Antioxidative Effect of Crude Saponin Fraction Prepared from Culture Product of Basidiomycota cultured with Fresh Ginseng as Substrate

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(1)KOREAN J. FOOD SCI. TECHNOL. Vol. 37, No. 1, pp. 67~72 (2005). ©The Korean Society of Food Science and Technology. >âj 8î ¶ V·b¦V ªÒ Ò

(2) ò~ Öz Î" ;Ò**Á*Ò&1Áæ2Á"*3Á;ÿ* Ï"&v ®«", 1(")æ, 2ÏÓ& ÚÞ;&Ò", 3ö*ú:JI("). Antioxidative Effect of Crude Saponin Fraction Prepared from Culture Product of Basidiomycota cultured with Fresh Ginseng as Substrate Jae-Hyun Jeong*, Jae-Joon Wee1, Ji-Young Shin2, Ju-Hyun Cho3, and Dong-Hyun Jung Department of Food and Biotechnology, Chung Ju National University 1 KT&G, 2Department of Health and Diet, Chung Cheong University 3 Fen Biotech Co., Ltd Antioxidative activity of crude saponin fraction (CSF) prepared from Basidiomycota cultured with fresh ginseng (Panax ginseng C.A. Meyer) as substrate was investigated by analyzing CSFs for ginsenoside and phenolic compounds. On TLC chromatogram, ginsenosides such as Rg2, Rg3, and Rh1, which were rare in fresh ginseng, were identified. CSF of Phellinus linteus culture product showed the highest total phenolic content and electrondonating ability (EDA), suggesting phenolic compounds contribute to EDA. In vitro lipid peroxidation was inhibited most by CSF of Ganoderma lucidum, indicating that the highest EDA does not imply highest inhibition against lipid peroxidation. Tyrosinase was also inhibited mostly by CSF of G. lucidum. These results suggest culture of Basidiomycota with fresh ginseng has more active substances than fresh ginseng alone. Key words: DPPH, Antioxidative, Fresh ginseng, Phellinus linteus, Ganoderma lucidum, Hericium erinaceum. B. . ö & ;K ÛBÎ" 5 α-α-diphenyl-β-picrylhydazyl (DPPH)² Wj ¾æêb æî"Öz¢ ÛB~ z Oæö Î"'¢ ©¢ > ® (13). Þ, ªU~~ ãÖöê · ¢ Û &æ ÒW ¸f ²Ò > ® . 7 §«ªU (Hericium erinaceum)f J¾*¦V Ï 5 £Ïb Ï>Ú zb z 5 VË Ã;, òW*", Úî£ ö £Ò ÎË ® rJæ ® (14). "öº §«ªUb ¦V ~ ~òB Ï &Ë bî ªÒ>Ú ræ C&rb, $ §«ªU > ºÂ~ z^ Ã ÛBÎ" 5 Sacorma 180 ^ö & «· Î"ê >î (15). ÏîêªU~ ãÖöº Maeda (16)ö ~ z W 96.7%¢ ~& . Chung (17)f Ïîê ªU ÒÚ~ W Ú& Sacorma 180 >z 5 ;zj Î"'b ÛB ~& . Þ Han (18)f 'æªU (Ganoderma lucidum) ÒÚ~ ^ãöB ºÂ >ÏW ~ β-glucanW ganderanj Sarcoma 180 ^"& B v ö R~ 94%~ «· ÛBN ¾æêj : ®b, Chung(19)f 'æªU ¶ Ú ºÂb¦V Öz b îj ¦ï~ Â ÖzB~ Ï &ËWj ~& . >â, ÏîêªU, 'æªU 5 §«ªUöº z, , Öz Î"¢ &ê Wªj ~ &æ · ÒWbî F>Ú ® . V¢B, öBº ¶ j âö V· ãÖ ;zB FÏ bîj Ö > ®. â(Panax ginseng C. A. mayer)f , ¢, 7 æö B 'Úê 2000j ç &Ë æ £Òò ÒÏ>Ú z. . â~ zWª" £ÒÎËf *& "' ¢ Û Òò "º FÎWªb C&r, £Ò·Ïbº 7 ºã ÛB·Ï(1), Wî W /ê·Ï(2), VË .· Ï(3), ¶Ö FÒ·Ï(4), ë·Ï(5), Bz" z·Ï(6) >î . Òòf ¦(glycone)f j¦(aglycone) WB VÚB bêö 6Ò ª~ ®º, â Òòf j¦& dammaraneê triterpene 6 ßû' PD 5 PTê ¾*Úæ *Òræ 30« ç~ z & C&^ ® . âªöj ör ºÂ~ áÚæ² >º 1-2%~ æÏW Wªöº ¾¦ê zb, Òj^2, r¢ (7,8) 5 rÂ(9,10) B . 7 ¾¦ê zb ~ ãÖ, sâb¦V Öz Î"¢ ¾æÚº FÎ Wª ªÒB ¾(11) &~ &ç >Ú >â" Wâb¦V ô f Öz bî ªÒ>î (12). Öz bîf æî"Öz *Corresponding author: Jae-Hyun Jeong, Department of Food and Biotechnology, Chung Ju National University, 123 Geomdan-ri Iryumyeon Chungju, Chungbuk, 380-702 Korea Tel: 82-43-841-5248 Fax: 82-43-841-5240 E-mail: [email protected] 67.

(3) ®"²æ B 37 ² B 1 ^ (2005). 68. j ©b Òò>Ú, >âö ~ ¶j BÎ V·~ áf ÒÚ V·b¦V Òòj ªÒ~ Öz Wj ¦Æ~& .. µLO silica gel TLC plateö 6'~& . *BÏ º CHCl3/ CH3OH/H2O(13 : 7 : 2, ~[)¢ ÒÏ~&b, Bï£f 30% H2SO4¢ ªZ~ 110oCöB 10ª* Bï~& .. Òò 5 O». ¾¦W zb ;ï ò 10 mgj 50% öêR 1 mg/mL~ ³ê C~ AOAC~ Folin-Denis»j ¢¦ æ;~ jï ;ï~& . ò 0.2 mL(1 mg/mL)ö Na2CO3¢ 2.0 mL &~ 2ª* NöB O~ ê 50% Folin-Denis £j 0.2 mL &~ b~. NöB 30ª ;~ ê ª77êê(UV-Vis spectrophotometer, UV-1601, Shimazu) 750 nmöB 7ê¢ G;~& . &b î chlorogenic acid~ ³ê¢ Ò~ B ê &F j ·W~& .. þÒò þöB " ÒÏ ÏîêªU(Phellinus linteus), 'æªU(Ganoderma lucidum) 5 §«ªU(Hericium erinaceum) ÒÚº 7"³ëVFööB ª·Aj Ï"& v z þ öB potato dextrose agar(PDA, Difco, USA) Væö 15¢ *Ïb ê&V·(multi room incubator, VS-1203, Vision)~ &~B ÒÏ~& . ' "¢ 7« Vî ÒÏ~V * ÏÎ .Ö â ËöB 5j" >â( òÒ 45-50 g, φ2.5Ü27 cm)j «~& . âÒò &® ginsenoside Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2, Rg3, Rh1 10 «f KT&G 7ööB ªÒ ©j ÒÏ~& . "ê V· 5 «B Væº mushroom complete media(MCM) bVæ¢ ÒÏ~ &b, ÏîêªU 5 'æªUf 30oC, §«ªUf 25oCöB 15¢* V·~& . ï6ö Ú V·Î "¢ 8 mm FÒ&b ¦~ 100 mL MCMÚVæö 5BO 7 «~& . ÏîêªU 5 'æªUf 30oCöB 100 rpmb 10¢* êû V·(shaking incubator, SI-4000R, Jeio Tech)~&, §«ªUf 25oCöB 140 rpmb 15¢* V·~& . Ú V· ©j îV(Waring blender, BL91, Waring) îz~ 10 mLO 100 mL~ MCM ÚVæö 7«~ 7¢ * êû V· ©j «b ÒÏ~& . >âj 8î ¶ V· >âj b } ^¿ r ÚÝ~² îªÎZ ¢÷ j ÚÚ 48* ÿn çNöB Î ê 121oCöB 2* Î r Oï ©j Vî ÒÏ~& . Vî(>â òÒ)ö ''~ «j 20 mLO 7«~ ''~ V·Nêö B 20¢* BÎ V· ÒÚ V·bj −80oC .&NïÿV (deep freezer, VX490E, Jouan)öB 24* ÿÖÎ ê ÿÖ V(freeze dryer, FT8512, Ilshin)öB 72* ÿÖ Î © j îê~ ò ÒÏ~& . >â 5 >â V·b¦V Ò

(4) ò ºÂ 5 B Shin (20)~ O»ö V¢ ªöò 100 gö 70% zêR 900 mL¢ Î&~ ~~& ¦OB æ(80oC)öB 24*O 3² > ~~ ºÂ~& . ºÂf 60oCöB 6{ ³»V(rotary vacuum evaporator, N-N series, Eyela) ³»~ Ï 100 mL& F rræ zêRj B ê Vö bj &~ 200 mL ;Ï~& . ò 200 mL¢ Diaion HP-20(φ2.5Ü35 cm, Mitsubishi chemical, Japan)>æö O8 . O ê Ã ~>(1500 mL) >æ¢ Ïª® ^¿ r 90% zêR(500 mL) ÏÂ~ Òò ª³j áî . Òò ª³j ê³»V ³»~ zêRj B ê ÿÖ(−80oC, 9 mmHg)~ Òò ò ÒÏ~& . Ginsenoside~ TLC ªC ªÒ Òò 1 mgj 1 mL~ 50% öêRö Ï* 10. DPPH free radical ²»ö ~ *¶Ë ºÂb~ *¶Ë(electron donating ability, EDA)f Blois (21)~ O»j æ;~ ªC~& . ' òÏ 1 mLö 4Ü 10−4 M~ DPPH 1 mL¢ I v> ê 30ª* Nö O~ r ª77êê 520 nmöB 7ê¢ G;~& (1) ö V¢ EDA¢ ÖÂ~& . ·g EDA(%) = (1 − -------------------------------------- )Ü100 8ÆÂ/·g. (1). æî"Öz ÛB ·Ï Mitsuda (22)~ O»ö V¢ þ~& . VîÏ(linoleic acid)j B ê VîÏ 10 mLö 0.1 M phosphate buffer (pH 7.0) 9.6 mL 5 ºÂò Ï 0.4 mL¢ I 40oCöB 24 * ÿn >w8 . >w 2 mLö 35% trichloroacetic acid(TCA) 1 mL 5 0.75% thiobarbituric acid(TBA) 2 mL¢ b ~ 95oCöB 40ª* Bï>w 8 . NöB ï' ê acetic acid 1 mLf chloroform 2 mL¢ Î&~ ^N² z. r 3,000 rpmöB 5ª* öªÒ ê çj ª77êê 532 nmöB 7ê¢ G;~& (2)ö V¢ æî"Öz ÛBNj ÖÂ~& . Inhibition effect of lipid peroxidation(%) = ·g (1 − -------------------------------------- )Ü100 8ÆÂ/·g. (2). Tyrosinase & W /; Tyrosinase &W G;f tyrosinase~ ·Ï Ö" W>º DL-β-3.4-dihydroxyphenyl alanine(DOPA) chromej jï»ö ~ G;~º Masamoto (23)~ O»ö &~ þ~& . V îB 5 mM DL-DOPA solution 0.2 mL, 0.1 M phosphate buffer (pH 6.8) 0.2 mL 5 òÏ 0.5 mL~ bö tyrosinase (250 unit/mL) 0.1 mL¢ Î&~ 35oCöB 2ª* >wÎ. r 475 nmöB 7ê¢ G;~ (3)ö V¢ tyrosinase & Nj ÖÂ~& . ·g Inhibition ratio(%) = (1 − -------------------------------------- )Ü100 8ÆÂ/·g. (3). Ûê¾Ò Ö"º ïb ¾æÚî þ *~ jvªC f SAS(Statistical analysis system)¢ Ï~ ANOVA ªC ê Duncan’s multiple range test¢ ~& ..

(5) >âj Vî ¶ V·b¦V ªÒ Òò~ Öz Î" Table 1. Yields of crude saponin prepared from fresh ginseng and culture products of Basidiomycota Sample. Table 2. Total phenolic contents in the crude saponin prepared from fresh ginseng and culture products of Basidiomycota. Content (%, w/w). 1). b. 4.0Û0.2 4.3Û0.2a 4.6Û0.2a 4.2Û0.2b. FG GP2) GG3) GH4) a-b. Mean values (3 replication) with the different letters are significantly different by Duncan's multiple range test (p<0.05). 1) FG: Fresh Ginseng. 2) GP: Ginseng-Phellinus linteus. 3) GG: Ginseng-Ganoderma lucidum. 4) GH: Ginseng-Hericium erinaceum.. Fig. 1. TLC profiles of crude saponin prepared from fresh ginseng and culture products of Basidiomycota. Abbrebiations are the same as in Table 1.. Ö" 5 V V·b 7 Ò

(6) ò~ ºÂ >N >â 5 >âj Vî ÏîêªU(Phellinus linteus), ' æªU(Ganoderma lucidum) 5 §«ªU(Hericium erinaceum) ÒÚ V·b~ Òò ºÂ >Nf Table 1" ? '' 4.0, 4.3, 4.6 4.2% >âj Vî 'æªU ÒÚ V·b~ ºÂ>N &Ë ¸~ . Ginsenoside~ TLC ªC >â 5 >âj Vî ' ÒÚ V·b¦V ªÒB Òòj TLC ªC Ö"º Fig. 1" ? >âj Vî ' ÒÚ V·b~ Òò ÎvöB >â Òò" ? f *~ö ²ï~ ginsenoside& Îj ®rj r > ®î . ß ® ÏîêªU 5 'æªU ÒÚ V·b ÒòöBº Rg2f Rh1 *~öB spotj " > ®îb, §«ªU ÒÚ V·böBº Rg3f Rh1" ?f *~öB spotj " > ®îº, º >âj sâb B~º "; 7ö W>º bî >âVî~ ;j ~B WB ©¾(24) ¶ BÎ"; 7ö WB ©b " > ® . f ?. 69. Sample. Total phenolic content (%). FG GP GG GH. 1.04Û0.02d 1.11Û0.03c 1.45Û0.03a 1.29Û0.01b. a-d. Mean values (3 replication) with the different letters are significantly different by Duncan’s multiple range test (p<0.05). Notes are the same as in Table 1.. >âj Vî ¶ ÒÚ V·böBº â Òò º~~ ®rj r > ®î . Ginsenoside Rg2º VÛÛË K~ Ë¢ BFÊº ÎË(25) rJê: ®, ginsenoside Rg3º z^~ V&ï Jj F~~² &æ~º Wj ¾ æÞ (26) >î . "öº ginsenoside Rh1º Ò² F Oz ^ MCF-7 ^öB estrogen >ÏÚ¢ Wz Î. >î (27). C¾¦W zb ;ï Han (28)f mouseö "ï~ öêRj R~&j r ¢V >º æî"Öz 5 æO*ö &~ ¾¦Wª Î"& ®î b B> ÒòöBº "ÖzÛBÎ"& ì ~&. . Mei (29)f â Òò .& Úb^~ Öz' ¶ çOÚÎ"& ® ~&b, Chen (30)f " "Özæ î ÛBf SODWj Ã&8 ~& . $ Kim(31) f ÏîêªU~ ¾¦W zb 7öº WÖ² ², æ W Öz ÛB 5 .²6 w ÛB& ~& . . þöBº >âVî V·b~ Òò ª³7 Öz Î" f &N ¸f ©b rJê C¾¦W zb ïj G; Ö" Table 2f ? >â, ÏîêªU, 'æªU 5 §«ªU ÒÚ V·b ÒòöB 1.04, 1.11, 1.45, 1.29% ¢ ¾æÚÚ >â º >â~ ÒÚ V·b Îv ¸f ¾¦W zb ïj ¾æîb, 7 'æªU ÒÚ V· böB &Ë ¸f ¾¦W zb ïj ¾æÚî . Shin (20)f HP-20>æ¢ Û" * B â~ Òò 7öº ¾¦ï 2.2%¢ ¾æÚî ~ Ö" º. ² ¸~ . Kim(31)f Ïîê ªU~ ¾¦W zb~ ï f ªU~ zêR ºÂböB 33.3 mg%, >ºÂb 20.7 mg% 5 ª ºÂb 0.2 mg%¢ ¾æî ~& . $ Kim(32)f 'æªU ör 5 ¦êR ºÂböB '' 11.5, 15.5 mg%¢ ¾æÚî ~&º Ö"fº ç ^ jv~V& ¦ 6 ® . Fº Kim(32)~ G;~ º ªU ¶ Ú~ zêR ºÂböB ç7 ¾¦ïj G; © öBº >âj Vî ¶ ÒÚ V· b ¦V ºÂ 5 ªÒ Òòb G;~&V r^ . V¢B b ÿ¢ þ öB >âj Vî ¶ ÒÚ V·b" ªU V·b*~ ¾¦W zb ïj ç ^ jv *b >â V·b~ ¾¦W zb ïö ~º 'Ëj ÚÚ¶ . DPPH free radical ²»ö ~ *¶Ë DPPH ª¶Ú î²º £² *¶¢ Aj ªb ¾~ ÿ f ¶ïj ©ÚªÒº Wîj <º . *¶ ·Ïf æî"Öz~ ê>wö &~º ÖzW W free radicalö.

(7) ®"²æ B 37 ² B 1 ^ (2005). 70. Table 3. Electron donating ability (EDA) of crude saponin prepared from fresh ginseng and culture products of Basidiomycota Sample BHA1) CA2) AA3) FG4) GP5) GG6) GH7). EDA (%) 0.1. 1. 10 (mg/mL). 85.7Û1.9b 94.3Û2.7a 95.4Û0.7a 23.9Û0.1d 31.1Û0.6c 28.6Û0.2c 25.5Û0.4d. 86.5Û1.2d 95.8Û1.7a 95.9Û0.8a 90.4Û0.2c 92.8Û0.8b 83.9Û0.1e 86.5Û0.5d. 93.9Û1.7c,d 95.8Û0.9a,b 97.8Û1.7a 93.3Û0.4b,c 94.9Û1.2b,c 94.4Û0.1b,c 91.9Û1.1d. a-d Mean values (3 replication) within a column that followed by same letters are not significant different by Duncan’s multiple range test (p<0.05). 1) BHA: Butylated hydroxyanisole. 2) CA: Chlorogenic acid. 3) AA: Ascrobic acid. 4) FG: Fresh ginseng. 5) GP: Ginseng-Phellinus linteus. 6) GG: Ginseng-Ganoderma lucidum. 7) GH: Ginseng-Hericium erinaceum.. *¶¢ ~ Öz¢ ÛBÊº ¿ê& B . W free radicalf Ú ÚöB '« î÷" z¢ ¢bÊV r^ö " >w~ Wj ²Êº ËKj æò ÖzB ·Ï > ®º îÚ bîj öï jºW ® . >â 5 >âj Vî V·b¦V ªÒ Òò~ *¶ Ëf Table 3ö ¾æÚî . >â 5 >âj Vî V· b~ Òò~ ³êê *¶ËKj 0.1 mg/mLöBº Ôf *¶ËKj ¾æÚîb¾ 1 mg/mL 5 10 mg/mLöB º ¾ rJê Â 5 W ÖzBf Ò*ò ¸f *¶ ËKj ¾æÚî . ¯, 1 mg/mLöB BHA, chlorogenic acid, ascorbic acidº '' 86.5, 95.8, 95.9%¢ ¾æÚîº, >â 5 >âj Vî ~ V· ÏîêªU, 'æªU 5 § «ªU ÒÚ V·b Òòf '' 90.4, 92.8, 83.9 5 86.5%¢ ¾æêb >â 5 V·b Òò 7ö Öz W bî ï FNj r > ®î . Þ, 10 mg/mLöB º '' 93.3, 93.9, 94.9 5 91.9%B BHA, chlorogenic acid, ascorbic acidf ? ¸f >&~ *¶ËKj ¾æÚî . >âj Vî V·b Òò 7öº ÏîêªU Ò Ú V·b~ ãÖ 1 mg/mL 5 10 mg/mLöBº '' 92.8, 94.9% &Ë ¸f *¶ËKj ¾æÚîº, º Table 2 ö ¾æÂ C¾¦W zb ï~ Ö"f ¢~ . Kim(31)f ÏîêªU ¶ Ú zêR ºÂb~ *¶ËK 1 mg/mL öB 78-88%ªj ~&b, Chung (33)~ öBº V· ÏîêªU b ºÂböB 1.2, 10 mg/mLöB 8.3, 15% Ô f *¶ËKj ¾æÚîº, þöBº >âj Vî ÒÚ V·b~ Òò 1 mg/mLöB 92.8%B Kim(31) " Chung(33)~ Ö" ¸f *¶ËKj " ® . ©f >âj Vî ÒÚ V·b¦V Òòj º Â 5 ªÒ~º ";öB ÒÚ $º >â 7ö FB ¾¦ Wª þ ºÂ 5 ³»Nb Òò 7ö ¾¦ ï zêR $º b ºÂb ¸V r^ ©b ÒòB . æî"Öz ÛB·Ï >â 5 >âj Vî ' ÒÚ V·b~ æî"Öz. Table 4. Inhibition effect of crude saponin prepared from fresh ginseng and culture products of Basidiomycota against lipid peroxidation Sample BHA CA AA FG GP GG GH. Inhibition effect (%) 10 (mg/mL) 71.4Û0.17a 57.1Û1.11b 11.9Û0.87e 7.10Û0.20f 16.2Û0.26d 29.5Û0.60c 11.9Û0.34e. a-e. Mean values (3 replication) with the different letters are significantly different by Duncan’s multiple range test (p<0.01). Notes are the same as in Table 3.. ÛB Î"º 0.1, 1 mg/mLöB æî"Öz ÛB ·Ï ìîb , 10 mg/mLöBº Table 4öB º :f ? '' 7.1, 16.2, 29.5, 11.9% >â ÒÚ V·b~ æî"Öz Û B Î"& ±f ©b ¾æÒb ¶~ö V¢ N & ®rj r > ®î . ß® 'æªU ÒÚ V·b Ò ò~ Î"º 29.5% >â~ Òò¾ ÏîêªU Ò Ú $º §«ªU ÒÚ V·b Òò ¸f æ î"Öz ÛB Î"¢ ¾æÚî . *¶Ë~ ãÖöº > âj Vî ÒÚ V·b Òò W $º Â ÖzBf FÒ >&~ Wj ¾æÚîb¾, æî"Öz Û BöBº W ÖzB BHA $º chlorogenic acidö j R ÎÚæº ©b ¾æÒ . º *¶Ë B > æî"Öz ÛBW ¸æ prj ~ . ß® * ¶Ëö ®ÚBº ÏîêªU ÒÚ V·b Òò &Ë ¸² ¾æÒb¾ æî"Öz ÛBö ®ÚBº 'æªU ÒÚ V·b ÒòöB &Ë ¸² ¾æÂ 6ê f ? f £¢ ÒòB . Þ, *¶ËöBf ? æî" Öz ÛBWöBê >âj Vî ¶ V·b Ò ò >â ë~ Òò W ¸² ¾æÂ ©b j >âj Vî ~ ¶j V·b ;zB Wbîj B > ®rj & . Tyrosinase &W /; Tyrosinase &Wj G; Ö" 1, 10 mg/mLöB 0.1 mg/mLöB ¸f tyrosinase & W ¾æÒb, Fig. 2ö B º :f ? 0.1 mg/mLöB >â 5 >âj Vî ÏîêªU, 'æªU 5 §«ªU ÒÚ V·b ÒòöB '' 61.2, 61.9, 65.6 5 59.6%¢ ¾æÚîº, º 66.3%f 63.8%¢ ¾æÞ BHAf ascorbic acidf & > & . ß®, >âj Vî 'æªU ÒÚ V·b~ Òò~ tyrosinase &W 65.5%B V·b 7 &Ë ¸ ~ . Park (34)f ÏîêªUb¦V B Ö, ª, öj^rÞ, ¦êR 5 b ºÂ 1 mg/mL ³êö B 29.8, 100, 95.6, 19.6, 13.7%~ tyrosinase &Wj ¾æ î ~&º, WG; ò B O»" ÒÏ ³ê & þ" ¢ ç^ jvö ÚJÚ 6 ® . ß®, Kim (31) ÒÏ ÏîêªU~ ª" öj^rÞ º ÂböB Ö ; tyrosinase &W ¾æÂ 6 ßû' . ö b þöB ÒÏ ÒÚ V·bê .

(8) >âj Vî ¶ V·b¦V ªÒ Òò~ Öz Î". Fig. 2. Tyrosinase inhibition effect of crude saponin prepared from fresh ginseng and culture products of Basidiomycota. a-e Mean values (3 replication) with the different letters are significantly different by Duncan’s multiple range test (p<0.05). Notes are the same as in Table 3.. ª $º öj^rÞ ºÂbj B~ tyrosinase & Wj G; " jº& ® ÒòB . Tyrosinase & W bî öïf zË® Öë~ WÎ" 5 ®Öë~ . æz Oæö ®ÚB Ö 7º ¦ª F > ®b ß® ¢ò Wj ÛB > ®º zË® BBö ÏF ©b V&B .. º. £. >â 5 >âj Vî V·b¦V Òòj ªÒ~ TLC¢ { Ö" ÏîêªU 5 'æªU ÒÚ V ·b ÒòöB Rg2f Rh1 *~öB spotj " > ®îb , §«ªU ÒÚ V·böBº Rg3f Rh1" ?f * ~öB spotj " > ®î . V·b~ C¾¦W zb ï f >â, ÏîêªU, 'æªU 5 §«ªU ÒÚ V ·b ÒòöB 1.04, 1.11, 1.45, 1.29% ¾æ¾ >â. º >â V·böB ¸~b, ß® 'æªU ÒÚ V·bö B &Ë ¸f ¾¦W zb ïj ¾æÚî . >â V·b Òò~ *¶Ë~ ãÖ ÏîêªU ÒÚ V·b 1 mg/mL 5 10 mg/mLöB '' 92.8, 94.9% &Ë ¸~ . æî"Öz ÛBÎ"º 'æªU ÒÚ V·b Òò 29.5% ÏîêªU ÒÚ¾ §«ªU ÒÚ V· b ¸~ . Tyrosinase &Wf 'æªU ÒÚ V· b Òò 65.5% &Ë ¸~ .. 6Ò~ º 2002-2004j ÏÓ§ê ÎbÖë BBÒ ëö >¯B Ö"~ ¢¦ ö 6Òãî .. ^. ò. 1. Benishin CG, Lee R, Wang LCH, Liu HJ. Effect of ginsenoside Rb1 on central cholinergic metabolism. J. Pharmacol. 42: 223-229 (1991) 2. Lee KY, Park JA, Chung E, Lee YH, Kim SI, Lee SK. Ginsenoside Rh2 blocks the cell cycle of SK-HEP-1 cells at the G1/S boundary by selectively inducing the protein expression Pkip1. pp. 88-101. In: Korea-Japan Ginseng Symposium October 4, Novotel Ambassador Hotel, Seoul, Korea. Korea Ginseng & Tobacco Research Institute, Seoul, Korea (1996). 71. 3. Lee YS, Kim YR, Kim KM. Effect of Panax ginseng on morphine-induced immune suppression. J. Appl. Pharmacol. 3: 177181 (1995) 4. Elma ZT, Ilian EZ, Christina IH. Effect of ginsenoside Rg1 on insulin binding in mice liver and brain membranes. Phytotherapy Res. 5: 46-48 (1991) 5. Kim HS, Lee MK. Effect of ginsenoside on the development of morphine-induced torerance and physical dependence in mice. Korean J. Pharmacol. 20: 123-127 (1989) 6. Kikuchi Y, Sasa H, Kita T, Hirata J, Tode T. Inhibition of human ovarian cancer cell proliferation in vitro by ginsenoside Rh2 and adjuvant effects of cisplatin in vivo. England Anticancer Drugs 2: 63-67 (1991) 7. Park JD, Kim MW, Yoo SJ, Wee JJ. A thiazole and two β-carboline constituents from Panax ginseng. Arch. Pharmacol. Res. 11: 52-55 (1988) 8. Park JD, Kim MW, Yoo SJ, Wee JJ. Chemical studies on the ether soluble alkaloidal fraction of Panax ginseng. Isolation of 1carbobutoxy-β-carboline and 1-carbomethoxyoxy-β-carboline. Arch. Pharmacol. Res. 10: 197-199 (1987) 9. Jeong BS. Studies on the components of Korean ginseng (II). Korean J. Pharmacol. 7: 41-45 (1976) 10. Kim MW, Choi KJ, Wee JJ. Volatile flavor components of fresh ginseng. pp. 185-190. In: The 4th International Ginseng Symposium September 18, Daejeon, Korea. Korea Ginseng & Tobacco Research Institute, Seoul, Korea (1984) 11. Han BH. Studies on the anti-oxidant components of Korean ginseng. pp. 13-17. In: The 2nd International Ginseng Symposium September 7, Seoul, Korea. Korea Ginseng Research Institute, Seoul, Korea (1978) 12. Wee JJ, Park JD, Kim MW, Lee HJ. Identification of phenolic antioxidant component isolated from Panax ginseng. Korean J. Applid Bio. Chem. 32: 50-56 (1989) 13. Kim MW, Choi KJ, Cho YH, Hong SK. Study on the components of the antioxidant activity of Panax ginseng. Korean J. Agric. Chem. 23: 251-255 (1984) 14. Ahn DK. Medicinal fungi in Korea. Korean J. Mycology. 20: 154-166 (1992) 15. Kawagishi H, Shimada A, Hosokawa S, Mori H, Sakamoto H, Ishiguro Y, Sakemi S, Bordner J, Kojima N, Furukawa S. Erinacines E, F and G stimulators of nerve growth factor synthesis from the mycelia of Hericium erinaceum. Tetrahedron Lett. 37: 7399-7402 (1996) 16. Maeda YY, Ishimura K, chihara G. Antitumor polysaccharides and host defence against cancer; a new way for cancer immunochemotherapy. Protein Nucleic Acid Enzyme 21: 426-436 (1976) 17. Chung KS, Kim SS, Kim HS, Kim KY, Han MW, Kim KH. Effect of Kp, an antitumor protein-polysacchride from mycelial culture of Phellinus linteus on the humoral immune response of tumor bearing ICR mice to sheep red blood cells. Arch. Pharmacol. Res. 16: 336-338 (1993) 18. Han MD, Jeong H, Lee JW, Back ST, Kim SU, Yoon KH. The composition and bioactivities of ganoderan by mycelial fractionation of Ganoderma lucidum IY009. Korean J. Mycology. 23: 285-297 (1995) 19. Chung DO. Studies on antioxidative substances of Ganoderma lucidum. Korean J. Food Sci. Technol. 24: 497-503 (1992) 20. Shin JY. Choi EH, Wee JJ. New methods for separation of crude ginseng saponins. Korean J. Food Sci. Technol. 33: 166-172 (2001) 21. Blois MS. Antioxidant determination by the use of a stable free radical. Nature. 26: 1198 (1958) 22. Mitsuda H, Yasumoto K, Iwami K. Antioxidative action of indol X compounds during the antioxidation of linolic acid. 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