901353日本クリニック微量栄養素26/++20‐n‐2‐ …jtnrs.com/sym26/26_096.pdfwild...

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Page 1: 901353日本クリニック微量栄養素26/++20‐N‐2‐ …jtnrs.com/sym26/26_096.pdfWild herbs Sun-dried Extracted with hot water Filtered Fresh vegetables and fruits Washed

������������ ������������������������������� �!"#$%!&'�!"(#%)*���+,��-.�/��1�4012����+,����3���45�6��"2���7��89:��45��;�9<=>?�@A��B�C:�DEFG5�70� ��������������HIFGJJ��1K�����LMNOP�Q��R�ST� UV�89:��WX��Y)��7���Z[��89:���U\�]^_��R���`2��ab�cWXd)�7����801e��3�eGd�LM��f��"3�M�7g[cdhUG/iR�jk9&l��ml!no!�p��q�rst)���fNO����uv�801

wxy!k�jkz{|}PELY0��jk�&lNO�Y2[~��U����&���$����/^�:�9�8cdz{|������eGdz{|���������cJwxy9k��Rjk!��F�� ���jk�����1����/�Z�Z�) ��45�����������7��12����}Crassostrea gigas0�����)����/�dG/iR�������/7���[��FG/��90��PELY���3��� �jk��7��������������tf�100��/�dG/��1�c��������� )R�¡¢�����Z[�£��jk�¤ ����¥�!¦§FG9B[)�/��¨��/�����110�©vdG/��7��e�ª�«¬��2­�®�) �7Z�12��PELYi�/�jk¯°�rst)LM±²!� �

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Studies on the Plant-based Extract Fermented by Lactic Acid Bacteria and Yeast

Shinsuke KUWAKI10, Nobuyoshi NAKAJIMA20, Kohji ISHIHARA30, Shuangquan40and Hidehiko TANAKA1010Functional Food Creation Research Institute Co., Ltd, Okayama 701-1145, Japan,

20Department of Nutritional Science, Faculty of Health and Welfare Science,Okayama Prefectural University, Okayama 719-1197, Japan,

30Department of Life Science, Okayama University of Science, Okayama 700-0005, Japan,40College of Food Science and Engineering, Inner Mongolia Agricultural University,

Inner Mongolia 01008, P.R. China

SummaryA plant-based diet is thought to be better for the prevention and treatment of lifestyle-related diseases. A plant-based

extract fermented by lactic acid bacteria and yeast (PELY) was made from various plant materials and was fermentedproduct applying to traditional food-preservation technique, that is, fermentation and sugaring. In this study, the charac-terization and physiological function of PELY were examined.PELY contained 58.8 % carbohydrate, 3.7 % protein, and 1.3 % lipid. It contained 18 kinds of amino acids and vitamin A,

B1, B2, B6, B12, E, K, niacin, pantothenic acid, and folic acid. It also did a lot of dietary fiber and phytochemicals (polyphenol,terpenoid, i .e .).Moreover, it was determined that PELY had several physiological functions such as antioxidant, antihypertensive, an-

tibacterial, prompt digestive and anti-tyrosinase activities. As the results, PELY possessed antioxidant, antihypertensive,antibacterial, and anti-tyrosinase activities in vitro .

Trace Nutrients Research 26 : 96�104 (2009)

� 96�

Page 2: 901353日本クリニック微量栄養素26/++20‐N‐2‐ …jtnrs.com/sym26/26_096.pdfWild herbs Sun-dried Extracted with hot water Filtered Fresh vegetables and fruits Washed

Wild herbs

Sun-dried

Extracted with

hot water

Filtered

Fresh vegetables

and fruits

Washed with water

Extracted with sugar

Filtered

Blended

Fermented and matured

Filtered

Product (PELY )

The other materials

Grinded

���������� ���������������PELY�������� !"#$%&�'()*�"+,-./012�34'56178�9:;<�=>?�>��@AB�C��DE>�FGH>IPELY���1�JK�;LMN�

6O�������>�P"Q�RS�TU���V�VWXYZ in vitro �X[�� !"#$%&�'()\]N�E@@^6�

����

1������ ���PELY����PELY��_�`(6abcI Table 1�d^6@Xe

>���75fg��f�h1iQ�ac;j`^6�klRI Table 2�d^6 15fg�iQ�klR;��mnono@^)pq^6�r6�Fig. 1� PELY�_stuvm�wx;d^6�yz{@5|1}�ac;~�����C���;��N�E@1���V;��^�DC�;�������^6ac@@*�����^�klR����Ye������6*�;�c@^6�1�����������!0�200�$�X[�����R�RS���� XYZ pH�� ;�¡�6O��¢£�Y¤1¥¦;`(6�klRSI�u§¨�©oª«otªqt�o¬­®�¬¯°±²!¢£BCP@³´$;`()�37µ> 72�¶±

Plant name

Sugar Brown sugar, Oligosaccharide

Fruits

Prune, Ume, Citron, Strawberry, Apple, Iyo-orange, Grape, Fig,Persimmon, Kiwi, Mandarin, Lemon, Kumquat, Akebi, Wild vineArtubus, Winter strawberry, Blueberry, Blackberry, Raspberry,Burmese rosewood, Peach, Pear, Goumi

Vegetablesand

Wild herbs

Pumpkin, Carrot, Mugwort, Cabbage, Kale, Spinach, Japanese radish,Eggplant, Perilla, Tomato, Pimento, Cucumber, Bitter gourd,Komatsuna, Turmeric, Japanese mallotus, Plantain,Young leaf of barley, Striped bamboo, Burdock, Field horsetail,Loquat leaf, Broccoli, Jew�s mallow, Japanese moutain ginseng, Parsley,Japanese parsley, Celery, Lotus root, Mitsuba, Japanese ginger,Asparagus, Ginger, Qing geng cai, Vitamin-na

Mushrooms Shiitake mushroom, Lucid ganoderma, Jew�s-ear, Maitake mushroom

Seaweed Kombu, Wakame, Fucus, Kombu root, Hijiki

Pulse and Cereals Soybean, Cocoa, Sweet corn, Rice bran, Unpolished rice

RodsLactobacillus acidophilusLactobacillus brevisLactobacillus fermentum

Lactobacillus acetotoleransLactobacillus buchneriLactobacillus kefiranofaciens

Lactobacillus amylovorusLactobacillus caseiLactobacillus plantarum

CocciLactococcus lactisPediococcus damnosus

Leuconostoc mesenteroidesPediococcus pentosaceus

Pediococcus acidilacticiPediococcus urinae-equi

Table 1 Food materials of PELY*

* Plant-based extract fermented by lactic acid bacteria and yeast.

Table 2 Lactic acid bacteria added as a starter in PELY*

* Plant-based extract fermented by lactic acid bacteria and yeast.

Fig. 1 Manufacturing process of PELY*.* Plant-based extract fermented by lactic acid bacteria and yeast.

� 97�

Page 3: 901353日本クリニック微量栄養素26/++20‐N‐2‐ …jtnrs.com/sym26/26_096.pdfWild herbs Sun-dried Extracted with hot water Filtered Fresh vegetables and fruits Washed

����������� ������������PDA�������� 30�� 6�������� ! "��#$%&��'�(��)���*��� CFU/g�������pH� PELY+,���-"�.�/0�10��w/w�12��3"� pH4�5��6����2������PDA#$%&��'�(�����1%�v/v�Triton

X-100�78�9:;$<=�99��10�:��DNA�ITS1F primer : GTAACAAGGT�T/C�TCCGT>?@ITS1R primer: CGTTCTTCATCGATG����28S rRNAA�B"C � PCR$?=D����"EF��$FG��������HA�B" PCRD���94��3�"I:J�94��30��55��1��72��1�"KLMN� 30OP=���QJ$ 72��7��RSTU��VW���A�B"EF����ITS1F primer� BigDyeTerminator FS ver. 1.1 Cycle Sequencing Kit�AppliedBiosystems����X�����Y �EF���BLASTZ[��C\�"3W�3]��^����^����

2�����1� ���C�_�`�ab"�c����d���e��f_��12�$gW��0��hij kl$?Th!�$?=6����5mnMb�o"pNq�N�$?W��!��rs!tYu%���vb�wxyNx�N$?Tz{M�|�}%�$?=�!���~��#�~���550��$?=�!�������������������������m�m��B���$?=�!����N���� ICP$��c�$?=�!����m��������m����%�$?=�!���&� o -���m���m���%�$?=�!����|m����%�$?=�!���e'���NDF���PELY� NaOH��(J�AACC�13�$gW��!���2�����������c���"?�$VW����ym�4y (m��PELY�¡¢���J 6 NE��0.04% β-4N�£�x5��N7)�� 150��20¤�j0�¥J��¦�M����¦�§�������¨*�c)©���6������£��ªm� PELY�0�������y wxy|m��'�N7)�� 110��12¤�j0�¥J�]«;9M����¦�§��HPLC��6�����"¬"������N¢(m��wm�­�ywm���(N�¦(m�y��m��L�m�Lz�L�m���m��¦(m����m�£��m��N5�m����m��| (m���n¦¢m����PELY� 6 NE��0.04%β-4N�£�x5��N7)�� 100��24¤�j0�¥J��¦�M����¦�§�������¨*�c)©���6����

3��������®5�m+"�c��d���e��f_��12�"��

$gW��®5�m A�|y��N���ODS¯�¦��0-45��N°;$?T±² �³Z%-HPLC���!���α-��m�β-��m�M�£� Kmym��ODS¯�¦��M��´N�-45��N,;$?Tµ¶ �³Z%-HPLC�$?=�!���®5�m B1�y��m���ODS¯�¦��45��N�0.01 mol/L�m�-0s������0.15 mol/L¡Es������°;$?T�.����¦��"�����m������"/·$?T��Z%�HPLC�$?=�!���®5�mB2��¸�¦®m���ODS¯�¦��45��N�¹�º»;$?T��Z%�HPLC�$?=�!���®5�m B6��Saccharomyces cerevisiae ATCC9080$?T0¼'½1�!�$?=�!���®5�m B12��Lactoba-cillus delbrueckii subsp. lactis ATCC7830$?T0¼'½1�!�$?=�!����L��m��Lactobacillus plan-tarum ATCC8014$?T0¼'½1�!�$?=�!���®5�m C��¾¿À�¦��¹�Án- KmÁ¹�xyN°;$?Tµ¶ ��Z%�HPLC�$?=�!���®5�m D��ÿÀ�¦�����(��NÁ45��N°;$?T�<]«;9M����¦��"Ä��¾¿À�¦�� 2-£�n��NÁn-Å Km°;$?T±² �³Z%�]«;9M����¦��$?=�!���®5�m E��¾¿À�¦��¹�Á 2 -£�n��NÁn-Å Km°;$?T��Z%�HPLC�$?=�!���®5�m K��ÆÇ�¦��ODS¯�¦��x5��NÁ45��N°;$?T��Z%�HPLC�$?=�!���nm�m���Lactobacillus plantarum ATCC8014$?T0¼'½1�!�$?=�!���2���Lactobacil-lus rhamnosus ATCC8014$?T0¼'½1�!�$?=�!���4����������������N7)!��È�m�(��14�$��+W

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3� !"#$�%&1�'�()*Myagmar� Aniya"��15��ESR-�Ëm�¦{£��$gW��PELY�/0� 1Ì1}%��50Éx5��N$,¥��ESR6�ÍÎ�JEOL, JES-FR 30 ES������HPX-XOD�­ KmymÏ Kmym  �q�ÐÑEC 1.1.3.22Ò�¯$?W�$¼ST��n�  KL�" 5,5-w4yN-1-Ë��m-N -  ���DMPO�3j9" ESR�,M�N�6����ÓÔ�H2O��4Õ���"hÖtY¦w�N×ØÙÚ�u%���ÛY$Ü5������n�  KL��§��5�Ð�SOD�$?TZ!Ý�Þ_��SOD6ÙÚ�������

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Page 4: 901353日本クリニック微量栄養素26/++20‐N‐2‐ …jtnrs.com/sym26/26_096.pdfWild herbs Sun-dried Extracted with hot water Filtered Fresh vegetables and fruits Washed

2��������Okamoto����16�����PELY� PBS 11�� ����� ����������� �Hippury-Histidyl-Leucine�HHL����� ������� �!"#$�ACE��%&'(�)*�+#$�,�� �-./0������� �!"#$�1�+3���23�4�5 �3��� 6789:��;�<=�Table 3�+>?3�Lactobacillus acidophilus NBRC13951�L. brevisNBRC3960�L. fermentum NBRC3071�L. plantarum THT030701�Lactococcus lactis NBRC12007�Enterococcus fae-calis NBRC3971�� Streptococcus mutans NBRC139558MRS@A�B�OXIODCD��Helicobacter pylori JCM12036� Porphyromonas gingivalis JCM122578EF@A�B�Vibrio parahaemolyticus NBRC127118 LB@A�B�GHIJK� 1.0%�w/v��#�LMN 0.5%�NaCl 1.0O�@A 1.3%�pH 7.0��P�Q�R38�S@A�B����B� �1�+T��U�B� PELY� 5%�w/v�VW�NaOH pH)X �Y���4�B� �+P�Z�� [\]�^ �J]\K�_` �3a��3bcdef �UJ]\K� 50 µLghi�j\��k� �+P���lm�n ��;�� ����B�o`p��iqr\s� 100t1000 CFUuJ]\K�vef����4�B�o`p��iqr\�wW�23(�x�� �+4�� ���α-�yj\z'(8�α-�yj\z'(�{M|K

�M|i\}����1�~�J����� ��� �+40��pH 5.0�;1� 30��� 1O~�J��F 1 mL��-$���s�� 670 nm���� 10 mm 66O������W=T���='(� 1 U� �+Jq��\z'(8��,���{�17��,���� ��q ��� ��� �+�z���>D����� ��?(Jq��\z838��pH 3.0��(Jq��\z838��pH 6.0����H(Jq��\z8 38��pH 9.0�;�

=����� 1��� 1 µg� L-�q �����=�����(�� ¡\��������¢V�Y���'(� 1 U� �+KHJ �'(8�Hirano����18��£¤��¥`��N-benzoyl-DL-arginine-p-nitroanilide�BANA��1��� �+H�\z'(8�H�\zM|K S�¦§�D�¨©ªC��1�« ?�¬��Jq\��BALB����� ��� �+5�����������Nihei����19��£¤��#$8mushroom��q

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� �

1�PELY��� !"�#$%�&'pH�( Fig. 2� PELY�>?3w�±3w�"#����²^�n �+>?3;�³±3�U3w8 3§#�´¦�µ��P�Z��5.8 log�CFU/g���7.4 log�CFU/g��16§#T¶·¸�¹º �+30§#�8��3.3 log�CFU/g��� 3.9 log�CFU/g��µ��P�»Y>?3�#��3w8¼½v��¾�²^ �¿pµ¢¹�2t4 log�CFU/g�À�$�Á�s%�¸�µ¤�+pH8 0§#8�pH 5.4<¤�s�"#���Â: Ã� 90§#�8 4.3TÂ: �+T��PELY�"#N�\�\� �ÄÅÆ�ÇV8 �1µ1s�"#����#��È��%�¸��=�É��U.�JH�ʧ�;�= PELY��3wËÌ�;1�PDAl��Í �iqr\�Î3 �±328S rRNAÏÐÑ¢&Jj�}\�1� PCR�®¤�+PDAl��Í �iqr\���8�PCR¢&Ò�s���µ1Ó¥s<��R3�iqr\YÔÕ �1��ÖW���+Á��É��{×Ø<¤�iqr\w8Ù��8ÚWµ1s�Saccharomyces Û�Candida Û�#�sÜ� �Ë�p��'Ý�ÖW���+�Y�(Þv� �Saccharomyces cerevisiae � Candida apicola � PDA��

Strain Temperature Incubation time Condition

Lactobacillus acidophilus NBRC13951Lactobacillus brevis NBRC3960Lactobacillus fermentum NBRC3071Lactobacillus plantarum THT030701Lactococcus lactis NBRC12007Enterococcus faecalis NBRC3971Escherichia coli NBRC3301Pseudomonas aeruginosa NBRC3080Salmonella typhimurium NBRC12529Staphylococcus aureus NBRC3060Staphylococcus aureus IID1677Vibrio parahaemolyticus NBRC12711Providencia rettgeri NBRC13501Streptococcus mutans NBRC13955Helicobacter pylori JCM12036Porphyromonas gingivalis JCM12257

37�37�37�37�37�37�37�37�37�37�37�35�30�37�37�37�

36 h36 h36 h36 h36 h36 h18 h18 h18 h18 h48 h24 h18 h48 h96 h96 h

AnaerobicAnaerobicAnaerobicAnaerobicAnaerobicAnaerobicAerobicAerobicAerobicAerobicAerobicAerobicAerobicAerobicMicroaerophileAnaerobic

Table 3 Microorganisms used for antibacterial effects

� 99�

Page 5: 901353日本クリニック微量栄養素26/++20‐N‐2‐ …jtnrs.com/sym26/26_096.pdfWild herbs Sun-dried Extracted with hot water Filtered Fresh vegetables and fruits Washed

Fermentation period (day)

8

7

6

5

4

3

2

1

0

7

6.5

6

5.5

5

4.5

4

3.5

30 50 100 150 200

Bac

teria

l cou

nt

log(

CFU

/g)

pH

LABYeastpH

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2�������1����PELY%�fgh.b7ij�k1/�Z�l,mP�

5n�op5�!W�8Xqr��&'st?u/25n�v�/+,?u/�Z�w6�8Xx��yz7{|=}~n�{��?u���Table 4�2PELY7&'e+�7 pH7����?$��������7()A/&'?u/�Z����t7 pH% 4.1��P?u��23��

�bP�������#�/�Z������X�3#"��h���?u/������=�=$� ?¡/���¢������2G�!�sb£q$<¤¥~���}¦�����§¨>7sb£q�!©ªA/ 1.9 g/100 g�3#��22��� ��PELY7�«¬�­+�®¯���°�±²�«¬�³´?�X�,µ¶·��� 18LM7�«¬�%A¯#�3#"��¸¹+,��#B�/º |«=�d��}¦»=�7�f�������R����Table 5�23��������PELY!�3/¼|«=M?½¾¿���%¨<�À=������?u���Table 6�2¼|«= C%EFG����2¼|«= C%��G:A�Pn?u/�����&'ÁÂ?��G��«¬��ÃÄA/������«¬� ÅÆ ÇÈ�É���Ê���21��01��2¨<�À=�%���7ËÌÍÌ�ÎÏstÐÑ+,Ò12���ÓFG/Ô%Õ0.7 mg/100 g?u/�����&'!()A/���d'D�.-�#�/�01��24�����������PELY%&'Ö×!ÈØ#�Ù��Ú¬� �f��i

ArginineLysineHistidinePhenylalanineTyrosineLeucineIsoluecineMethionineValineAlanineGlycineProlineGlutamic acidSerineThreonineAspartic acidCystineTryptophan

0.09 g0.11 g0.04 g0.14 g0.07 g0.23 g0.13 g0.04 g0.15 g0.15 g0.15 g0.20 g0.53 g0.14 g0.12 g0.45 g0.04 g0.04 g

Water contentProteinLipidCarbohydrateAshSodiumPotassiumCalciumMagnesiumPhosphorusZincIronManganeseSeleniumDietary fiber (NDF**)

34 g3.7 g1.3 g58.8 g2.2 g

137 mg619 mg131 mg68 mg97 mg11 mg3 mg1 mgNone1.9 g

Fig. 2 Time course of LAB*, Yeast, and pH in PELY**.* LAB : Lactic Acid Bacteria.** Plant-based extract fermented by lactic acid bacteria and yeast.

Table 5 Amino acid content of PELY* (per 100 g)

Table 4 Nutritional compounds in PELY* (per 100 g)

* Plant-based extract fermented by lactic acid bacteria and yeast.** Neutral Dietary Fiber. * Plant-based extract fermented by lactic acid bacteria and yeast.

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Fermentation period (year)

SO

D lik

e a

cti

vit

y (

U/g

)

600

500

400

300

200

100

0

0-year PELY 1-year PELY 3-year PELYPolyphenol content (µg/mL)

SO

D-l

ike a

cti

vit

y (

U/g

)

600

500

400

300

200

100

0

0-year PELY

1-year PELY

3-year PELY

200 210 220 230 240 250

r = 0.9844

��������Table 7��PELY3 �����������������246 mg/100 g�� �!"#$%&&'�����������()*�+,�PELY3 �����!"#$� 1.7-�&&'� 0.17-�� �

3��������1��� �PELY./0�12345SOD678��9:;<

3�=>.��Fig. 3���� ?@ABC�.DEF�12345�GH��I(@JKL.C �M�����������(12345(�NOFig. 4��PQ(IR�����������(12345.�S�NO@TKE�2������� �PELY�UVWXY�.Z[GH45����\]^$_

'$`ab$c$<def.Og���hACE��ij��XY@�MEFk�LlmL�PQ�Table 8.��nm.�PELY�opLKqXr�sq@�&$t

u��()*�F 61.4vijwE�M�PELY.5kFk�xy� 1z�5kF ACEij78�{6.|}��)*�(IR�PELY�~&$({��� ACEij78���I(@��wE�3�� �PELY����"#$������Cl�B�����({6.���e���E���B�����qr@��_������8@��@���;C��9�.n��qr��%�PELY��� ���(¡¢KE�£2¤%�¥@qs�1¤rp.n¦�§¨©¤���@ª�wEFk�(¡¢KE�«I��PELY�1¤45�PQ(IR�£2¤L. acidophilus�L. brevis�L. fermentum�L. plantarum�L. lactis�E. faecalis�%¬­¤E. coli�.®�F�1¤8��wCL @�¯°c�$±8²³´µ~¶¤MRSA�S. aureus IID1677��·¸¤P. aeruginosa��¹uº»$c'¤P. rettgeri�.®�F�¼k1¤8���Table 9��wK.½u�¤H. pylori�%¾¿À¤P. gingivalis��­Áº´�aV. parahaemolyticus�.®�FÂ1¤8���I(@JKL(C �M�ÃÄ

Polyphenol content(per 100 g)

Caffeic acid equivalent

PELY* (0-year fermentation)PELY* (1-year fermentation)PELY* (3-year fermentation)Red wineCocoa

212 mg236 mg246 mg145 mg1,440 mg

ACE inhibitory activity (%)

PELY*Bitter gourdJew�s mallowTurmericParsley

61.499.068.663.040.7

Vitamin A (retinol equivalent)α-caroteneβ-caroteneCryptoxanthinVitamin B1 (thiamin)

B2 (riboflavin)B6B12

Niacin (nicotinic acid equivalent)Vitamin CVitamin DVitamin E (α-tocopherol)

(β-tocopherol)(γ -tocopherol)(δ-tocopherol)

Vitamin KPantothenic acidFolic acid

20 µg25 µg192 µg77 µg0.05 mg0.07 mg0.27 mg0.03 µg1.69 mgNoneNone0.4 mgNone0.2 mg0.1 mg30 µg0.19 mg1 µg

Table 7 Polyphenol content of PELY*Table 6 Vitamin content of PELY* (per 100 g)

* Plant-based extract fermented by lactic acid bacteria and yeast.

Table 8 ACE inhibitory activity of PELY*

* Plant-based extract fermented by lactic acid bacteria and yeast.* Plant-based extract fermented by lactic acid bacteria and yeast.

Fig. 4 Correlation between polyphenol content and antioxidantactivity in PELY*.R is pearson product-moment correlation coefficient.* Plant-based extract fermented by lactic acid bacteriaand yeast.

Fig. 3 Time course of antioxidant activity in PELY*.* Plant-based extract fermented by lactic acid bacteriaand yeast.

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������S. typhimurium���� ���S. aureusNBRC3060�������������������� !"�#�$%!&� '(��)*+&(����,-.+� PELY/01�����23425�(67�89�:+ '(�� !�;<=>?@�AB�CD!&(E�&!,FG !�PELY,"�?@,�HIJK�LMNO#PQ#,RS,T�4U!&(E�V��WXYZ[�\]���^��_�"�?@/`a���b! ��'(EPELY�cd&B"�#[e�f����gh�ih�j�klmhn,WohpqrIstlum�vw&BT��x'�Zy+&Bz�?@/{|��("�?@/}fHj��AB��~m�Ie�CD!&BE�&d4��PELY�cd&B"�[e,#e�����{����"�#�����l�JI����121��15�������"�#��������#/W������� 1,000��,���*�4AB��d4� '��������{4ABE4�����������{� PELY�cd&B�q~m�Ie/��

������B��� ¡+&(�� !���P¢*/£( PELY��¤#,¥�¦&!,�§�¨Vcd&�PELY/©ª���«¬�(67�­,©ª,®¯P°±

/²³B?@/�´�� PELY��³Bµ¶®¯�§¤#/·9�¸(E� ����P¢*/£( PELY,¹ºy[���»�M�¼�j�s»�¼�Jtjum/c½��t��¼,�¾&,®¯�§¤#�$%!&� '(�Table 10�Ed(�Jtjum�t��¼�¤#¯�x¿ÀX,�¾& ,?@/WaB*��cd&B /·9(���§¤#�Á¯�� '((%�¦,xÂ�*��cd&�����CD!&(E5���� ������uÃ�¼¤#,Ä�FG/ Table 11�`�(EPELY��³B��uÃ�¼ÀX¤#, IC 50/ÅÆaB��58.5 mg/mL4A'(EPELY,Çt�ÈÉ�KcW�� 0.25 mg/mL4A'(,4�Çt�ÈÉ�KcW�

Total Activity Lower limit

α-amylaseAcid-ProteaseNeutral-ProteaseAlkaline-ProteaseTripsinLipase

n.d.n.d.n.d.n.d.n.d.n.d.

50 U/g20 U/g20 U/g20 U/g1 U/g

50 BALB Unit

Additive amountof PELY*

Colony account(CFU/plate)

Antibacterialactivity

L. acidophilus NBRC139510 % 908

�5 % 1097

L. brevis NBRC39600 % 517

�5 % 577

L. fermentum NBRC30710 % 140

�5 % 132

L. plantarum THT0307010 % 148

�5 % 125

L. lactis NBRC120070 % 139

�5 % 138

E. faecalis NBRC39710 % 546

�5 % 408

E. coli NBRC33010 % 112

�5 % 110

P. aeruginosa NBRC30800 % 290

+5 % 0

S. typhimurium NBRC125290 % 40

Growth-inhibition5 % 20

S. aureus NBRC30600 % 289

Growth-inhibition5 % 191

S. aureus IID16770 % 201

+5 % 0

V. parahaemolyticus NBRC127110 % 162

+5 % 0

P. rettgeri NBRC135010 % 294

+5 % 0

S. mutans NBRC139550 % 235

+5 % 0

H. pylori JCM120360 % 191

+5 % 0

P. gingivalis JCM122570 % 716

+5 % 0

Table 9 Antibacterial activity of PELY*

* Plant-based extract fermented by lactic acid bacteria and yeast.

Table 10 Activities of digestive enzymes in PELY*

n.d.: not detected.* Plant-based extract fermented by lactic acid bacteria and yeast.

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� IC 50������ 0.086 mg/mL��� ���������������������L-���� ��!"������#�PELY�!$�%&'�(��)*�+,�-.�/0*12%��3456�7�%-� �-89:���

� �

;<!=>?#�PELY�@A$�%B�� !CD����EFG�H?#�G�� ����I��-7���:JKJA��L�M��N�O�7�%��P�Q�MR�� �-89:���PELYH�S�L�M��T�UVW�XW��Y+���ZP���L�M��@[! T\��\]^�F�\)�-_J�5`#�"�;aHbcdH�eHfg���hijk\l�Fm�+�WAno !L�p��q#rs�56A622(t�%u�&'���)*��Q�vw�(b PELY�H�x�y�� �-��zHA��$�%&'�(��)*�+,���mE{ �m�+{ F^|l�}~�An�!4�:�H�"�#!�<�>�m�i��{ F��{ An![A! �$t��8� �z�������+���Q�%����&�An�45b'(:�56�23(�PELY�@)!$�%B�� 656�!��$�%&'�(��)*�+,�b*���A6������-�������u�!+,��P56x�O���J��PELYH_J��t�%u�H�65��-!�./A�GQ�Hbx_J�5`#�\)�! 1@�������z��01�,u:�����\�����\|���j��/�!?�A'��\�F�2$�F���H�x�3456� �-4��56��\)�! 1@�� γ-�\)5�&GABA(-_J��7�%b������:�H�o ��61H�y�����\)������5O� D-�\)�-��7!�� ¡!�8H96�>�F:¢�Hb"!�y-£;:�5`#�¤jAn!<�2���"!� F�$/t�-'(:�24(�¥y¦§-�P��56�� ��!t�%u�- PELYHb_J�56�7�%-��������P� ��!t�%u�H�65b����P� �b��!¨=���t�%>©!=>�PELYH��ª«?4�}+GM!

2%�¬�45`�­�}+@�{ H®-�4¯�A��Az��J�� ��!}+GM��3��{ b;P��Az��-!GM2%H�65b°OBOC��DE-��F�±�PELYH`65GM2%-;P���7�%!�GM�45��i���T*1-²³����i���T*1������$��2���x���H�345`#�´u�!µi��b2%-¬��� �-G¶:�56�25(��ª�PELY�m�+{ �·¸<¹H�{ �mE{ �.�/0*1�3{ �!�� �-8:���m�+{ H`65���G!�ºH95'���)*�-I�45`#�'���)*���xH�^»)i¼�XOD��3�� �-G¶:�56�26( �z���½¾¿H`À�m�+{ !<¹��½Á! SOD2%!I��'���)*�&�^»)i¼�(H?� XOD!�3! 2�!EÂH?�b!�������:�H�PELY�mÃÄ{ !J/GM��COX&/V�ÅD/Æ0*1(F�m����*{ !J/GM�� LOX&�'D/Æ0*1(HK���3{ �Ç� �-LM/A=>���-89:�5`#�m����*{ FmÃÄ{ H®-��$t��!456�7�%b#�¥y ��!!�%H�65b«?����4�-5�PELY���L�M!�NÈÀ�Ax��2ÉÊNLO�(/�4�t�%Q�MR�45!���7�%�Ç��GQ����������

����

1(Egusa G, Murakami F, Ito C, Matsumoto Y, Kado S,Okamura M, Mori H, Yamane K, Hara H, Yamakido M(1993) Westernized food habits and concentration ofserum lipids in the Japanese. Atherosclerosis 100 :249�255.

2(PË�ÌQ�R BS�TU Í�VÎ�Ï�ÐWÑÒ��XYÓ�ÔZÕY[&1999(\]�Ö3��×0�j^ØÙ^i��Ti�!Ú_+�6zHÛÜ��zÝ2`Þ+ 26 : 295�300Ý

3(Imazu M, Yamamoto H, Toyohuku M, Watanabe T,Okubo M, Egusa G, Yamakido M, Kohno N (2001) As-sociation of apolipoprotein E phenotype with hyper-tension in Japanese-americans : date from the Hawaii-Los Angeles-Hiroshima Study. Hypertens Res 24: 523�529.

4(Egusa G, Watanabe H, Ohshita K, Fujiwara R,Yamane K, Okubo M, Kohno N (2002) Influence of theextent of Westernization of lifestyle on the progres-sion of preclinical atherosclerosis in Japanese Subjests.J Atheroscler Thromb 9 : 299�304.

5(McCullough ML, Fesknanich D, Stanmpfer MJ, RosnerBA, Hu FB, Hunter DJ, Variyam JN, Colditz GA, Willet

IC 50 (mg/mL)

PELY* (water soluble ingredients)PELY* (polyphenol content equivalent)

58.50.086

ScopoletinAnisic acidBenzoic acidUmbelliferoneL-Ascorbic acidAnisaldehydeArbutin

0.240.0910.0780.0680.0580.0540.011

Table 11 Anti-tyrosinase activity in PELY*

* Plant-based extract fermented by lactic acid bacteria and yeast.

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8������1991������ ��������pp. 2�4�

9����������������������2006���Crassostrea gigas� !"#$%&'()*+,�-.�,/012,�-3,45�6 7�89: 23 : 89�92�

10�;< =8>=��2008�?@ABC� 1239D�24�11�EF�G��HIJK������2003�LCMNOPQR>STUVWX=��Y�*+Z,[\]^_�`�4�a����pp.88�

12�[bcdefghi�2000�jk `���lm.�n�opqrst����

13�Insoluble Dietary fiber. AACC Method : 32�20.14�AOAC OFFICIAL METHOD 952. 03, 955. 25 (1965).15�Myagmar BE, Aniya Y (2000) Free radical scavengingaction of medicinal herbs from Mongolia. Phytomedi-cine 7 : 221�229.

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24�ÆÇ­È�ÉÊË­�!�Ì��2006�"ÍÎϽÐ�, D-�¸¹�,Ñ �y1W�ÎÒ� D-�¸¹�,�Ó.§Ô�6 7�89: 23 : 1�4.

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