TRNG I HC KHOA HC B MN KHOA HC S SNG

TIU LUN:
NG DNG CA ENZIM V PROTEIN
Nhm 5: Dng Th Kim Th Nguyn Th Thu Nguyn Th Huyn Trang Quc Vit Nguyn Quc Nam

Thi Nguyn, thng 11 nm 2009

NG DNG CA ENZYME I. ng dng ca enzim amylase

1. THUC, HO CHT CHA HOT TNH ENZYME AMYLASE PHC V TRONG NUI TRNG THY SN * ng dng: - Enzyme amylase thng c b sung trong thnh phn cc hp cht ha hc * i tng - C n thc vt v c n ng vt * C ch -i vi c n thc vt -i vi c n tht ng vt 2. NG DNG ENZYME AMYLASE TRONG SN XUT CHT TY RA Cht ty ra bao gm nhng cht kim, sodium silicate, sodium bicarbonate, sodium tripolyphosphate - Mc ch: loi b cc cht v c, hu c bm vo qun o nh : protein, lipid, carbohydrate v nhng cht mu Enzyme -amylase ca vi khun l mt trong nhng enzyme thng c ng dng trong cng nghip sn xut cht ty ra

3.NG DNG ENZYME AMYLASE TRONG Y HC V DC PHM *VAI TR CA ENZYME AMYLASE TRONG TIU HA V BIN DNG CARBOHYDRAT S tiu ho tinh bt amylase Tinh bt Dextrin + maltose + glucose Thy phn -1,4 ca amylose

-1,6 glucosidase
Dextrin Thy phn -1,6 ca amylospectin glucosidase (maltase) Maltose + glucose

Maltose
glucosidase (lactase) Lactose

2 glucose

Glucose + galactose

fructofuranisidase (sucrase)
Suctose Glucose + fructose

4. NG DNG ENZYME AMYLASE TRONG CHUN ON VIM TU CP TR EM

Da vo cc c tnh biu hin ca enzyme amylase ngi ta tin hnh nghin cu phng php chun on bnh vim tuyn tu *i tng: - enzyme s-amylase - p- amylase - LIPRE - S-AMYLASE v LIPRE - P-AMYLASE v LIPRE Theo nghin cu ngi ta nhn thy: Vi im ct: S-AM =400 u\ l; P-AM=150 u\ l th gi tr ca enzyme S-amylase, P- amylase, lipre mu trong chun on vim ty cp tr em ng tin cy v t gi tr cao nht. * Kt lun: - Khi chn la im ct thch hp v phi hp c 2 enzyme samylase + lipre hoc p-amylase + lipre trong chun on vim ty cp tr em th gi tr chun on chnh xc cao nht.

5. NG DNG ENZYME AMYLASE TRONG CNG NGHIP THC PHM *NG DNG ENZYME AMYLASE TRONG CNSX M CHNH - Nguyn liu s dng ch yu : Tinh bt sn, r ng ma A-amylase Tinh bt Dextrin + maltose + glucose Thu phn A-1,4 amylose B-amylase Tinh bt Maltose + B dextrin (glucogen) *NG DNG TRONG SN XUT GLUCOZA V MT Chng ta bit t tinh bt c th thu c cc phm vt ng khc nhau khi thy phn tinh bt bng acid cng nh bng Enzym amylase s thu c mt. Mt glucoza hay mt maltoza thng c dng trong sn xut bnh ko v trong sn xut cc sn phm n kin cho tr em v ngi bnh

* NG DNG ENZYME AMYLASE TRONG CNSX BIA

Nguyn liu s dng: Ng cc, hoa Houblon, nc, Nm men, cht ph gia Cc chng vi sinh vt s dng : -Saccharomyces cerevisizae ( ln men ni) -Saccharomyces cerevisidae (Ln men chm) Trong cng ngh sn xut bia, ngi ta thng s dng emzyme amylase c trong mm i mch.

* NG DNG ENZYME AMYLASE TRONG CNSX CN * NG DNG TRONG SN XUT SIRO

Qu trnh chuyn ha tinh bt thnh siro fructose

* NG DNG AMYLASE TRONG CNG NGH SN XUT BNH M Trong sn xut bnh m, ngi ta s dng enzyme nhm gii quyt mt s vn sau: +lm tng nhanh th tch bnh +lm mu sc ca bnh p hn +lm tng mi thm cho bnh. Trong sn xut bnh m, ngi ta s dng c hai loi Enzyme a -amylase v _amylase, cc loi enzyme ny tham gia thy phn tinh bt de tao thanh ng. Nh nm men Saccharomyces cerevisiae s d dng chuyn ha chng thnh cn, CO2, lm tng th tch ca bnh v to ra mu sc, hng v tt cho bnh. *NG DNG AMYLASE TRONG SN XUT BNH KO Mc ch + Lm tng mi v v bnh, khi ch bin bt thnh cc loi bnh quy cc Enzyme protease v amylase ca bt hot ng lm tng hm lng cc amino acid t do v lm tng lng ng kh. + ng kh v cc amino acid t do c trong khi bt s cng tham gia vo cc phn ng oxy - ha kh v kt qu to cho bnh quy c mi, v mu hp dn.

II. ng dng ca enzyme trong y hc

1.ng dng ca enzyme trong cha bnh: Enzym nh cht cho thm vo c th cha bnh km tiu ha i vi mt s ngi . Enzym c s dng nh cht lm sch vt thng v lm lnh vt thng . Enzym c s dng trong cc phn ng min dch . 1.1 Enzyme c nh : Ngy nay ngi ta tm ra hn 120 bnh v ri lon chuyn ha ngi , a s cc bnh ny l do thiu mt loi enzyme c bit no . V d : bnh phenylcetone niu (phenylketonuria) do mt khuyt tt bm sinh v chuyn ha protein , to ra qu mc amino acid phenylamine trong mu , lm tn hi h thn kinh v a n chm pht trin tr tu nghim trng . Nhng ngi b mc bnh ny thng thiu enzyme chuyn i phenylalanine . Mt enzyme c chc nng tng t khng c ngun gc t c th ngi khng th a mt cch trc tip vo c th bi v n s gy ra mt p ng min nhim c hi cho c th . gii quyt vn ny ngi ta c lp enzyme trong cc vi ht , si hay gel . Khi , enzyme c th khng gy ra p ng min nhim c hi no trong khi c cht ca n c kch thc nh c th i xuyn qua gel , cc l trn si hay mng ca cc vi ht

1.1 Enzyme c nh :
Ngy nay ngi ta tm ra hn 120 bnh v ri lon chuyn ha ngi , a s cc bnh ny l do thiu mt loi enzyme c bit no . V d : bnh phenylcetone niu (phenylketonuria) do mt khuyt tt bm sinh v chuyn ha protein , to ra qu mc amino acid phenylamine trong mu , lm tn hi h thn kinh v a n chm pht trin tr tu nghim trng . Nhng ngi b mc bnh ny thng thiu enzyme chuyn i phenylalanine . Mt enzyme c chc nng tng t khng c ngun gc t c th ngi khng th a mt cch trc tip vo c th bi v n s gy ra mt p ng min nhim c hi cho c th . gii quyt vn ny ngi ta c lp enzyme trong cc vi ht , si hay gel . Khi , enzyme c th khng gy ra p ng min nhim c hi no trong khi c cht ca n c kch thc nh c th i xuyn qua gel , cc l trn si hay mng ca cc vi ht

Cng theo nguyn tc ny ngi ta c tng lm thn nhn to .Trong thit b nhn to ny , urease v ht resin hp th hay than ch c kt thnh nang vi nhau .Khi urea s b urease phn hy to thnh ammonia , ammonia s c hp th trong cc vi nang : khuch tn urease hp th ln serin UREA UREA HCO-3 + NH4 Vo vi nang hay than ch Trong s cc th nghim lm sng quy m nh v enzyme c nh , ngi ta tin hnh th nghim trn qu trnh chuyn i steroid . Cortisol l mt loi thuc hu dng dung trong iu tr bnh vim khp , cortisol c th c sn xut t mt loi tin cht rt r tin l 11-deoxycortisol nh ct enzyme c nh 11--hydroxylase Trong mt tng lai gn enzym s c s dng rng ri trong ngnh y t lm u d cho cc thit b phn tch y t v cha bnh. Ngai ra cn c mt s ng dng : -Vi tiu cu cha catalaza c th thay th mt cch hiu qu cc catalaza cn thiu trong c. - a vi tiu cu c gn enzyme L-asparaginaza vo c th , c kh nng c ch s pht trin ca mt s u c tnh bi s pht trin ca cc u ny ph thuc vo s c mt ca L-asparagin.

1.2 Enzyme t ngun thc vt: a/ Bromelin : C ch tc dng: -Vai tr h tr tiu ha. -Gim au v ph n sau phu thut, sau chn thng. -Lm lnh vt thng -Gim au nhc c -Chng ri lon tim mch. Ngoi ra cn c mt s nghin cu khc b/ Papain v Chymopapain: C ch tc dng : -Trong nha qu u c cha Papain v Chymopapain .Papain gn ging vi Chymopapain nhng c hat tnh mnh hn gp nhiu ln . -Papain gip km hm mt s vi trng gy bnh nh gram v tiu dit nhiu vi trng khc nh: staphilooccus, thng hn. -C kh nng gim c i vi toxin v toxanpunin. -Nha papain th dng iu tr ri lon tiu ha do thiu men, gip tiu ha tt cht m trong thc n , lm thuc ty nhiu loi giun. -C tc dng nga thai v gy sy thai do hot tnh ca n i vi progesteron ca thai ph. -Chit xut papain c dng trong phu thut ct sng nh l mt loi dao phu thut t nhin m a m ct sng. -c dng iu tr l lot , lm tiu gii trong bnh bch hu, chng kt dnh sau phu thut. -Khi tinh ch du gan c thng b sung papain tng hm lng vitamin A v D.

1.3 Enzyme t ngun ng vt: a/ Pepsin:

C ch tc dng : -Pepsin l mt enzyme phn hy Protein ,phn b trn cc phn khc nhau ca d dy . Hat ng trong dch v ca ng vt c v , chim , b st v c. heo enzyme tp trung nhng t bo ca phn y bao t .c ng dng cha bnh km tiiu ha . -Pepsin phn hy c cht , lm gim acid trong d dy .Nh hat ng phn hy protein , khi lng thc n gim v kh nng tiu ha protein tng ln .

Sn phm : ch phm enzyme tn ti dng bt v nh hnh , trng hay vng nht , hay mnh nh , trong hay hi c , mi c bit ging mi nc tht , v hi chua . b Trypsin va Chymotrypsin :

-Trypsin v Chymotripsin l nhng protease kim tit t tuyn ty ca ng vt mu nng . Cc enzyme ny khi mi c tit ra u dng tin enzyme (proenzyme) bt hot ( trypsinogen v chymotrypsinogen ), sau chng c hot ha v tr thnh dng hot ng l trypsin v chymochypsin trong t trng . Cc enzyme ny thuc nhm enzyme phn ct cc lin kt amide , lin kt peptide . Ngoi ra chng cng c th ct c lin kt carbon carbon. -Tuy nhin vai tr chnh ca enzyme ny trong c th l thy phn lin kt peptide trong sut qu trnh tiu ha protein rut non. -Trypsin cng c ng dng cha bnh km tiu ha .

2.ng dng ca enzyme trong chn on bnh

2.1 ng dng enzym trong xc nh nng c cht c xc nh theo 2 phng php : a/Phng php xc nh im cui :
+Phng php xc nh urea

Urea b thy phn bi urease (EC.3.5.1.5) 0,7 UI/ml. V amoniac c to thnh khi cho enzym glutamate dehydrogenase (EC.1.4.1.3) c hot tnh 6,2 UI/ml tc ng. Urea + H2O 2NH3 + CO2 2-Cetoglutarate + 2NH4+ + 2NADH 2L-glutamate + 2NAD+ +2H2O +Phng php xc nh glucose vi glucose-oxidase Trong phn ng u tin, glucose b oxy ha bi glucose oxidase (EC.1.1.3.4), to thnh peroxide hydro theo phng trnh sau: Glucose-oxidase 10 UI/ml Glucose + O2 + H2O gluconate + H2O2 Trong phn ng th hai peroxide hydro, di tc dng ca enzym horse-radish peroxidase (EC.1.11.1.7) s to mu theo phn ng sau: H2O2 + chromogen mu + H2O horse radish peroxidase Trong phn tch ny, 2,2-azino-bis (3-ethyl 2,3 dihydrobenzothiazol sulfanote (ABTS) c s dng nh chromogen.

b. Phung php ng hc : (kinetic methods) Phng php ny ch xc nh nng c cht di gi tr Km . + Phng php xc nh glucose : phn ng xy ra trong phng php ny nh sau: D-glucose + ATP D-glucose -6-phosphate + ADP D-glucose -6-phosphate +NAD.P+ D-glucose-- lactone 6-phosphate + NADPH + H+ phn ng u , glucpse c phosphoryl ha bi hexokinase (EC .2.7.1.1 ) Sau d , glucose -6phosphate b hydrogen ha bi tc ng ca glucose-6-phosphate dehydrogenase (EC.1.1.1.49 ) .S to thnh NADPH s c xc nh bng my quang in + Xc nh triglyceride : Cht bo c thy phn bng lipase ( EC .3.1.1.3) v carbpxylesterase (EC.3.1.1.1) . Glycerol sau s c phosphoryl ha bi glycerol kinase (EC .2.7.1.30) . ADP c to thnh s tip tc c phosphoryl ha n ATP vi phosphor enol pyruvate v pyruvate kinase 9EC.2.7.1.40) . Cui cng pyruvate c hydrogen ha bi L-lactate dehydrogenase ( EC.1.1.1.27) v NADH s gim dn . Triglyceride + 3 H2O glycerol + 3 acide bo Glycerol + ATP glycerol 3 phosphate + ADP ADP + phosphor enol pyruvate ATP + pyruvate Pyruvate + NADH + H+ L-lactate +NAD+
2.2 Xc nh hat tnh enzyme : Xc nh hat tnh ca alkaline phosphatase (EC.3.1.3.1) 4-Nitro phenyl phosphate + H2O phosphate + 4-nitro phenolate pH ti u 9,8 sn phm s phn tn v tc phn ng s tng theo hp th bc sng 405nm . Xc nh hat tnh ca creatine kinase (EC .2.7.3.2) Creatine phosphate +ADP creatine +ATP ATP c to thnh trong phn ng ny nh xc tc ca enzyme creatine kinase

2.3 Thc hnh min dch :

Enzyme c s dng y xc nh hn hp khng nguyn khng th , to thnh trong phn ng min dch . Ta c th s dng my so mu quang in , hynh quang xc nh phn ng . +Enzyne alkaline phosphatase ( EC .3.1.1.3) Enzyme ny c ng dng trong phn ng min dch . Ngi ta thng s dng enzyme ny vi hat tnh 2500UI /mg , nhit 37oC . xc nh hat tnh alkaline phophatase c th s dng my hynh quang vi 4methylumbelliferyphosphatase lm c cht . + galactosidase ( EC .3.2.1.23) Ngi ta thng s dng enzyme ny c hat tnh ring ln hn 250UI/mg vi c cht l 4-nitrophenyl--D-galactosidase , 37o C . Hat tnh ca -galactosidase c o bng my quang in vi 4-methylumbellifery- galactosidase . Ngai ra ta c th s dng 5-bromo-4-chloro-3-indolyl- -galactosidase. +Horseral peroxidase (EC .1.11.1.7) Enzyme ny cha 2 n 3 nhm hat ng aminE trong phn t v cha 12-14,5% carbphydrate . Xc nh hat tnh enzyme ny bng my quang in . C cht thng s dng l chromogen-2-2azinobis [3-ethybenzothiazoline-sulfonate] (ABTS) v peroxidase c hat tnh ring l 100u/c 25oC. Trong min dch ngi ta thng s dng c cht l 3,3; 5,5-tetramethyl benzidine.

III. ng dng Enzyme trong phn tch thc phm

1.Xc nh carbohydrate Trong thc phm, carbohydrate chim khi lng ln v ng vai tr quan trng trong dinh dng. Cc loi ng nhng i tng c phn tch thng xuyn. *Glucose Glucose c xc nh bng phng php Enzymehexokinase. Phn ng ca qu trnh xy ra nh sau : Hexokinase D-glucose + ATP ADP + glucose 6-phosphate Glucose 6-phosphate-dehydrogenase Glucose-6-phosphate + NADP+ D-gluconate -6phosphate + NADPH + H+

* Fructose
Enzyme hexokinase cng tc ng ln fructose. Ta c th xc nh fructose sau khi xc nh glucose. D- glucose + ATP ===== > ADP + Fructose -6- phosphate Glucose-phosphate -isomerase Fructose 6- phosphate glucose-6-phosphate

* Galactose
D-galactonic acid + NADH + H+ D- galactose + NAD+

* Manose

hexokinase

D-manose + ATP

ADP + Mannose-6- phosphate Phosphonase-isomerase Mannose-6- phosphate Fuctose-6-phosphate

*Saccharose
Saccharose thng khng c trong t bo ng vt. Xc nh saccharose theo phng trnh sau : Saccharose + H2O D-glucose + D-fructose

*Maltose
Xc nh theo phng trnh sau : Maltose + H2O 2-D glucose

*Lactose
Lactose thng c trong sa. Xc nh lactose theo phng trnh phn ng sau : Lactose + H2O D- galactose + D-glucose

*Rafinose
Rafinose c trong c ci ng. Xc nh rafinose theo phng trnh phn ng sau : Rafinose + H2O D-galactose + saccharose

*Tinh bt
Tinh bt c nhiu trong thc vt v c c mt s loi vsv. Xc nh tinh bt theo phn ng sau : Tinh bt + ( n 1 ) H2O n D-glucose 2. Xc nh trong acid hu c
Acid hu c v mui ca chng c nhiu nguyn liu v sn phm thc phm. Chng ng vai tr rt quan trng trong sinh l ngi , ng vt , thc vt v vsv. Chng cn c to ra do qu trnh ln men.

*Acetic acid
Acetic acid thuc nhm acid bay hi . Ngi ta thng s dng enzyme xc nh acetic acid. Phn ng trong phng php ny c trnh by nh sau : Acetyl-CoA.synthetase Acetate + ATP + CoA Acetyl-CoA + AMP + Pyrophosphate

Citrate-dehydrogenase
Acetyl-CoA + Oxaloacetate + H2O L-malete dehydrogenase L-malate + NAD+ H+ Phn ng sau cng c xem nh phn ng ch th . Acetate c nhiu trong vang oxaloacetate + NADH + citrate +CoA

*Ascorbic acid
Ging nh vitamin , ascorbic acid ng vai tro sinh hc ln trong sinh l ngi v ng vt. Chng c s dng nh cht ph gia thc phm. Ngi ta s dng enzyme xc nh ascorbic acid theo phng trnh sau: L ascorbic acid ( XH2 ) + MTT dehydro ascorbic acid ( X) + fomazan + H+ MTT : 3 ( 4.5 dimethylthiazolyl -2 )-2.5-diphenyltetrazolium bromide Ascobacte c oxy ho tip : L-ascorbic acid + ehydro ascorbic acid + H2O Ngi ta thng xc nh ascorbic acid trong nc qu , trong rau , qu , trong sa , trong sn phm tht . Dehydro ascorbic acid + dithiothreitol L-ascorbic acid + dithiothreitol

*Aspartic acid
Aspartic acid c nhiu trong nc to v c xc nh theo phn ng sau: L-Aspartate + A- oxoglutarate Oxaloacetate + L-glutamate Oxaloacetate + NADH + H+ L-malate + NAD+

*Citric acid
Citric acid ng vai tr rt c bn trong trao i cht vi sinh vt. Chng c nhiu trong tri cy , trong sa. Xc nh citric acid theo phng trnh sau: Citrate Oxaloacetate + acetate

*Formic acid
Acid Formic l sn phm trao i cht ca vi khun v ca nm si. Acid ny c xem nh cht bo qun nhiu thc phm. Tuy nhin, vic s dng acid ny trong bo qun thc phm phi tun theo lut an ton v v sinh thc phm. Ngi ta xc nh nng lng acid formic theo phng trnh sau :
Formate + NAD+ + H2O Hydrogencarbonate + NADH + H+

*Glucomic acid Ngi ta thng s dng enzyme gluconate kinase xc nh acid glucomic, phn ng xy ra nh sau :
Gluconate-kinase D-gluconate + ATP D-gluconate 6-phosphate + ADP 6.PGDH D-gluconate-6-phosphate+NADP+ D-ribulose-5-phosphate + NADPH+ + H+ + CO2

*Glutamic acid
Ngi ta s dng enzyme glutamate dehydrogenase xc nh glutamic acid. Phn ng xy ra nh sau : Glutamate-dhydrogenase L-glutamate + NAD+ + H2O A-Oxoglutarate + NADH + NH4+

*Acid Isocitric
Ngi ta thng s dng enzyme isocitrate dehydrogenase xc nh isocitric acid. Phn ng xy ra nh sau: iso-citratedehydrogenase D-isocitrate + NADP+ A-Oxoglutarate + NADPH + CO2 +H+

*Acid Lactic
Acid Lactic c to ra nhiu trong qu trnh ln men. Ngi ta xc nh lactic acid bng lactate dehydrogenase. Phn ng xy ra nh sau : L-lactate-dehydrogenase L lactate + NAD+ Pyruvate + NADH + H+ L-lactate-dehydrogenase D lactate + NAD+ Pyruvate + NADH + H+

*Acid Malic
Acid Malic c nhiu trong nho , trong rau , qu khc. Ngi ta xc nh Malic acid bng malate dehydrogenase v NAD+ , phn ng xy ra nh sau: Malate-dehydrogenase L-malate + NAD+ GOT Oxaloacetat + NADH + H+

Oxaloacetate + L-glutamate

L- aspartate + A-oxoglutarate

*Acid Oxalic
Acid Oxalic , ng vai tr quan trng trong hp th calcium c th ngi. Ngi ta s dng enzyme oxalate dehydrogenase xc nh oxalic acid. Phn ng xy ra nh sau: Oxalate-dehydrogenase Oxalate Formate + CO4

*Acid Pyruvic
Pyruvic acid l mt acid c bn trong chu trnh chuyn ha mi c th. Ngi ta s dng enzyme L-lactate dehydogenase. Phn ng xy ra nh sau: L-lactate-dehydrogenase Pyruvate + NADH + H+ L-lactate + NAD+

*Acid Succinic
Acid Succinic cng l mt acid quan trng trong chu trnh tricarboxylic acid. Ngi ta xc nh succinic acid bng enzym succinyl CoA-synthetase Phn ng xy ra nh sau: Succinyl-CoA.synthetase Succinate + ITP + CoA IDP + succinyl-CoA + P Pyruvate-kinase IDP + PEP ITP + pyruvate

3. Xc nh alcohol:
a. Xc nh ethanol:
Ethanol l sn phm ln men ng bi nm men. Ngoi nhng phng php bnh thng, ngi ta con s dng enzyme xc nh ethanol. Phn ng xy ra nh sau:
ADH Ethanol + NAD+ Acetaldehyde +NADH + H+

b.Xc nh glycerol:
Glycerol ph bin nhiu trong thin nhin v c nhiu trong qu trnh ln men. Ngi ta xc nh glycerol bng enzyme glycerol kinase v pyruvate kinese. Phn ng xy ra nh sau: Glycerol-kinase Glycerol + ATP Pyruvate -kinase ADP + PEP ATP + Pyruvate glycerol-3-phosphate + ADP

*Xc nh alcohol ng

Ngi ta xc nh sorbitol bng enzyme sorbitol dehydrogenase phn ng xy ra nh sau : Sorbitol-dehydrogenase D Sorbitol + NAD+ D-fructose + NADH + H+ Tng t , ngi ta cng xc nh xylitol bng enzyme sorbitol dehydrogenase. Phn ng xy ra nh sau : Sorbitol-dehydrogenase Xylitol + NAD+ xylulose + NADH + H+

C. Xc nh cc thnh phn khc

*Xc nh cholesterol Cholesterol l mt steroid c ngha rt ln trong sinh l ngi v ng vt. Ngi ta xc nh cholesterol bng enzyme cholesterol oxidase v catalase. Phn ng xy ra nh sau: Cholesterol-oxidase Cholesterol + O2 Cholestenone + H2O2 Cholesterol-oxidase H2O2 + methanol Formaldehyde + 2H2O Formaldehyde + NH4++ 2acetylacetone Lutidine + 3H2O

*Xc nh triglyceride Ngi ta xc nh triglyceride bng esterase v lipase. Phn ng xy ra nh sau: esterase v lipase Triglyceride + 3H2O glycerol + 3 acid bo *Xc nh acetaldehyde y l cht taoi mi cho bia v yauort v cc loi nc gii kht. Ngi ta xc nh acetaldehyde bng enzyme acetaldehyde dehydrogenase. Phn ng xy ra nh sau : Acetal dehydrogenase Acetaldehyde + NAD+ + H2O acid axetic + NADH + H+ *Xc nh Amoniac y l cht cha nitrogen n gin nht. Ngi ta s dng enzyme glutamate dehydrogenase xc nh amoniac. Phn ng xy ra nh sau: Glutanate dehydrogenase Oxoglutarate + NADH + H+ + NH4 L-glutamate + NAD+ + H2O

*Xc nh Nitrate Ln u tin s dng enzyme xc nh nitrate. Enzyme c s dng xc nh nitrate l nitrate reductate. Phn ng xy ra nh sau: Nitrate reductase Nitrate + NADPH + H+ Nitrite + NADP+ + H2O *Xc nh Sulfite: Xc nh sulfite bng enzyme c bt u t nm 1983. enzyme c ng dng xc nh sulfite l sulfite oxidase. Phn ng xy ra nh sau : Sulfite oxidase SO32- + O2 + H2O SO42- + H2O2 NADH-peroxidase H2O2 + NADH + H+ 2H2O + NAD+ *Xc nh creatin v creatinine Hai cht ny c trong c. Enzyme c s dng xc nh creatinine l creatiminase , xc nh creatine l creatine kinase. Phn ng xy ra nh sau : creatiminase Creatinine + H2O creatine creatine kinase Creatine + ATP creatine phosphate + ADP

*Xc nh lecithin Lecithin l mt phospholipide quan trng. Ngi ta xc nh lecithin bng phn ng v nhng enzyme sau: Phospholipase - C 1.2diglyceride + phosphorylcholine Alkaline-phosphatase Phosphorylcholine + H2O choline + Pi Choline - kinase Choline + ATP phosphoryicholine + ADP *Xc nh urea Ngi ta s dng urase xc nh urea. Phn ng xy ra nh sau: urease
Urea + H2O 2NH3 + CO2

Lecithin + H2O

IV.MT S NG DNG TIU BIU TRONG CHUN ON BNH

1. ng dng ca enzym Raffinase c nh: S pht trin ca enzym Raffinase (Galactosidase) ph hp vi cc ng dng thng mi l mt thnh cng trong lnh vc cng ngh enzym +Chng sinh vt c kh nng sn xut - Galactosidase: l chng nm mc Mortierella vinacea var: +Raffinoseutilizer, c th p ng c cc yu cu ny. Chng nm mc ny c nui cy trong nhng dng ht, thu nhn nhng dng ht ny, sy kh v s dng trc tip nh mt loi enzym c nh. Ch phm ny c khuy trn vi nc chit c ci ng trong mt Reactor c cnh khuy theo tng m. Khi qu trnh loi b Raffinase hon tt, ngng khuy v bm nc chit c ci ng ra khi cc h enzym lng y enzym b mt i s c b thm vo tip tc tin hnh phn ng vi m iu kin Baz trong giai on u ca qu trnh tinh sch nc c ci ng v khng gy ra bt k tr ngi no trong sut qu trnh Sucrose c ti to. Qu trnh ny lm tng thm 3% hiu sut v lm gim ng k vic gy nhim do thi b rc ng. Raffinase c nh c th cng c s dng loi b Raffinose v Stachyose t sa u nnh nh l mt loi sa thay th trong mt ch n king c bit.

2. ng dng ca enzym Invertase c nh Invertase l loi enzym c s dng trong qui m ln, di dng enzym c nh ===> Sn xut Sir vng bng enzym Invertase ca nm men. Cch tin hnh: - Cc t bo nm men s t hy v sn phm ca s th hy s c lm sch bng cch iu chnh pH v 4.7, sau lc qua lp Calcium Sulphate v hp th trong than xng. - Mt lp than xng c cha Invertase c thm vo trong than xng ang c s dng kh mu ca Sir. Ch phm ny rt n nh, cc yu t gii hn thng l s ngoi nhim vi sinh vt hoc kh mu khng tt, t khi l mt hot tnh enzym, sn phm khng c mi v tinh t nh qu trnh s dng nguyn liu c thy gii bng acid ( S dng enzym c nh). - Khi s dng enzym s trnh c nhng vn m phng php Acid thng gp phi l mu m, nng mui v tro cao, hiu sut chuyn i tng i thp. Nu Invertase t do c th c s dng (thi gian phn ng khong 1 ngy) th vic s dng enzym c nh (thi gian phn ng khong 15 ngy) lm cho qui trnh sn xut tr nn cnh tranh hn c th sn xut 16 tn Sir nghch o bng cch s dng 1 lt enzym ht.

III.NG DNG CA H PROTEASE

1.Trong sn xut Ph mai Tu loi phmai m lng enzyme rennin cho vo c khc nhau. V d nh trong qu trnh sn xut phmai Blanc th vi hot tnh ng t sa 1:10.000, lng ch phm rennin cn s dng trung bnh 5-10g/100kg sa, rennin c ho tan vi nc theo t l 1:10 ri cho vo bn ng t. Nhng vi quy trnh sn xut phmai Bleu des Causses th vi hot tnh ng t 1:10.000, hm lng rennin s dng l 25-35ml cho 100 lt sa nguyn liu. 2. Lm mm tht *Cc phng php lm mm tht bng enzyme: -Ngm tht vo ch phm enzyme. -Trn bt c enzyme vi tht. -Tim dung dch ch phm enzyme vo h tun hon ca ng vt trc khi git m. Phng php ny thc hin ln u vo nm 1960 v c pht trin nhiu vo nhng nm gn y. -Tim ch phm enzyme vo tht sau khi git m. 3. Cng nghip da Enzyme thng dng l cc protease kim ca vi khun, nm si. Lng enzyme cho vo khong 50-200g protease c hot tnh 100.000 tch mt tn lng. Thi gian ngm ko di khong 6-12 gi. Cch tin hnh: 500% nc c nhit 280C; 0,5% surfactant; 0,6% enzyme t Bacillus Subtilis, nm si Aspergillus Sojae, Pacreatic, c tng hot tnh l 4000; 1,5-2% soda; thi gian 12-18 gi; pH 9,3-9,6; nhit 27-280C.

4.SN XUT BNH M Ngi ta s dng enzyme protease nhm lm gim nht ca bt nho do gluten gy ra nh bt nho c iu kin thun li nht cho s pht trin ca nm men v qa trnh to ra CO2 va n nh va m bo cho vic to thnh cc phn ng melanoidin trn v bnh m. Nu s dng protease c hot tnh qa mnh th cu trc gluten s b ph hu v khi kh nng gi CO2 s b gim, cu trc bnh m d b thay i, bnh m n ra nhng b teo li khi nhit tr v bnh thng. Cc ch phm protease nm si c bn rng ri trn th gii gm c Amano A ca hng AMANO, Fungal protease ca hng Miles lab. 5. SN XUT CHT TY RA Cc cht carbonhydrate v lipide d dng ho tan vo mi trng kim ca bt git, cn protein th kh b loi hn cc cht khc. Do ngi ta s dng enzyme protease kim. Cc protease ng dng trong sn xut bt git phi p ng c nhng nhu cu sau: + Hiu qu ty sch cao. + C kh nng hot ng trong mi trng pH 9-11 v phi chu c nhit khong 950C. + C kh nng gi c hot tnh ca cc cht tham gia thnh phn bt git. Nhng yu cu trn ch c serine protease ca vi khun l p ng c. Lng ch phm protease a vo thnh phn ca bt git ch khong 0,5-1%.

NG DNG CA PROTEIN
I.ng dng k thut in di protein SDS-PAGE ci thin v nng cao phm cht cc ging la -in di protein SDS-PAGE (Sodium Dodecyl Sulfate PolyAcrylamide) l k thut ph bin trong lnh vc cng ngh sinh hc, c th ng dng trn thc vt, ng vt, vi sinh vt. --Trong chn to ging la, k thut ny gip pht hin nhanh nhng tnh cht ni bt nh mi thm, protein, amylose... cc nh khoa hc chn lc c nhng dng, ging c phm cht tt. Mt s ging la c sn c ci thin phm cht thnh cng v nhiu ging la trin vng ra i bng k thut ny. -K thut in di protein SDS-PAGE c tin s V Cng Thnh tip thu t Trng i hc Cng nng Tokyo (Nht Bn) v ng dng vo cng tc chn to cc ging cy trng t nm 1997 n nay.

II. Protein c th c dng lu tr d liu s

Gio s Tetsuro Majima thuc trng H Osaka (Nht Bn) cho bit Protein cn c th c s dng lu tr d liu my tnh in t. Khng nhng th kh nng lu tr d liu ca Protein cn vt qua mi cng ngh lu tr t tnh v quang hc hin ang c ng dng rng ri. Theo kt qu nghin cu ca gio s Majima, lu tr d liu bng Protein khng ch c n nh cao p ng tt yu cu ca cc sn phm ng dng thng mi m cn c th gip tng cng hiu sut vn hnh ca ng dng ln tng i cao. Ngoi ra thit b ng dng cng ngh lu tr bng Protein cn hon ton min dch trc nh hng ca t tnh - nguyn nhn thng xuyn gy nn nhng s c hng hc hoc mt d liu trn cc loi a cng hin nay. Minh chng cho kt qu nghin cu ca mnh, cc nh khoa hc Nht Bn s dng mt loi Protein hunh quang (fluorescent protein) c bit in d liu ln mt mt knh - y c gi l protein patterns. Qu trnh in d liu ko di trong khong mt pht. S dng kt hp nh sng v cc ho cht cc chuyn gia nghin cu c th d dng c li hoc xo b cc d liu tng t nh chc nng ca cc loi b nh hin nay. Tuy nhin, yu im ca cng ngh lu tr bng Protein nm ch cc thit b ng dng cng ngh ny khng th c lu tr trong iu kin mi trng bnh thng m phi lun c gi trong trng c nhit di 4 OC nhm bo m trnh s xm nhp ca cc loi vi khun ph v cu trc Protein.

III. S dng protein nh bt HIV khi ni n np

-

- Cc chuyn gia thuc H Thomas Jefferson va pht hin ra mt loi protein c tn interleukin-7 (IL-7), c kh nng nh bt mt s dng HIV ra khi ni n np. -TS Roger Pomerantz v ng nghip ly mu ca mt s bnh nhn HIV dng tnh ang ung hn hp cc loi thuc chng HIV c tn gi liu php chng retrovirus tch cc . Khng ai d c mc HIV trong c th h l bao nhiu. Nhm nghin cu kim tra tc ng ca cc loi thuc v protein IL khc nhau xem loi no kch thch virus HIV tt nht. Kt qu cho thy, IL-7 mang li hiu qu cao nht, c bit i vi cc dng HIV khng phn ng vi IL-2. -Pomerantz hy vng hai nm ti, nhm ca ng s c php s dng interleukin nh mt liu php trong vic iu tr cho bnh nhn HIV. -Mt nhm nghin cu Trung tm Y t VA San Francisco (M) c th ngn chn s pht trin v lan rng ca cc t bo ung th trn chut, bng cch s dng mt protein c trong c th ngi ( thay i cu trc) khin cho cc t bo ung th khng c kh nng bm dnh. -Protein ni trn c tn l galectin-3. Bnh thng, n ng vai tr quan trng trong vic gip cc t bo ung th bm dnh. Nh , cc t bo ung th c th tch ra t mt khi u, bm vo cc c quan khc v li pht trin thnh cc khi u c tnh trong c th. Tuy nhin, sau khi c thay i cu trc, cc t bo ung th ny khng cn c kh nng bm vo v pht trin cc c quan khc sau khi tch ra khi khi u gc v khng c s tr gip ca galectin-3. -Tuy nhin, mt s nh khoa hc cho rng, th nghim mi ch tin hnh trn chut, v cn phi rt lu na, liu php ny mi c th chng minh c tc dng ca n i vi c th ngi.

IV. S dng protein chng di cn ung th

V. S dng protein pht quang xanh lc lm du trong vic gim st vi khun hu ch Bacillus S11 tm s, Penaeus monodon.
Bacillus S11 (BS11) trong c th sng, cc t bo gc cha thun (wildtype) c cy vo protein pht quang xanh lc (GFP) biu hin trong huyt tng k hiu l pAD44-12, protein ny mang mt an gen vi tn gfpmut3a cha an gen hat ha UW85 cu thnh vi khun Bacillus cereus v huyt tng bnh thng khng c cy protein pht quang ca loi vi khun ny. Cc t bo c bin i pAD44-12 (BS11-GFP) v khng bin i (i chng, BS11-pAD) biu hin kh nng pht quang nhng vi mc khng cao.
-T thi xa xa, dng cht Placenta c gim st vi khun hu ch

VI. Protein Placenta: Dng cht k diu cho ln da

-S dng nh thnh phn chnh trong cc loi tho dc v thuc b qu dng ln cho vua cha. Hoc Ty u, ngi ta cng tin rng cc m nhn Cleopatra v Marie Antoinette cng s dng dng cht ny nh mt loi dng da t nhin. -Ngy nay, chng ta cng nghe n nhiu ng dng ca Protein Placenta trong ngnh y hc trong vic cha tr bnh vim gan, x gan, ri lon kinh nguyt, triu chng d ng hoc bnh da liu nh mi , chng da b sng ha, chng vim da, mn v tn nhang

phm Ashiyaroyal. Placenta c kh nng ngn chn s hnh thnh hc sc t trn da (melanine) bng cch ngn chn hot ng ca tyrosine enzyme sn xut melanine. Bn cnh , n c kh nng ngn chn qu trnh oxy ha 1 qu trnh ph v cu trc da khin da lo ha sm. Placenta gip kch thch trao i cht, tng vng lun chuyn ca da, loi b nhng t bo c v chun b cho lp sng chuyn ha thnh da. Vi lng axit amino t nhin khng l, Placenta c th cung cp mt lng m ti a ngay c ln da mt nc trm trng nht

-Cng dng ca Protein Placenta c pht huy ti a trong dng sn

VII.Dng protein t rau bina gip ko di tui th ca pin

-Cc nh nghin cu M tm c cch khai thc nng lng dng trong tin trnh quang hp bin nh sng thnh nng lng. H s dng tin trnh ny gia tng tui th ca pin trong in thoi di ng, my tnh xch tay v cc thit b in t khc. -Trong khi nhng n lc trc y to ra nhng dng in ch ko di khang vi gi, th nhm nh khoa hc M to c mt dng in ko di ba tun. Shuguang Zhang, ph gim c Trung tm Cng trnh Y sinh hc ti Vin K thut Massachusetts c th to ra mt lai cht ty peptide t axit amin l th lm n nh protein v cho php n truyn nng lng. -Cc nh nghin cu chn rau bina v rau ny cha nhiu dip lc t, th lm cho n c mu xanh m, v gi li r.