TRUONG DAI HOC BACH KHOA TP. HO CHi MINH KHOA Ki THUAT HOA HOC | BO MON CONG NGHE THUC PHAM BK BAO CAO MON HQC CNCB THT & THUY SAN Dé wai UNG DUNG AP SUAT CAO TRON QUAN VA CHE BIEN THIT GVHD: Th.S Nguyén Thj Hién SVTH : HCO7TP NAM HQC 2010- 2011MO DAU Theo xa hudng toin cu héa, ci nha sin xuit cée sin phim thit dang phii canh tranh gay git vii nhau, Bé gitt ving va ming cao vi tr cia ho, cée céng ty thye pham va tht edn quan tim dén sw thay di trong thdi quem ma sim va ida thu sin phim cia nguai tga ding, cding nhur quan diém va nhu cd cia ho. Nhu ciu cia ngwdi tiéu ding thay déi lign tye nhumg mét vai quan diém chinh khéng thay déi. Nhin chung, ngwai tiéu ding Iudn d&i hoi chat hegng cao va sy tign dung cita cée san pham thit, voi mii vi ty nhién, Bén canh d6, ho con yéu cia ‘sy an toan va cde sin phim phai ty nhién khong b6 sung phy gia nhu e« quan, chat gitt am... Dé dip img tit ca ning you clu nay ma khong lam gidm d@ an toan thye phim cin phii 4p dung cic cong ngh¢ méi trong cong nghigp thye phim néi chung va trong cong nghigp thi cstn phim tht 4éng g6i chin khong i rigng. Hom nita, ¢ dung lit ming va dang w6p la nhing ding sin phim co nu clu gia ting rit cao trong vii nim gin day, Nhitng sin phim nay e6 kha nang nhiém khuan trade ki ng géi. Vi thE, gidi php cln thidt 1 cng dung céc ky thugt m6i cho cée ding sin phim trén, Hign nay, mot vai cng nghé mOi dang duge nghién ctu trong dé dp suit cao li mot phirong php cé tiém ning img dune rit cao ft hita hen d6i v6i cde sin phim thit va nd cho Ap sult cao 18 mot ky thu thy tiem nang trong v phat trién cée ding sin phim méi tiéu thy it nang hrgng,1. Lich sie phat trign: Gio su PW Bridgman (1914), mat ngudi tgn phong trong vat ly &p suit cao, bio céo vé albumin long tring tig va long d6 bi dng ty dud dp suit thay tinh cao 500-600 MPa mi khdng 6 si phin rf ita lap v6. Bi ring &p suét ny cho th thiy tinh eao (cao dp) la mét edng ow hit ich cho ché bién thge phiim thay vi xir Ij nhigt, Tuy nhign img dung cia ép suit cao d@ ché bién thye phim da gin nb bo qua cho Xhi sy khii du eda dy én "Phat tién dp suit cao Ten men sit dung Dense Mass" duge hi try boi BG Nong nghigp, Lim nghigp va Thuy sin (1989) tai Nhat Bin , Bie bit, cée nha khoa hye ve thit & Ue datién hah ép dyng dp sut cao ké tr iu nhtng nim 1970 (Macfarlane 1973; Bouton, Ford, Harti, Macfarlane, O'Shea 1977). KE tir khi Khoi phat ci dy an tgi Nhgt Ban, vige 4p dung ép suat ché bién thye pham 43 tha hit ahiéu si chi @ Nat Ban va Chiu Au béi vi nhimg thay d6i trong dc tinh cia nguyén ligu thye phim gay ra béi dp suat trong cic phiong thie khie nha ti nhigt (Cheftel 1992 ; Hayashi 1992; Johnston 1995; Knorr 1996). Mét s6 loai thye phim duge ché bién bing vic ‘mat trén thj trudmg, (Suzuki 2002). sir dung ap suat da e6 Nhin chung dp suit cao sir dung trong khodng 100-600 MPa tai mht &9 phéng. Ap suit cao lim pha hily cae té bao sinh duéng cia vi khudn va v6 hoat enzyme, nhung khong lam thay doi die diém goai cia sin pham va gitt lgi mot 86 vitamins. Tuy nhién, kha nang chju dyng ca vi sinh vat rét khde nhau phy thude vio giéng va log tht dupe xi. Anh hung cia xir Ij ap sudt cao eting pha thape vio dp suft sr dang, nhigt d9 va thi gian, Sir dmg dp suit cao o6 thé gay ra nhamg bigs ddi dc bigt v8 cfu trie eta sin phim vi kha ning niy e6 thé duge sit dung dé phat trién mot ding sin phim méi hoge gia ting chic néng cia mét s6 thinh phan nao d6. vith 16 thé ap dung ky thudt nay trén quy mo cong nghigp voi cde nghién citu tir phong thi nghiém cdn tién hanh: Thiét lap cae digu kign vir ly tot nhat cho méi loai sin phim, K&t hop ap suit cao véi cdc hé thong déng géi méi, cée chat khang kin 6 ngudn gée te nin, enzyme.2. Anh hwréng cia ap suit cao trong cdng nghigp thit Anh hung cia ép suit cao Ién cdc think phan dinh dung trong thit Thit li moi tnrdmg t8t cho sy phittrién eda vi sinh vat, thinh phan héa hoe ‘cba thit chit yéu 1A made, protein (15-21%), chat béo (0,5-25%), fe vitamin (gidu vitamin nhém B) va 4c oligonutrient, Theo quan diém Ij, khi gia ting dp su sé 6 mt inh hurimg vit If 1én cde phan ti Lim ching tién igi gin nhau bom dn d&n sp chayén pha, sy chuyén pha nay od thé dio nguge Iai sau khi giém ép. Dida nay la hing ei 8 nay ra vi nue vA chit bo. Theo quan diém hia hoe, dp sult cao tbe dong nh hon so voi nhigt d>. Cée lién két cOng héa tr} Khong bj phi vo nbung ete Tign két yéu nhw lién két hydro va lién két ky nude 06 thé bj bién G61 bat thugn nghich (Cheftel, 1995). Anh huring ci ap suit lin nuse chi yéu bao gdm sy gidm nbift d néng chay vit gia ting sy fon hoa din dén sy giém pH. Nhimg bién déi may ta thud nghich theo ap suit, Nhung ching gép phin lam bién d6i nhimg die diém cia sin pham dirge xir ly ép sudt cao, Calpastatin bj tre ché tgi dp suit tir 200MPa tré én trong khi calpain bj thoai hia & ép suit tin 400MPa, Tai ép suit thip hon 200MPa cic lysosome bi pha v9, kha ning tr phin gia ting vi thit mém hon. Cathepsin H_ va aminopeptidase bj vo host & dp suit ti 200MPa tré 1én va cathepsin D bj v6 hoat khi 4p suat dat t6i SOOMPa (Montero & Gomez-Guillen, 2002), Cie vitamin va dudmg trong thit khdng bj bign di boi ip suit cao nhumg cée polysaccharide o6 thé bj bién Adi. Nhin chung, st hinh thinh gel bi ate ché bat dp suit cao vi ép suit cao o6 thé bién d6i nhigt d6 chuyén pha tir sol én gel, Sw ding Iai 6 thé dusge tgo ra bai dip suit vi sau 6 gel tao thanh sé mém vi sing hon... Clu trie chinh eta protein bj anh hutimg nhe bai ip sudt eao, sw bién dBi fe Hign két yéu o6 thé lam bién tinh protein hoe nguge Tgi lim hoat héa enzyme. Cée anh huéng nay rt khée nau phy thude vao logi protein va digu kign ela qua trink xi Ap suit cao mang Igi sy chuyén pha thuga nebjch cho lipid tr Ling thinh rin din dn sg dong Ini, Néu li mt hin hop lipid, dp suft cao o6 thé tgo ra sy phan tich té bao membrane. Cheah va Ledward ede pha khéc uhau bing vi (1996) cling nghién cina c pha bly cd inh hing cia p suit vé qui trinh oxy hia chit béotrong co hip bam nho. Trén ca si dya vio s6 do cia acid thiobarbituric (TBA), ho chi raring tr gid TBA khong ting trong oo bp bam nho tigp xtc vi dp sudt cao len 200 MPa, sau 46 hor tang Léa khi fp sudt ka 300 MPa, vi ting 16 16t khi 800 MPa. Ap suat cao trén 300 MPa dén 400 MPa sé lim gidm myoglobin va ‘oxymyoglobin, Fe myoglobin tr6 thinh Fe" metmyoglobin va protein globin bi biéa tinh, tir dé am gia tang qua trinh oxy héa lipid. Ngoai ra, nguyén nban cia sx gia fing cdn c6 thé do cae kim loai trong thjt nim trong cic mudi phospholipids hoe mudi acid hu co véi kim log, ViAhE trong qué tinh 4p suit cao, ci ion kim Jogi (cé thé La Fe va Cu) da gidi thot ra khéi cdc phite chat nay va xie te phan img, oxy hoa chit béo. Hoe sy gia ting oxy héa chit béo trong thit & ap suat cao c6 1 do sy thay ddi edu tg0 ciia hemoprotein lim be Id ede nh6m heme xiic tie oxy héa chit béo, Newoe Iai, Orlien va Hansen (2000) da bio edo qué trinh oxy héa lipid & fp sult cao khdng lign quan dén vige gidi phéng céc nonheme sit hay host tinh xe tie cia metmyoglobin, shumg c6 thé lign quan dén sy pha hiy membrane. Theo Cheftel va Culioli (1997), sy oxy ha gay ra béi Ap suat 6 thé durge hhan ché eae qua trinh cng nghé cho cde sin phim thit, bing ech déng g6i sin phim hay sit dung efe chit chéng oxy héa. Logi bi oxy hoje thém dioxide earbon (CO.) trade Khi xit yp suit oxo d& ngin chin qué trinh oxy héa lipid Anh hurimg ciia 4p suat eao aén 49 mém thit Khi mot con vat duge giét mé, qué trinh co cig sé phit trién trong vang vai gid véi sy co cia cdc sgi co va kim tang 46 nhét ca thit. Miéng thit sau do sé mém ‘hung huang vj bi giém sit ding ké, va tht d6 thi khéng phit hgp cho néu an vi ché bién vi dp nhét cao va kha ming git nude thip. Néu thit duge gid 6 nhigt 46 th trong mét vii ngiy, tht tro nén mém mai vil gid trang thii 46 trong vai tin tip theo. Vi vay, qua trinh sit dung rng rli nhit cho thit vii edi tién cia hug vi vi mii duge goi la conditioning va aging. Néa nhiimg migng thit ds dt bigt ltr nhimg con dng vit gia c6 thé diege im mém bai ap suét cao thi 4p suit cao o6 tim nang Im trong vige tn tha nguén nguyén ligu tir thit ca cde con vat gid, trink Hing phi.Mat thir nghigm dé ting a mim cia tit bari ép suit cao Tan dit tign durge thye hign boi Macfarlane (1973) & Uc. Mét diéu rit quan trong la phai Iya chon thai ‘gian nio eda thit sau Khi mé thich hgp cho vige dp dung ép suit cao. 2.2.1. Anh hung ciia 4p suat cao Ién co trrde giai doan té eiing: Macfarlane (1973) da thye hig cae phép do Khe nhau trén fc co Biceps femoris gi 100 MPa trong 2-4 phit, Két qui cho thiy ec ea nit ngin khong 35% 0 véi cic co khing duge xit ly dp suit. Tuy nhign cic phép do Ie ct chi ra ring dp suat lim ting ¢ mém cia thit. sit dung p suit cao ing trong vai phit & nhigt @b moi tnrimg 48 tam gidm Ive edt len oo tude t8, Phuong Kt qua nghién cira cia Macfarlane cho thi phép lim tang 49 mém cho thit béi ép suat cao cing di duge bio céo trong cdc tai ligu cia Macfarlane (Macfarlane, Mckenzie, Tumer, vi Jones nim 1981; Macfarlane va Morton 1978) va nhing ngubi khée (Elgasim va Kennick 1982; Kennick, Elgasim, Holmes, va Meyer 1980; Riffero va Holmes 1983), 2.2.2. Anh husng cin ce qua trinh xit If nhigt vi Ap sudt cao Ten co sau doan t@ cirng Mie dit vige sit dung dp suat tie dng lén co trude t@ cig 18 mot eéch higu qua trong vige xirly d6 dai cia thit. Qua trinh con eo tiém nang dp dung cho thit sau 8 ctmg. Bouton et al. (1977) chimg minh rang co sau té cimg ctia trau, bd sé khong dot due bign dé nr vay khi sit dang Ipc oft tomy, te khi sit dung ap sult cao la 150 MPa & 60°C trong 30 phi li cin thidt 6 nhigt 9 cao. Hp n6i ring khi dp 8 cai thign gid tr cit. Locker va Wild (1984) cling cho ring dp suit- nhigs (PH) «6 higa qui trong vige gid mém thit sau mt chi gian dng ké tgi mot nhigt d cao. Macfarlane (1985) a8 tinh bay mt dé én bao gm vige dp suét Kim phi gi ‘cae protein va mic d@ mém thit bing cach két hgp dp suit véi nhigt do. Trong dé fn cia minh, cic protein bj phan giai boi ap suit cao 1 bién tinh va khong thé két hop ing cich xir ly mht, két qua la lim mém hit. Vige sir dung PH thi higu qué trong vige khds phue d6 dei. Tuy nhign, qua trinh nay co inh hucmg khong tt cho thit do iu nu tao ra bai dp suft va mig2.2.3. Cai thign 4§ mém cia co sau t@ cimg bing céch sir dyng dp sudt cao. Te gée d@ cits eée ting dung thuong mai Khi sit dung ép suét cao, 4@ mém cha co sau t8 clmg thi quan trong hon cq truée 18 img. Suzuki, Kim, Honma, Ikeuchi, va Saito (1992) do 46 cig va d} din cia eo bip vai sau 12 cimg thu duge tit mot con bo sita gid khi sit dung dp suat cao tir 100-300 MPa trong 5 phit bing miy Rheo Meter (Fudoh Céng ty, Nhat Bin, Két qua cho thiy khong c6 sir hae bigt ding ké ao vé d@ din hbi, Diéu niy cho thay co sau 18 cimg o6 thé duge im mém bing ép suét cao ma khong edn xir Ij nhigt, Vige sir dang P-H trong thi gian dai duge Macfarlane a2 xudt (1985), cdn nhimg ngudi khée (Bouton et al 19775, Riffero vi Holmes 1983) thi cho « 1g khong cin thiét phai két hgp voi nhigt 4 trong vige lim mém og sau t8 cimg néu dp suit sic dang cao hom so vei trong thi nghi¢m. 2.2.4, Co ché Lim mbm va sy gia ting nhing bidn 48: cia thit khi si dung dp sult cao Ta biét ring thit ca con vt sau khi chét sé bj mém sau mgt khong tha gian hit dink do nhiing thay d3i xiy ra trong co, chi yéa la do tée ding cia cite protease ni sinh ~ Su suy yéu cia tuong tie acti Phan minh cba to eo than cfc phn dogn ngin do Z-line tan r& Se thoi héa cia cic soi din hi bao gm comnectn (cin go atin) ~ Suy yéu tia mé lign ket. thit hoge vige ting t3e cde bién 481 1a dp sudt lam mi ca tit, ec d8i trong sau dy dA dye xem xe: ~ dps nh hueéng Ién twong tc cia actin-myosin ~ Ap sudt anh hurdng dén sy phan minh cia ta eo, - Ap suat inh hnséng d&n sy chayén d&i a-coomectin than f -connectin ~ Ap sudt anh hurdng lén m6 ign2.2.4.1. Céc ah hudng tir si trong tac eita actin-myosin, Qui trinh twong tic eta actin-myosin va ca cla eta cée sgt ov duge bién 48% trong thi gian con vit sau khi chét dige minh ching bing nhimng thay ddi trong hoat tinh ATPase cta to eo. Ouali (1984) bio céo ring hogt tinh ATPase gia ting tsi ‘cudng d6 ion thap (khoang duéi 0,2 M KCl), trong khi né gi 6 nhing cusmg d6 ek cao hon (0,3 M hoje cao hon) khi gia ting thdi gian bio quan. On lugn ring gid tri nghiéng (i ‘chi thj chinh xa tri xe dinh sy nhay cim véi cudng do ion) c6 thé 1a mot thong s6 (© d9 Io héa cla cfu tric cde sgi oo va duge thé cc chi s sinh héa eda Miofibrilar Aging (BIMA), ‘Nishiwaki, Ikeuchi, va Suzuki (1996) do hogt tinh cia ATPase (cudng 6 ion 1a khodng 0,06-0,32 M_ KCl) cia cée oo durge nit bing dp sudt cao (30-300 MPa, 5 phit) va & 4°C trong 7 ngiy. Sy thay déi gid tri BIMA duge hién thi trong hinh thei 8.1, Déi véi co duge xir ly nhw ten, gid tr} BIMA ting din véi sy gia ting igian Iu trit va dat khodng 2,5 ln so véi ede cv eta con vat chét (hinh 8,1), Gid tri BIMA cia cde to co gia ting khi tang dp suat lén 200 MPa va dat dén mite giéng nh cia cae to co d8 6 4°C trong 7 ngiy, Tuy nhin, Khi ép suit cao hom (300 MPa) sé Lim sut giam dang ké gid tri cla BIMA. Su thay d6i clu tnic cin cic soi ming gay ra bei p suit 1a yeu tb chinh sinh +huéng dén gid tri BIMA thu duge trong cae sgi co khi sir dung ap suat cao trong mot thai gian ngén (5 phit), Nhing thay di cu tric manh mé dage quan sit tr nhimg sgi co duge xit ly ap suat , nhimg thay doi d6 khong quan sat thay trong e sgi co e20 cho ede khong xir bing dp suit, Két qua niy gg ¥ ring vige ép dung dp sui sgi co 1 nguyén nan gly ra nhing thay 46i trong host tinh eiia ATPase vi gift tr BIMA ciaHinh 1: Anh huréing cia &p suit cao va thdi gian len gid tr] BIMA 2.2.42. Anh huréng dén si phn minh cia to co fing héa thi ngin hon va tgo ra mhing, Cac to co khi sit dung cae diéu kign ke co ngin hon la ita mhimg ming thit sau khi giét md (Takahashi, Pukszawa, vi Yasui 1967) vi su pha va cic to co ti Z-line thi lgn quan dén sr gia ting 46 mém thit (Davey va Gilbert 1967; Fukazawa via Yasui 1967; Takahashi etal 1967). Do dé sy phan minh cde sgi to co duge coi 1a hiu dung dé dy dodn sy mém cia thit (Calkins va Davis nim 1980; Olson, Parrish, va Stromer 1977). Suzuki, Watanabe, Iwamura, Theuchi, vi Saito (1990) cho thdy mize 46 phan man trong to co tit cée bap thit tréu, bo vai 4p sudt (100-300 MPa, S phit) trong hinh 8.2. Mite d@ phin minh duge thé hign nhw ty 1g phin trim cia 86 cg to doan gom 1-4 sarcomeres (khiic co) dé tong $0 to co duéi kinh hién vi twong phan pha. Mite d@ phn manb, thi it hon 10% so véi cde eo bap khong duge xit ly 4p suit, da duge ting tbe bing céch tang ap lén hon 30%, 70%, 80%, va 90% tuong tng tai 100, 150, 200, va 300 MPa, Mite d@ phan manh, 80% dén 90%, trén mite t6i da chaTir ic phin minh ciia to eer trong ede co ty whi ikét qui cila sw phn manh nay li swt tiép wie agin cia ede co sau t@ cimg trong dita kign ap suds cao 6 vé la Initu ich cho qué trinh lim iém thit. Hinh 2: Anh huéng cia &p sult cao 1én mite a9 phi vo to eo Cée yéu 16 ae tring trong nbiing bio céo cia Suzuki et al. (1990) dé 18 tae dong cia 4p suit cao dén edu trie myofibrillar (sg: ev 10) sau t2 cing trong ede bip tht triu, bd, Trong to co duge xit ly tg1 100 MPa, gui trinh co cia ce sarcomere duge qua sit, va sy Khic bigt tong m@t d6 gidta cde A-band vi F-band tr nén dng nhait so véi ede méu binh thong (khong qua xi ly), Dinh dau sy vo cia edu trie dang sgi cla L-band va sy bién mit cia che M-line duge quan sét trong to cv cia céc co bap tai tai ap suat 150 MPa, Trong to eo ca eae co bap tai 200 MPa, mach lién te ofu tne ca sarcomere d& gin nur hon toan bj mit, voi tdm Ava Tsgi ta rOng trén ce khise oo nay, Sy bién mit ea M-line va sy diy Ién eta Z-line c6 thé Ia do su co rit ota Ifilament, d3 durge quan sit. Sw phin tach cia cée A-band, thém vito 446 nhiga thay di da durge d& cap va duge quan sit trong to co cia céc ea bap tai dp suat 300 MPa, Chidu dai cita céc sarcomere(khic co), duge xit Iy tai 100 MPa,dung nhur da din din phyc hdi véi sr pia ting ap sudt, vinhimg mit mat ngay cng niga cia kt cfu Tign tue. Nhur da d& c@p, phan manh cia to ea 6 ngudn gée tr st dist doan cita to ca tai Z-line, trong khi cde Z-line tir to eg bj phiin manh trong ede eo bap duge xtr ly dp suit van cin nguyén ven. Mie di vige nit ly dp suit cho cde co sau té eng din ra trong thd gian ngin (5 phi) va nhigt d6 thap (khoang 10°C), thi sy thay d6i sigu edu trie (ultrastructure) cha to co phi hgp véi nhing bio cio cla Macfarlane va Morton (1978) va Locker va Wild (1984). Vige ép dung dp suat cao duge biét li anh huéng ciia tinh trang két hop cia ca hai actin vi myosin, 46 la nhtng thanh phin chi yéu cia to co, Mét 38 bbio cho m6 té su khir polymer cia actin-F (Ikkai vi Ooi 1966; Ivanov, Bert, va Lebedeva 1960; O'Shea, Horgan, vi Macfarlane 1976), myosin polymer (Josephs vi Harrington 1966, 1967, 1968), vi actomyosin (Ikkai vi Ooi 1969) dudi dp suit cao di duge cong bé. Mac dit né khong ré ligu sit khir polymer ciia actin-F xay ra trong co va sit suy thoi cia cde sg, (tit 1, si khit polymer cite F-actin do ap suit cao) 6 thé 1a m@t trong nhing nguyén nhin cia sy phan manb, Mot kha nang lim tang tée sy phin minh eda ty eg va dong ty cta cde myofibrillar protein tg noi phan tich 44% duge Macfarlane d nghi (1985) c6 thé lim ting 49 mém cia thit Khi st dung &p suit cao. Ti nhimng quan sit vi phn tich ulrastrictral SDS-PAGE ea to oo (4 gu khong durgehign thi, Suzuki otal 1990), Ca ché cho sy rt logn trong edu tre tue eba to cw gy ra bai ip suit trong ee dia kign Khe nf 22. Anh huréng tir sy chuyén ai c-Connectin think -Connectin Nehign citu gin day cho thiy mot céch r8 ring ring mot protein, thug dirge {0 li connectin (cbn goi la titin) duy ti tinh dn hbi va tinh én dinh oo hoe cia oo sarong. Lie chét, cée a-comnectin (khong 3.000 kDa) tin tai cing véi mét hong anh cae phan doan phy cia nd, B-connectin (khong 2.000 kDa; Maruyama, Kimura, Yoshidomi, Sawada, va Kikuchi 1984; Wang, MeClure va Tu 1979).Hinh 3: Biéu dién két qua chay dign di trong gel 2% polyacrylamide chira 0.1% SDS va 0,5% agarose. Chi thich =.= a-conncetin = B= B-connectin ~1200k = 1200 kDa peptide anebulin -M faudi myosin gm Boi vi khi sir dung 4p suat cao khong tgo nén ee chudi peptid , do dé né diye quan tim 48 gidi thich mhing nghi ng® ring tgi suo vige chuyén 46; a-connectin thiah B-connectin duge giy ra béi ap suat. Co hai ly thuyét ve co chiconnectin; m@t la sw phan hiy protein boi calpain (protease lam ting hoat tinh Ca") va cathepsin D (Kim et al 1993, 1995;. Suzuki, Kim, va Ikeuchi 1996), va hai la ede phan img trac tiép eta ion Ca? (Tatsumi, Hattori, va Takahashi nlm 1996). Béo cio cia Kim et al. (1993) cho thay ring higu qui fp suat cao Ién ede connectin trong isolated myofibrils (co riéng bigt) wong ty nhir cia connectin trong co bip, duge ‘quan sét nhimag phan tich cia SDSPAGE, Ho tim thy hai loai chit te ché protease, 1 mM leupeptin va 1 mM E64, hodn toln ngin chin su suy thodi eta comnectin (100-400 MPa cho 5 phit), me di cie tw ca bén trong déu o6 sy ign dign eta 3mM CaCl, Nhu vay gid thiét vé sy suy thoai ciia connectin do tic déng trye tiép cla cic jon canxi dip suit cao nbs wy li khSng thé xay ra, K€t qué nay chimg minh sy tham gia cia mét s6 protease ni sinh, dic biét la calpain, trong vige chuyén d&i a connectin thin B-counectin, Diéu nay cé thé duge gidi thich bing cach gia sit ring sy mim cim cia connectin 46i véi calpain duge ting 16 rt béi vie dp dung cée ép suit, mg kha ning cia calpain ¢é thiy phan connectin duge gidm din v6i ving dp suit, No da Auge chimg minh ing ap suit cao 100 MPa hoc cao hom thi sé lim tang tinh mam im dén qué trinh proteolysis (phan hiy Protein). Bai vi ndng d@ ion canxi trong dung dich sarcoplasmic Ia gn 15i mu d8 kich hoat calpain do vigehinh hank tir hi sarcoplasmic trong Khi sit ding ap suit cao (Suzuki, Okamoto, Tkeuchi, vi Saito1993, 1994), mite 46 chuyén doi cha a-connectin thanh B-connectin duge cho 1a chit yu lién quan dén sir phy thuge dp suit cia cic thay 461 cfu tric ota a connectin va sy v6 hogt cba calpain. Co ché cho vige chia connectin dudi ap sudt cao c6 12 14 giéng nhu trong co sudt qué trinh xir ly (Kim et al 1995). Sy gia ting cia nhting hyp chat durge trich ly tt connectin c6 thé cho thay duge nhiing thay déi vé chét krmg cia chu trie connectin trong co gay ra béi dp suét (Suzuki ct al 1987). 2.2.44, Anh huéng cia che mé lién két bg mém cua thit thi phu thuge vio hai thinh phin khée nhau: d6 dai va d déo dai cia actomyosin, BH déo dai cia actomyosin Ki do cée protein myofibrillar, trong Kkhi 6 dai do sy hign din eta che m6 lin két. N6i chang, né duge chip nhin ringnim thay di cia ede mé lién két trong qua tink bio quin thit thi thép hom so véi nnhig bién d6i trong myofibrillar protein Mot vai bio céo mo ta nhiing inh hardng cis fp suit én mé lign két fing mht 1a dbi véi protein to ca. Ratcliff, Bouton, Ford, Harti, va Macfarlane (1977) cho thay ring mfe di sr dung P-H c6 higu qua trong vige log bd duge tinh dai cia to eo (d9 dai actornyosin), do mém ciia cée miu duge xir ly thi duge han ché bei do dat ‘cia cde mé lign két, Macfarlane et al. (1981) eiing cho théy mot sy chuyé thiéu ving cia ede actin-F, nhung ma do cdc mé lién Két 43 Khong duge thay 46i trong thermograms (bidu d3 nhigt d9) cla co 43 duge sit Wy bing ap suit, Beilken, Macfarlane, vi Jones (1990) cho ring ap suét diéa tr 6 ee nhigt 46 khée nhay, tr 40 C* dén 80°C o6 rit it hoe Khong c6 bign di 16 r8t vé sy déo dai hom Ta dé ting ahigt d6 bing xir ly uhigt lim gitm 46 dai, Suzuki, Watanabe, Tkeuchi, Saito, va Takahashi (1993) béo céo ring khong 6 sy khée bigt ding ké trong sigu efu trie (ultrastructure), ign di, nhigt d hia tan, va thermogram (bieu dd nhigt &6) eta phin 4quét calorimetry (do nhigt lugmg) (DSC) phan tich cia cae collagen bap phan lap tit mot con bd sta gid di duge quan sat trong s6 ce mau eg khong duge xit ly ap sudt va cée co duge xit IY Ap (100-400 MPa, § phit). An hurémg cia 4p suat vio nhigt 45 biga tinh va enthalpy theo thermogram duge thé higa tong bang 1, Gin day Nishimura, Hattori, va Takahashi (1995) cho ring sy suy yéu cua ede mo lign két trong co, endomysium (v6 ndi co) va perimysium (mang co), gay ra trong qué trinh thi trong quan véi sy mém tht bing tte dé qué - Ueno, Ikeuchi va Suzuki (1999) m6 lign két trong ee eo va xit ly ap sudt bing kinh hién vi quét dign ti, Trong suét thai gian xirly, sy suy vyéu viu trie cia cde endomysium va perimysium va sy phd vi ede edu tric t6 ong a xuit hign. Trong co duge xir ly 4p sudt, bidn dang cia cau tric té ong cia endomysium durge ting lén vdi sy gia ting cla ép suit téc dong lén cic co bap, va mé rong mang luéi cua endomysium di duge quan sit trong ede mau co tai 400 MPa, Hign nay, két ludn d6 khong phai li chée chin ring eéc ap suit gay ra nhimg thay di co cfu trong ete mo lign két din dén mot gu img ding ké dén & mim ‘cha thit, Nhumg néu muén higu r thém thi phai nghién edu su bon ita dé lam 10 van dé nay.Bang 1: Anh huéng eda dp sudt len tinh trang bién tinh cia collagen trong co Nhigt 49 bign tinh (°C) T, T, Te AH (mJ/mg) Control 62.401 1.03 65.65.1067 68.701 0.65 15.00 1 1.40 100 MPa 6213 £0.66 65.35 40.25 68.95 + 0.64 15.53 + 1.65 150 MPa 6258 41.03 66.05 +088 6880 0.64 15.78 + 1.80 200 MPa 6280 £1.65 66.03 £099 68.73 £0.72 16.93 + 1.10 300 MPa 62.68 40.63 65904123 68954083 15.50 + 1.65 nhigt 46 biém tinh nhigu nhé thie bién tink, AH = enthalpy cia bién tink, mi/mg khdi lrgng, kh, Neu: Suzuki, Wantanabe, Tkenchi, Sato, and Takahashi (1993), Anh hong cia ép suit exo len vi sinh vit Téc dO va dong hoe v6 hoat vi sinh vat phy thuge vo m@t vai thong s6 nhu: Jogi vi sinh vat, cudng 49 Ap st . thdi gian, nhigt d9, pH, host tinh made va thinh phan thye phim. Nhi é chung, sy Ge 2.3.1. Syphi hity cia Sp suit Ten vi sinh vats Sy te ché vi sinh vat bai dp suat cao 6 1é li do tac dong cua nhiéu yeu to két hop tai. Ap suat eno ki ne te ché ode ph hy mét v tr nao 46 trong té bao hove mot chife ning cia 1 bao., ma sy chét cit 18 bao 18 béi nhimg hu hong nhigu hode tich tu bén trong t8 bio (Simpson & Gilmour, 1997). Mang té bao la mue tigu chinh ova sy phi hy, cha yéu qua vige lim ri foan sw van chayén cée chit cia ming té bao do hu qua cita sir két tinh phospholipid Cée chite ning khie cia t§ bio bj anh hugng béi dp suit bao gbm: si thay d&i tra0 4% ion, thinh phn acid béo, hinh thi hoe cta ribosome, hinh thai hoc té bio, big tinh protein va te el boat tinh enzyme, sip mit n dish cia v hinh thinh khong bio...Si khae nhaw sao chép DNA, sit 8 tinh chit cia mang té bao o6 thé la mot nn 16 quan trong trong vige xc dinh anh hug cia ép sudt hoe stress Len vi sinh vit. Trong mét vai hé vi sinh vit, nga ta cho ring sy pha hiy li do 16i loan si wn chuyén protein qua ming 18 bio Gancle, 2001). Day 6 thé fogel, Molina-Guiterrez, Ulmer, Winter, & co ché chung lim cho 18 bio chét. Nhin chung, cfc t8 bio chét gia ting khi ting 4p suét nhumg né khéng tuaa theo dng hoe bac 1.visinh vat 2.3.2. Kha niing chju 4p sudt cao Nhin chung, cée vi khuin gram duong chi duge ép suit cao hon vi khan gram im, Hinh théi 8 bio cing anh hug dén tic ding cia 4p suit, vi Khun hinh que bj anh hurimg nhigu hom hinh céu, Ci 1€ bio & pha phat tidn d bj anh bung hon cic té bao & pha on dinh. Bang 2: Sy titu digt vi sinh vat bi dp suit Tt min cam Dé min cam Bao tir bao sinh dugg Gram duong Gram am . € bao trong pha on dinh Té bao trong pha phat trién Bacillus, Clostridium, Staphylococcus, Yersinia, Vibrio, Salmonella, parasites Listeria, Escherichia coli O1S7:H7 a, thap cao pH acid Nhigt d9 cao khang sink Nguén: Masoliver va Grebol. Nhiét d6 oé vai t quan trong trong vige it ché vi sinh vat bing dp suét cao Tai nhigt d6t6i wa cho sy phittign, sy ite ch sé thép hom s0-v6i cée mhigt 6 khdc. i wu, DG chay ming c6 thé bi phi va d8 ding hon tai nhigt €6 cao hon nhigt €} Khi cde té bao duye lam se lanh tage khi ding ap sudt cao, chimg 66 kha ning chia dp tt hon. Bigu niy duge gidi thich 18 do sr duy tr d6 chiy ming ca té bao & nhigt d9 thap. Sy thich nghi cla mang 6 nhigt 46 thip duge thye hign bing vige thay 46i 49 phén nhinh vi lim giim chigu dai cia cae acid béo ming, vi thé lim cho vi sinh vét chiu duge dp suit t5t hon (Wemekamp-Kamphnis, Karatzas, Wouters, & Abee, 2002), Cae protein tao ra trong qui trinh sc lanh 6 thé bio bao trade dp suat cao. Sip Khéc nhau vé anh hung eta dp sudt len céc gidng ely bénh cla mot vai Todi (Listeria monocytogenes; Staphylococcus aureus; Escherichia coli va Salmonella typhimurium) €& duge bio efo bei mot vai tée gid (Alp, Kalchayanand, Bozoglu, Sikes, Dunne, & Ray, 1999; Simpson & Gilmour, 1997)2, Anh hurdng cia thye phim Ién kha ning chju dp sudt cao cia vi sinh vat ‘Theo Archer (1996), 2 nh hung chinh cia thye phim lén d@ an toin vi sinh tis anh huring cia thinh phan thye phém trong suét qué trinh xir lj va anh hucimg sau xit ly trong sudt qué trink phye hdi cla vi sinh vat, Kha ning chiu dp suét cia vi sinh vat e6 thé bi tie dng bai nhidu yu t6 mdi traimg vi bén trong, ban chét cia méi tnréng la quan trong mbit. Két qua dat duge trong méi tnzimg tng hgp hoe moi tramg dém khong thé két luin ngay tite khic vé tinh trang eda thinh phin hye phim, Nhw da bi, thinh phin thye phim 66 46 acid thap nhu thjt (bang 1) va sita (Garcia-Graells, Masschalck, va Michiels, 1999), 6 khuynh huréng bio vé vi khan chéng Igi nh hung cia dp suit cao so vi moi tnrng dgm phosphate, Bang 3: Sy gidm lung vi khudin sau khi xir ly Ap sudt S00 MPa, 40°C trong 10 phat Giong Dung di Waa thit Phosphate Carnobacterium pisciola 5.79) 467 LMG2739 6.79 4.67 Enterococcus faecium CTC492—— 7.23 3.97 Escherichia coli CTC1007 53 5.54 Escherichia coli CTC1O18 5.78 431 Escherichia coli CTC1023 7.01 3.98 Lactobacillus sakei CTC494 6.33 4.22 Lactobacillus sakei CTCTA6 633 391 Leuconostom carnosum CTC747 7.92 455 Listeria innocua CTC1O14 3.84 Pediococcus acidilactici F 1.29 Staphylococcus camosus LHT2012 ‘Nguon: Garriga (chwa xuat bin) Kh ning chiv ép suit cao cia vi khudn 08 thé duge Bi ding dang ké Ki duge xt Wy trong moi tring gidu dinh dung chia céc chit aku carbohydrate, cle ch iy c6 thé bio vé vi Khun koi sy hu hai (Hoover etal, 1989). Sy c6 mt cia carbohydrate va protein trong thé nh tromg thye phim lim gia tang kha nang chi 4p suit cia mot vii vi sinh vit (Simpson vi Gilmore, 1997), Sau khi so sinh sy te ché vi khudn trong cdc sin pham thjt 6 cdc hogt tinh nude khae nhau (bang 2). Dya trén bing ta thy mie d9 chét cia Staphylococcus aureus va vi kinuin acid lactic "7trong thit di lyn khé 1a thap nha néu so vi thit bd wip va thit Abi lyn nu chin ‘Trong thy t8, kha ning chiu ap sudt cao cia vi sinh vit s& gia ting khi giém host tinh nude. Bang 4: So snh sy tigu digt vi khudn Staphylococcus aureus va lactic a bacteria (LAB) biing 600 MPa, 31°C trong 6 phiit Toit ‘Sin phim durge dong gol chin Khong Thit bo udp. Jambon néu chin Tambon mudi hun Khoi Ban @iu ding HHP Ban dau ding HHP Ban dau _ ding HHP Staph, aureus 3.62 0953.70 258 2.74 2.19 BG giim 267 12 0.55 LAB 494 0.95 5.63 1.06 4.23 2.69 BS gid 3.99 457 158 Aw 0.985, 0.987. 0.890 Nuon: Ganga etal Vi khudn 48 nhay cm v6i pH gin t6i wu sau khi xit I nhigt hode 4p suat, vi thé, khéng chi pH thdp lam gia ting sy te ché trong qua trinh xir ly ma odn ite ché ‘sy phat trign té bio bj tn throng bei nhigt hoe ap suit, Ngoai anh hung ciia pH thi khong 68 aah huréng dng ké nio gay ra bai cdc acid hitu co. Bigu nay o6 thé 18 do ip sudt chi anh hung chi yéu Ién sy ion hoa va ede avid hu eo 1é nhimg chit re chi de thi dui dang Khong hoa tan, Acid sorbic he dong gidng uh acid how ca, hime né cling tae dng dén mang té bio cia vi khuin, va tae dng s& ahiu hom khi két hop véi ép suit (Mackey, Foresticre, va Isaacs, 1995), 2.3.4, Anh huréng ci logi thie bj Ten higu quit ire ch vi sinh vat cia dip Cée thidt bj ap sudt va moi trréng khée nhau c6 thé tao ra nhiing két qu khée nhau v8 nbime vi khuin séng s6t trong sudt giai down bao quin, Hom nifa, sie Khe nhau v8 thai gian, nhigt 4 dp suét trong qué tinh thye 48 tai ede vi ti Khe hau ci vi sinh vit khéng ding nbit ti cde vj tri khie nhau 3, Tri vi qua trinh thanh tring hay tgt tring ob dign, diém ti han cia ép suit cao 1a gin vs tht bi, bai vist traydn hit xiy ra gta v6 thigt bi va khbi chat ling, din dén su gin nhigt 46 sin phim nim & gin vé thing. Anh hung cita thai gian ting dp, tdc d@ xa dp vi nhigt d6 cia qué trinh vin chua duge 46 gidm 4p chm c6 thé lam gidm higu qua nghién etru mot cich day di. Mot t6 ‘qué trinh va do dé vige gidm ap phai due thy hign nhanhSin phim tht di lym 8 chin vi bao gi duye xi y bing dp suit tai 6O0MPa trong 6 phit tei 31°C trong 2 thidt bi kha nhau (A vi B), s6 kung vi kuin lactic trong suot giai dogn bio quan lgnh da duge dénh git (hinh sau). Hinh 4: Dong hoe phat trién ciia vi khudin lactic trong jambon chin theo thi gian Trong méu cia thiét bj A, sO lygng vi sinh vat lun nim dudi him Ingng phét hign trong suét thoi gian ru mu (120 ngiy, 4°C). Cée mau trong thidt bi B duge gitt duéi diém phat hign trong 30 ngiy bao quiin, Khi két thi thoi gian Iwu tri g. Tron, thi sé lugng cia ching khoang 10°ef not thi nghigm tuong ty (bing 3) so sénh anh Tung cia ap sult Yen cde miu abi lym khd duge eby Listeria ‘monocytogenes, vai trong 2 thiét bj khae nhau (A va C) tai 600MPa, 31°C trong 6 hit ing 5: So sinh sy tigu digt Listeria monocytogenes trong 2 thiét bj khée haw Thai gan THETA THebIC ‘Trrée khi ding HI 27 27s" I ngay sau khi qua HHP | 0/25 373% 30 ngay sau khi qua HEP | 0/3” 33" 60 ngay sau khi qua HHP | 0/3" 33" 120 ngay sau khi qua HHP | 0/3" 23" Neudn: Garriga et al logis cfu g', * So Iugng miu 6 Listeria monocytogenes trong 10 g/sb lugng miu thir ban dau,(Qua thi nghigra thay ring miu trong thiét bj A da v6 hoat hod toan Listeria ‘monocytogenes (trong 10g) trong subt giai doan tin tt (120 ngay). Trong khi @thigt bj C thi mim bénh van hign dign trong suot théi gian ton tri. Su khéc nhau nay khong phii 1 do miu vi ede miu duge ly tir mot ngudn vi duye chudn bj gidng nhau nhumg chi khac thit bj. Do d6 6 ¥ kin cho ring cin thiét Ip ede each thite chuan bj mau cho cée thiét bj ap suat truée khi kiém tra thi nghigm. Anh hung ciia ap suat cao dén cde chu tir hwong. hur ta da bit tit 6 nog vi tuong tw ahur huyét thanh ode hayét, khi higt dé thay di vé gan uring dn hong vi (thrmg o6 inh hide tt Mii vj cia bip thit ndu chin thi phong phi hon Ia cde bp thit cé mii ty mhign. Ha hit céc ofu tr go nén huang vi vi "meaty favor" bao gdm mot rong dudmg khit (thuéng 1a glucose), m6t nguon axit amin va peptide, va hgp chat kim tang huong vi (taste enhancer)(vi dy nbu ino: nic avid; Baines vi Mlotkiewiez 1984), Noi chung 6 thé chép nhdn rng hi hét cfc thinh phin nay Him cho efe m6 co bap tang thi igian bao quinn, do phiin dng héa hge cia eée think phan nhét djnb cia eo, Rat it thong tin vé cée anh hudng cia dp suit cao dén cée tit iu ti tgo mii va huong vj cho bao cdo cha Suzuki, Homma, Fukuda, Hirao, Uryu, va Tkeuchi (1994) cho thiy ring s6 lugng cit c peptide vi axit amin dugg nu ting len khi ta xit1y 4p sudt cao dén 300 MPa (5 phit) d6i véi bp tit thd dorsi Longissimus, name sx Ikhée nhau gia cfc lin xit ly bing dp suit cao di khong ¥ nghia thong ké. Khi cic co bip duge lu tr tai 2°C trong 7 ngiy, s6 hung PPM (phenol reagent positive materials) ting Ién ding ké (khong 140-150%) d4 duge Khao sét ci déi véi cae co bp durge xi ly Hin khong duge xt ly bing ap suat cao, $6 hugng PPM tt ee co bp duge xit ly 4p sudt cao thi cao hon nhing tir cée co bap khéng duge xir ly, Nhung PPM lai gidm gi 400 MPa, so voi 300 MPa, c6 thé 18 do mot gidm nhg ci hoat

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