446 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 (Chen et l., 11; Qin et l., 8), (Xu et l., 3), (Himy et l., 12) in vitro. (Hippompus dominlis) (Syngnthif

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1 한수지 49(4), , 16 Originl Artile Koren J Fish Aqut Si 49(4), ,16 빅벨리해마 (Hippompus dominlis) 유래펩신가수분해물의 In vitro 와 In vivo 에서의항산화효능 김현수 신병옥 김서영 Lei Wng 이원우 김윤택 1 노섬 1 조문제 2 전유진 * 제주대학교해양의생명과학부, 1 한국해수관상어센터, 2 제주대학교의학전문대학원 Antioxidnt Ativity of Pepsin Hydrolyste Derived from Edile Hippompus dominlis in vitro nd in Zerfish Models Hyun-Soo Kim, Byeung-Ok Shin, Seo-Young Kim, Lei Wng, WonWoo Lee, Yoon Tek Kim 1, Sum Rho 1, Moonje Cho 2 nd You-Jin Jeon* Deprtment of Mrine Life Siene, Jeju Ntionl University, Jeju 63243, Kore 1 Center of Ornmentl Reefs nd Aquriums, Jeju 63354, Kore 2 Deprtment of Mediine, Medil Shool, Jeju Ntionl University, Jeju 63243, Kore Sehorse Hippompus dominlis mrine teleost fish, hs long een used s one of the essentil mterils in trditionl Chinese mediine. However, the uses of sehorse hve een limited due to its high ost, despite its enefiil iologil tivities. Sehorse hs not een widely explored for its iofuntionl properties nd tive omponents. In the present study, the enzymti hydrolystes of sehorse were prepred y using two digestive enzymes (trypsin nd pepsin) nd five food grde enzymes (neutrse, protmex, llse, kojizyme, nd flvourzyme). The enzymti hydrolystes indited higher hydrolysis yields thn its wter extrt. Among them, the distilled wter-pepsin hydrolyste (DP) whih ws otined y distilled wter extrtion followed y pepsin hydrolysis, showed the highest yield nd protein ontent s well s the highest lkyl rdil svenging tivity. Also, it provided protetive effets ginst oxidtive stress indued y AAPH in vero ell nd zerfish. Further frtiontion sed on the moleulr weight ws rried out to identify it s tive omponents, nd < 5 kd (less thn 5 kd) moleulr weight frtion ws onfirmed to hve the highest ntioxidnt tivity. In onlusion, this study suggests tht DP of sehorse hs ntioxidnt properties, nd might e novel nd useful mteril from the mrine origin for helthy funtionl foods nd osmetis. Key words: Sehorse, Hippompus dominlis, Enzymti hydrolysis, Antioxidnt tivity, Zerfish 서론 (Syngnthiformes), (Syngnthide) (Hippompus) 32, 7% - (Myers, 1979; Vri, 1982; Lourie et l., 1999). (Hippompus orontus), (Hippompus histrix), (Hippompus kud), (Hippompus mohnikei) (Hippompus trimultus) 5 (Kim nd Lee, 1995).,, (Vinent, 1996; Choi et l., 6). (Zhu, 5), (She et l., 1995; Zhng et l., 1994), (Xu nd Xu, 1997), (Ryu et l., 1; Ryu et l., 1), Koren J Fish Aqut Si 49(4) , August 16 This is n Open Aess rtile distriuted under the terms of the Cretive Commons Attriution Non-Commeril Liens ( whih permits unrestrited non-ommeril use, distriution, nd reprodution in ny medium, provided the originl work is properly ited. Reeived 3 June 16; Aepted 11 July 16 *Corresponding uthor: Tel: Fx: E-mil ddress: youjin@jejunu..kr Copyright 16 The Koren Soiety of Fisheries nd Aquti Siene 445 pissn: , eissn:

446 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 (Chen et l., 11; Qin et l., 8), (Xu et l., 3), (Himy et l., 12) in vitro. (Hippompus dominlis) (Syngnthifromes), (Syngnthide).,, (Woods, 7),.

2 446 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 (Chen et l., 11; Qin et l., 8), (Xu et l., 3), (Himy et l., 12) in vitro. (Hippompus dominlis) (Syngnthifromes), (Syngnthide).,, (Woods, 7),. (Hippompus kud Bleeker), (Hippompus guttultus), Short-snouted sehorse (Hippompus hippompus). 2 (trypsin, pepsin) 5 (neutrse, protmex, llse, kojizyme, flvourzyme),,,,,,,, (Bekmn et l., 199; Cho et l., 12; Heo et l., 5; Ahn et l., 7; Prk et l., 4). 실험시약및재료 재료및방법 (CCORA) m (Fig. 1),. Fig. 1. The photo of different sizes of sehorses Hippompus dominlis. 2 (trypsin, pepsin) Sigm Chemil Co. (USA) 5 (neutrse, protmex, llse, kojizyme, flvourzyme) Novozyme Co. (Denmrk). 2,2-zois-(2- midinopropne) dihydrohloride (AAPH), -(4-Pyridyl-1- oxide)-n-tert-utylnitrone (4-POBN), 3-(4,5-dimethylthizol- 2-yl)-2,5-diphenyltetrzolium romide (MTT), 2,7-dihlorofluoresein diette (DCF-DA), sodium dodeyl sulfte (SDS), dimethyl sulfoxide (DMSO), ovine serum lumin (BSA) Sigm-Aldrih (St. Louis, MO, USA). Piere BCA protein ssy kit Thermo Sientifi (Rokford, IL, USA) RPMI 16 fetl ovine serum (FBS) Gio/BRL (Cnd).. 해마의일반성분분석 AOAC (Helrih, 199) , Kjeldhl, Soxhlet. 해마의단백질효소가수분해물의제조 Ko et l. (13), 7 (distilled wter, DW). 1:1, ph 24. DW 24 pepsin trypsin 12 DW-pepsin (DP) DW-trypsin (DT). 1 1 ph 7.. pore size 7 m 7. 단백질효소가수분해물의수율및단백질함량측정 1. 7 Piere BCA protein ssy kit, BSA.

3 빅벨리해마효소가수분해물의항산화효능 447 Alkyl rdil 소거능측정 lkyl rdil Kng et l. (13)., µl, DW µl, mm AAPH µl, mm 4-POBN µl 37 3 pillry tue ; Eletron spin resonne (JES-FA ESR, JEOL, Tokyo, Jpn) signl. DW. Power: 1 mw, Amplitude: 1, Modultion width:.2 mt, Sweep width: 1 mt, Sweep time: 3 s, Time onstnt :.3 s 세포배양 Vero ell 1% FBS, 1% ntiioti RPMI 16. Cell ulture flsk 37, 5% CO Vero ells 에서의독성평가 Ko et l. (12) MTT ssy. Vero ells (1 1 5 ells/ml) 96 wellplte 19 µl, 37, 5% CO mg/ml MTT 5 µl dimethylsufoxide (DMSO) 12 ELISA reder 5 nm. Vero ells 에서의 ROS 생성측정 ROS Ko et l. (12) AAPH ROS vero ells 96 wellplte 18 µl, mm AAPH 1 µl, % DCF-DA 1 µl 37 1 ELISA reder. Vero ells 에서 AAPH 에의한산화적손상에대한보호효과측정 AAPH Ko et l. (12) AAPH vero ells 96 wellplte 18 µl seeding, mm AAPH 1 µl mg/ml MTT 5 µl. 37 3, 37 3 DMSO 12 ELISA reder 5 nm.. Zerfish emryo 에서의 AAPH 산화보호효과측정 Zerfish (Dnio rerio) (Kim et l., 14). 7-9 hpf (hours post-fertiliztion) zerfish emryo 12 well pltes 15 emryo medium 1.8 ml (25, 5, 1, µg/ml) 1 µl mm AAPH 1 µl.. 통계분석. SPSS (SPSS In. Ver. 12.) Onewy ANOVA-test Turkey s multiple rnge test P<.5. 해마의일반성분 결과및고찰 Tle 1,,,, %. Tle 1. Chemil ompositions of sehorse Hippompus dominlis (%, w/w) Composition Moisture Ash Crude protein Crude lipid Hippompus Adominlis

4 448 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 해마의단백질효소가수분해물들의수율및단백질함량, DW (Tsi et l., 6). DW 24 1 g mg, 7,. DP mg, DT mg (Tle 2). 5. DP neutrse llse, mg, mg (Tle 2). 1 g DW mg, DW 24 2 (Tle 2). DP mg, 5 (Tle 2). DW DP DT, 5. 해마단백질효소가수분해물들의 Alkyl rdil 소거능 Alkyl rdil. IC 5 (mg/ml) DP.15. mg/ml, DT.15. mg/ml, Neutrse mg/ml, DP DT lkyl rdil (Tle 2).. DP,, lkyl rdil DP. DP 의세포독성및 AAPH 산화보호효과 MTT ssy MTT (suinte dehydrogense) (formzn), (Yng nd Boo, 13). DP Fig. 2A. 5 g/ml, 1 g/ml, g/ml, 4 mm AAPH. DP AAPH (Fig. 2B), AAPH AAPH ROS, AAPH Tle 2. Yields nd lkyl rdil svenging tivity of enzymti hydrolystes prepred from sehorse Hippompus dominlis y seven proteinses Yields (mount tined from 1 g of rw smple) Rdil svenging tivity (IC 5, mg/ml) Hydolystes ontents (mg) Protein ontents (mg) Alkyl DW 374.5± ±3.4.38±.6 Trypsin 41.5± ±2.79 <.25 Pepsin 445.± ± ±.6 DT 1 5.± ± ±. DP ± ±7.2.15±. Neutrse ± ± ±.16 Protmex 67.± ±6.21 <.25 Allse 78.± ±3..73±.9 Kojizyme ± ± ±. Flvourzyme ± ±1.56.7±.3 1 Distilled wter-trypsin hydrolyste (DT). 2 Distilled wter-pepsin hydrolyste (DP).

5 빅벨리해마효소가수분해물의항산화효능 449 ROS (Fig. 2C). DP AAPH ROS. AAPH ron rdil, ron rdil ron rdil peroxy rdil, peroxy rdil,, (Niki, 199). DP 및분자량별분획물의 SDS PAGE 를이용한분석및각분자량분획물의 Alkyl rdil 소거활성 DP. DP 5 kd 1 kd 1 kd, 5-1 kd, 5 kd, Fig. 3A SDS-PAGE., 5 kd lkyl rdil IC mg/ml (Fig. 3B). DP lkyl rdil DP. (Ko et l., 12, Je et l., 5, Ren et l., 8, Zho et l., 9),. DP 분자량별분획물의세포독성및 AAPH 에의해산화된 Vero ells 의보호효과 DP Fig. 4A. MTT ssy 1 kd, 5-1 kd, 5 kd 5 g/ml, 1 g/ml, Vero ells. 4 mm AAPH 5 g/ml, 1 g/ml Vero ells, Vero ells., 5 kd Frtion 1 g/ml % (Fig. 4B). 4 mm AAPH 5 g/ml, 1 g/ml, Vero ells Totl ROS, 5 kd 1 g/ml Totl ROS %. 5 kd Totl ROS Cell viility (%) (A) (B) (C) Cell viility (%) d 1 8 ROS prodution (%) 1 8 d CON 5 1 CON 5 1 CON 5 1 DP (µg/ml) 4 mm AAPH + DP (µg/ml) 4 mm AAPH + DP (µg/ml) Fig. 2. (A) The ytotoxi effet of distilled wter-pepsin hydrolyste (DP) on viility in vero ells. Cells were treted with DP t the indited onentrtions (5, 1 nd µg/ml). (B) Protetive effet of DP on AAPH-indued oxidtive dmge in norml ells. The viilities of ells on AAPH tretment ws ssessed y MTT ssy. (C) Inhiitory effet of DP on the intrellulr ROS generted in AAPHindue oxidtive ells. Experiments were performed in triplite, nd the dt re expressed s men ± SD. - P<.5 indites signifintly different in eh groups.

45 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 (A) 75 kd 63 kd 48 kd 35 kd 25 kd 17 kd 11 kd Mrker DP 1 kd 1-5 kd 5 kd Alkyl rdil svenging tivity (%) 1 9 8 7 5 3 1 (B) IC 5 vlues (mg/ml).125 mg/ml.

6 45 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 (A) 75 kd 63 kd 48 kd 35 kd 25 kd 17 kd 11 kd Mrker DP 1 kd 1-5 kd 5 kd Alkyl rdil svenging tivity (%) (B) IC 5 vlues (mg/ml).125 mg/ml.25 mg/ml 1 kd 1-5 kd 5 kd 1 kd 1-5 kd 5 kd.24±..21±..15±.1 Fig. 3. (A) SDS-PAGE ptterns of moleulr weight frtions from DP. (B) Alkyl rdil svenging tivities of the moleulr weight frtions prepred from DP. Alkyl rdil svenging tivities of the three smples (.125 mg/ml,.25 mg/ml) were mesured y ESR spetrometry. - P<.5 indites signifintly different in eh groups. 1 1 (A) 5 μg/ml 1 μg/ml 1 1 (B) d 5 μg/ml 1 μg/ml 1 1 (C) d 5 μg/ml 1 μg/ml Cell viility (%) 8 Cell viility (%) 8 ROS prodution (%) 8 CON 1 kd 1-5 kd 5 kd CON 1 kd 1-5 kd 5 kd 4 mm AAPH CON 1 kd 1-5 kd 5 kd 4 mm AAPH Fig. 4. (A) The ytotoxi effets of the moleulr weight frtions from DP on viility in vero ells. Cells were treted with the moleulr weight frtions from pepsin digestive hydrolyste t the indited onentrtions (5, 1 µg/ml). After 24 h inution, ell viilities ws ssessed y MTT ssy. (B) Protetive effets of the moleulr weight frtions from DP on AAPH-indued oxidtive dmge in norml ells. The viilities of ells with AAPH tretment ws ssessed y MTT ssy. (C) The intrellulr ROS ws deteted y DCFH-DA ssy. Grph showed the intrellulr ROS svenging tivity. Experiments were performed in triplite, nd the dt re expressed s men±sd. -d P<.5 indites signifintly different in eh groups. (Fig. 4C). ESR lkyl rdil, DP in vivo 5 kd zerfish emryo. DP 5 kd 이하분획물의 AAPH 에의해산화된 Zerfish emryo 의보호효과 Zerfish., emryo,, zerfish

7 빅벨리해마효소가수분해물의항산화효능 451 in vivo (Kng et l., 15; Ko et l., 14). 7% epioly zerfish emryo 15 mm AAPH zerfish emryo, 3 DP 5, g/ml, 24, 48, 72. g/ml 5% 5 kd AAPH. in vivo zerfish emryo DP AAPH., lkyl. DP Vero Cells ROS, AAPH. DP, lkyl, 5 kd, vero ells ROS AAPH DP Survivl rte of zerfish (%) CON mm AAPH + DP 5 kd frtions (μg/ml) 3 h 24 h 48 h 72h Elpsed times (h) Fig. 5. Survivl rtes fter tretment with AAPH or o-tretment with 5 kd moleulr frtion from DP. The emryos were treted with 15 mm AAPH with or without 5 kd moleulr frtion from DP t the different onentrtions (25, 5, 1, µg/ml).., 5 kd AAPH zerfish emryo., DP,. 사사 15 " " Referenes Ahn GN, Kim KN, Ch SH, Song CB, Lee J, Heo MS, Yeo IK, Lee NH, Jee YH, Kim JS, Heu MS nd Jeon YJ. 7. Antioxidnt tivities of phlorotnnins purified from Ekloni v on free rdil svenging using ESR nd H 2 O 2 - medited DNA dmge. Eur Food Res Tehnol 226, Bekmn JS, Bekmn TW, Chen J, Mrshell PA nd Freemn BA Apprent hydroxyl rdil prodution y peroxynitrile implitions for endothelil injury from nitri oxide nd superoxide. PNAS 87, org/1.173/pns Choi YU, Rho S, Jung MM, Lee YD nd Noh GA. 6. Prturition nd Erly Growth of Crowned Sehorse, Hippompus orontus in Kore. N Am J Aquult 19, Chen CB, Yun XH, Chen Z nd Yi MH. 11. Study on extrtion nd ntioxidnt tivities of liquefied protein of threespot hippompus. Chinese Tropil Mediine 11, Cho CH, Lee BH, Kim HY, Kim YC nd Kim DO. 12. Effet of feedstuffs on minerl omposition, ntioxidnt pity, nd protetion of neuronl PC-12 ells of deer tlers. KSBB J 27, Helrih K Offiil Methods of Anlysis of AOAC Intl. 15th ed.; Assoition of Offiil Anlysis Chemists, Wshington, DC, USA. Chpter Himy SWA, Ryu BM, Qin ZJ nd Kim SK. 12. Peonol from Hippompus kud Bleeler suppressed the neuro-inflmmtory responses in vitro vi NF-kB nd MAPK signling pthwys. Toxiol In Vitro 26, org/1.116/j.tiv Heo SJ, Prk EJ, Lee KW nd Jeon YJ. 5. Antioxidnt tivities of enzymti extrts from rown seweeds. Bioresoure Tehnol 96, ioteh Je JY, Prk PJ nd Kim SK. 5. Antioxidnt tivity of peptide isolted from Alsk Pollk (Thergr hlogrmm) frme protein hydrolyste. Food Res Int 38,

8 452 김현수ㆍ신병옥ㆍ김서영ㆍ Lei Wng ㆍ이원우ㆍ김윤택ㆍ노섬ㆍ조문제ㆍ전유진 Kim IS nd Lee WO First reord of the sehorse fish, Hippompus trimultus (Pises: Syngnthide) from Kore. Kor J Zool 38, Ko JY, Lee JH, Smrkoon K, Kim JS nd Jeon YJ. 13. Purifition nd determintion of two novel ntioxidnt peptides from flounder fish (Prlihthys oliveus) using digestive proteses. Food Chem Toxiol 52, org/1.116/j.ft Kng N, Ko SC, Smrkoon K, Kim EA, Kng MC, Lee SC, Kim J, Kim YT, Kim JS, Kim H nd Jeon YJ. 13. Purifition of ntioxidnt peptide from pepti hydrolystes of Mideodeok (Styel lv) flesh tissue. Food Si Biotehnol 22, Ko SC, Kim D nd Jeon YJ. 12. Protetive effet of novel ntioxidtive peptide purified from mrine Chlorell ellipsoide protein ginst free rdil-indued oxidtive stress. Food Chem Toxiol 5, org/1.116/j.ft Kim EA, Lee SH, Ko CI, Ch SH, Kng MC, Kng SM, KO SC, Lee WW, Ko JY, Lee JH, Kng N, Oh JY, Ahn G, Jee YH nd Jeon YJ. 14. Protetive effet of fuoidn ginst AAPH-indued oxidtive stress in zerfish model. Crohydr Polym 12, Ko SC, Kng N, Kim EA, Kng MC, Lee SH, Kng SM, Lee JB, Jeon BT, Kim SK, Prk SJ, Prk PJ, Jung WK, Kim D nd Jeon YJ. 12. A novel ngiotensin I-onverting enzyme (ACE) inhiitory peptide from mrine Chlorell ellipsoide nd its ntihypertensive effet in spontneously hypertensive rts. Proess Biohem 47, dx.doi.org/1.116/j.proio Kng MC, Kim SY, Kim EA, Lee JH, Kim YS, Yu SK, Che JB, Choe IH, Cho JH nd Jeon YJ. 15. Antioxidnt tivity of polyshride purified from Anthopnx korenum Nki stems in vitro nd in vivo zerfish model. Crohydr Polym 127, rpol Ko JY, Kim EA, Lee JH, Kng MC, Lee JS, Kim JS, Jung WK nd Jeon YJ. 14. Protetive effet of quultured flounder fish-derived peptide ginst oxidtive stress in zerfish. Fish Shellfish Immun 36, org/1.116/j.fsi Lourie JR, Vinent ACJ nd Hll HJ Sehorses: An Identifition Guide to ther Worlds s Speies nd Their Conservtion. Projet Sehorse, London, 214 pp. Myers GS A freshwter sehorse. P Dis 32, Niki E Free rdil inititors s soure of wter-or lipid-solule peroxyl rdils. Meth Enzymol 186, Prk PJ, Shhidi F nd Jeon YJ. 4. Antioxidnt tivities of enzymti extrts from n edile seweed Srgssum horneri using ESR spetrometry. J Food Lipids 11, Qin ZJ, Ryu BM, Kim MM nd Kim SK. 8. Free rdil nd retive oxygen speies svenging tivities of the extrts from sehorse, Hippompus kud Bleeler. Biotehnol Biopro E. 13, s Ryu BM, Qin ZJ nd Kim SK. 1. SHP-1, novel peptide isolted from sehorse inhiits ollgen relese through the suppression of ollgenses 1 nd 3, nitri oxide produts regulted y NF-kB/P38 kinse. Peptides 31, dx.doi.org/1.116/j.peptides Ryu BM, Qin ZJ nd Kim SK. 1. Purifition of peptide from sehorse, tht inhiits TPA-indued MMP, inos nd COX-2 expression through MAPK nd NF-B tivtion, nd indues humn osteolsti nd hondroyti differentition. Chem-iol Intert 184, org/1.116/j.i Ren J, Zho M, Shi J, Wng J, Jing Y, Cui C, Kkud Y nd Xue SJ. 8. Purifition nd identifition of ntioxidnt peptides from grss rp musle hydrolystes y onseutive hromtogrphy nd eletrospry ioniztion-mss spetrometry. Food Chem 18, org/1.116/j.foodhem She M, He GX, Chen H nd Jin ZJ An experimentl study of five speies hloios on nti-ging tivity. Chinese J Mr Drugs 53, Tsi JS, Lin TC, Chen JL nd Pn BS. 6. The inhiitory effets of freshwter lm (Coriul flumine, Muller) musle protein hydrolystes on ngiotensin I onverting enzyme. Proess Biohem 41, org/1.116/j.proio Vri RP Fishes of three western North Atlnti, Sufmily Hippompine. The sehorses, Sers Foundtion for Mrine Reserh, Yle University Report 1, Vinent AC The Interntionl trde in Sehorses. TRAF- FIC Interntionl, Cmridge, U.K., Woods C. 7. Aquulture of the ig-ellied sehorse Hippompus dominlis Lesson 1827 (Teleostei : Syngnthide). Ph.D. Thesis, Vitori University, Meloume, Austrli. Xu DH nd Xu SB The extrt from Hippompus trimultus Leh: its effet of ntithromosis on rts nd ingredients nlysis. Chinese J Mr Drugs 61, Xu DH, Mei XT, Li BJ, Lin ZI nd Xu SB. 3. The phrmologil effets of Hippompus psule on enhning sexul funtions of rts. J Chinese Med Mter 26, Yng AY nd Boo HO. 13. Phenoli ompounds, ntimiroil effets nd tyrosinse inhiition tivities of uumer grown greenhouse ording to ultivrs nd growth stges. Koren J Plnt Res 26, kjpr Zhu AM. 5. Phrmologi reserhes on ethnol extrts

9 from Hippompus. Chinese Phrmeutil Affirs 19, Zhng ZH, Xu GJ, Xu LS nd Wng Q Inhiitory effets of Hippompus SPP. extrts on L-glutmi id indued C influx in rts neurons. Chinese J Mr Drugs 52, 6-9. Zho Y, Li B, Dong S, Liu Z, Zho X, Wng J nd Zeng M. 9. A novel ACE inhiitory peptide isolted from Audin molpdioide hydrolyste. Peptides 3, 빅벨리해마효소가수분해물의항산화효능 453

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05 (01-10-OK).hwp J Koren So Food Si Nutr 한국식품영양과학회지 43(5), 661~667(14) http://dx.doi.org/1.3746/jkfn.14.43.5.661 천년초선인장줄기의항당뇨및항산화효과 이지나 김형은 김용석 전북대학교식품공학과 Anti-dieti nd Anti-oxidtive Effets of Opunti humifus Cldodes Ji-N