Chng 6 - ENZYME LIPASE

1 Gii thiu chung Lipase (E.C. 3.1.1.3) c ng dng ch yu xc tc thy phn acylglyceride hoc mt s ester khc nh thiol, polyol hay polyacid ester. Ngai ra lipase cn c th c s dng lm cht xc tc trong cc phn ng tng hp cc acylglyceride hay ester c gi tr ng dng cao trong i sng. Lipase c bn cht l mt globulin v c th c sn xut t tuyn ty v mng d dy ca ng vt. Tuy nhin sinh tng hp lipase t vi sinh vt mang li thun khit v hiu sut cao hn. Hin ti ngi ta ng dng rng ri lipase trong cng nghip dc phm, ha m phm, dt da, cht ty ra, thc phm hoc tng hp cc cht hu c khc nhin liu sinh hc (bio-fuel). Cng nh a s cc enzyme thy phn khc, lipase khng c nhm ngai m ch c trung tm hat ng vi cc acid amin c bit nh imidazol ca histidin v nhm hydroxyl ca mt trong s cc gc serine.

Lipase thy phn lin kt ester ca lipid nm ti b mt phn gia lipid v nc. Trong nc, trung tm hat ng cha serine b kha li bi vng xon helix.

Khi lipase gn vo b mt phn pha gia c cht lipid v nc th vng xon helix di chuyn cho php c cht gn vo trung tm hat ng.

C ch tc dng ca enzyme lipase: do cu trc bc 3 ca enzyme m nhm hydroxyl ca serine v vng imidazol ca histidin tip xc nhau v to thnh lin kt hydro. Kt qu l oxy ca nhm hydroxyl thu c tnh cht i nhn. V tm i nhn ny ca enzyme s tng tc trc tip vi mt trong hai nguyn t tch in dng ca lin kt b thy phn. V do lin kt ny d b ct t v s thy phn thun li hn. Mt c ch khc l enzyme to thnh lin kt tng ng vi c cht nhng v tr lin kt gn gi vi lin kt thy phn. Kt qu l lm tng s khuyt electron vn tn ti trc , v phn b electron trong phn t c cht thay i theo mt chiu hng cn thit. Nh m vic ct t lin kt c d dng, hoc lm cho mt trong hai nguyn t ca lin kt tng tc c vi cc tc nhn i nhn. Xc tc enzym lipase c chia lm 2 lai nh sau: thy phn v tng hp. Phn ng thy phn: RCOOR + H2O RCOOH + ROH 79

Phn ng tng hp: C th c chia thnh 4 nhm nh sau: a) Ester ha (Esterification): RCOOH + ROH RCOOR + H2O b) Ester ha tng h (Interesterification): RCOOR + RCOOR* RCOOR* + RCOOR c) Ru phn (Alcoholysis): RCOOR + ROH RCOOR + ROH d) Axit phn (Acidolysis) RCOOR + RCOOR RCOOR + RCOOH Trong 3 nhm b, c v d thng c gi chung l l transesterification. 2 u th xc tc lipase so vi phng php s dng xc tc ha hc Hu ht cc ng dng ca lipase ngy nay tp trung thay th cc phn ng xc tc ha hc ci bin hc to dn xut ca cc lai du m. Hin ti cc triacylglyceride mch trung bnh c sn xut bng cch ester ha acid bo vi glycerol p sut v nhit cao (200-250oC) bng cc cht xc tc v c. Cc xc tc v c ny thng khng hiu qu v i hi phi cung cp ngun nng lng rt ln. Sn phm to thnh cn phi c tip tc em tinh luyn tip bng cch ra kim, tinh luyn bng hi nc, chng ct phn t, siu lc, hoc s l bng carbon hat tnh v.v Mt qui trnh sn xut cng nghip quan trng hin nay l thy phn du thc vt (du liu, du da, du u nnh) sn xut cc acid bo v glycerol nhm ng dng rng ri trong cng nghip cht ty ra, ha m phm, dc phm v thc phm. Cc qu trnh sn xut hin ti c iu kin phn ng cao: p sut 3000-5000 kN/m2 v nhit 250oC. Trong 2h c th thy phn c 96-98% du thu c 12% glycerol. Cc acid thu c khng th em s dng ngay m cn phi tip tc em i chng ct ty mu v lai b cc sn phm ph. Qu trnh ny sn xut kiu ny i hi tiu tn nhiu nng lng v to ra rt nhiu cc phn ng khng mong mun nh l cc phn ng trng hp ca cc acid bo nhiu ni i v to thnh cc ketone v hydrocarbon. Chi ph u t cho qu trnh tch bng hi nc rt cao bi v ct chng tch phi c cu to c bit chu c nhit v p sut cao cng nh kh nng chu n mn acid. Trong khi , do xc tc enzyme c tin hnh p sut v nhit trung bnh thp (40 - 60oC) nn c th tit kim c chi ph nng lng. So vi phng php sn xut truyn thng, chi ph sn xut bng enzyme cng c gim bt nh cc bnh phn ng khng cn phi chu n mn cao, cng nh cc iu kin mi trng khc nghit khc. Cc sn phm ca qu trnh xc tc sinh hc cng cho mu sc v mi v tt hn, cng nh tinh khit cao (c rt t hc khng c phn ng ph). V nh ngi ta cng gim thiu c chi ph sn xut do khng cn phi tin hnh cc qu trnh tinh luyn tip theo nh trong phng php xc tc v c. Ngai ra nhit phn ng thp cng gp phn lm gim thiu s phn hy nhit ca sn phm. V do lipase c s dng cho tt c cc mc ch trn. 80

3 Cc ng dng c th ca cc loi Lipase 3.1 Thy phn Thy phn nhm tch du ra thnh cc acid v ru. Sn phm mong mun c th l mt loi acid c bit hoc l mono, diglycerides, ru... Mt s nghin cu, ng dng lipase thy phn mt s c cht c trnh by trong Bng 1. Bng 1. Xc tc thy phn ca Lipase Lipase Rhizopus delemar + Pencillium + Rhizopus niveus Canida rugosa, Aspergillus nige C. rugosa C cht Du u nnh Du c Du u nnh M b, m heo Mc ch Tng tnh hiu qu thy phn bng cch kt hp nhiu lai lipase Lm giu cc PUFA Thy phn kt hp vi h thng mng lc Thy phn trong dung mi hu c nhit thp hn nhit nng chy ca m Lm giu acid Docosahexaenoic (DHA) Sn xut monoglyceride

C. rugosa

R. niveus R. delemar

Du c Du c

3.1.1 Trong phn gii lipid Nh vo kh nng lipase thy phn du m thnh glycerol v cc acid bo vn c ng dng quan trng trong cng nghip. Hin ti, cc acid bo c s dng sn xut x phng. Cc enzym x dng cho mc ch ny c th thu t Candida rugosa v Pseudomonas fluorescens (quy trnh c cng nghip ha). Cc phng php enzyme ny hiu qu hn cc phng php ha hc v mt chi ph v sn phm thu c c mu sc v mi v tt hn. Do cc phn ng enzyme xy ra nhit v p sut thp hoc trung bnh nn chng c th c s dng sn xut cc lai acid bo t cc lai du d bin tnh (cha nhiu acid bo khng no hoc c nhiu ni i lin hp). Thng thng iu ny rt kh t c vi cc phng php sn xut trc y v nhit v p sut cao d dn n oxt ha du m. 3.1.2 Trong cng ngh thuc da Trong cng nghip thuc da, mt trong nhng bc quan trng l lai b cc phn tht v m vn cn dnh trn da v lng. S dng cc quy trnh ha hc nh ty kim thng khng hiu qu. Ngy nay ngi ta ang dn s dng lipase v protease t c mc ch trn. Ly v d nh ngi ta s dng lipase t Rhizopus nodosus x l ty nhn lng cu. So vi phng php x l bng dung mi truyn thng th cht lng sn phm x l bng enzyme co cht lng ng nhau, nhng ch r hn v t c hi hn. 81

3.1.3 Trong x l nc thi Lipase c s dng trong cc bn hat tnh v cc quy trnh x l cht thi hiu kh m trong lp lipid mng trn b mt bnh hiu kh c lin tc lai b cho php oxy xm nhp vo ( duy tr iu kin sng cho VSV hiu kh). V d nh Lipase-MY t Cadida rugosa do cng ty Meito Sangyo, Nht Bn sn xut c s dng trong cc nh my x l nc thi M. Trn thc t vic phn hy cc cht thi rn, lm sch v ngn chn lipid lm tc nghn hay to thnh lp trong cc h thng x l nc thi, cc bnh cha rt quan trng trong nhiu nh my cng nghip thc phm, thuc gia, git m gia cm. 3.1.4 Trong sn xut cht ty ra gia dng Cng nhm vi hai nhm ng dng bn trn, lipase c ng dng cng nghip sn xut cc cht ty ra nh bt git, nc ra chuyn dng Lipase sn xut t Aspergillus oryzae c dng trong bt git do c kh nng chu kim cao. Hng nm khang 1000 tn lipaza c cho vo 13 triu tn bt git, vi tng gi tr vo khong 30 triu USD. 3.1.5 Trong sn xut hng v cc sn phm t sa Cc acid bo t do, cc peptide ha tan v cc amino acid to nn mi v v cu trc cho cc sn phm b, phmt, margarine. Bi vy cc lipase v protease ng s dng rt ngn thi gian lm chn ph mt v sn xut cc mi v c trng khc. Mi v to thnh ph thuc vo lai enzyme s dng. Hin ti cc PGE (pregastric esterase) c s dng lm ph mt c v tiu cay . Enzym e t Penicillium roqueforti th c dng to cc lai phmt xanh (blue cheese). Cc sn phm phmt sn xut nhanh bng enzyme (EMC, Enzyme modified cheeses) ngy cng xut hin rng ri (v d nh lipase t Mucor meihei, Aspergillus niger, A.oryzae). Bn cnh lipase cn c s dng to hng cho cc sn phm bt kem c ph, ko b sa cng, scla, snack, soup.... 3.1.6 Trong sn xut cc thc phm khc y l mt lnh v y tim nng ng dng lipase. Phn gii lipid bng enzyme c ng dng vo trong sn xut cc lai tht, c khng cha m. Thy phn mt phn triglyceride lm tng hm lng monoglyceride bng cc cho lipase vo trong bt bnh m cho php lm chm i qu trnh thai ha tinh bt. S to thnh mono v diglyceride cng cho php ci thit tnh to xp ca lng trng trng g. Lipase cn c ng dng trong sn xut sa u nnh, c v tht xng khi, hay ngay c trong sn xut thc n cho vt nui king nh ch, mo. 3.1.7 Trong iu tr bnh Bn cnh cc cht hydradcarbon v protein, lipid l thnh phn khng th thiu trong c th con ngi. V vy, nu xy ra cc sai st trong qu trnh thy gii, hp th hay tng hp lipid u dn n kh nng gy bnh cho ngi. Do lipase c s dng nh l mt cht tr gip tiu ha. Hm lng lipase trong serum mu l mt ch nh chn an tt cho cc bnh tn thng v tuyn ty. Ging nh nhiu lai enzyme khc, lipase cng c th c c nh trn cc in cc pH hoc oxy ch to cc sensor sinh hc nhm xc nh hm lng triglyceride v cholesterol trong mu. PGE cng c s dng cho cc bnh nhn khng th hp thu c lipid. 82

3.1.8 Trong phn tch cu trc triglyceride Lipase c th c phn lai da trn tnh c hiu ca c cht. C th bao gm: - Enzym c hiu v tr sn-1,3 (Rhizopus oryzae, Humicola lanuginosa, Rhizomucor meihei) ; - mt loi axit bo c trng nh oleic acid (Geotrichum candidum) hoc linolenic (Brassica napus v Mucor miehei); - hoc hon ton khng c hiu (Candida antarctica, Candida. rugosa, Pseudomonas cepacia). Nh vy s dng tnh c hiu ca Lipase s gip chng ta phn tch c cu trc ca triglyceride. 3.1.9 Trong sn xut cc acid bo Sn xut du n giu cc acid bo nhiu ni i (PUFA, polyunsaturated fatty acid) nh docosahexaenoic acid (DHA) v eicosapentaenoic acid (EPA) l mt trong nhng ng dng ht sc c quan tm hin nay ca lipase. V d: Sn xut hn hp giu acid bo omega-3 t du c mi bng lipase (T. OKADA and M. T. Morrissey - B mn Cng ngh thc phm, i hc Oregon State USA). Trong vng 30 nm gn y, cc nghin cu cho thy cc acid bo omega-3 c nh hng tt n sc kho, v rt nhiu loi sn phm cng c tung ra th trng. Du c mi th trc tin c ty mu bng carbon hot tnh (4%), kh mi bng cch chn ct chn khng v c n nh vi 200 ppm TBHQ. C th s dng cc loi lipase thng mi nh t Candida Rugosa, Candida cylindracea, Aspergillus niger hay Mucor javanicus. m pH bng phosphate t 6.5 7.0 ty thuc vo pH ti u ca mi loi enzyme. Nghin cu cho thy i vi c mi th 500U lipase t Candida Rugosa c th thy phn 82.5% du c trong 9 gi. Phn tch thnh phn acid bo bng phng php sc k kh cho thy so vi du c mi ban u thnh phn EPA v DHA c ci thin. Thnh phn acid bo EPA DHA Du c mi ban u Thy phn bng 500U/g lipase t Candida cylindracea trong 6 gi 35,8 19,23 Thy phn bng 250U/g lipase t Candida Rugosa trong 6 gi 29,75 14,85

19,44 7,62

Nh vy nghin cu cho thy x dng lipase c th dng sn xut cc hn hp lipid giu cc acid bo omega-3 t du c . 3.1.10 Cc ng dng khc: Ngoi ra Lipase cn c ng dng ty mc in trn giy, x l t tm, hay trong sn xut nc sc ming kh mi.

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3.2 Tng hp ester Kh nng ca lipase nhm xc tc cc phn ng nghch ca qu trnh thy phn c s dng sn xut cc sn phm mong mun. Hn hp ester ha thng ch cha cc c cht v enzyme. Nc ch l sn phm ph ca phn ng v c lai b kh d dng nn y c coi l quy trnh tin tin tng hp cc cht. Bn cnh qu trnh transesterification thay v to ra nc s to ra sn phm ru, acid hay ester, do qu trnh ny s sinh li nhiu hn nu nh cc lai sn phm trn l sn phm chng ta mong mun t c. Trong ha hc phn ng ester ha ch yu ni n phn ng gia acid vi ru, trong khi lipase li c th xc tc nhiu lai c cht hn: ester ng, thiol esters, peptide

3.2.1 Tng hp ester c khi lng phn t thp hoc trung bnh Cc ester c khi lng phn t thp nh l isoamyl butyrate v benzyl propionate c s dng lm cht to mi. Hin ti bytyl laurate l thnh phn hng ch yu ca o v l, v thng c dng trong sn xut hng tri cy. Ethyl butyrate c bit n nh hng da ca n. Bytyl oleate v butyl laurate c s dng lm cht lm mm do, cht bi trn. V do chng c ng dng rng ri trong cng nghip du thm, m phm, x phng, thc phm Mt ng dng khc l s dng cc methyl v ethyl ester thay th cho cc lai nhin liu. Tt c cc ester trn u c th tng hp mt cch hiu qu bng cch s dng lipase t M. miehei, R.delemar, P. cyclopium, G. candidum v.v... 3.2.1 Polyester Lipase c s dng sn xut cc polymer ca dimethyl terphthalate, vinyl acetate, butyl acrylate c s dng lm cc cht do, sn v cht kt dnh. 84

3.2.2 Thc phm Lipase dng tng hp monoglyceride nh l monolaurin, ng ester v ester ca amino v acid bo vn c s dng nh l cc cht nh ha hoc cc cht nn trong cc mng bao n c. Hin ti mt sn phm quan trng c sn xut t lipase l cc sn phm thay th du m. Ngy cng c nhiu nhu cu thay th cc cht du m giu nng lng. Hn th na cc sn phm thay th li c tnh n nh nhit rt cao. V d nh sn phm Olestra, hn hp ca hexa-, hepta- v octaester, c hnh thnh bng cch ester ha c sucrose vi cc acid bo mch di do Proter & Gamble ang gi bn quyn hay sn phm ester ha ca glycerol vi propoxylate ca ARCO Chemical. Cc sn phm ester trn c s dng rng ri trong ch bin cc nc st, mayonnaise, hay kem n. 3.2.3 Dc phm v ha m phm Mono-, di- v triglyceride ca octanoic v decanoic acid c th c s dng lm nn cho cc cht mu v mi trong cc sn phm ha m phm v dc phm. Ester ca ng v sorbitol vi lauric v stearic acid c dng trong thuc chng khi u. Oleyl monooleate c s dng trong du tm, cc sn phm dng tc, dng da. 3.2.4 Cc sn phm t sa v bnh ko Mt trong nhng ng dng d thy nht trong lnh vc ny l s dng lipase xc tc cc lipid tng hp cc triglyceride c tnh cht mong mun. V d nh s dng lipase t C. viscosum, M. miehei, v P.fluorescens ch to cht thay th b cacao t du c nhm s dng lm scla v bnh ko, hay nu sn xut t du olive v ht bng vi th s dng lipase thng mi Lipozyme (sn xut t M. miehei).

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Ngoi ra ngi ta cng dng lipase ci bin du c thay th cht bo trong sa m.

Hin ti, Lipases ch chim khong 5-10% sn lng enzyme s dng (protease chim 60-70%, carbohydrase chim 25%). iu ny c l gii bi vic s dng lipase cn gp mt s mt hn ch. Trc ht l lipase thng xc tc trong mi trng nc, trong khi c cht lipid th vn rt t tan trong nc. Bn cnh tnh n nh ca lipase i vi nhit , pH cha cao. V sau cng l kh thu hi v ti s dng lipase nn dn n chi ph sn xut cao. Tuy nhin, cng vi s pht trin ca khoa hc k thut, cc c im hn ch trn ca lipase ang tng bc c gii quyt. Ngy nay cc lipase c th hat ng trong cc dung mi hu c hoc cc lu cht trng thi siu ti hn. Bng 2 trnh by mt s lipase thng mi ang cung cp cho cc ngnh ngh trn th trng.

Bng 2. Mt s lipase thng mi ang cung cp cho cc ngnh ngh trn th trng. STT 1 2 3 4 5 6 7 8 9 Tn Lipopan Lipozyme PalataseTM Clear-LensTM LIPO Greasex NovoCorTM AD LipolaseTM LipoPrime Novozym 871 86 Phm vi ng dng Bnh ko Ha du m Sn phm t sa Ty ra Thuc da Thuc da Bt git Bt git Thc n cho vt nuigia sc

4 Lipoxygenase 4.1 Gii thiu chung Lipoxygenase (E.C. 1.13.1.13) xc tc s oxi ha ca cc acid bo khng no nhiu ni i nh linoleic, linolenic, arachidonic v cc ester ca chng. Lipoxygenase rt quan trng trong cng ngh thc phm v chng c th nh hng n mu sc, mi v v gi tr dinh dng ca sn phm. Lipoxygenase c mt nhiu loi m thc vt v ng vt. c bit l s c mt ca lipoxygenase trong cc ht mang du (v d nh u nnh) c th c xem l nguyn nhn chnh hnh thnh cc peroxide trong qu trnh trch ly du thc vt.

Hnh: Cu trc (a) v trung tm hat ng (b) ca lipoxygenase u nnh Lipoxygenase thng tn ti nhiu dng isozymes khc nhau v pH hot ng ti u cng nh l sn phm v tnh c hiu ca c cht. Ly v d nh t u nnh ngi ta c th tch c 4 loi isozyme khc nhau. - Isozyme u nnh 1: c pH ti u l 9.0. Loi ny ch tc dng trn cc loi acid bo t do c nhiu ni i. nhit phng, loi enzyme ny hnh thnh peroxide v tr Cacbon s 9 v 13 theo t l 1:9. - Isozyme u ny 2: c pH ti u l 6.8. Loi ny c th xc tc trn triglyceride cng nh l acid bo t do c nhiu ni i v hnh thnh peroxide v tr 9 v 13 theo t l 1:1 nhit phng. - Isozyme u nnh 3: tng t nh isozyme 2 nhng hot tnh ca n d b c ch bi ion Ca2+ - Isozyme u nnh 4: tng t nh loi isozyme 3 nhng c th c tch bi phng php sc k hay in di Ngoi ra cc isozymes cn thng c phn thnh loi 1 (pH ti u > 7 v c hiu vi cc acid bo t do) v loi 2 (pH ti u = 7 v oxi ha c carotenoids). Kh nng lipoxygenase loi 2 c kh nng ty mu carotenoids ngy nay c ng dng trong vic sn xut bnh m trng (bng cc thm bt u nnh vo bt m b sung lng isozymes loi 2). Lipoxygenase ng vt v thc vt thng c hat ha khi cc m t bo b tn thng hoc b ph v. Kt qu l cc acid nhiu ni i b oxi ha thnh cc hydroperoxide. Sau s phn hy thnh nhiu sn phm bay hi khc nhau. 87

Xt v phng din sn phm ca qu trnh xc tc, Lipoxygenase to ra cc sn phm bay hi tng t nh sn phm trong qu trnh t oxy ha kh, mc d t l ca thnh phn sn phm khc bit nhau ty vo tnh c hiu ca enzyme cng nh iu kin phn ng (Bng 3). Bng 3. So snh thnh phn ca sn phm bay hi ca phn ng oxy ha xc tc bi lipoxygenase u H Lan (a) vi s t oxi ha kh (b) C cht: Linoleic acid Sn phm % (a) % (b) n-pentanal 7 0,7 n-hexanal 41 67 n-hept-2-enal 15 6 n-oct-2-enal 10 18 n-nona-2,45 0,4 dienal 20 5 n-deca-2,4-dieanl C cht: Linolenic acid Sn phm % (a) % (b) Acetaldehyde 5 Propanal 27 n-but-2-enal 3 1 n-penta-2-enal 22 10 n-hex-2-enal 2 1 n-hex-3-enal 14 n-hepta-2,4-dieanl 38 50

4.2 C ch phn ng Phn t lipoxygenase c cha mt nguyn t st. Nu nguyn t st ny chuyn t dng Fe2+ sang Fe3+ th lipoxygenase s chuyn t dng th ng sang dng hot ha xc tc phn ng. Qu trnh xc tc c din t trong s 1. u tin Fe2+ c th b oxi ha thnh Fe3+ nu c tip xc vi acid bo peroxide hoc l OOH. Do nu c cht ch n thun l acid bo t do th qu trnh oxy ha din ra kh chm. Lipoxygenase c kch hot s t t tch nguyn t hydro t nhm methylene nm gia ca acid bo nhiu ni i. Trong iu kin hiu kh, phc hp enzyme -gc alkyl s c oxi ha bng phn t oxy tr thnh phc hp enzyme-gc alkyl peroxide. Sau in t s c truyn t nguyn t st sang nhm peroxide. V nh vy s thm vo v tch proton t enzyme to ra sn phm hydroperoxide. Trong iu kin ym kh th gc alkyl s c tch ra khi phc hp enzyme-gc alkyl, ri t s hnh thnh hn hp cc sn phm nh cc cht nh trng, ketones v epoxides. 4.3 Hn ch v bt hot lipoxygenase Vic lipoxygenase gy ra mi i trong cc sn phm cha du m l mt trong nhng vn ang c quan tm. Hin ti c rt nhiu cc nh nghin cu ang lm vic ti u ho cc iu kin lm bt hot lipoxygenase trong cc sn phm cha du m. Cc phng php thnh hnh ang c nghin cu v p dng l b sung cc cht chng oxi ha, hiu chnh pH v nhit ... Lipoxygenase c th b hn ch bi rt nhiu loi cht chng oxi ha (v d nh pyrocatechol, homocatechol, propylgallate, nordihydroguariaretic acid, resorcinol, phioroglucinol, hydroquinone, butylated hydroxyanisole v nhiu loi flavonoids cng nh cc hp cht lin quan khc.Ngy nay cc cht chng oxi ha c ngun gc t thc vt ang c cc khoa hc gia ht quan tm nghin cu v khai thc.

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S 1: Qu trnh xc tc ca lipoxygenase X l nhit c th lm nh hng n tnh tan ca protein cng nh lm thay i cc tnh cht chc nng ca cc sn phm protein theo chiu hng bt li. V d nh ngi ta hiu chnh u xanh xung m 16,3% vi m pH 9,8 sau un nng trn 90oC trong vng 10s. Bng cc ngi ta c th lm bt hot 99% lipoxygenase, nhng bn cnh cng ch gi li c 70% tnh ho tan ca protein. Ngi ta cng thy rng lipoxygenase l nguyn nhn chnh gy nn hin tng tr mi u H lan xanh. Ngoi ra lipoxygenase cng chu trch nhim cho vic lm bin i mi v ca chui. Mc bn nhit ca cc loi lipoxygenase trong cc loi thc phm rt khc nhau. Ngi ta nhn thy rng lipoxygenase trong u H lan v u xanh chiu nhit km hn peroxidase 60oC trong 10 pht. Do vy v hot cc loi u trn th cng ch cn x l nhit mc khng qu cao. Trong khi mt nghin cu khc, ngi ta li nhn thy lipoxygenase trong mng ty li bn nhit hn so vi peroxidase. Bi vy cho d x l nhit nhit 70oC trong vng 10 pht c th lm v hot peroxidase nhng li khng vo hot lipoxygenase X l nhit bng microwave c th hn ch rt hiu qu hot ng ca lipoxygenase. Ngi ta nhn thy x l u nnh bng microwave c th va lm tng tnh ht nc ca ht li va c th lm v hot lipoxygenase v cc cht c ch hp thu trypsin. Do ngi ta c th x l u nnh (8,7% m ) bng sng siu vi trong 4 pht trc khi em i sn xut sa u nnh. Nu u c em ngm nc trc th thi gian x l nhit lm bt hot enzyme s cng ngn hn.

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4.4 Nhng xu hng pht trin trong tng lai Hin nay ngi tiu dng ngy cng quan tm n nhng sn phm c cha nhiu DHA v EPA. V v th cc sn phm ny rt d bi oxi ha bi lipoxygenase. iu ny i hi cn c nhiu nghin cu hn na v cc cht chng oxi ha, cc phng php vt l ngn chn s h hng do oxi ha. Cc phng php s ch du n (c bit l loi du cha nhiu acid bo c nhiu ni i) trc khi a vo ch bin thc phm c ci tin sao cho lm gim thiu vic to nn cc peroxide.

5 S i ha sinh hc ca lipid 5.1 Gii thiu chung cc phn trc, chng ta cp n ng dng ca thy gii lipid trong cc ngnh cng nghip khc nhau. Trong cng ngh thc phm ni ring vic thy gii lipid cng vi s oxi ha cc acid bo cn ng mt vai tr rt to ln i vi cht lng ca sn phm trong qu trnh ch bin v bo qun. Trong cc sn phm ng vt v thc vt, hai nhm lipid chnh l phospholipid v triglyceride. Phospholipid l thnh phn khng th thiu trong thnh phn mng t bo bi vy chng c gi l lipid cu trc. Cn triglyceride thng c gi l lipid d tr nng lng, v thng c cha trong cc t bo m c bao bc bi cc mng phospholipid. Trong thi gian ch bin v bo qun, lipase v phospholipase s thy gii triglyceride v phospholipid. Cng vi cc acid bo lin kt, cc acid bo c to ra trong qu trnh trn (c bit l cc acid bo nhiu ni i) s d dng b oxid ha bi cc tc nhn ha hc hoc enzyme lipoxygenase. C ch oxy ha do lipoxygenase:

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5.2 Phospholipases Phospholipase A2 (EC 3.1.1.4) l phospholipase u tin c cng nhn v ng vai tr rt quan trng trong cc ng dng ca phospholipase. Enzyme ny xut hin rt nhiu trong t nhin v c mt hu nh tt c cc lai t bo. Phospholipase A2 (PLA2) l mt thnh phn ca thc phm t ngun ng thc vt v v vy lun c a vo c th ngi thng qua ng n ung. PLA2 ng vai tr tch cc trong vic ci thin cc tnh nng ca lecithin (u nnh, lng trng), lai gum trong qu trnh tinh luyn du..., nhng bn cnh n cng l nguyn nhn gy nn mt s h hng trong cc sn phm. Ngy nay ngi ta khm ph c rng nguyn nhn thy phn phospholipid trong tht c ch yu l do PLA2. Mt s nghin cu v PLA2 trong c quan trng nh: PLA2 c Hi c pH ti u l 7 v khng i hi ion Ca2+ hat ha. PLA2 trong c Tuyt khng ph thuc Ca2+ v pHopt >7. PLA2 ny c kh nng thy phn phospholipid trong c tuyn tng dn trong 8 tun tr lnh u tin 30oC, sau s gim dn n tun th 12. V chng ta cn lu rng hat ng ca PLA2 gng lin vi vic c ca c tr nn cng hn do tc ng ca cc acid bo vi cc c protein. PLA2 trong c Plc th i hi s c mt ca Ca2+. Khc vi PLA2 trong c Tuyt, pHopt cua PLA2 trong c Plc c tnh kim 91

Tuy nhin ngi ta ngi ta vn c th tm thy cc hat ng thy gii phospholipid acyl khc c. V d nh c ng Euthynnus pelamis, PLA1 li l nguyn nhn to ra lysophospholipid trong tht c. PLA1 ny khng ph thuc Ca2+ v c pHopt vo khang 6,5 7,0. Mt nghin cu khc cho thy c thu i Ty Dng cng c loi PLA 1 khng ph thuc Ca2+ vi pHopt khong 4,0. Nh vy c ni chung, nhng tn hi m khi b nh bt hoc do stress s lm tng hot tnh ca PLA1 v PLA2, v kt qu l nh hng n qu trnh bo qun ca sn phm. Phospholipase A2 c xc tc cc phn ng thy phn phosphoatidylcholine to thnh acylglycerophocholine. Phospholipase A2 c s dng thy phn lecithin ci bin cc tnh cht nh ha ca n. Cc lecithin thng mi l mt hn hp phosphatide ca choline, ethanolamine v inositol vi mt lng nh cc lipid khc v c s dng rng ri trong nhiu lai thc phm khc nhau. Cng dng ca lecithin nh l mt cht nh ha trong ch bin thc phm th rt ph bin, tuy nhin cc lai lecithin cha c ci bin th tnh cht nh ha ca chng rt hn ch trong cc thc phm du m. Phospholipase A2 thy phn lin kt ester gia glycerol v acid bo ti v tr sn-2 ca lecithin nhm to ra lysolecithin v mt phn t acid bo. Kt qu l lysolecithin to thnh c kh nng to nh ho cao hn trong nhiu loi thc phm khc nhau.

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