty, such date is the date on which the microorganism was received by the international depositary authouity.

BUDAFEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSE OF PATENT PROCEDURE ATESTATION CONCERNING THE LATER INDICATION OR AN' AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION pursuant to Rule 8.2 O Yeunhec Ice Departme.ot of Biology, Seoul womb's University, Seoul 139-774, Korea The enctosed communication) MS been received by this Intcmationa). Depositary Authority onIun. 2. L200L INDUSTRIA).. OBFOSITARY AUTHORITY a<fte : Korean Culture Ceni. M' of Microorganisms.,.,.. Snniature (s) of persons) ha-vrng the power-to Atldrcss. 1-uula Yurim. $/D rcprcsent the : l. nLerniiiion ;'il 'thrity 'Hongje i. Ollg, or authori7. ed o l (s) SeoctaelYItm-gu, g Ira c : ; aiw 1, C : puilic. of Kora ; , . t -- Enclosure: Communication of the later Indication or an amendment of the scientific description proposed taxonomic designation purusant to Rule 8.1 BUDAFEST TREATY ON THE INTERNATIONAL, RECOGNITION OF THE DEPOSIT OF MICRORGANISMS FOR THE FURPOSE OF APTENT PROCEDURE COMMUNICATION OF THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED TAXONIMIC DESIGNATION<BR> pursuant to Rule 8.1 TO. KCCM 361-221, yurin B/D Hongje-1-dong Seodaemun-gu Seoul. 120-091 <BR> <BR> <BR> ReDuMic of Korea<BR> <BR> <BR> <BR> <BR> <BR> L I-MPNTIFTCAtiON OF THE MICROORGANISM Accession numb given by lEe b3 [NA1IONA : L DEOSITAR 0KE : l (CCM 1 Zj () KLCCM) 020 II. SCNTC DESCRIPTION AM/OR PROPOSED TAXONOMC DmO-NATtON Lj' Scieiltific desoliptil) n- Dtot pxewduig wientific descriptioii (if my) : 22. Proposed taXnoaomia ti nalion : LatoTKtCiHus PL 9005 df Last ptecedia pPOpcsed taxenomic desigattoa (if) my) : lLactobacillus PL 9-5 Mark with a cross if additional information is given on an attached sheet.

Mark with a cross the applicable box or boxes. Cl. KBQST FOR ATTESTAnoN Thx ; imdersite$ 11 i 03 teçesls doer,tot rlquest theattetiMi jfiMMd to m R. e S. 2 W. DPOSZOR, -uw-i N, tle : Ycoitffl 1, ; 'tu ; '. ;'A.. c.. t AdRe. ss l) eptErlrAcnt of B,. iajogy, Seou women's Uversity, Seful 39-774, Me : Jtmc. 21. 2001 Korea 2.Mark with a cross the application box.

4. Where the signature is required on behalf of a legal entity, the typewritten name(s) of the natural person(s) signing on the legal entity should accompany the signature(s). INDICATIONS RELATING TO DEPOSITED MICROORGANISM OR OTHER BIOLOGICAL MATERIAL (PCT Rule 13bis) A. The indications made below relate to the deposited microorganism or othel biological material retbned to in the description on page 4, line J. $. B. IDENTIFICATION OF DEPOSIT Funher deposits are identified on an additional sheet Name of depositary institution Korea Culture Center of Microorganisms Address of depositary institution (including postal code and countr/) 361-221, Yurim B/D Hongjr-1-dong, Seodaemun-gu SEOUL 120-091 Republic of Korea Date of deposit Accession Number Mar. 2.2001. KCCM-10251 C. ADDITIONAL INDICATIONS (leave blnrk if not applicnble) This ioformation is continued on an additional sheet D. DESIGNATED STATES FOR WHICH INDICATIONS ARE MADE difthe invSications are lzotfor all designaJed States) E. SEPARA, TE FURNISHING OF INDICATIONS (leave blanlr if not oppliccrble) The indications listed below will be submitted to the Intemational Bureau iater (speci the genercrl nahwe oftthe indica6oru e, g.,'Accessior Atzmaber of Deposit'7 For receiving Office use only For lntemational Bureau use only This sheet was received with the international application a This sheet was received by the International Bureau on : Authorized officer Authorized officer BUDPEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR : THE PURPOSES OF PATENT PROCEDURE INTERNATIONAL FORM To. Yeonhee Lee Department of Bblogy, Seoul Women's University, Seoul 139-774, Korea L RECEIPT IN THE CASE OF AN ORIGINAL issued pursuant to Rille 7.1 by the INTERNATIONAL DEPOSITARY ALITHORITY identified at the bottom of the page I. IDENTIFICATION OF THE MICROORGANISM I. T. DE. LF MON OF T, HE MICROORGANISM . Tden. l : catI. olt reference gi, ven bY l : he Accesslon. number $iven by the DEPOSITOR : INTERNATIONAL DEPOSITARY AUTHORITY : ZocM PL 9-6 KCCM-10251 n ; SCIENTIFiC DESCRIPTION AND/OR PROPOSED TAXONOMIC DESIGNATION Me micrnorganism idenfflied under I above was accompallied by : Q a scieuiific description . a proposed t'axonomic desIgnaHon with a cross where applicable) IH. RECEIPT AND ACCEPTANCE lE Itllernati. ons tepositary Authoui. tY acceyts fLe microorganísm identi. 6cd lmder I ahove, which was received by M oji Mar. 2* zou1. (date of the original IV. MTERNATIONAI. DEPOSITARY AUTHORITY -|-- Name : Korean Cture Center o'fMicfooraanIsm8 Si) Mturp (s) of persona) a. v ! n the power to repfesent the International Depoaitaty liongje-l-dong, AuffioBty of of auNoulzed ( (k Ioyje-1-aong,-ft',. y Hone-1-dong, BH Seotlaemun-gu ¢ SEO11I O-ORl Date : Maw 01 BE F RepuMc <? f Ka! f) f Republic ( ? f Korea 1 Where Rule 6.4 ( 1) applies, such date is the date on which the status of international depositiary authority was acquired : where a deposit Made outside the Budapest. Treaty after the acquisition of the status of 1 nternational depositary authoriy is converted into a deposit under the Budapest Treaty, such date is the date on which the microorganism was received by the international depositary authouity.

BUDAEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FORR THE PURPOSE OF PATENT REOCEDURE ATTESTATION CONCERNING THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESCRIPTION AND/OR PROPOSED IAXONOMIC DESIGNATION pursuant to Rule 8.2 Yeonhee lee Department of Biology, Scout woman's University, Seoul 139-774, Korea The enclosed communication ha. s been j'ccei'Vt : () by this JntematMnaJ Depositary Authority on Jun. 2 tNDUSTRIAL DEPOSITARY AUTHORITY w Name : horean Cnlterre Cent. er Of Microorgallism6 ttdlCB$ :. u7-21. L1T7111 B, mJ Adi-cs6 : 36 urim Bm. ... repfesent tllß llstert1. ati) nal atHA. ulhority or authorized officioLi. (s) Ui' SEOUL 1. 20-091 : U'UL 7. 20-91 Uat; Jun, 25_ 2 (, 1f, 11. , . RepubLic of Ko-ea -< 'iB Enlosoure: Communication of the laster indication or vu amfndmmt of the scientific descriptio proposed taxonomic designation par to Rule 8.1 BUDAFEST TREATY ON THE INTERNATIONAL RECOGNITION OF THE DEPOSIT OF MICROORGANISMS FOR THE PURPOSE OF PATENT COMMUNICATION OF THE LATER INDICATION OR AN AMENDMENT OF THE SCIENTIFIC DESORPTION AND/OR RPOPOSED TAXONOMIC DESIGNATION pursuant to Rule 8.1 TO. KCCM 361-221 Yurim E/D Hongje-1-dong Seodaemun-gu Seoul. 120-091 Republic of Korea I.IDEXCAEUN o'rSlSM I. ITaIT'IATIc') 7hT '', f.: fEl: IVII'": LLii; iltiAN'1"''. YVI. Accession wumtxa givc by fhe INTERNA'nONAL DEOSn-ARY AUTHORirY : KCCM 10251 n. SCIBNTMC DBSCRjmON AND/OR. PROPOSED TAXONCMC DESIGNAnON D' ( ScientiiHe desohption : Lj Lost pMeedms scKaitifie < ! < : scdptiON (if any) : 1'12 P. ropose monomiiJ designation : Lactocilltts PL <M) 06 Cj'Last preening pfoposed tasmomi' ; desitaOB (if an, y) : : tobacilJu$ I9X. SX zu Mark with a cross if additional information is given un an attached sheet.

2. Markwith a cross the applicable box or boxes.

Form 39/7 (first page) ni. ROQUES FOR ATTBS'CATtON 1] ddZ ßd 3 requests does sot request atte-4Ation referxd to in Rule 8. 2 1V.DEPOSITOR ---8 L ' ! ' A44reog : Depwtm (. nt of Biology. . Seoa vomea'3 Univets S ; oul 139-774, Diate} me. 2 001 Kot'ea 1, Mark with a cross the application box.

'. tvhcre the signature is reqired on behalf of a legal entity, the typewritten nature(s) of the natural person(s) signing on the legal entity should accompany the signature(s).