Patricia Yuca Hamaguchi

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1 1 Nippon Shokuhin Kagaku Kogaku Kaishi Vol. /,, No.2, -,/-,3 (,**/) 325 Patricia Yuca Hamaguchi Film-forming Mechanism of Biodegradable Films Prepared from Fish Myofibrillar Proteins Yusuke Shiku, Patricia Yuca Hamaguchi, Weng WuYin and Munehiko Tanaka Department of Food Science and Technology, Tokyo University of Marine Science and Technology,./1 Konan, Minato, Tokyo +*22.11 In our previous study, semi-transparent and flexible biodegradable films were successfully prepared from myofibrillar proteins of blue marlin meat. The objective of this study was to elucidate the mechanism of film formation. E#ect of ph on the amount of surface hydrophobicity and surface SH group in the film-forming solution, solubility of film proteins in various protein denaturant solutions, SDS-PAGE, and addition e#ect of protein denaturants in the film-forming solutions were investigated. At first, the necessity of unfolding of protein molecules was revealed for the formation of self-standing films. Secondly, the important chemical bondings involved in the protein-protein interactions were confirmed to be hydrophobic interaction for acidic condition, hydrogen bondings for neutral condition, and hydrophobic interaction and disulfide bondings for alkaline condition. (Received Feb. +*,,**/ ; Accepted May +,,,**/) / *** / -* +, CMC -. +*22.11./1 corresponding author mune@s.kaiyodai.ac.jp / ph / ph,- 1+, ph.0 ph 1 ph ph + Makaira mazara 0 /,* mm EDTA *.+ M Tris-HCl ph 1.0

2 326 /, 2,**/ 2 2 Laboratory Blender, Model -2B0, Waring Prod. Co. 3,**g, -*.,,/, + /* + M HCl + M NaOH ph, +, Hybrid Mixer HM-/** ANS ; ANS 2 *.*+ ph,+,. ml *.*. ANS *.. ml. +* +/ RF-+/** -0/ nm,.1* nm ph ph 1 +. SH SH, DTNB ; 3 *.*/ ph,+, / ml +* mm ph 1.* *.*+ M DTNB *.*-- ml. 0* + 1**g, +* UV-+0*.+, nm + g SH *. M + cm + /. g /.* cm/.* cm Environmental Chamber H++*K--*DM ;,/*./ /*/,. 0 SDS-PAGE ph,+,, SDS 2 M,* mm Tris-HClpH 2.2,, SDS-PAGE Laemmli +* 1./ NPU-1./L, ATTO *.+ R-,/*-* +* - -* +* 1,* 1* +,/ ++,, +, 1* + 2 Perez-Mateos +-. ph 1 *.0 M NaCl S+ *.0 M NaCl+ M S, *.0 M NaCl2 M S- *.0 M NaCl2 M *./ M, S.,/* mg / ml -*,. Advantec No. /C, Bradford +.. S+S. 3 ph ph, 1 +,, *.**/+.* M *.*++.* M *.*+ *./ M TS EAB 0,/*./ /*/ 1, TS EAB RE---*/ ;,* mm,./ mm *./ mms + TS MPa N m, EAB +* +* Excel SPSS Statistical Software, USA Duncan s multiple-range test +/ p*.*/ + SH ph,+,

3 : 327 SH Fig. + ph 1 ph SH ph 1 ph ph.0, SDS- PAGE SDS-PAGE ph SDS- PAGE Fig., ph SDS-PAGE ph, - +0 -. S+S. Table + S+ ph, -,,.. ph 1+, -.-1 S, S, S+ S,-S+ S,-S+ Fig.

, SDS-PAGE patterns of myofibrillar protein based films M+ : High molecular weight standard (,3,*/ kda). M, : Low molecular weight standard (,300 kda).

3 3 : 327 SH Fig. + ph 1 ph SH ph 1 ph ph.0, SDS- PAGE SDS-PAGE ph SDS- PAGE Fig., ph SDS-PAGE ph, S+S. Table + S+ ph, -,,.. ph 1+, -.-1 S, S, S+ S,-S+ S,-S+ Fig. + E#ect of ph on the amounts of surface hydrophobic group () and surface SH group () of myofibrillar protein film-forming solutions Fig., SDS-PAGE patterns of myofibrillar protein based films M+ : High molecular weight standard (,3,*/ kda). M, : Low molecular weight standard (,300 kda). Table + ph Protein solubility () of myofibrillar protein based films in protein denaturant solutions* +,, Protein denaturant solutions S+ /, /- S., * ++ +,,,4.-4+ a,,4.-4, a b -14/.4, b b b -.4/-41 b b,24/.4- a,14-.4, a /04.-4/ b /34,/4, b c c.,4*/4* c -24./42 d /141/4+ a 0*4//4- a 0-40/4* b 014,/43 b 0/4/04+ b b 0-4+/4, b / a / a 0,4//4- a , b 1*4+041 b 0/43/40 a 0243/4/ ab 1,4+041 b 0/4104. a * + S+ : *40M NaCl, S, : *40M NaCl+4/M urea, S- : *40M NaCl24*M urea, S. : *40M NaCl24*M urea*4/m,-mercaptoethanol4 *, Meansstandard deviation4 Any two means in the same column followed by the same letter are not significantly di#erent (p*4*/)4

4 328 /, 2,**/ 2 4 Table, E#ect of additives on the mechanical properties of myofibrillar protein based films Additives Urea Guanidine hydrochloride,-mercaptoethanol Concentration (M) * *4*+ *4+ *4/ +4* *4*+ *4+ *4/ *4**/ *4*+ *4+ +4* TS (MPa) ph, ph 1 ph +, +.4, * / /4/ , +*4. 04, / EAB () ph, ph 1 ph +,, ,,24* -*4,,/ ,.42,14*,+41,,4* +142,.42,/40,.4-, : Unable to measure4 : Unformed4 S- /1.101., S- S, S--S, S. /3.31,.+ S.-S- / Table, ph +, +.* M, *.+ M TS, EAB ph, *./ M *.+ M ph 1 *.+ M, *.*+ M, ph +, *.+ M ph, +.* M ph 1 *.**/ M TS ph 1 ph 0 Fig. + ph +1 unfolding Kim +2 ph SH ph ph 1 Fig unfolding SH,* S+S. Table + S. +**-S.

5 5 : 329 SHSS +2 ph ph,- ph +++, 0 SDS-PAGE Fig., Table,, ph 1 ph 1 unfolding ph unfolding +, - Krochta, J.M., Proteins as raw materials for films and coatings : Definitions, current status, and opportunities. In Protein-based Films and Coatings, eds. Gennadios A., (CRC Press, New York), pp. +.+,**, Kester, J.J. and Fennema, O.R., Edible films and coatings, A review. Food Technol.,.*,.1/ Gennadios, A., McHugh, T.H., Weller, C.L. and Krochta, J.M., Edible coatings and films based on proteins. In Edible Coatings and Films to Improve Food Quality, eds. Krochta, J.M., Baldwin, E.A. and Nisperos-Carriedo, M., (Technomic, Lancaster), pp.,*+-* Park, H.J., Rhim, J.W., Weller, C.L., Gennaidios, A. and Hanna, M.A., Films and coatings from proteins of limited availability. In Protein-based Films and Coatings, eds.gennadios, A., (CRC Press, New York), pp. -*/-,1,**, / Iwata, K., Ishizaki, S., Handa, A. and Tanaka, M., Preparation and characterization of edible films from fish water-soluble proteins. Fisheries Sci., 00, -1,-12,*** 0 Shiku, Y, Hamaguchi, P.Y. and Tanaka, M., E#ect of ph on edible films based on fish myofibrillar proteins. Fisheries Sci., 03, +*,.+*-*,**- 1 Shiku, Y, Hamaguchi, P.Y., Benjakul, S., Visessanguan, W. and Tanaka, M., E#ect of surimi quality on properties of edible films based on Alaska pollack. Food Chem., 20,.3-.33,**. 2 Kato, A. and Nagai, S., Hydrophobicity determined by a fluorescence probe method and its correlation with surface properties of proteins. Biochem. Biophys. Acta, 0,., +-,* +32* 3 Habeeb, A.F.S.A., Reaction of protein sulfhydryl groups with Ellman s reagent. Method Enzymol.,/,./ , +* Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage T.. Nature,,,1, 02*02/ +31* ++ Haurowitz, F., Chemical reactivity of proteins. In The Chemistry and Function of Proteins, eds. Haurowitz, F., (Academic Press, New York), pp. +/.+/ , Lugg, J.W.H., On the use of mercuric salts and nitrous acid in the colorimetric determination of tyrosine and tryptophan present in solution. Biochem. J., -*, +.,, Perez-Mateos, M., Lourenco, H. and Montero, P., Borderias, A. J., Rheological and biochemical characteristics of high-pressure and heat induced gels from blue whiting (Micromesistius poutassou) muscle proteins. J. Agric. Food Chem.,./, Bradford M., A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 1,,,.2,/ / Stell, R.D.D. and Torrie J.H., Principles and procedure of statistic. A biometrical approach. (McGraw-Hill New York), p. 20, +32* +0 Cuq, B., Aymard, C., Cuq, J.L. and Guilbert, S., Edible packing films based on fish myofibrillar proteins : formation and functional properties. J. Food Sci., 0*, / +1 pp.,+., Kim, S., Park, J.W. and Choi, Y.J., New approaches for the e#ective recovery of fish proteins and their physicochemical characteristics. Fisheries Sci., 03, +,-+ +,-3,**- +3 Orban, E., Quaglia, G.B., Casini, I., Caproni, R. and Moneta, E., Composition and functionality of sardine native proteins. Lebensm. -Wiss. u. -Technol.,,/, ,,* Okamoto, S., Factors a#ecting protein film formation. Cereal Foods World,,-,,/0,0, , +* +1 / +,

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