1. 1 Sephadex G210 Sephadex G215 (Sigm a ) ; Am berlite CG CHEM ICAL JOU RNAL O F CH IN ESE UN IV ERS IT IES :

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1 V ol. 25 N o CHEM ICAL JOU RNAL O F CH IN ESE UN IV ERS IT IES , 1, 1, 2, 3, 1, 1 (1., ; 2., ; 3., ),, ;, L eu2a sp2tyr2glu;. ; ; ; ; Q 503 A (2004) , [1 3 ]. : : ( : 2001BA 960C). 2.,, 2 3, [4, 5 ]. [6 ]., [ 7 ], 52. [8, 9 ].,, L eu2a sp2t yr2glu,, Sephadex G210 Sephadex G215 (Sigm a ) ; Am berlite CG2120 ( ) ; A lcalase (N ovo ) ; (, 60. 6% ) ; ( ) ;. [ 10 ]. SHA 2C ; ; Am icon M 2000 ; HD ; Beckm an J222m ; H itach i ; W aters ; F innigan M A T LOQ ; H itach i , 1 molgl N aoh ph 8. 0,, 50, ph 8. 0., m in, (7 000 rgm in, 15 m in), (M w = ),. [11 ]., Sephadex G215 : m g 1 ml molgl, Sephadex G215 (2. 0 cm 66 cm ) molgl, 0. 6 ml gm in. 220 nm,,,. : 100 m g 2 ml 0. 1 molgl (ph 2. 5), Am berlite CG2120 (N a ) (3. 5 cm 40 cm ) molgl : (1942 ),,,,. E2m ail: zhangxz@m ail. jlu. edu. cn

2 N o. 3 : 2, ph 2. 5, 3. 0, 4. 0, 4. 5, 5. 0, 6. 1, 6. 7, 8. 3, ml gm in,,,.. Sephadex G210 : 60 m g 1 ml, Sephadex G210 (210 cm 80 cm ), 0. 5 ml gm in,. H PL C : 1050 R P2H PL C, T SK OD S C 18 ( 5 Λm, 7. 8 mm 300 mm ) ; 220 nm, 1 ml gm in, 30, 20 m g (0. 8 ml ), A : 70% ( 0. 1% T FA ) ; B: 0. 1% T FA ; A B (35% 65% 50% 50% ).,,,,, (PR ) [12 ]. (M S), , 4 h, 30%. ( ), 4 h Sephadex G215 Am berlite CG2 120 (N a ) Sephadex G210, H PL C, ( 1),., m in, H PL C F ig. 1 Elution prof ile of the ma in fraction from the Sephadex G-10 column by a prepara tive reversed-pha se HPLC ( ), 1. 4, n (Glu) n (T yr) n (L eu) n (A sp) = Table 1 Am ino ac id pa ttern s of the corn an ti-ox ida tive peptide Ratio of Am ino acid Ratio of Am ino acid Am ino acid Θ(m g ml - 1 ) Am ino acid Θ(m g ml - 1 ) M aagm tot Comp. M aagm tot Comp. Glu A sp Tyr O ther L eu (M S) H PL C, , (M w ) ( 3), M w M w = 230, M w = ( 4). M w = 392 ( 5), M w = , M w = ( 6), M w = M w = , = M w = , = 162. M w = , = Sek i [13 ] ( Schem e 1 ), B C, Y N : 1, C 1 (17), N 1 467

3 V ol. 25 F ig. 2 Full M S of corn an tiox ida tive peptide F ig. 3 Secondary M S2 of peptide fragmen t(m w= 538) F ig. 4 Secondary M S3 of peptide fragmen t(m w= 230) F ig. 5 Secondary M S2 of peptide fragmen t(m w= 392) (1). 1 n (Glu) n (T yr) n (L eu) n (A sp) = ; M w = N C. C, = , M w, ; N, = ,. M w = 162, = 163, ; = 180.,, M w , 1, = 265,, F ig. 6 Secondary M S3 of peptide fragmen t(m w= 230) Scheme 1 Sek imethod for a peptide bond ana lysis ;,., ( )., L eu, L eu2a sp2t yr2glu L eu2a sp2t yr2glu, T yr, [14 ],, T yr A sp Glu, T yr,, T yr. T yr,., N L eu,,.,.,,,, 468

4 N o. 3 :,, [9 ].,,, [14 ].,,. [ 1 ] Park K. J., J in H. H., Hyun C. K.. P rocess B iochem. [J ], 2002, 38: [ 2 ] Hook V. İ H., Burton D., Yasothornsrikul S. et al.. B iochem. B iophyṡ Reṡ Co. [J ], 2001, 285: [ 3 ] L iepke C., Zucht H. D., Forssm ann W. G. et al.. J. Chrom atogṙ B: B iom edical Sciences and App lications[j ], 2001, 752: [ 4 ] Yam aguchim., Takada M., NozakiO. et al.. J. N utṙ Sci. V itam inol. [J ], 1996, 42: [ 5 ] Tanabe S., Tanimoto S. Y., W atanabe M. et al.. A gric. B iol. Chem. [J ], 1991, 55: [ 6 ] U chida K., Kaw akishi S.. J. A gric. Food Chem. [J ], 1992, 40: [ 7 ] M arcuse R.. N ature[j ], 1960, 186: [ 8 ] M urase H., N agao A., Terao J.. J. A gric. Food Chem. [J ], 1993, 41: [ 9 ] Chen H. M., M uramoto K., Yam auchi F. et al.. J. A gric. Food Chem. [J ], 1996, 44: [ 10 ] YOU Xin ( ), Compositive U tilization and Further P rocess of Corn ( ) [M ], Beijing: China L ight In2 dustry P ress, 1995: [ 11 ] GUO M in2l iang ( ), CHEN Jun ( ), J IAN G Yong2M ing ( ) et al.. Food Science ( ) [J ], 1992, (10) : 1 3 [ 12 ] ZHAN G Ying ( ). Food Science( ) [J ], 1997, 18 (5) : [ 13 ] Seki S., Kam bara H., N aoki H.. O rg. M ass Spectrum [J ], 1985, 20: [ 14 ] Chen H. M., M uramoto K., Yam aguchi F.. J. A gric. Food Chem. [J ], 1995, 43: Prepara tion and Structura l Character iza tion of a New Corn An ti-ox idative Peptide XU L i 1, L I X iang2l u 1, WU X iao2x ia 1, WAN G H ua 2, HOU R ui2zhen 3, HUAN G Yi2B ing 1, ZHAN G Xue2Zhong 13 (1. K ey L aboratory f or M olecular E nzym olog y and E ng ineering of the M inistry of E d ucation, J ilin U niversity, Chang chun , China; 2. China2J ap an U nion H osp ital of J ilin U niversity, Chang chun , China; 3. Colleg e of Chem ical E ng ineering, Chang chun U niversity of T echnolog y. Chang chun , China) Abstract A k ind of new corn anti2oxidative pep tide w as p repared v ia p rotease hydrolysis of zein catalyzed by alcalase, an alkaline p ro tease under a certain condition, fo llow ed by iso lation and purification. T he hy2 drolysis tim e w as fixed on the basis of the dependence of antioxidative activity on hydrolysis degree. It w as found that the hydrolysis for 4 h gave the h ighest antioxidative activity. T he isolation and purification step s include ultrafiltration w ith a m em brane of cutoff M w 1 000, successive ch rom atography on Sephadex G215, Am berlite CG2120 (N a) ion exchange, Sephadex G210 and p reparative H PL C colum nṡ T he am ino acid se2 quence of the antioxidative pep tide w as characterized to be L eu2a sp2t yr2glu by am ino acid analysis and electro sp ray ion trap m ass spectrom etry. W e also discussed the po ssible an ti2ox idative m echan ism of the antioxidative pep tide on the basis of its structure. Keywords Corn pep tide; Enzym atic hydrolysis; A ntioxidative activity; Ch rom atography; M ass spec2 trom etry (Ed. : H, J, Z) 469

5 461 M olecular Recogn ition Study of Som e A l ipha tic Ch ira l Enan tiom ers and D yes with Β-Cyclodextr in and Its D er iva tives ZHAN G Q iang, L IU Yu 3 Chem. J. Chinese U niv., 2004, 25 (3), 458_ L ow-va len t Titan ium Induced Reductive Cycl iza tion of o-n itrobenzam ides and Ketones SH ID a2q ing 3, WAN G Ju2X ian, RON G L iang2ce, ZHUAN G Q i2ya, HU Hong2W en Chem. J. Chinese U niv., 2004, 25 (3), 462_ Prepara tion and Structura l Character iza tion of a New Corn An ti-ox ida tive Peptide XU L i, L I X iang2l u, WU X iao2x ia, WAN G H ua, HOU R ui2zhen, HUAN G Yi2B ing, ZHAN G Xue2Zhong 3 Chem. J. Chinese U niv., 2004, 25 (3), 466_ Hydroxyl M acrocycl ic Polyam ines Coordination System s In teraction with D NA and The ir An titum or Activ ities in vit ro L IAN G Feng, W E I Jun, L I Zhao2Yang, WU Cheng2T ai 3, ZH EN G Cong2Yi, SUN M eng2x iang Chem. J. Chinese U niv., 2004, 25 (3), 470_ Investiga tion s on the Adsorben ts for Urem ic M iddle M olecular Tox in s(g ) Crossl inked Ch itosan Ba sesm odif ied by Ac idic Am ino Ac ids WAN G Hong, ZHAN G M ing2m ing, YAN G M ei, L g U J ian, YUAN Zh i 3, H E B ing2l in, L IU B in, ZHAO Cheng2M ei, SH EN B in Chem. J. Chinese U niv., 2004, 25 (3), 474_ Surface Sta te Studies of Nanocrysta ll ine TiO 2 with H igh D ielectr ic Con stan t RUAN Sheng2P ing 3, DON G W ei, WU Feng2Q ing, WAN G J ing, ZHAN G X in2dong, ZHAO M u2yu Chem. J. Chinese U niv., 2004, 25 (3), 484_ 487 g