TRUONG DAL HQC Y HA NOL HOA SINH (SACH DAO TAO BAC Si Y KHOA) Chi bién: GS. BS. TA THANH VAN PGS. BS. BANG THI NGQC DUNG (In lan thir ba cé sita chita va b6 sung) NHA XUAT BAN Y HOC HA NOI - 2022GS. BS. TA THANH VAN PGS. BS. DANG THI NGOC DUNG THAM GIA BIEN SOAN GS. BS. TA THANH VAN PGS. BS. TRAN HUY THINH PGS, BS. PHAM THIEN NGOC PGS. BS. BANG THI NGOC DUNG TS. BS. TRAN KHANH CHI TS. BS. NGUYEN THI THANH HAT TS. BS. NGUYEN THI NGQC LAN BSNT. NGO THI THU HIEN THU KY BIEN SOAN ‘TS. BS. TRAN KHANH CHI Hoa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungLOI MO BAU Hoa sinh 1a mt nginh khoa hoc nghién ctru vé cae qua trinh phan ing héa hoe dign ra trong co thé s6ng ¢ diéu kign binh thong cing nhw bénh ly trong méi tuong tac qua lai véi méi trudng xung quanh, Day a linh vuc khoa hoe giao thoa véi nhiéu nginh Khoa hoe khée nur Y hoc lim sang, Sinh ly hoe, Héa hoc, M6 phdi hoc, Dinh duéng.. Nhting hiéu biét méi nhat trong linh vye Héa sinh da gép phn quan trong trong viée cham séc sire khoe cua con ngudi dic biét trong linh ve Y hoc kim sing, Dinh dung va Duge hoc. Cuén gido trinh Héa sinh duge bién soan lan thir 7 vao nam 2007 v tai bn vai mOt s6 chinh sita, bo sung nhiing kién thire khoa hoc cap nhat. Sach duge bién soan dua trén nhimg kinh nghiém ging day bac dai hoc, sau dai hge va nghién ctu khoa hye cing nhu cae kinh nghiém trong linh vue héa sinh lim sing cia nhieu thé hé cae thay c6 b} mén Héa sinh, Trudng Dai hoc Y Ha N6i, Cudn sich bao gém 2 phan chinh: é, Cdu tao va chuyén hoa chdt; ii. Héa sinh té bao, mé va co quan: Phan Cau tao va chuyén héa chat trinh bay vé céu tao, tinh chit va vai tro cla cic chat sinh hoc co ban trong co thé séng, cae qué trinh chuyén héa va diéu hoa chuyén hda chat trong co thé séng. Nhiing kién thire khoa hoc nay li nén tang giup ngudi dc 6 thé hiéu duge cae phan ké tiép theo trong cudn sich nay ciing nhur céc chuyén nganh y hoe chite nang. Phin Héa sinh té bio mé va co quan trinh bay qué trinh chuyén_héa chat xay ra trong té bio song, 6 cdc m6 va co quan cita co thé trong diéu kign binh thudng va cing nhur digu kign bénh ly dé tir d6 dura ra cdc img dung trong Y hoc lam sing. Trong timg churong, céc tic gia da cd ging ling ghép céc théng tin khoa hoc cap nhat vé Héa sinh Kim sing, Y sinh hc phan tir va té bio nhim ting tinh hip din va logie cdc ndi dung khoa hge ciia cudn séch Sach Hoa sinh nay di duge Truéng Bai hoe Y Ha NOi thm dinh va ban hinh lim gido trinh day va hoe chinh thie ctia Nha truéng. Ching tdi hy vong rang cudn gido trinh nay sé dap img duge nhiing kién thie co ban va cp nhat trong Tinh vue Hoa cia sinh vién cdc Truéng Dai hoc Y, cing nhu nhu cau tham khio cia cde ban doc Tinh vue khoa hge nay. Thay mat cde téc gid, t6i xin chan thinh cém on céc dng nghiép 43 dinh thoi gian bign soan, doc ban thio va g6p ¥ chi tiét vé ni dung cling nhur cach trinh bay cho Jan tai ban lan thir nhat nay. Loi cam on cing danh cho cae can bd cua BO mon Hoa sinh Trudmg Dai hoc Y Ha Noi, Nha xudt bin Y hoc da gitip a0 cdc tac gia trong qua trinh hoan thién cuén sach nay. Thay mat cdc tac gia GS. BS. TA THANH VAN Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungMUC LUC Loi mé dau Ta Thanh Van Myc lye PHAN I: CAU TAO VA CHUYEN HOA CHAT Chuong 1. Enzy Pham Thién Ngoc 1. Cfch ggi tén va phan loai enzym 2. Mot sé die tinh eita phan tir enzym 3. Cu trac va chite ning ctia ede coenzym 4. Co ché xtc tic cia enzym 5. Déng hge enzym 6. Cac yéu t6 anh huéng dén hoat dng cia enzym Chuong 2. Ning long sinh hgc Pham Thign Ngge 1. Ban chat su h6 hap té bao 2. Su phosphoryl-oxy héa 3. Chu trinh acid citric Chuwong 3. Héa hoc carbohydrat Tran Khanh Chi, 1. Dai cuong Trin Huy Thinh 2. Monosaceharid 3. Oligosaecharid 4. Polysaccharid 5. Glucid lign hyp Chuong 4. Chuyén héa carbohydat Tran Khanh Chi, 1. Tiéu héa va hap thu carbohydrat D§ng Thi Ngoc Dung 2. Su thodi héa ciia glucose 6 té bao va m6 . Chuyén héa cia cic monosaccharid khac 3. 4, Con dung tin tao glucose 5 . Chuyén héa earbohydrat 6, Digu hoa chuyén héa carbohydrat 7. Mot so bénh ly rdi loan chuyén héa_ carbohydrat Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungChung 5. Héa hge lipid Ng6 Thi Thu Hién, 2. ‘Thanh pl tao ctia lipid Dang Thi Ngoc Dung Phan loai lipid Chuong 6. Chuyén héa lipid va van Ding Thj Ngge Dung cl 1 2. 3. huyén lipid trong mau . Thoai héa lipid 6 té bao . Tong hgp lipid 6 té bao . Chuyén héa cholesterol 4. Van chuyén lipid trong mau Churong 7. Héa hge acid amin, protein va Nguyén Thi Ngoc Lan, hemoglobin Tran Huy Thinh 1 2. 3 4. . Acid amin Peptid . Protein Hemoglobin va myoglobin Chuong 8. Chuyén héa acid amin Ng6 Thi Thu Hién, 1 2. 3 4, a 5. ‘Thuy phn protein thanh acid amin Ta Thanh Van Sw thodi héa acid amin Tang hop acid amin . Tong hop mot s6 chit c6 hoat tinh sinh hoe tir cid amin Bénh ly acid amin Chuong 9. Chuyén héa hemoglobin Neguyén Thi Ngoc Lan, 1 Sy thoi héa hemoglobin va y nghia lim Tran Huy Thinh sing 2 Ting hop hemoglobin Chuong 10. Sinh ting hop protein Tran Huy Thinh 1 2 3. 4, . Sinh tng hop protein 6 té bao nhan so Sinh téng hop protein & té bao nhan that Sw hoan thién protein sau tong hop Dicu hda sinh tong hgp protein Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungChuong 11. Héa hgc acid nucleic Ta Thanh Van ‘Thanh phin héa hoc ciia acid nucleic Deoxyribonucleic acid (DNA) 3. Ribonucleic acid (RNA) Chuong 12. Chuyén héa acid nucleic Ta Thanh Van 1. Chuyén héa nucleotid 2. Chuyén héa acid nucleic Chwong 13. Héa sinh hormon Pham Thign Ngoc 1. Phan loai hormon . Co ché tac dung cua hormon Tac dung sinh ly cua hormon . Hormon la din xuat acid amin 2. 3 4. Nhimg hormon protein, polypeptid 5. 6. Cac hormon steroid PHAN II: HOA SINH TE BAO, MO VA CO QUAN Chuong 14, Héa sinh mang té bio Nguyén Thi Thanh Hai ‘Thanh phan cau tric mang té bao Dang Thi Ngoc Dung . Van chuyén chat qua mang 3. Bénh hoc mang té bao Chuong 15. Trao di mudi nuse Tran Khanh Chi, 1. Nude trong co thé Ta Thanh Van 2. Cac chat vé co 3. Trao déi mudi nude 4, Diéu hda trao dbi mudi née 5. R6i loan nude - dign giai Chuong 16. Khi mau va thing bing aci Tran Huy Thinh base Su van chuyén khi Su thing bing acid - base Rdi loan thang bang acid - base Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungChuong 17. Héa sinh gan Pham Thign Ngoc 1. So luge giai phiu gan 2. Thanh phan héa hoc eda nhu mé gan 3. Chite ning chuyén hoa glueid, lipid, protein cia gan 4, Chite nang tao mit 5. Chite nang khir déc 6. Mot sé xét nghiém héa sinh vé gan Chuong 18, Héa sinh than va nude tiéu Nguyén Thi Ngoc Lan, 1. Than Trin Huy Thinh 2. Nude tiéu va céc tinh chat cia nude tiéu Chuong 19, Héa sinh miu Ta Thinh Van 1. Cac chite ning sinh ly eita mau 2. Tinh chat ly héa ctia méu 3. Thanh phan cia mau Chuong 20, Héa sinh eo Dang Thj Ngoc Dung 1. Dic didm thanh phan héa hoc va chuyén Nguy€n Thj Thanh Hai héa trong co 2. Cu trite cla co van 3. Sweo co van 4, Su-co co trom 5. Su chuyén déng ctia té bio khic 6. Mat sé bénh ly chuyén hia cita co Chwong 21. Héa sinh than kinh Ngo Thi Thu Hién, 1. Cfu tao héa hoe ctia té chite thin kinh Ta Thanh Van 2. Chuyén héa chat trong mé than kinh 3. Su dai & dn kinh 4, Cae chdt din truyén thin kinh Chuong 22. Hoa sinh dich sinh vat Trin Khinh Chi, 1, Dich nao tay Pham Thién Ngoe 2. Dich vj 3. Bach huyét Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungPhanI CAU TAO VA CHUYEN HOA CHAT Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Bang Thi Ngoc Dung 9MUC TIRU HQC TAP I. Trinh bay duge ctich gpi tén va phan logi theo quoc té ctia enzym, cho duge vi du méi logi. 2. Trinh bay dieoc céu tao, edu trite ctta phan tit enz 3. Ké tén va vai tr6 ctic loai coenzym, co ché hoat ding ctia coenzym NAD* vit FAD. 4. Trinh bay diegc co ché hoat déng ctta enzym, § nghia hing 86 Km, phuong trinh Lineweaver-burk. 5. Phan tich duge ctic yéu t6 anh hong dén hoat dng ctia enzym. Sy séng Ia qué trinh dng, 14 qua trinh bién ddi chuyén hoa khong aging ciia chat dién ra trong cic 16 bao, mo co thé sng. Thue chat etia cia céc qua trinh bién a6, chuyén héa la cdc phan tmg héa hoe lign tiép, phite tap nhung dién ra 6 moi trudng rit dic bigt: 2/3 1a nude, nhiét 46 37°C, pH trung tinh, 4p suat 6n hoa. Ngoai ra, cae phan img héa hoc, bién déi chuyén héa cc chat dién ra trong té bdo phai dién ra nhanh chong, manh mé va dac biét durge kiém soat, diéu hoa chat ché pha hop véi nhu cau co thé. Dé lam duoc didu nay, té bao séng san sinh ra chat xtc téc sinh hoc dic bist duge goi [A enzym. Enzym ciing ¢6 day du dic tinh eta chat xtc tée thong thudng: - Cae enzym khdng bj tiéu hao hoje duge sinh ra thém trong qué trinh phan tng, - Cfc enzym chi kim ting téc d6 phn img ma khdng tao ra phan img, khGng lim thay d6i hang s6 can bang cia phan img, khéng lim thay d0i chiéu cia phan img. Tuy nhién, ngoai céc tinh chét néu trén, enzym cén c6 nhiing tinh chit khdc voi tinh chat ctia cae chat xtc téc héa hoc théng thudng: - Hau hét enzym cé ban chat la protein (mt s6 it enzym 6 ban chat la RNA), - Enzym cé higu lye xe tac 1én va thong lén hon nhiéu so véi cae chat xuc tae v6 co hoje tng hgp. Théng thuéng ting téc 4 phan img enzym dén 10% 10! lan, vi dy enzym catalase (c6 & cde mé khac nhau, dic biét c6 nhiéu & gan) cé6 thé xiie tée phan huy 40.000.000 phan tir H202 thanh H2O va O2 trong 1 gidy - Enzym cé tinh dic higu co chat va phan img, mdi enzym chi xiic tée phan img cho mt hode mét nhom co chat hoac loai phan img nhat dinh, - Hau hét cc enzym hoat déng & ving nhiét 46 6n hoa va pH trung tinh, - Dic biét phan ing ma enzym xite tac duge té bao kiém soat, diéu hoa chat ché thong qua kiém soat hoat tinh enzym. 10 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungBang 1.1. Tém tat cdc dac diém ctia chat xtc tac v6 co va enzym Dac aiém Chit xtic tac v6 co Chat xtic tac hiru co (Enzym) Nguén géc u Do té bao san xuat Ban chat héa hoc at vo Chat hivu co (Protein co thé RNA) Tang téc dé phan wng | 10? - 10° lan 10° - 10" lan Diéu kién phan ting: Méi trun ngoai co thé: Mi truéng trong co thé: Nhigt d6 100°C, hoac cao hon Thap (36-45°C) pH Acid/kiém manh pH sinh ly (7,4) Ap suat Cao (vai atm) Ap suat khi quyén (1 atm) Thay di céu trac Khong C6 thay déi, tre lai cdu trac ban dau khi két the phan Ung Tinh dae higu Thap Cao C6 thé n6i enzym cé vai trd rat quan trong trong moi qua trinh héa sinh hoc dién ra trong co thé nhw xite tac chudi phan ing héa hoc ké tiép nhau trong thodi hoa cic phan tir dinh dudng, tich lay va chuyén dang nang lung héa hoe va xite tac tao nén cde dai phin tir phite tap tir cdc tién cht don gian. Nghién citu enzym ngoai nhiing kién thite khoa hc co ban cén e6 gid tr thye tign lam sang. Viée do hoat d§ mét sé enzym trong huyét tong, héng céu, miu mé gitip cho chan doan va diéu tri mOt sé bénh. O mét s6 bénb, die biét rdi loan di truyén do sir thiéu hyt, thm chi khéng cé mgt hose mot sO enzym. Mot sé bénh khac lai cé sy ting cudng hoat dong ciia mt enzym nao d6, Nhiu loai thuéc hoat dong thong qua tuong tie voi enzym, Nhiéu enzym ciing la cfc céng cy thye hanh trong céng nghé héa sinh y hoc, céng nghé thye pham va néng nghiép. 1. CACH GOI TEN VA PHAN LOAI ENZYM. 1.1, Cach ggi tén enzym: enzym cé 4 cach goi tén: 1.1.1. Tén co chét va thém tiép vi ngi ase. Vi du: co chit 1a ure tén enzym li urease, tén co chat li protein tén enzym la proteinase,... 1.1.2. Tén téc dung va thém tiép vi ngiF ase. Vi du: tac dung oxy héa, enzym ki oxidase, tic dung trao déi amin enzym li amino transferase, tic dung khit nhém COs, enzym la decarboxylase, 1.1.3. Tén co chat, tac dung va thém tiép vingé& ase. Vi dy: co chat la lactat va téc dung li khit hydro thi t8n enzym la lactat dehydrogenase, co chat la tyrosin va tie dung la khir nhom COs thi tén enzym la tyrosin decarboxylase... i: cach goi tén nay khong c6 tiép vi ngit ase. Vi dy: pepsin, Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung1.2. Phan loai enzym Khi s6 trong enzym duge tach chiét, tinh sach va xdc dinh de tinh Jen dén con s6 hang nghin, viée goi tén va phan loai trérnén phitc tap. Dé chuan héa cach goi tén va phan loai enzym, higp hi enzym quéc té (Enzym Commission: EC) 43 dua ra ech phan loai enzym nim 1961 (chufin hoa lai nm 1972 va 1978) dua theo phi mg ma ching xite tac Theo cach phan loai nay, enzym duge xép thinh loai (class), theo thir ty tir | dén 6, mdi loai lai durge chia thanh nhém (subclass) dura trén co chat xéic téc, mdi nhém lai durge chia thanh céc phan nhém (sub-subelass) dua trén coenzym tham gia, mdi phan nhom gom mét s6 enzym. Nhu vay, mdi enzym déu duoc ky higu bang mét ma sé EC chita 4 chit $6, cach nhau bai cac dau cham thap phan, Chir sé thir nhat chi loai enzym, chit s6 thit hai chi nhém, chit s6 thir ba chi phan nhém va chit s6 thir tur chi tén cua ban than timg enzym rigng biét trong nhém. Vi dy: enzym hexokinase c6 ky higu la EC 2.7.1.1 1a enzym thuge loai 2, nhém 7, thude phan nhém | va e6 s6 thir ty clia enzym trong phan nhém Li 1. Séu loai enzym duge sip xép theo thir ty sau: 1.2.1. Enzym oxy héa khié (oxidoreductase): la loai enzym xic téc cho phan ting oxy héa va phan img khir, nghia la cc phan ting ¢6 su trao doi H hode dién tir theo phan imng tng quat sau Al, +B A+ BH: Loai enzym oxy héa khir g6m cde duéi lap: - Cae dehydrogenase: sir dung eae phan tir khong phai oxy (vi dy: NAD") lim nh§n dign tit. Vi du: lactat dehydrogenase, malat dehydrogenase, ~ Cae oxidase: sit dung oxy nhur mot chat nbn dign tir nhung khéng tham gia vao thanh phin co chit. Vi du: cytochrom oxidase, xanthin oxidase, - Cac reductase: dura H va dién tir vao co chit. Vi du: B-cetoacyl-ACP reductase. - Catalase: xtic tae phin img: HO: + H202— O2 + 2H20 - Cée peroxidase: xitc tac phan tng: H202 + AH: > A + 2H;0 - Cac oxygenase (hydroxylase): gin mOt nguyén tir O vao co chat. Vi du: cytochrom P-450 xixe tae phin tmg: RH + NADPH + H’ + O2 > ROH + NADP* + HAO, phenylalanin hydroxylase, .. 1.2.2. Enzym van chuyén nhém (transferase): la loai enzym xtc tac cho phan ing van chuyén mt nhém héa hoc (khéng phai hydro) gitta hai co chat theo phan tng téng quat sau: AX+B—A+BX Loai enzym van chuyén nhém gém cée duéi lop: - Cae aminotransferase: chuyén nhém -NH2 tit acid amin vao acid cetonic. Vi du: aspartat transaminase, alanin transferase,... - Transcetolase va transaldolase: chuyén don vi 2C va 3C vao co chat. Vi du: transcetolase, transaldolase,.. 12 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung- Cac acyl-, metyl-, glucosyl-transferase, phosphorylase: chuyén cée nhém trong ting vio co chat. Vi dy: acyl Cod-cholesterol acyl transferase (ACAT), glycogen phosphorylase, .. - Cae kinase: chuyén gdc -POs’ tir ATP vao co chit. Vi du: hexokinase, nucleoside diphospho kinase, PEP carboxykinase, .. - Cée thiolase: chuyén nhém CoA-SH vao co chit. Vi du: acyl-CoA acetyltransferase (thiolase), ... - Cée polymerase: chuyén cde nucleotid tir cée nucleotide triphosphat (NTP) vao phan tir DNA hoc RNA. Vi du: cde DNA polymerase, cdc RNA polymerase. 1.2.3. Enzym thiy phan (hydrolase): li loai enzym xite tée cho phan ting ign két cia chat héa hoc bing cach thiiy phan, nghia la phan img c6 sy tham gia cla phan tir nudc, theo phan ig tng quat sau: AB +H20 > AH + BOH Loai enzym thiy phan gdm cae dui lop: - Cae esterase: thiiy phan lién két este. Vi dy: triacylglycerol lipase. - Cac glucosidase: thiy phan lién két glycosid. - Cée protease: thity phan lién két peptid trong phan tir protein. - Cac phosphatase: thity phan lién két este phosphat, tach géc POs kh6i co chit. - Cée phospholipase: thay phan lién két este phosphat trong phan tir phospholipid. - Cac amidase: thiy phan lién két N-osid. Vi dy: nucleosidase - Cae desaminase: thiy phan lign két C-N, téch nhém amin ra khéi co chit. Vi du: adenosin desaminase, guanin desaminase, - Ce nuclease: thiiy phan ef lién két este phosphat trong phan tr DNA hoae RNA. 1. Enzym phan cat (lyase): enzym tach nhom, la loi enzym xtic tac cho phan tmg chuyén di m6t nhom héa hgc khoi m6t co chat ma khéng c6 sy tham gia cia phan tir nude. Phan ting tong quit nhu sau ABSA+B Loai enzym tach nhém gém cac dudi l6p: - Cae decarboxylase: tach phan tir CO: tir eo chat, Vi du: pyruvat decarboxylase, glutamate decarboxylase, - Cac aldolase: téch mt phan tr aldehyd tir co chit. Vi du: aldolase xtc téc phan img tach fructose 1,6-diphosphat thinh glyccraldchyd phosphat (GAP) va Dihydroxy acetone phosphat (DHAP) - Cée lyase: téch doi mgt phan tir ma khOng ¢6 sy tham gia cla phan tir H2O. Vi dy: arginosuccinase. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung 13- Cfe hydratase: gin m6t phain tir H2O vo mot phan tit co chat. Vi dy: fumarase, ~ Cfc dehydratase: téich mot phin tir HzO khoi mét phan tir co chat. Vi du: B-hydroxyacyl-ACP dehydratase, B-hydroxyacyl-Cod dehydratase, . - C4e synthase: gan hai phan tir ma khng can su tham gia cua ATP dé cung cp nang lung. Vi dy: ATP synthase, citrat synthase, glycogen synthase, acid béo synthase, B-levulenat synthase, ... 1.2.5. Enzym déng phan (isomerase): li loai enzym xiic tac cho phan ing bién d6i giita cdc dang déng phan ciia chat héa hoc. Phan ting tong quat nhur sau: ABC > ACB Loai enzym ding phan gdm céc duéi lép: - Cc racemase: chuyén dang déng phan gitra day D va day L. - Cae epimerase: chuyén dang déng phan epi. Vi dy: ribose 5-phosphat epimerase. - Cac isomerase: chuyén dang gitta nhém ceton va nhém aldehyd. Vi du: phosphopentose isomerase. - Cie mutase: chuyén nhém héa hoe gitta cic nguyén tir trong mot phan tit 1. Enzym téng hop (ligase hoac synthetase): 1 loai enzym xiic tac cho phan (mg gan hai phan tir voi nhau thinh mét phan tir lin hon, sir dung ATP hoe cée nucleosidtriphosphat khac dé cung cap nang lugng; phan img tong quat nhur sau: ATP ADP +Pi A+B NZ AB Loai enzym ting hop gém céc due - Cée synthetase: gin hai phan tir véi sy tham gia cua ATP dé cung cap nang hrgng, - Cie carboxylase: gin CO vio phan tir co chat. Vi du: pyruvat carboxylase, ~ Ligase: str dung cho vige gin 2 doan nucleotid véi nhau. Vi du: DNA ligase. 2. MOT SO BAC TINH CUA PHAN TU ENZYM. 2.1. Cu tao va cau tric cla enzym Cac enzym la céc protein 6 khéi hrong phan tir tir 12.000 dén hang trigu don vi dalton (Da). Cang nhw cae protein, vé thinh phan cu tgo, enzym cing duge chia Lim hai logi: enzym thuan va enzym tap. Enzym thuan la cae enzym ma phan tit chi do cic Nhiing enzym nay khéng doi hoi céc nhom ngoai (coenzym) cho hoat dng cia ching la enzym mot thanh phan). Vi dy cae enzym thi phn: amylase, proteinas 14 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungEnzym tap li cée enzym ma ngoai thinh phan protein (acid amin), phan tir enzym cdn c6 chat cdng tae (cofactor) la cae ion nhu Fe®, Mg”, Mn®*, Zn’, ... hogic 1a mot phan tr chat hitu co hoie phite hop hiu co kim loai, céu tgo nén, Néu chit eGng tic (cofactor) la chat hitu co hoie la phite hyp hiu co kim logi thi duge ggi li coenzym. Mét s6 phan tit enzym doi hoi cd coenzym va ion kim logi cho hogt dong cua ching, Trong phan tir enzym tap (con goi 1a holoenzym), phan protein duge goi 1a apoenzym, phan chat cong tac duge goi la cofactor: Holoenzym = apoenzym + cofactor Phén apoenzym mang nhimg dic tinh co ban cia enzym, trong khi phin coenzym hod ion kim logi 14 chit phéi hop ciia enzym, c6 vai tro bo sung kha nang phan img va kha nang xitc téc cho phan tir enzym, Coenzym thuéng cé trong thnh phan céc enzym thuge loai oxy héa khtt va logi enzym van chuyén nhom; thigu coenzym, enzym loai nay khéng hoat dong. Céc cofactor (coenzym) thuéng li cac vitamin va din xudt cia chung. Mét s6 cofactor gin chat vao phan tir enzym, kh6ng thé tach ra duge goi la nhdm phu (prosthetic group). Nhiéu coenzym trong thinh phan cau tao ¢6 chita vitamin. Nhiing enzym chita kim logi hog d0i hdi kim logi cho hoat dng eita né duge goi 1a enzym kim loai (metalloenzym). Vai tro ciia kim loai trong thanh phan cia enzym kim logi la: - Tham gia tryc tiép vao phan tg xtc tac ctia enzym, - Hoat dong nhu mét chit oxy héa khir, - Tao thinh phire hop véi co chat Bang 1.2. Cac ion co chtre nang la chat cong tac cla enzym ton Enzym Cu Cytochrome oxidase Fe** hoac Fe* | Cytochrome oxidase, catalase, peroxidase K Pyruvate kinase Mg Hexokinase, glucose 6-phosphatase, pyruvate kinase Mn?" Aginase, ribonucleotide reductase Mo Dinitrogenase Nit Ureaso Zn?" Carbonic anhydrase, alcohol dehydrogenase, carboxypeptidases A va B 2.2. Trung tam hoat déng ciia enzym va tinh dic higu co chat Trung tim hoat dong hoie vi tri hoat dong (active site) ca enzym Ki mot ving dic bigt ciia enzym e6 téc dung gin véi co chat dé xc tée cho phan tmg lam bién déi co Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung 15chit thinh sin phim, M8i enzym c6 thé c6 mét_hofe vai trung tim hoat dng. Trung tm hoat déng cia enzym gdm nhing nhém héa hoc va nhiing lién két tigp xite tryc tiép véi co chat hoc khong tigp xtc truc tiép véi co chat nhung c6 chite ning tryc tiép trong qua trinh xtc tac. Vé thanh phan céu tao, trung tam hoat dng thudng bao gbm céc acid amin chita cdc nhém héa hoc 6 hoat tinh cao nhu serin (cé nhém -OH), cystein (cd nhém -SH), glutamic (¢6 nhom 7-COO), lysin (co nhom ¢-NHs*), histidin (c6 nhém imidazol’), tryptophan (c6 nhém indol’),... 14 nhing nhém phan cye hoac ion héa, c6 kha ning tao ign két hydro hoc ion véi co chit. Hinh 1.1 minh hoa sw gan ciia glucose vio trung tm hoat déng cia glucokinase. Chinh nhé trung tim hoat dng ma enzym thé hién tinh dic higu co chit: glucokinase khéng xtc tic (gin) véi galactose. A Asp 205 INN cy 229 Hinh 4.1. Vi tri gn glucose ctia glucokinase. A. Glucose dugc gn vao trung tam hoat déng béi cac lién két hydro gitta céc nhém OH cla cac acid amin phan cyc & cac vj tri khdc nhau. Sy twong tac phic tap lam glucose thay déi cau hhinh gn dung vao trung tam hoat dong. B. Tinh dac higu enzym duoc minh hoa béi c4u hinh glucose va galactose. Galactose khac glucose chi mét vj tri nhém OH vi vay khéng gan duge vao trung tam hoat dong va can mot enzym khdc chuyén héa galactose la galactokinase Vé quan hé gitta trung tm hoat déng va co chat, c6 hai gid thuyét duge dua raz - Thuyét “6 khéa va chia kha”: Fischer. E (1894) di dua ra thuyét “6 khéa va chia khéa” (“lock and key") vé tac dong ciia enzym. Theo thuyét nay, tong tac gitta enzym (B) va co chat ($), nghia la sy gin gitta enzym va co chat dé tao thinh phite hgp enzym - co chat (ES) cing gidng nhu quan hé gitta “6 khda” va “chia khoa”, nghia li enzym nao thi chi xtic tae cho dang co chat dé, Thuyét nay chi gidi thich duge tinh dic higu tuyét dbi ciia enzym nhung khéng giai thich duge tinh dc higu tuong déi cua enzym. ~ Thuyét “nd hinh cam img khong gian™: dé giai thich tinh dic higu tong déi cita enzym, Koshland D (1958) 4a dura ra thuyét “md hink cam img khong gian” (“induced 16 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dungfit"). Theo thuyét nay, trung tém hoat dng eiia enzym e ¢6 tinh mém déo va linh host, 6 thé bién d6i vé cdu hinh khdng gian trong qua trinh twong tac véi co chit sao cho phi hgp v6i cau hinh khdng gian cua co chat, dé e6 thé tao thanh phire hp enzym - co chat (ES). Thuyét niy giai thich duge tinh die higu trong di ciia enzym va da sd hign nay chap nhin thuyét nay. Co chat == ~ Gs ) )) Vittri hoat déng Chia khéa (co chat) (enzym) Phitc hop 6 khéa - chia khéa ritcingp eraynvcerchtt: “S hinh 6 khéa - chia khéa — . wy \ od X Phitc hp ES Eneym b. Mé hinh cam tg khéng gian Hinh 1.2, Mé hinh “6 khéa’ va “chia khéa” (a) cla Emil Fischer va md hinh “cam tang khong gian” ctia Koshland (b) 2.3. Cc dang cAu tric cla phan ti enzym 2.3.1. Enzym don chuéi va enzym da chudi Enzym cé thé do m6t chudi hojc nhigu chudi polypeptide tgo nén. Enzym don chudi (monomer) la enzym chi do m6t chudi polypepid cau tao nén, vi du: ribonuclease a, lysozym, lipase, pepsin, chymotrypsin, .. Enzym da chuéi (oligomer ho§e polymer) la enzym do hai hofe nhiéu chudi polypeptid céu tao nén, vi du: aspartat transaminase (AST): 2 chudi, alkalin phosphatase (ALP): 2 chudi, creatin kinase (CK): 2 chudi, hexokinase (HK): 2 chudi, lactat Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dungdehydrogenase (LDH): 4 chudi, RNA polymerase: 5 chudi, ATP synthetase: 12 chudi, glutamat dehydrogenase (GLDH): 40 chudi. 2.3.2. Enzym dj lip thé (Allosteric enzym) va diéu héa hoat tinh enzym Enzym dj lap thé la logi enzym ngoai trung tim hoat déng con m9t hoge vai vj tri dj lap thé; trung tim hoat dong déng vai tro tigp nhin eo chat dé xiie tic cho phan img enzym trong khi vi tri dij lip thé tip nbiin yéu t6 di lap thé dé digu chinh hoat dong xte tac clia enzym, Vé cdu tao phan ti, enzym di lip thé ¢6 thé 14 enzym don chudi hode enzym da chudi, Phan tir enzym dj lap thé c6 thé cé loai vi tri dj lap thé duong, loai vi tri dj lap thé am hode c6 ea hai Khi vi tri di lap thé duong tigp nhan yéu t6 di lap thé duong A (chat hoat hoa: activator) thi cu hinh enzym thay 46i theo huréng thuan I9i hon cho vige gin co chat, 4i le enzym véi co chit ting Ién, sy tao thinh phife hop enzym - co chat tot hon, tée d6 phan img ting lén. Co chat = Trung tam hogt dng Trung tam hogt dng wv ~~ Chat ire ch. dj Ip thé Vj tri dj lap thé (a) We ché Allosteric Be chit Trung tim hogt dong X ‘Trung tam hoat dong a ‘Vi trl dj gp thé Chat if héa dj lap thé (b) Hoat héa Allosteric Hinh 1.3. Tac dung ctia yéu té di lap thé am va duong lén hoat déng cia enzym (a) VOi enzym dj lap thé khi cd sy gan chat dj lap thé am, cdu hinh enzym va trung tam hoat dong thay déi khéng thuan loi cho sy gan co’ chat dé xuic tac va lam gidm téc do phan ting (b) Tac dung cla yéu t6 dj lap thé duong lam thuan Igi cho sy gén co chat va tang téc do phan tng, 18 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungKhi vi tri dj lap thé am tiép nhjin yéu t6 dij lap thé am I (chit ite ché: inhibitor) thi ciu hinh enzym thay déi theo huéng kh6ng thuin Igi cho vige gin co chit, enzym bi te ché, ai lye enzym véi co chat giim nén téc d6 phan ing giam di. Thong thudng, nhiing chat hoat héa di lap thé 1a nhimg chat dig trude co chat trong chudi phan img, trong khi nhimng chat tie ché di lap thé li nhimg chat dimg sau chudi phan tmg hoge 1a sin phém cudi cing cia chudi phan img. Vi du: trong con duing dung phan, enzym phospho fructokinase la mot enzym di lip thé, duge hoat hoa béi yeu t6 di lip thé duong la ADP va AMP, nhung bi tre ché boi yeu t6 di lap thé am la ATP va citrat. Hinh 4.4. Trong chudi phan ting tir chét A dén F. 84 didu chinh téc 46 ctia oa qua trinh rt nhigu phan ting khi sn phdm F da dap Ging dui nhu cu té bao; thay vi diéu chinh kiém soat cc sn pham trung gian, san phém cudi cling F tre ché ngay enzym & phan ving dau tién Ga qua trinh. Cac enzym allosteric c6 dic tinh khic voi cae cae enzym khéng ¢6 vai tr diéu hoa khac. Ngoai trung tim hogt déng, enzym allosteric cé mét hoke nhiéu vj tri dj lap thé (allosteric) dé gin chat dij lap the (chat diéu héa) Hinh 1.3. Trung tam hoat dong dé gin co chat, vj tri dj lp thé dé gan chat dj lap thé. Mdi vj tri dj lap thé gin voi mot chat di [ap thé, Mot enzym allosteric c6 thé c6 vai vi tri di lap thé khéc nhau dé gin véi cae chat dj lip thé khac nhau. Trong cac enzym dong dang (homotropic enzyms), trung tim hoat ddng va vi tri di lap thé I gidng nhau. Cac enzym allosteric thing Ién va c6 cau trie phite tap thug gdm nhiéu tiéu don vj. Vi dy enzym Aspartate transcarbamoylase xic tic phan img két hgp earbamoyl phosphat va aspartat dé hinh thinh carbamoyl aspartat. Bay la phan img dau tién trong, qué trinh tong hop cae nucleotide pyrimidine, Aspartat transcarbamoylase c6 12 chudi polypeptit duge 16 chic thanh 6 tigu don yj xtc tac (2 phic hop trimer) va 6 tigu don vi dieu hoa (3 phic hgp dimer). Trong qué trinh chuyén héa cac chat 6 té bao, cac nhom enzym hoat déng cing nhau trong cdc phan ting lién tiép, tuan ty dé thye hién mét qué trinh chuyén hoa, vi du nhw qua trinh phan huy glucose tao lactat hodc sy tong hop qua nhiéu phan ting tao mot amino acid tir tien chat don gian. Trong cac hé thong enzym nhwr vay, san phim ctia phan tg truéc li co chit ctia phan tmg ké tiép. Hau hét cdc enzym trong mdi qua trinh trao déi chit déu tun theo cic mé hinh d9ng hoe di trinh bay, Tuy nhién, mdi con duéng bao gém mgt ho%e nhiéu enzym cé vai tro lin hon dén téc 46 chuyén hoa cua toan bd qué trinh, Nhting enzym diéu hoa nay thudng 1a cdc enzym allosteric c6 hoat tinh xuc tac ting hodc gidm rat nhanh d4p img véi cae tin higu nhat dinh theo nhu cau Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung 19ciia 18 bio. Thay 46% téc d6 phan tmg xtc tée boi cdc enzym digu hoa nay s& lim thay d6i nhanh chong téc d6 ton b9 qué trinh chuyén héa, Digu nay cho phép té bao dap ng nhanh chéng nhu cau thay déi vé ning Iugng va cae phan tir sinh hoc cn thiét cho sur phat trién va stra chira, Sur phat trign va sur séng cia té bao phy thuge vao viée str dung higu qua céc ngudn nguyén ligu, va higu qua nay duge thyc hign bai céc enzym diéu hoa, C4n hru y la, ngoai enzym diéu héa la enzym allosteric, trong té bao con c6 co ché digu hoa quan trong khéc la thay déi déng héa tri cia mét hoc nhiéu géc acid amin trong phan tir enzym. Hon 500 loai thay déi déng héa tri da dugc tim thy trong phan tt protein-enzym. Cc nhém chite ning thay déi thuéng li nhém phosphoryl, acetyl, adenylyl, uridylyl, metyl, amid, carboxyl, myristoyl, palmitoyl, prenyl, hydroxyl, sulfat va cae nhom adenosine diphosphate ribosyl. Phosphorylase ‘enase 2a0P ie ZITAT Phosphorylase o Phosphoryase 2, Phosphatase 24,0 Hinh 1.5. Sy thay d6i hoat tinh glycogen phosphorylase co bang thay aéi Bing héa tri phosphoryl hoa M6t sé enzym thay di hoat tinh chuyén tir dang khéng hoat déng sang dang ho dong bang cach thiy phan lign két peptid loai bo doan peptid tir tien chat enzym. Nh enzym protease clia dréng tigu héa duge hoat héa theo cach nay: chymotrypsinogen, trypsinogen, pepsinogen hoat héa thinh chymotrypsin, trypsin, pepsin. Sau khi loai bo doan peptid, cduhinh enzym thay déi va be 1p trung tm hoat dng cua enzym. Neoai ra, m6t s6 céc enzym protease tham gia vao qué trinh déng méu cing dirge hoat hoa theo cach nay. 2.3.3. Céc dang phan ti cua enzym (isoenzym hoac isozym) Trong cing mot loai, cing mét co thé, c6 nhiing enzym tuy ciing xite tae mot loai phan img héa hoc nhung Iai tn tai duéi nhimg dang phan tir khie nhau, e6 nhiing tinh chat vat ly va héa hoc khée nhau. Céc dang phan tir khae nhau ctia mét logi enzym duge goi Id isoenzym hoac isozym. vi dy: phan tir enzym lactat dehydrogenase (LDH) do bon tigu don vj, mdi tigu don vj la mét chudi polypeptid cau tao nén. céc chudi nay gom 2 20 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungJogi, do hai gen khae nhau téng hgp nén: chudi ngudn géc tim (H) va chudi ngudn géc co (M). Vi enzym LDH li loai enzym tetramer do bén chudi polypeptid edu tao nén, cho nén sy t6 hop gitta hai loai chudi polypeptid da tao thanh nim dang phan tr (isoenzym) cua LDH khae nhau, LDH do 4 chudi H tao thanh: HHHH LDEP do 3 chudi H va | chudi M tao thinh: HHHM LDH} do 2 chudi H va 2 chudi M tao thanh: ~HHMM LDH, do 1 chudi H va 3 chudi M tao thanh: HMMM LDHs do 4 chudi M tao thanh: MMMM Vi vay, LDH ; durge goi la isoenzym kiéu tim va LDH s duge goi la isoenzym kiéu gan, Cie isoenzym niy c6 hing s6 Michaelis (Km) va tc d6 phan tng t6i da (Vmax) khdc nhau. (CK) do 2 chudi polypeptid cu tgo nén: mot chudi c6 (B) vi mot chudi c6 ngudn géc eo (M), do dé tgo thinh 3 logi isoenzym: Vige hiéu biét vé cae isoenzym c6 y nghia trong thyc té lam sing do céc dang isoenzym khac nhau t6n tai céc mé khéc nhau. Khi ton throng m6 nao thi chi cb dang isoenzym dé tang trong mau va khi do hoat 46 riéng dang isoenzym 46 sé tang va ting tinh dc higu chan doan. Vi du khi nghi ngo bénh nhan bi nhdi mau co tim, ngudi ta do hoat dé CK-MB (cé nhiéu 6 co tim- duge goi enzym tim) huyét thanh c6 gid tri hon nhiéu khi do hoat 6 CK ton phan (gdm ca 3 dang isoenzym). 2.3.4. Cac tién chat cua enzym Mét sé enzym sau khi duge tng hgp con 6 dang chwra cé hoat tinh (dang khong hoat déng) durge goi la cae tien enzym (proenzym hoe zymogen). Cac tién chat nay khi duge bai tiét vio mdi trudng khdc nghigt ca co thé sé chju tic dyng thiiy phan cia mdi truéng, bj thiy phan cit di m6t doan polypeptid vén che lip trung tam hoat dong dé bio hoat dong, lam cho enzym duge hoat héa, tré nén dang enzym chinh thie enzym cé tén tip vi ngit 1a ogen. vi du: cae tién enzym cia during tiéu chira c6 hoat tinh nhu pepsinogen, trypsinogen va chymotrypsinogen sau khi duge 6a sé bj thuy phn, loai bot m6t doan peptid dé tro thinh cic hoat tinh tuong ting ki pepsin, trypsin vai chymotrypsin; tigm enzym cé thé cé tiép dau ngit “pro”, vi du: tién enzym cila thrombin li prothrombin. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 21Gp cvs Peptia y g wo) Hinh 1.6. Chuyén dang chua hoat déng (Zymogen) sang dang hoat déng ctia enzym 2.3.5. Phirc hop da enzym Phite hgp da enzym Li mét phite hyp gdm nhigu cde phan tit enzym khée nhau nhung cé lién quan v6i nhau trong mt qué trinh chuyén hoa nhat dinh, két ty voi nhau thinh mét khéi nhigu enzym. Khéng thé tach riéng timg enzym trong phite hop da enzym boi vi néu bj téch ra, cde enzym riéng bigt trong phite hop da enzym sé bj bién tinh va mat hoat tinh. Sy ket ty cdc enzym tao thinh phite hop da enzym cé tée dung tang cudng sy cong téc ca ce enzym khac nhau trén m§t qua trinh hoge chudi chuyén hoa gém nhiéu phan img, kim ting higu lye va higu qua xtc tac. ‘Vi du: phite hop da enzym pyruvat dehydrogenase xtc tie cho chudi phin tmg bién pyruvat thinh acetyl CoA (hinh 1.7). Chudi phan ting nay gdm bin phan img voi sw tham gia cua phite hop da enzym pyruvat dehydrogenase gom ba enzym pyruvat dehydrogenase, dihydrolipoyl transacetylase va dihydrolipoyl dehydrogenase voi bon coenzym [a thiamin pyrophosphat (TPP), acid lipoic, coenzym A va NAD’. Pyruvate TPP Pyruvate co, Denydrogenas Per ieone, Dilbydeoliposl Acyl-TPP Transaceylase Dinydrolipost Debyelrogenase Acetyl-CoA NADH + H* Hinh 4.7. Phuc hop da enzym pyruvatdehydrogenase gém 3 enzym xac tac chuyén pyruvat thanh acetyl CoA 22 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung3. CAU TRUC VA CHUC NANG CUA CAC COENZYM Corehit -~- Enzym khOng hogt héa on Enzym hoat hia Corchit gin vio tru tim hoat dong Hinh 1.8. Vai trd cla Coenzym Coenzym tham gia tao trung tam hoat dng: trong hinh trén, coenzym thé hién vai trd tham gia hinh than trung tam hoat déng, sau khi gan voi coenzym, trung tam hoat dong méi od kha nang gan v6i co chat Cée coenzym cé chite ning la tham gia cing enzym trong qué trinh xtie tac. Coenzym thudng c6 ai le voi enzym cing tuong tw nhu di lye cua enzym voi co © vi vay, coenzym c6 thé duge coi nhur mot co chat thir hai. Trong céc truémg hop khic, cae coenzym duge gn déng héa tr) véi enzym va c6 chite nang nhu hode gin nhw vi tri hoat dong trong qua trinh xtc tic. Mot sé coenzym duge tng hop tir ede vitamin nhém B. Vitamin B6, pytidoxin cin bién dai chit it da c6 thé chuyén thanh pyridoxan phosphat la dang coenzym hoat déng. ‘Trong khi d6, niacin cn mOt sy bién doi co ban bdi té bio déng vét mdi cé kha ning tac dong nhw m§t coenzym. 3.1. Cac coenzym oxy héa khir 3.1.1. Cac coenzym niacin (nicotinic acid: vitamin Bs): NAD* va NADP* Niacin li acid pyridin 3-carboxylic, c6 thé duge bién déi thanh 2 coenzym chit yéu tham gia vao logi enzym oxy héa khir, Hai coenzym nay 1i nicotinamid adenin dinucleotid (NAD*) va nicotinamid adenin dinucleotid phosphat (NADP*). C4u tric cua coenzym NADP * khic véi coenzym NAD* & ché cé thém m6t géc phosphat & vi tri 2° cia ribose trong phan tir adenosin monophosphat, Ca hai coenzym nay déu c6 chire nang la van chuyén 2 dign tir va mot H" gitta chat cho va chat nhan H trong phan img oxy hoa khir xtc tac boi enzym dehydrogenase. Tuy nhién, c6 enzym dehydrogenase can coenzym NAD”, cé enzym dehydrogenase can coenzym NADP * trong qua trinh xtc tée Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgocDung 23.adenin ‘ eH e207 > I 1 moose —p—p—roose reese —p—p— hose NAD" (dang oxy héa) NADH (dang khi7) Hinh 1.9. Céng thirc cdu tao va co’ ché phan teng cia coenzym NAD". 3.1.2. Céc coenzym flavin (vitamin B2): FMN va FAD ssenn “ retel pp tse arn DoD FAD (dang oxy hés) FADH: (dang kh) Hinh 1.10. Cdng thtrc cau tao va co ché hoat déng cia coenzym FAD C6 2 dang coenzym cita riboflavin la flavin mononucleotid (FMN) va flavin adenin dinucleotide (FAD). Vitamin riboflavin chita mét di vong, isoalloxazin (flavin), noi qua nguyén tir N-10 dén mot aleol 14 ribitol, FMN c mot goc phosphat 6 vj tri S? cua ribitol trong phan tir riboflavin. FAD co cau tric tuong ty nhu NAD‘, nhung co. adenosin lién ket qua pyrophosphat gin véi di vong riboflavin. Ca FMN va FAD déu cé chire nang li tham gia vio phan tmg oxy hoa khir bing cach trao d6i 2 dign tir va 2H” & vong isoalloxazin, 3.1.3, Cae porphyrin Fe** (cin goi la coenzym hem): coenzym hem Ii coenzym cla hé théng cytochrom, cia enzym catalase, peroxidase, monooxygenase va dioxygenase Vai trd ciia cde coenzym hem la van chuyén dign tir nhd kha ning bién di thuan nghich gitia Fe" va Fe’: Fe’ -¢<> Fe Cée phan ting durge xite tac boi cdc loai coenzym hem: ___ Hai dign ti duge van chuyén tir cytochrom b sang cytochrom e trong chudi hé hap 16 bio nhur sau: 2Cyt b Fe” + 2 Cyt c Fe* < 2Cytb Fe* +2 Cyt ¢ Fe? 24 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungPhin tg phiin huy HsO2 duge xtie téc béi catalase (ed coenzym hem) nhur sau: 2HO, —Caelase, 210 +O Phin img phan huy H202 duge xite tie bii peroxidase (e6 coenzym hem) va ddi hoi kém theo m6t co chat dang khir nhur sau: 2H:O; + AH, Perens, 2 OFA Phin tg oxygen héa mét co chat (thurdng 1a thuée hoac cae chat xenobiotic) duge xite tée boi cée enzym monooxygenase (thudc ng _cytochrom P-450) 6 coenzym li Cytochrom p-450 1 mt loai coenzym hem va doi héi mot coenzym dang khir li NADPH +H! nbur sau: RH + NADPH +H" +02 _Monooxygenase (eytochrom p-450), ROH+ NADP* + H20 Phan ing nay noi chung c6 tie dung bién mét chat dc, it tan trong nude thinh mot chat khéng ddc hoje it déc hon va tan trong nude nhiéu hon dé c6 thé dao thai Khoi co thé Cae enzym dioxygenase la loi enzym cé coenzym hem, e6 tic dung xtc tae phan img peroxy héa mot co chit. Phin (mg duge thé hign nhur sau: RH +02 Dioxygenasg R-O-O-H 3.1.4. Acid lipoie: acid lipoic 14 m6t acid béo chia 2 nhém sulfur (-SH) c6 tén khoa he Ia acid 6,8-dithio-octanoic. Acid lipoic cd phé bién trong cae chat tw nhién, NO la mét chat quan trong trong chuyén héa chit, tham gia vao phitc hgp enzym khtr carboxyl oxy héa ciia acid pyruvie va acid a-ceto glutaric cig véi céic coenzym khic nh TPP, coenzym A, FAD va NAD+. 3.2, Cac coenzym van chuyén nhém 3.2.1. Thiamin pyrophosphat (TPP) van chuyén nhém COz. Trong thinh phin cia TPP cé thiamin i vitamin By. TPP la coenzym cia che enzym cé vai tro tich nhém CO> cia cde acid a-cetonie nhut acid pyruvic hoge acid a- cetoglutaric. Si thiéu hyt thiamin (vit.Bi) lam cho enzym chuyén héa acid pyruvic khéng hoat dong, gay i dong acid pyruvic trong mau va cée md, dic biét ce mé no, than kinh anh hudng cha yéu dén hé than kinh ngogi bién, dudmg tigu héa va hé théng, tim mach, Vi vay, bd sung Thiamin c6 gid tri trong diéu tri cde bénh nhu beriberi (bénh 16- phi), viém than kinh do rugu, viém than kinh do thai nghén, Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 25Vong thiazolium 4 i * ch AEF a 3 CHa—CH2—0—P—0—P—0" aw re Thiamine pyrophosphate (TPP) Hinh 1.11. Cong thire cu tao clia TPP 3.2.2. Coenzym A van chuyén nhém acyl Coenzym A (viét tt 1a CoA-SH) g6m acid pantothenic (vitamin Bs) néi voi mot thioethanolamin tao thanh pantethein va noi vi mét gdc phosphat va voi m6t nucleotid la adenosin monophosphat qua lién két pyrophosphat. Coenzym A c6 vai trd trong chuyén hoa cic acid béo, thé cetonic, acetat va cic acid amin. Vi du: coenzym A ket hop vi acetat dé tao nén “acetat hoat ding” la acetyl CoA, chat nay c6 thé két hgp voi acid oxaloacetat dé to thanh acid citric, mé dau cho chu trinh acid citric, c6 thé tham gia vio qué trinh sinh tong hop acid béo, sinh tng hgp cholesterol va cée hormon steroid, Nie ° Hy ° 9 l 1 R ergot ntl —crycty=ht ~gi-f—oro—to- OH CH, ° EE ‘Acid Pantothenic Coenzym A Hinh 1.12. Céng thire cAu tao cia Coenzym A 3.2.3. S-adenosyl-methionin S-adenosyl-methionin cé tac dung vn chuyén nhém methyl -CHs. 3.2.4, Acid tetrahydrofolic (FH4) Acid tetrahydrofolic e6 vai tro van chuyén nhém 1 nguyén tir carbon, 26 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung3.2.5, Biotin Biotin c6 chite nang nhur mot coenzym ciia enzym carboxylase, enzym xiie te cho sur gin CO2 (goi Ia syr carboxyl héa). Vai trd héa sinh quan trong nhat cita biotin 1a tham gia cde phan (mg carboxyl héa. oO ye “ HE—CH ul US Ns New —CH: ~CH;~CH: ~COOH Hinh 4.13. Céng thirc cu tao cia Biotin 3.2.6. Pyridoxal phosphat Pyridoxal phosphat la din xuat cia pyridoxin (vitamin B6). Pyridoxal phosphat la coenzym cia enzym trao déi amin, cé vai trd vin chuyén nhém amin cia acid c.-amin | cho mét acid a-cetonic 2 dé bién thanh acid a-cetonic 1, con acid u-cetonic 2 nhan nhém amin dé bién thanh acid a-amin 2. Hai enzym trao déi amin quan trong vi cé nhiéu 6 gan str dung coenzym nay 1a la AST, ALT. Rr-GH-COOH NH ° Acid amin 1 Acid «t- cotonic 1 o z \, CH:—NH2 4 Ho a0 oN -o-0-® Ho, x Sy Ny Transaminase Pyridoxal ©) Pyridoxamin -® Ri-GH-COOH Ri- GH-COOH NH ° > Acid amin 2 Acid -cetonic 1 Hinh 1.14, Co’ ché hoat dng cita enzym transaminase Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 27Ngoai vai trd tham gia vio thanh phin cia cic enzym trao déi amin, pyridoxalphosphat cén cé vai trd li coenzym ca céc enzym khir carboxyl cita mt s6 acid amin nhu tyrosin, arginin, acid glutamic va mt s6 acid amin khac. Sin phdm cua su khtr carboxyl cia acid amin ki céc amin ¢6 hoat tinh sinh hge. Vi dy: sy khté carboxyl cia acid glutamic sé tao thanh y-amino butyric acid (GABA) li mot chat tre ché hoat dGng ciia than kinh, sy khir carboxyl cia histidin sé tao thanh histamin la mt hormon cia m6, lim ting tinh thim thanh mach, gay hign tuong di img, sw hydroxy héa cing véi sur khir carboxyl ciia phenylalanin va tyrosin sé tao thinh nor-epinephrin va epinephrin 1A nhimg hormon quan trong ciia tily thugng than, Bang 1.3. Mét sé vitamin c6 trong cdc coenzym va enzym tuong Ung Loai phan ong | Hau qua cua thiéu dien hinh hut Vita Beriberi (gidm cn, dau tim, ri loan chure nng than kinh) Cheliosis va Flavin adenine. ‘ . stomatitus géc (t6n dinucleotide (FAD) Oxy héae bhi thuong cua miéng), viem da Tram cam, nha 1&n, co giat ‘Acid nicotinic | Nicotinamide adenine sta Pellagra (viém da, (niacin) dinucleotide (NAD*) Oxy hoa-khir tram cam, tu chy) Acid Pantothenic Coenzym A Chuyén nhém Acyl Cao huyét ap Phat ban vé léng may, dau co, mét méi (hiém) Ch a Su mau Et Sager Se | ds au tf bon: tng h than kinh trong sy ‘on, tong hop phat trién| thymine | ‘Chuyén cdcnhom | Thiéu mau, thiéu mau Bi S=Deowyadenosy! | methyl; sy sap xép | ctinh, nhiém toan lai ndi tuyén, methylmalonic Suy than (Sung phi va C (acid ascorbic) Chat chéng oxy hoa | chay mau, xuat huyét dui da) Vitamin Coenzym Thiamine Chuyén nhom: Thiamine (B81) pyrophosphate aldehyde Riboflavin (B2) Pyridoxin (Bs) | Phosphate Pyridoxal | Chuyén nhém amin Phirc hgp biotin- Chuyén nhom Biotin lysine (sinh hoc) carboxyl Acid folic Tetrahydrofolate 4. CO’ CHE XUC TAC CUA ENZYM 4.1. Sy bién thién nang lugng ty do (AG) va nang Iugng hoat hoa Nang lugng ty do cia mét hé théng phan img 1a nang lurgng cé thé tgo ra cng cd ich, Nang long tyr do duge ky higu li G. Mt phan ting héa hoe chi c6 thé xay ra theo chiéu ning Iuong ty do gidm, bién chat co ning lugng ty do cao thanh chat ¢6 mite nding lung thap hon. Diéu nay co nghia li diéu kign can cia mOt phan tng héa hye la bién thién nang hrong ty do phai am (AG < 0): 28 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungGi>G. > AG=G:-Gi<0 V6i: Gr ld nang Iugng ne do cia chat tham gia Gr ld nding long ne do ctia sin phém tao think Tuy nhién, do vat chat c6 site y vé mat hoa hoe nén mét phan img di c6 AG < 0, van chura thé tr xay ra duge. Vat chat thudng cé stte y vé mat ha hoc. Sire y vé mat hoa hoc ctia vat chat Ki do cdc yéu t6 sau gay nén: - Yéuté vé entropy (su chuyén ddng hén loan ctia ede phan tir vat cha) - Lép do nude can tré va cé thé Lim mat hoat tinh cua co chat. - Hinh thé khng gian cng kénh cita co chat. - Su sip xép chua dinh hudng cita cde nh6m chite ning trén phan tir enzym Vi vay, mot s6 phan ing hoa hoe mie dit c6 digu kign edn la kha nding xay ra theo huéng nang luong thip hon (AG < 0), nhung phan mg van khong xy ra duge. Muén phan img xay ra phai o6 thém diéu kign di, nghia ld phai cung cap cho hé thong phan img mot nang hrong dé thing durge site y vé héa hoc ctia vat chat. Nang lugng cin cung cip ay duge goi li nding leong hoat héa Trang thai chuyén tiép do) Cachat (s) Tien tinh phan ung Hinh 1.15. M6 ta nang lugng ty’ do va trang thai chuyén tiép cla phan ung Nang luong hogt hoa (AB-Activation Energy) la nang lugng cin thiét dé dua tit ca cde phin tir cda 1 mol co chat & nhigt dO nhat dinh lén trang thdi chuyén tiép (.ransition state) 6 dinh cia hang rio nung luong, dé phan tig enzym c6 thé xy ra, 6 trang thai chuyén tiép, mdi phan tit co chat c6 thé san sang tham gia vao sy tao thanh san pham phan tng. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 29.4.2. Co’ ché tac dung cla enzym Nhu da trinh bay 6 trén, dé phan ing c6 thé xay ra can ¢6 2 diéu kign: bién thi ning lugng ty do cia phan tg phai am (AG <0) va eo chat phai duge cung cap lugng hoat héa dé thing duge sire ¥ hoa hoc (dat téi trang thai chuyén tiép). Co ché tée dung cia enzym li enzym ldm gidm mite nding rong hogt hoa cua phan img dé cdc co chat dé dang dat duge mite ning lvong dé dura phan tg vao trang thai chuyén tiép, tir 46 phan tg c6 thé xay ra. Tée 49 cua phan tmg phu thudc vao sé cae phan tir co chit vugt qua hing rao ning long dé phan img vio trang thai chuyén tip. Vay enzym a gidm ning hrong hoat héa cia phan img bing cach nao? That su, enzym lam giim nang long hoat héa cita phan img bing cich két hop vi cor chat tao thanh phite hop enzym- co chat (E-S) theo phan tng qua 2 bude sau: E+S@ES>E+P (a) (b) O day, E li enzym, S li co chat (substrate), ES la phite hop enzym-co chat va (P) 1a sin phim ciia phin img (product). Nhu vay, enzym c6 tie dung bién mot phan img héa hgc mét bude thanh mét phan ing héa hoc qua 2 bude gdm mot phan ting lién phdn ti (a) vi mot phan ting n6i phén née (b) nhis go thainh phite hgp enzym-co chit, hai phan tng nay déu déi hoi nang lwgng hoat héa thap hon rit nhiéu so véi phan img khdng c6 sy xite téc eta enzym M6t phan quan trong cla nang ltgng duge sir dung lim ting tbe a} phan ding enzym xuat phat tir cde tong tac yéu (twong tac hydro, tuong tic ky nude va tong tac ion) gitta co chat va enzym. Trung tam hogt d6ng cua enzym duge eau trite dé ede trong tic yeu xay trén trong trang thai chuyén tigp, Su cn thiét c6 nhiéu tuong tac yéu La mt trong nhiing Iy do lam cho enzym eé kich thuée ln, Nang luong gan dura trén cdc nhém héa hoe cing li co sé quyét dinh tinh dic higu enzym. Cfch lam giam nang long hoat hoa cia enzym 1a enzym két hgp voi co chit dé tao thanh phitc hop E-S qua trang thai chuyén tiép E-S1* bing nhiing tong tac, tao ra cdc lién u nhdy mOt nang lung hoat héa thap, ding thoi giii phéng ra nang Inong tu do. Nang lugng tw do due giai phong nay lai g6p phan hoat héa phire hop E-S dé dua phite hyp nay vao trang thai chuyén tiép E-S* véi nding Iugng hoat héa cing rat ap dé tao sin pham P va E ty do, dong thoi ciing giai phong nang Iugng ty do. Nhu y, bing cach tao ra phite hgp E-S, enzym chi can nhiing ning lugng hoat héa rit nho cling c6 thé thie day phan img xay ra, Ciing chinh vi vay, ede phan mg enzym dé dang xay ra trong diéu kign nhiét a9 sinh ly cia co thé. 30 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungTrang thai chuyén tiép Nang long tw do (G) AGI: AE khi khong c6 xic tae enzyn ‘AG2: AE khi o6 xtc tae enzym Tién trinh phan ing, Hinh 1.16. Co ché tac dyng cla enzym Enzym lam giam nding long hoat héa cla phan (eng bang céch tao thanh phic hop enzym- co chat (ES) voi mirc nang lugng hoat héa thdp hon nhiéu so véi khi khéng cé enzym xtic tac dé dua phan Ung vao trang thai chuyén tiép, tir 46 tao thanh phic hop enzym san phadm (EP) va phan tng tgo thanh san pham (P) va gidi phéng (E) dé dang xay ra. 5. DONG HOC ENZYM 5.1. Téc 46 phan tng enzym va don vi do Téc dd phan img cia mét enzym a lugng co chat bj bién ddi duéi tae dung cia enzym Ay trong mt phiit & nhiét d6 25°C va cdc diéu kign duge chuin héa. Don vj hoat dé enzym duge thé hign bang: Don vj quéc 16 (international units, TU hoje U) va duge dinh nghia li hrong enzym Lim bién déi | pmol co chit thinh sin phim trong | phit 6 25°C va cae digu kign da durge chun héa. Don vj Katal (Kat) li long enzym lam bién déi 1 mol co chat thinh sin pham trong | gidy 6 25°C va céc diéu kién da duge chuin héa, InKat = 6010 Téc dé phan tng ban déu (Vo) Toc do ban dau cuia mét phan tng enzym (duge ky higu la Vo) c6 nong do enzym, ndng d6 co chat, 6 mét nhigt dO va pH nhat dinh, la toc d6 phan img enzym 6 nhing phat dau tién cia phan img, khi ma téc d6 phan mg chua bj anh huéng bai sy thay doi cita nhigt d6, pH, néng d9 sin phim phan img,... t6c dO ban dau ting lén mt cach tuyén tinh, sau dé khong tng tuyén tinh nira. Hoat d6 enzym chi duge do mot cdch chinh xde 6 téc dO ban dau, nghia la do trong khoang 5 phitt dau tién cia phan ting. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 31Téc dé cye dai (Vmax) Voi mot ning a6 enzym thich hop, nhigt 46 va pH thich hop, khi ndng 46 co cha tang len thi téc d6 phan tng ting len. Khi cée phan tir enzym déu bao hoa co chat thi téc 4 phan ting dat téc d6 toi da (Vmax), 5.2. Thuyét Michaelis-Menten Nam 1913, Michaelis va Menten da dé ra gid thuyét vé vai tro cita ndng 46 co chat trong vigc hinh thanh phire hp enzym - co chat (ES). Sw ligm quan néi chung gitta enzym, co chat va sin phim phan img dugc thé hign bang phwong trinh sau: E+s <4 Es <& E+P ka ka Gia thuyét cia Michaelis-Menten vé sur lién quan gitra téc dé phan img va ning 449 co chat duge tinh todn theo phuong trinh Michaelis-Menten: GdayV —=t6e d6 phan img Vinx = t6e d6 tdi da [S] = ndng d6 co chat Ky = hing sé Michaelis ctia enzym d6i véi co chat. Khi ndng d6 co chat thap hon Ky rit nhiéu, nghia li véi s6 lvong enzym vugt qua s6 lung co chat, trong phuong trinh Michaelis-Menten, ta co thé bo [S] & mau s6, phuong trinh tré thanh dang V=Vnex [S/Km, day li phuong trinh tuyén tinh dang y ax, nghia la téc d6 phan img chi phy thuge vao ndng dé co chat [S]. Lac nay phan ing 4a phan Ging ding hoe bf 1 bai vi tbe d6 phan img ty 1g thudn véi néng 45 eo chit Khi ndng 46 co chat ting lén dén mie dat gid tri bang gia tri Km thi phyong trinh Michaelis-Menten tré thinh dang V = Vnu/2, c6 nghia la téc d6 phan tg bing 1/2 téc d6 tdi da. Khi néng d6 co chat én hon Ky, rat nhieu thi ta co thé bo Km @ mau sO cia phuong trinh Michaelis-Menten, va phuong trinh tro thanh dang V = Vina, cO nghia li tc d6 phan img dat téc d6 t6i da (Vax). Lic nay tat ca cde phan tir enzym déu bao hoa phan dng dat dong hoc “bac khong”, bé i j c dd phan img eiing khong thay déi va I ndng a9 enzym, 32 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung< (Téc 46 ban dau) Nang d6 co chit [S] Ss Hinh 1.17. 86 thj Michaelis-Menten vé sy phy thudc ciia téc dé phan tng vao néng dé co chat Km la néng d6 oo chat ma & dé t6c d6 phan tng bang 1/2 téc d6 tdi da ¥ nghia cita cdc gid trj Kn: - Ke li hing s6 tong hop ciia cae hing 36 t5e 46, c6 git tri bing ndng dé co chat can thiét dé toc dé phan tmg dat bang 1/2 toc do toi da. Nhu vay, Km durge tinh bang mol/L. - Ky la hing s6 de trung iia mi enzym déi voi mdi,co chat, né thé hign ai bre cita enzym di voi co chat: Kn cing nh, ai lye cia enzym d6i véi co chit cing Kin, boi vi chi clin m6t lugng co chat rit nho, téc dO phan ting da dat 1/2 tbc d6 t6i da, Km cng Idn, ai lure cla enzym déi v6i co chat cang nh6, boi vi phai can mét hrgng én co chat, téc dé phan tmg méi dat 1/2 tc dé t6i da. Bang 1.4. Hang sé Km ctia mét s6 enzym va co chat Enzym Co chat Km (mM) Hexokinase (brain) ATP 04 D-Glucose 0.05 D-Fructose 15 26 Carbonic anhydrase HCO; Chymotrypsin Glycyltyrosinylglycine N-Benzoyltyrosinamide 25 40 f-Galactosidase D-Lactose Threonine dehydratase L-Threonine 5.0 Muén dat duge Vmax, néng dé co chit phai > 100 lan Ku, ¥ nghia ctia Vmov: the 46 t6i da Vox thé hign sé vong quay (turnover number) cia Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Bang ThiNgoc Dung 33.mgt enzym, Hing sé ding hgc K2 duge goi la s6 vong quay. Sé vong quay cla mot enzym 1a sé phan tt co chat duge bién déi thanh sin phim trong m6t don vj thoi gian, khi enzym nay duge bao hoa day du véi co chat, Phuong trinh va do thi Lineweaver-Burk: D6 thi hyperbol cia Michaelis-Menten la dé thi chuan xéc va co ban vé dong hoc enzym vé phuong dign ly thuyet, nhung kho ap dung vé thuc hanh. Vi du, rat kho c6 thé duge xde dinh mét céch chinh xéc Vmax tir d6 thi hyperbol cua Michaelis-Menten, vi vay, Lineweaver-burk da cai tién phuong trinh Michaelis-Menten bang cach nghich dao phurong trinh nay va thu duge phuong trinh tuyén tinh dang y= ax + b nhw sau: 1 Kn 1 1 —_—-— + v Vmax [S] Vinax Hinh 1.18. Dé thi Lineweaver-Burk ¥ nghia ctia dé thi Lineweaver-Burk _ ~ Dé thi nay di bién d6 thi hyperbol thanh d6 thi tuyén tinh (dang thing), nhu vay, tir dé thj may 06 thé tim Km va Vinx mot cdch dé dang trén thyre nghiém. = Bb thi nay fi c6ng cy dé xac dinh pH va nhiét dQ ti wu ca phan tg enzym. - D6 thi nay ciing cdn la cng cu dé xée din loai chat tre ché la chat tre ché canh tranh hay khéng canh tranh di véi m6t enzym nhat dinh. 6. CAC YEU TO ANH HUONG BEN HOAT DONG CUA ENZYM Vi ce enzym cé trong huyét thanh véi nhiing lygng rat nho nén trong thye té lam sing ngudi ta thueng do hoat dé enzym chit khéng do néng 46 enzym. Trong nghién citu vé enzym, ngudi ta thuéng khao sat tc dd phan img enzym 6 cae digu kign khic nhau, Cac yéu t6 sau ¢6 thé anh huéng dén téc 46 phan ing enzym, 34 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung6.1. Nong d6 co chat [S] Su anh hudng cla néng 6 co chat dén hoat déng cila enzym da duge mé ti & phan déng hoc enzym véi phuong trinh va dé thi Michaelis-Menten. 6.2. Nong a6 enzym [E] Ngoai néng 46 co chat, néng dO enzym cing anh hudng dén téc 6 phan img enzym, Déi véi cing mot lung co chat, téc a} phan ing enzym ting khi ting ndng 46 enzym vi nguge lai. Tuy nhién, gid tri Km khong bj phy thudc vao néng 49 enzym. 6.3. Nhiét do Nhigt d6 tang thug lam ting t6e d6 cua mét phan ng hoa hge do lim tang sy chuyén dng cua cdc phan tir, lam ting s6 va cham higu qua ciia cde phan tir enzym va co chat va cing cung cp lugng cho phan ting, Tuy nhién, sau khi dat duge téc do t6i da, t6c do phan img gid din boi vi ban chat cua enzym la protein nén khi mhigt d6 tinh protein, lam mat hoat tinh xt tac cita ching. Hau hét cdc enzym cé mot ranh gidi nhigt d6 t6i uu gidng nhw diéu kign nhigt d6 sinh ly cua co thé, Su bién tinh bat dau xay ra & nhiét do tir 40 dén 50°C va sy bién tinh xay ra 6 nhing nhigt d6 cao hon, Théi gian tiép xtc véi nhigt dO cang anh huréng dén sy hoat dong cua enzym. Enzym ¢é thé chiu dung duge nhiét 46 cao hon trong mét théi gian ngan. N chung, & ranh gidi nhiét 46 enzym chua bj bién tinh, khi tang nhiét 46 Ién 10°C, toc d6 phan ing ting lén gap hai lan, nghia li gid tri hé sO nhigt do (temperature coe Q10 bang 2. Nhu vay, cdc két qua phan tich enzym phai duge néu 10 la duge thye ign 6 nhigt dO nao va phai higu chinh bang bang hiéu chinh hoat d enzym theo nhiét d6 néu can thiét, Cae mau huyét tuong c6 thé duge bio quan & nhiét d6 trong tt anh (0°- 4°C) hoge déng lanh trong mét thiri gian nhit dinh cho dén khi phan tich ma c khong bi mat hoat tinh. Tuy nhién, khong nén dong lanh rdi k bai vi diéu nay cé thé gay bién tinh enzym protein. v Téc d6 phan teng e Nhigt do Hinh 1.19, Anh hudng cia nhiét dé trén téc do phan ng enzym Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 35Ngoai ra, ngay nay, céc vi khudn sng 6 day bién néng hoge sudi nude néng ngudi ta di phat hign duge mot s6 enzym bén vi nhigt, e6 kha nding chju nhigt rat cao. Vi dy: cdc enzym Tag DNA polymerase, Tli DNA polymerase, Pfit DNA polymerase, Tth DI polymerase, Tma polymerase,... c6 nhigt d9 t6i wu khoang 75-80°C, 6 thé hoat dong 6 nhigt d6 95°C trong vai chye phit. Vi vay, cde enzym nay hién dang duge sit dung, rng rai cho phan img chudi polymerase (polymerase chain reaction: PCR). 6.4. pH méi trvdng Ban chat ciia céc enzym la protein nén ching mang ign. Cac mire 46 pH khic nghiét c6 thé gay bién tinh enzym hoje anh hyéng dén trang thai ion héa cua enzym, gay nén sy thay di eau trie hode thay déi dign tich trén ede géc acid amin 6 trung tim hoat dng. Vi é chi hoat dng trong ving pH nhat dinh va hoat dong t6i 6 mét pH thich hop. Ngoai vig pH nay, pH cao hon hoae thap hon hoat tinh enzym ‘ac phan (mg enzym sinh ly xay ra trong mét gigi han pH khodng tir uu ciia pepsin & ving acid (pH 2,0-4,0), cdn pH t6i wu cua aypsin 6 ving pH kiém (8-10). Digu nay hoan toan pha hgp tao kién cho qué trinh phan hiiy protein thie an tgo acid amin & duéng tiéu héa. Trong phdng thi nghigm, hoat 6 enzym phai duoc kiém soat mot cach chat ché & pH tdi wu bang nhimg dung dich dém thich hop. Amylase Trypsin 2 — > 8 & S 2 s = pH Hinh 1.20, Anh huéng cia pH dén téc dé phan tng enzym. a) pH téi wu cla pepsin b) pH t6i uu ctia trypsin 36 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung6.5. Cac chat hoat hoa Céc chat hoat héa (activator) la cac chat lim ting t6c d6 cua phan img enzym hoje la lam cho enzym 6 trang thai khéng hogt dng tré thanh trang thai hogt dong. Cac chat hoat héa thudng 1a dc phan tir nhé hodc cdc ion, Cac chat hoat hoa ctia enzym ndi chung thuing 1a cdc kim logi (Ca, Fe, Mg”, Mn, Zn? va K*) hoe 4 kim (Br va Cl). Co ché hoat dng ctia cac chat hoat héa 1a tao nén mot vi tri hoat dong tich dién duong dé 66 thé tae dong vao cic nhém tich dign am cua eo chat. Cac chat hoat hoa Kae ¢6 vai tro tim thay di cdu hinh khéng gian cia enzym, kam én dinh edu trie bie ba va bac bén cia phan tir enzym, lim enzym dé gan véi co chat, cing c6 thé o6 vai tr lign két co chat véi enzym hode véi coenzym, ho’c tao ra sur oxy héa hode sir Khu, Mét sé coenzym cé vai trd nhir mét chit hoat héa di mét sé enzym doi hdi c phan tir hitu co nay cho hoat tinh enzym day dii cia ching. Vi du: NAD* li mot cofactor c6 thé bj khir thinh NADH trong dé co chat thir nhat bi oxy hoa. 6.6. Cac chat tre ché Chat ire ché la nhing chat khi két hop véi enzym c6 tae dung te ché hoat dong ciia enzym, nghia la kim giam hodc kim mit hoat tinh cla nhimg enzym nhat dinh. Enzym xtc tac hau nhu tit cd cac phan img trong té bao séng, tuy nhién cing khéng ngac nhién ring nhiéu chat ite ché enzym Li nhimg thuéc quan trong da duge biét. Vi du, aspirin (acetylsalicylate) tre ché enzym xtic tée bude dau tién trong tong hyp prostaglandin, chat nay lién quan dén nhigu qué trinh, bao gdm qué trinh gay ra dau, Nghién ciru vé cdc chat ite ché enzym cing cung cap nhigu théng tin c6 gid tri vé co ché enzym, giap xac dinh mot s6 con dudng trao di chat. Cé hai loai chat ite ché enzym: tee ché thudn nghich (Reversible inhibition) va ite ché khéng thugn nghich (irreversible inhibition) 6.6.1. Uc ché thuan nghich (reversible 6.6.1.1. Uc ché canh tranh (competi Mt dang ire ché thudn nghich théng thuing li ste ché camh tranh. Su tte ché canh tran li si tre ché ciia nhimg chat c6 cau trie tuong tyr nhur phan tr co chat va vi vay 06 kha ning canh tranh voi co chat dé gin vao trung tim hoat dong cla enzym (hinh 1.21). Khi chat tre ché gin vio trung tam hoat dng ciia enzym tao phite hgp El sé ngan can su gin cia co chat vao enzym va khéng c6 phan tg xtic tac xy ra. Su tte ché canh tranh c6 kha ning thuan nghjch (dio nguge), vi vay c6 thé khie phue sw tre ché canh tranh bing eich ting néng d co chat. Khi co chat nhiéu hon, ching sé canh tranh vGi chat ire ché dé gin vao trung tim hoat d6ng. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 37; e* & ~@ Hinh 1.21. Uc ché canh tran Chat we ché (1) canh tranh voi co’ chat gn vao trung tém hoat dong cia enzym, ngn sy gan cila co chat vao enzym va phan tng xc tac khOng xay ra. Nhu duge chi ra & hinh 1.22 tir dd thi Lineweaver-Burk, gid tri Vinx ki khong thay déi nhung gid tri Km la lén hon, diéu nay chi ra ring cin mt ndng dé co chat lon hon dé dat dirge dng hgc bic “0” do eae anh hung cia chat tte ché canh tranh. ‘Trong trudng hop c6 chat ite ché canh tranh, phurong trinh Michaelis-Menten cé dang: {[s] Vax ———————— Km (1+ [I]/ Ki) + [S] ‘Va phuong trinh Lineweaver-Burk Ia: 1 + Hinh 1.22. Anh huéng chat ec ché canh tranh trén dé th) Lineweaver - Burk 38 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Bang Thi Ngoc DungVi du vé tte ché canh tranh: & phan img tir suecinat dén fumarat trong chu trinh acid citric, co chit 1a succinat. (OOC-CH»-CH:-COO), enzym li succinat dehydrogenase. Cac chit tre ché canh tranh v6i enzym succinat dehydrogenase li chat cé cdu tric gin giéng nh succinat nh oxalat (OOC-COO)), malonat (OOC-CH2- COO), hode glutarat (OOC- CH>- CHe- CH>- COO). M6t tng dung chat tre ché canh tranh duge sir dung trong y hoe 1 digu tri bénh nhan ngo déc methanol. Khi ng6 déc methanol, 6 gan e6 enzym alcohol dehydrogenase chuyén methanol thinh formaldehyde. Day 1a chit dde gay ton thuong mhi mat 1a két aut a thing ip ciia ng dée methanol, bai vi mit dic bigt nha nh iranh, vGi su khac biét la ethanol cing nih co chat cho alcohol dehydrogenase va néng d6 cita né sé gidm theo thai gian khi enzym chuyén héa né thanh acetaldehyde. Ligu phap cho nhiém d6c methanol la truyén cham trong tinh mach ethanol, v6i téc 46 duy tri nong 46 c6 kiém soat trong mau trong vai gid. Ligu phéip nay lam chim su hinh thinh formaldehyde, lam gidm tac hai do formaldehyde, trong khi methanol sé duge than loc va bai tiet nuée ticu thanh chat v6 hai. M6t vi dy khée cia vige img dung chit ire ché canh tranh trong vige ché tgo thuée diéu tri bénh gout. Bénh gout cé sy tt dong acid uric va dé dang tao tia dang mudi urat 6 cdc mé gay viém, dau khép. Vige tao acid uric tir xanthin nho enzym xanthin oxidase (xem chuyén héa acid nucleic). Mot loai thude diéu tri bénh gout li Allopurinol ¢6 céu tao tong ty xanthin (nhumg khée mt chiit 6 vi tri nito so 7 chuyén sang vj tri sé 8). Vi 6 cau tao tung ty xanthin, Allopurinol c6 thé canh tranh gin vao trung tam hoat dong enzym nhung khéng c6 phan tg tao acid uric, han ché tao thanh urat. Xanthin oxidase — HN ot N H Acid uric Xanthin oxidase Allopurinol Hinh 1.23. Co’ ché tac dung cia Allopurinol Allopurinol c6 cu tric tuong ty xanthin da canh tranh gan vao trung tam hoat dong xanthin oxidase va tre ché tao acid uric. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 39.6.6.1.2. Ue ché khéng canh tranh (uncompetitive inhibition) ché khéng canh tranh cing la mot dang ctia ite ché thudn nghich, Su ite ché nay xay ra khi mot chat tre ché chi gin vio phire hop enzym-co chat (ES) 6 mot vi tri khac véi trung tam hoat dng dé hinh thanh mot phite hop enzym-co chat-chat ite ché (ESI) ma khéng tao ra san pham (P): ES+I1=ESI Trong kiéu tee ché nay, chit dre ché I khong gin vaio enzym ty do (hinh 1.24), E+S— ES—>E+P pee @ = Hinh 1.24, Chat tre ché | gan vao phite hop enzym-co chat (khéng gn vao enzym tu do) va phan tng xe tac kh6ng xay ra Sy ting ndng d6 co chit thyc sy lam tang sy ite ché bai vi di cung cdp nhiéu phire hop enzym-co chat hon dé chat we ché 6 thé gin vio. Anh hwéng ctia chat ite ché khong canh tranh duge chi ra trén dé thi Lineweaver-Burk la lam giim gid tri Vinx do bat hoat enzym va Lim gidm gid tri Km (hinh 1.25). = (vis) vi, ile) Hinh 1.25, Anh hudng chat tre ché | khong canh tranh trén dé thi Lineweaver - Burk Hoa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung6.6.1.3. Uc ché khéng canh tranh kiéu hén hop (mixed inhibition) Sur tie ché xay ra khi chat tie ché nay gin vio enzym 6 mot vi tri khéng phai trung tam hoat dong. Su gin niy c6 thé xay ra voi ed enzym te do va vii cé ca phive hyp enzym co chat tao thanh phite hop EI va ESI (hinh 1.26): E+I=El ES +1=ESI Sy gin nay gay nén mét sy thay déi cdu hinh kh6ng gian ciia cdu trie phan tw enzym, lam cho trung tam hoat déng cing bi thay doi, khéng thé tiép nhan duge co chat, néu da tigp nhin co chat cling khéng thé bién d6i co chat thinh san phim, Sy ting néng d@ co chat khéng anh huong dén sy gan ciia ite ché khéng canh tranh vio phan tir enzym nén khéng thé khac phye duge tinh trang tte ché bing cach ting néng 46 co chat. Ets —ES—>E+P + + 1 1 eee El+S— ESI Ir Is G2 @:- Hinh 1.26. Uc ché hén hep: Trong kiéu tre ché hdn hep chat tc ché | gan vao ca enzym ty do va phtéc hop Enzym-co chat va phan ting xtic tac khéng xay ra. Do dé, anh hudng ciia hdn hop trén ddng hoc cia phin img 1a lam giim Vowas boi vi the 46 tdi da Khong thé dat duge do enzym bj bat hoat nhung gid tri Kn Khong thay déi. wal fen) Hinh 1.27. Anh hudng chat tie ché hén hop trén dé thi Lineweaver - Burk Hoa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungTrude day loai tte ché phi canh tranh (noncompetitive inhibition) li tuimg hop dic bigt cia loai te ché nay khi di Ie cia chit tre ché voi E ty do va phite hop ES 1a bang nhau (a= 0’). Thue t6, sur tte ché khéng canh tranh va tre ché hén hgp chi gp 6 cdc enzym xite tac phan tng cé 2 co chat hode nhiéu hon. 6.6.1. Uc ché khéng thugn nghjch (irreversible inhibition) Trong ite ché khOng thudn nghich, chat ite ché gin déng héa tri hode pha hy nhém chite nang can thict cho hoat dong xc téc cia phan tir enzym hoac tao phic khdng phai lién két dong héa tri nhung rat bén ving. Su hinh thinh lién két dong héa tri gitta chat tre ché va enzym tao ra su ire ché khdng thé dao nguoe (khéng thuan nghich), Cc chit tre ché khong thujin nghich 1a mt cng eu hitu ich dé nghién ctru co ché phan ting, Cac acid amin (véi cae nhém ehtic ning) tham gia cdu tao trung tim hoat dong cia enzym c6 thé xde dinh bing céich gdc nao, nhém chite nao lién két céng héa tri voi chat tie ché sau Khi enzym bj bat hoat, Sw tre ché khong thuan nghich nay cing gap trong thu té khi bénh nhan ng6 déc thude trir sau loai phosphor hiru co nhur hinh 1.28. Acetylcholine Choline Acetate Hyc~ Ero cH, -Chi= NICH), HO= CHC + |Ho=c% goes ne J 40 Enz -Ser A) Phan ting binh thudng ctia Acetylcholinesterase See H=c-0-B-0-¢-4 Cy JF |_Oy pes cy [8 Enz ~Ser B) Enzym bj tre ché boii chat tee ché phosphor hitu co. Hinh 1.28. Uc ché enzym Acetylcholinesterase béi chat tre ché phosphor hiru co. (A) Acetyicholinesterase binh thudng xuc tac phan hily lam bat hoat chat dan truyén than kinh acetylcholine baing phan tg thiy phan tao cholin va acetat. (B)_Khi bénh nhan nhiém doc chat phosphor hiiu co diisopropylphosphofluoridate (DFP) (mot cchat khi d6c than kinh va thuéc triv su loai phospho lam bat hoat Acetyicholinesterase do tao phtre hop déng héa tri voi trung tam hoat déng. Didu nay dan téi tang tich dy, W dong acetylcholine va gay d6c. Mt loai chat tre ché khong thudn nghich dic biét Hi chat bat hoat tur sat (suicide inactivators). Cac chat nay khong hoat déng cho dén khi gin vao trung tam hoat dng ciia enzym. Chit bat hoat ty sit trai qua mot s6 bude phan img héa hoe dau tién nur Hoa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dungphan phan tg enzym binh thuéng. Nhung sau dé thay vi duge bién déi thinh san phim théng thudng, chat bat hoat lai chuyén thinh mét chit cé hoat tinh cao gin khéng thudn nghich voi enzym. Phite hyp nay duge goi 1 chat bat hoat dya vao co ché phan ting (mechanism-based inaetivators) béi vi ching dya trén co ché phan (mg enzym binh ie ché enzym. Cac chat bat hoat ty sat c6 vai tro quan trong trong v Ac logi thuée digu tri. Céc nha héa hoc tong hgp cae thuée méi dya trén cae vé co chit, co ché phan tng va chat bat hoat ty sat la mé hinh dé tao ra cdc th AU HOLON TAP 1. Trinh bay each goi tén va phan loai enzym theo phan loai quéc té, cho vi du mi loai. 2. Trinh bay thanh phan cu tao, cu tric, trung tam hoat dng ca enzym. 3. Trinh bay trung tim dj lp thé enzym, ¥ nghfa trong chuyén héa chit 4. Trinh bay vai tro ca cdc eoenzym trong hoat dong enzym va co ché hoat dong cia coenzym NAD’ va FAD. 5. Trinh bay co ché hoat dong cua enzym, 6. Trinh bay phuong trinh va dé thi Michaelis-Menten, 7 nghia cua Km, 7. Trinh bay phuong trinh va d6 thj Lineweaver-Burk, ¥ nghia cua do thi, 8. Trinh bay anh huéng cia nhiét d6 va pH dén hoat déng ctia enzym. 9. Trinh bay anh hung cia cdc yéu té hoat hoa va tre ché dén hoat déng cua enzym. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 43,Chuong 2 NANG LU'Q’NG SINH HOC MUC TIhU HOC TAP 1. Trinh bay duoc ban chat va ¥ nghia chudi ho hdp té bao va co ché tao ATP G ty the. 2. Trinh bay dirge khéi niém ning leong sinh hoc, sw phosphoryl hoa va phosphoryl-oxy héa, § nghia ciia qué trinh nay 3. Trinh bay duoc chu trink acid citric: céc giai doan, céng thitc, enzym, ning heong, dae diém va ¥ nghia. Mi té bao, co thé s6ng déu can nang lugng cho sy hi ng, ton tai, phat trién cia minh. Khi thigu ning hegng co thé tré nén moi met va phai “nap” ning Iugng tir ben ngoai qua duong fin va udng. M6t ngudi truéng thanh nang 70 kg hang ngay can khoang 1920-2900 kcal, tity thuge vio hoat dng thé chat. Bong vat lon doi hoi it nang Iugng hon cho méi kg co thé, va déng vat nho hon can nang lugng nhiéu hon cho mdi kg co thé. Tré em cé nhu cau nang lugng cao hon cho su ting truéng, An trong thite An, nude uéng cé kha nang cung cap nang long cho co 1 glucid, lipid, protein. B4i véi con newdi, nhu cau nding rong lay tr glucid (40% -60%), lipid (ch yéu la triacylglycerol, 30% - 40%), va protein (10% -15%). Nhu clu ning hong theo nga twong di hing dinh; hogt dng thé lye trung binh lam ting t wuyén héa chit chi khodng 40% dén 50% so voi te d6 chuyén héa co s@ hoge nghi ngoi. Tuy nhién, ‘moi ngudi hap thu nguén ning long tir thite an hang ngay qua hai hoac ba bita an, vi vay edn ¢6 dy tit nguén nang hrgng carbohydrat (glycogen trong gan va co), lipid (triacylglycerol trong mé m6), va protein khéng bén trong giai dogn xa bira an. ng long sinh hoc hay str oxy héa sinh hgc hay cn goi i su hd hap té bio 1a qua trinh dét chdy cée chit hitu co glueid, lipid, protein (G, L, P) tao ning hrong cho cae hoat dong séng cia co thé. Khi vao co thé, té bio, cdc thinh pn hiwu co trén sé thodi héa cung cép ning tugng va sinh ra céc chat c&n ba (COs, NIB... Néu chi xem xét vé hong chat thai hoa va ning lugng, sin phim tgo ra thi vige shoal héa (dét chay) ce chat hiru eo dién ra trong va ngoai co thé [4 gidng nhau. Vi du sy dét chay mét phan tr ghicose trong va ngodi co thé déu cé thé biéu dién nhu phan img sau: 44 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc DungCeH206 + —_4 6CO2 + 6 H20 + Q calo Tuy nhién, qué trinh dign bién sy dét chay cae chat hiu co dign ra trong va ngoai co thé khae nhau vé ban chat. Sur dt chay cdc chat hitu co ngoai co thé xy ra nhanh, manh mé, cin ngon lita, oxy khong khi tac dung truc tiép, nhanh v6i carbon, hydro ciia chat hitu co; ning Iugng (Q) giai phong cing mét lik Sy dét chay cdc chat him co trong co thé trong diéu kién nhiét d6 khéng cao (37°C), moi trudng 2/3 la nude, lrgng nhiét tod ra khong due qué lén mét lic, oxy khong khi khong tiép xtic true tiép véi carbon vi hydro clia co chat. Vi vay, qué trinh dot chay céc chat xay ra tir tir, timg buéc, khong cé ngon lira, it ting nhiét do, ning lugng duge giai phong din, Nang lugng gidi phong trong qua trinh dét chay cae chat hifu co trong co thé duge tich tr gi duéi dang ning Ivgng héa hoc. Nang lgng nay sé duge sir dung trong cdc hoat dng séng cita co thé nhu: co co, dan truyén xung déng than kinh, hap thu, bai tiét, tng hop cdc chat can thiét,.v.v Sy thoai héa cac chat G, L, P tir thie an duge minh hoa 6 hinh 2.1 ra qua 3 buse: Thite an | lucia Lipid | Cae auong don Acid béo, glycerol Chu trinh acid citric NADH H* maIN O ATP NHs H:0 +——— cae cht dao that —_ Hinh 2.4. Khai quat sy dt chay cae chat hiru co’G, L, P trong co’ thé Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 45,Bude 1: sy thodi héa cde chit dén khi tgo ra cdc don vj edu tao. Thoai hoa glucid s@ tao ra don vi céu tao Ki cde dudmg don, Thodi héa lipid tao ra don vi cdu tao Hi acid béo, glycerol. Thoai héa protein tao radon vi edu tao la cae acid amin, Buéc 2: sy thoai hoa cae don vj edu tg0 dén khi tgo ra céc sin phim chuyén héa trung gian nhur acid pyruvic, acetyl CoA,.v.v. Bude 3: sur thodi hoa cée chit chuyén héa trung gian dén sin phim cudi cing la ce chat cn ba (CO2, NH) dao thai ra ngoai co thé. Sy thoai héa cdc chat 6 bude 1 va 2 dién ra theo nhiing qué trinh riéng, con duong riéng va sé duge trinh bay chi tiét 6 cde churong sau. Sy thoai hoa cae chat G, L, P 6 bude 3 la ging nhau va ning hong sinh hoc giai phéng khi thoai héa cic chat chtr yéu xay ra 6 butée nay. Vi vay, chuong nay trinh bay chi tiét giai doan 3 ciia qué trinh thoai héa chat to nang Ingng. Riéng thodi ha glucid 6 bude 2 ciing c6 to nang hrgng, nhung chi véi m6t lugng rat nh6 va co ché tao nang Ingng sinh hoc dy trit (ATP) la hoan toan khac véi co ché cha yéu tao ATP 6 buGe 3. 1. BAN CHAT CUA SU’ HO HAP TE BAO (OXY HOA KHU SINH HOC) Vé mit hoa hoe, oxy héa duge dinh nghia la sy cho dign tir (electron) va sy khtr la nh§n dign tir. Do d6, qué trinh oxy héa cia mét phan tir (chat cho electron) luén luén di kém véi vige nan dign tir cua m§t phan tir thir hai (chat nhan electron). Nguyén tic oxy hoa khir nay duge ap dung nhu nhau trong cae hé théng héa sinh va 1a mot khai nigm quan trong dé hiéu biét ve ban chat oxy héa sinh hoc. Luu ¥ ring nhiéu qua tinh oxy héa sinh hoc 6 thé dién ra ma khéng ¢é sw tham gia trye tiép ca phan tir oxy, vi du Khir hydro. Sur séng cia dng vt bic cao hoan toan phy thuge vio vige cung cap oxy cho qué trinh hé hap, qué trinh ma té bao tao duge ning lugng duéi dang ATP tir phan img van chuyén hydro téi oxy dé tao thanh nude. Khi dét chay cae chat hitu co tao nding Iuong, té bao can sir dung O2 va tao ra ip, va qué trinh sir dung O2, tao CO2, H20 di trong té bao nén cin goi ki hé hap té bio. 1.1, Qua trinh tao HzO va CO, n dai vé sur hé hap té bao giai thich mét cach toan dign, chinh xéc vi sur sit dung O2, gidi phong khi COs, tao thanh H20 trong qué trinh dot chay céc chat hitu co. - Khi CO: tao thinh do phan tmg khir carboxyl ciia phan tir chat hitu co nhé enzym xiic tae la decarboxylase: R-COOH ————+ RH+CO; Phan tng nay khong gidi phong nhieu nang lugng - FLO duge tao thanh nhé mét day chuyén phan ting bao gdm hang loat qua trinh tach, van chuyén Hp ra khoi eo chat va van chuyén Hz qua mét chudi dai cdc chat trung, gian cudi cling toi O2. Trong qué trinh nay, cd hydro va oxy déu dugc hoat héa chuyén 46 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dungthanh dang cic ion H’ va O®. Nhiing ion nay hoat déng manh nén khi gp nhau tao thanh HO. ~ Qua trinh van chuyén FH t6i O2 tao thanh HO giai phong rat nhidu nang long va duge tich tri lai cho co thé str dung. 41.2. Chuéi van chuyén dién tir 1.2.1. Thanh phan chudi Chudi van chuyén dign tir duoc hinh thanh bei 4 phite hop protein van chuyén dign tir va 2 chat van chuyén dign tir rigng biét - Phitc hgp I: NADH-CoQ Reductase. Trong phtre hop nay, cde dign tir duge chuyén tir NADH tude tién t6i FMN (flavin mononucleotid, c6 chita vitamin B2) va duge chuyén t6i mdt protein chita kim loai gin véi ru huynh (trung tam sit hru huynh). Sau d6 2c duge chuyén t6i CoQ tao CoQHb. Toan bé phan ting & bude nay cé thé viét NADH + H’ + CoQ. ————+ NAD! + CoQ Co ché van chuyén dign tir va hydro cia NAD" (Nicotinamid adenin dinueleotid) t6i phire hgp I dign ra thong qua nhin nicotinamid nhu 6 hinh 2.2. ° yt 9 yf 2 é Ass ° " 4 cy ers a ye Cy ™ note (Tm + a L ae bas J “NO Ht i | mata 4 mat NADH Dang kh? Dang oxy héa Dang khtr (NADH) D6 hap thy Dang oxy héa on on 0320 240 260 280 300 320 340 360 380 Buse séng (nm) Hinh 2.2. Co ché van chuyén dién tir va hydro clia NAD* NAD* dang oxy héa vi NADH (dang khir) cé su khée biét 16n vé d6 hap thy quang hoe & bude séng 340 nm nén duge ting dung nhigu trong cdc phép do (xét nghigm) vé hoat tinh enzym. Trong phite hgp I, dign tir va hydro duge van chuyén trude hét t6i FMN (thong qua nhan Flavin) nhu & hinh 2.3. Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgocDung 47> 4 b Hyc—c# So S07 SNH ee: B3_B_o— 2 FMNH . FMNH2 z oa ° Hinh 2.3. Co ché van chuyén dign tir va hydro clia FMN Tiép theo dign tw tir FMN duge chuyén téi trung tam sat - lu huynh. Co ché van chuyén dign ttr cua trung tam sét lu huynh 1a théng qua su thay déi héa trj cia ion sat. Co ba trang thai cau hinh cia trung tm sit -lru huynh twong img voi 2 trang thai hoa tri cia sit nhur 6 hinh 2.4. a—w Hinh 2.4, Ba dang trung tam sat - lwu huynh, Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Bang Thi Ngoc Dung+ Phite hgp I: Succinat-CoQ reductase. Trong phite hop nay, 2 €* durge chuyén tir succinat t61 FAD trude tién va co ché {n chuyén dign tir cla FAD duge chi ra 6 hinh 2.5. Ca FMN va FAD trong thinh phan 6 chifa vitamin B2 Vong isoalloxazin 4 “OO FADH2 Dang khir Flavin adenin dinucleotid (FAD) va flavin mononucleotid (FNIN) h 2.5, Co ché van chuyén dign tu va hydro cla FAD Dign ti tir FADH2 sau do duge chuyén t6i trung tim sit -Iru huynh va cudi cing t6i CoQ dé tao CoQH>. Torin bd phan img c6 thé viét nhur sau, Succinat +CoQ ——___» Fumarat + CoQH2 Co ché van chuyén dign tir ciia FAD va trung tam sat -lru huynh ciing giéng nhur di trinh bay 6 phitc hop trén. + CoQ hay Ubiquinon. CoQ la mét chat mang céc nguyén tir hydro (H* va e’). Quinon dang oxy héa ¢6 thé nhan 1 ¢ dé hinh thanh semiquinon va nhan tiép theo I e- va 2H" dé tgo ra dang hydroquinon. CoQHe chuyén 2 ¢* téi phire hyp CoQH-Cyte reduct: CoQ vi CoQH: ¢6 thé khuyéch tn tu do vio mang trong ty thé va la diém néi cla 2 phite hgp enzym dau va phite hop thir 3. Co ché vin chuyén dign tt cia CoQ duge trinh bay ¢ hinh 2.6. Héa sinh - Chii bién: GS. BS. Ta Thanh Van, PGS. BS. Dang ThiNgoc Dung 49.CHs (CH2—CH =C—CHa)10—H CoQ dang oxy hoa (Q) CoQ dang trung gian (semiquinon) (QH*) CHs OH ke ae CH30, R CoQ dang Khir (QH:) CH30 CH OH Hinh 2.6. Co ché van chuyén dién tir cla CoQ + Phite hop II: CoQH2-Cyte reductase. Phite hop ndy gdm 3 thinh phin: Cyt b, trung tim Fe-S, va Cyt cl. Nhé phite hop nay, 2e duge chuyén tit CoQH2 t6i Cyt c. 2e” duge van chuyén theo mot trat ty tir Cyt b, trung tim Fe-S va tdi Cyt cl, cudi cling toi Cyt ¢ dé tao Cyt ¢ dang khit. Dign tit v: chuyén 6 cdc cytochrom thong qua ion Fe’ va Fe’! va ion sit nam 6 nhan Hem. Fe lién phdi tri vai 4 nguyén tir N cua vong porphyrin (hinh 2.7), Hinh 2.7. C4u tric Hem c tham gia c4u tao cytocrom 50 Héa sinh - Chi bién: GS. BS. Ta Thanh Van, PGS. BS. Dang Thi Ngoc Dung