Cu trc phn t enzyme

Bn cht ha hc ca enzyme
T gn mt th k trc y, cc nh khoa hc x vo vic
xc nh bn cht ha hc ca enzyme. Cho n nay, c th ni
rng, ngoi nhm nh phn t RNA c hot tnh xc tc, tuyt i
a s enzyme c bn cht l protein v s th hin hot tnh xc
tc ph thuc vo cu trc bc 1, 2, 3 v 4 ca phn t protein v
trng thi t nhin ca chng. Thc t l bn cht ha hc ca
enzyme ch c xc nh ng n t sau khi kt tinh c
enzyme.
Enzyme u tin nhn c dng tinh th l urease ca u
tng (Sumner, 1926), tip theo l pepsin v trypsin (Northrop v
Kunitz, 1930, 1931). Sau nhng tc gi khc cng kt tinh
c mt s enzyme khc v c bng chng xc nhn cc tinh
th protein nhn c chnh l cc enzyme.
1

Cu trc phn t enzyme

Bn cht ha hc ca enzyme
Kt qu nghin cu tnh cht ha l ca enzyme
cho thy enzyme c tt c cc thuc tnh ha
hc ca cc cht protein v hnh dng phn t:
a s enzyme c dng hnh cu (dng ht). T
l gia trc di v trc ngn ca phn t vo
khong 1 - 2 hoc 4 - 6.
V khi lng phn t: cc enzyme c khi
lng phn t ln, thay i rt rng t 12000
dalton n 1.000.000 dalton hoc ln hn.
2

Cu trc phn t enzyme

Bn cht ha hc ca enzyme
Do kch thc phn t ln, cc enzyme khng i qua
c mng bn thm. Enzyme tan trong nc, khi tan
to thnh dung dch keo; chng cng tan trong dung
dch mui long, glycerin v cc dung mi hu c c
cc khc. Enzyme khng bn v d dng b bin tnh
di tc dng ca nhit cao. Enzyme b bin tnh th
mt kh nng xc tc. Mc gim hot tnh ca
enzyme tng ng vi mc bin tnh ca protein
trong ch phm. Kim, acid mnh, kim loi nng cng
lm cho enzyme bin tnh. Cng nh protein, enzyme
cng c tnh cht lng tnh.
3

Cu trc phn t enzyme

Thnh phn cu to ca enzyme
Cng nh protein, enzyme c th l protein n gin
hoc protein phc tp. Trn c s , ngi ta
thng phn enzyme thnh hai nhm: enzyme mt
thnh phn (enzyme mt cu t) v enzyme hai thnh
phn (enzyme hai cu t). Trng hp enzyme l mt
protein n gin gi l enzyme mt thnh phn. Trng
hp enzyme l mt protein phc tp ngha l ngoi
protein n gin cn c mt nhm ngoi no khng
phi protein gi l enzyme hai thnh phn.
4

Cu trc phn t enzyme

Thnh phn cu to ca enzyme
Phn protein ca enzyme hai thnh phn c
gi l apoprotein hay apoenzyme, cn phn
khng phi protein gi l nhm ngoi hoc
coenzyme. Phn khng phi protein thng l
nhng cht hu c c hiu c th gn cht
vo phn protein hoc c th ch lin kt lng
lo v c th tch khi phn protein khi cho
thm tch qua mng
5

Cu trc phn t enzyme

Thnh phn cu to ca enzyme
Coenzyme l phn khng phi protein ca enzyme trong
trng hp khi n d tch khi phn apoenzyme khi cho
thm tch qua mng bn thm v c th tn ti c lp.
Phn khng phi protein ca enzyme c gi l nhm
ngoi hay nhm prosthetic, khi n lin kt cht ch vi
phn protein ca enzyme bng lin kt ng ha tr.
Mt phc hp hon chnh gm c apoenzyme
v coenzyme c gi l holoenzyme.
Mt coenzyme khi kt hp vi cc apoenzyme to thnh
cc holoenzyme khc nhau xc tc cho qu trnh
chuyn ha cc cht khc nhau nhng ging nhau v
kiu phn ng.
6

Cu trc phn t enzyme

Thnh phn cu to ca enzyme
Coenzyme trc tip tham gia phn ng xc tc,
gi vai tr quyt nh kiu phn ng m enzyme
xc tc v lm tng bn ca apoenzyme i
vi cc yu t gy bin tnh.
Apoenzyme c tc dng nng cao hot tnh xc
tc ca coenzyme v quyt nh tnh c
hiu ca enzyme. .
7

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
Trong nhiu trng hp, cc chui polypeptide
c cu trc bc ba c th kt hp vi nhau to
thnh phn t enzyme c cu trc bc bn. Nh
vy cu trc bc bn l cch sp xp c trng
trong khng gian ca cc chui polypeptide
ring bit trong phn t enzyme. n nay ngi
ta xc nh rng s ln cc enzyme trong t
bo u c cu trc bc bn.
8

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
Cc enzyme c cu trc bc bn l enzyme olygomer
v polymer do nhiu n v nh cu to nn, mi n v
nh l do mt chui polypeptide. Cc n v nh trong
mt phn t enzyme c th ging nhau, nhng cng c
th khc nhau v cu to v chc nng, hoc cng c
th mt s ging nhau, mt s khc nhau. Nhng
enzyme do nhiu n v nh cu to nn cn c gi
l cc enzyme polymer v cc n v nh c gi l
protomer (cc n v nh cn c gi l cc mnh
hoc tiu phn di n v).
9

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
So vi cc enzyme monomer, cc enzyme c cu
trc bc bn c nhng im sai khc sau y:
- C trng lng phn t tng i ln, vo
khong hn 100.000
- Phn t thng cha mt vi trung tm hot
ng, c khi c n 3,4 trung tm hot ng.
10

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
So vi cc enzyme monomer, cc enzyme c cu trc bc
bn c nhng im sai khc sau y:

- Kh nng tng tc ca mt trung tm

hot ng vi c cht s ph thuc vo
trng thi chc nng ca cc trung tm hot
ng khc.
11

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
- L iu kin cn thit xut hin tnh cht
allosteric ca enzyme. Enzyme allosteric (enzyme
d lp th, d khng gian) l enzyme m cht trao
i c th lm nh hng (c ch hoc hot ha)
ln tc dng ca chng. Hnh nh hin tng d
lp th (allosteric) bt u xy ra trc ht cc
enzyme c xy dng nn t mt s tiu n v
v hiu ng d lp th c nh hng n bn
ca lin kt gia cc tiu n v ny (xem thm
phn enzyme d lp th).
12

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
- Gm cc tiu phn di n v: a s cc
enzyme c cu trc bc bn cha t 2 - 4
protomer, mt s enzyme khc cha t 6 - 8
protomer. V d enzyme catalase c trng lng
phn t 252.000, cha 6 mnh di n v, mi
mnh c phn t lng l 42.000.
Mt s enzyme cha n 12 protomer v d
nh arginine carboxylase, oxaloacetate
carboxylase.
13

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
- S sp xp ca cc mnh di n v
trong phn t enzyme thng c tnh cht
i xng cao.
C 4 kiu chnh, c biu th hnh di
y.

14

Cu trc phn t enzyme

Cu trc bc 4 ca enzyme
- Cc tiu phn tng tc vi nhau bng cc kiu
lin kt khc nhau. Trong a s trng hp nh
tng tc k nc, lin kt hydrogen, mi s
trng hp khc nh lin kt disulfide (v d
glucoseoxydase
ca Asp.niger)
hoc cu
polypeptide (v d nh enzyme leucin-s-RNAsynthetase). Cu polypeptide ny c vai tr quan
trng vi tnh c hiu ca enzyme, khi mt n s
thay i tnh cht phn ng.
15

Cu trc phn t enzyme

Trung tm hot ng ca enzyme
Ton b cu trc khng gian ca phn t enzyme c vai tr
quan trng i vi hot tnh xc tc ca enzyme. Tuy nhin,
hot ng ca enzyme lin h trc tip vi mt phn xc
nh trong phn t enzyme. Trung tm hot ng ca
enzyme l phn ca phn t enzyme trc tip kt hp vi
c cht, tham gia trc tip trong vic to thnh v chuyn
ha phc cht trung gian gia enzyme v c cht to
thnh sn phm phn ng. Trung tm hot ng bao gm
nhiu nhm chc nng khc nhau ca amino acid, phn t
nc lin kt v nhiu khi c c cofactor hu c
(coenzyme) v v c.
16

Cu trc phn t enzyme

Trung tm hot ng ca enzyme
cc enzyme mt thnh phn, trung tm hot
ng thng bao gm mt t hp cc nhm chc
nng ca amino acid khng tham gia to thnh trc
chnh ca si polypeptide. V d nhm - SH ca
cysteine - OH ca serine, threonine v tyrosine,
- NH2 ca lysine, -COOH ca glutamic acid,
aspartic, vng imidazol ca histidine, indol ca
tryptophan, nhm guanidin ca arginine.
17

Cu trc phn t enzyme

Trung tm hot ng ca enzyme
Cc nhm ny c th xa nhau trong mch
polypeptide nhng li gn nhau trong khng
gian, c nh hng xc nh trong khng
gian cch nhau nhng khong cch nht
nh sao cho chng c th tng tc vi
nhau trong qu trnh xc tc.
18

Cu trc phn t enzyme

Trung tm hot ng ca enzyme
Trung tm hot ng ca cc enzyme hai thnh
phn thng bao gm nhm ngoi (vitamin, ion
kim loi ...) v cc nhm chc nng ca cc amino
acid phn apoenzyme.
Theo quan nim ca Fisher th trung tm hot ng
ca enzyme c hnh thnh sn vi mt cu
to nht nh ch cho php c cht c cu to
tng ng kt hp vo. Do c th v s tng
ng nh kha vi cha kha
19

Cu trc phn t enzyme

Trung tm hot ng ca enzyme

20

Cu trc phn t enzyme

Trung tm hot ng ca enzyme
Thuyt ny tuy cng gii thch c mt s hin tng
nhng khng gii thch tha ng c nhiu kt qu thu
c trong thc nghim. V vy, Koshland a ra mt
gi thuyt khc hp dn v t nh hn. Theo thuyt ny th
c im ca vng trung tm hot ng l rt mm do v
linh hot, cc nhm chc nng ca trung tm hot ng
ca enzyme t do cha t th sn sng hot ng, khi
tip xc vi c cht, cc nhm chc nng trong phn
trung tm hot ng ca phn t enzyme thay i v tr
trong khng gian, to thnh hnh th khp vi hnh th ca
c cht. Cng v vy, ngi ta gi m hnh ny l m hnh
tip xc cm ng hoc khp cm ng.
21

Cu trc phn t enzyme

Trung tm hot ng ca enzyme
Gia c cht v trung tm hot ng to
thnh nhiu tng tc yu, do c th d
dng b ct t trong qu trnh phn ng
gii phng enzyme v sn phm phn ng.

22

Cu trc phn t enzyme

Cc dng phn t ca enzyme
Trong cu trc phn t ca enzyme, tnh cht tinh vi v phc tp khng
ch gii hn phm vi tng phn t, t thnh phn cu to v cc bc
cu trc cho n cu to ca trung tm hot ng cng vi vai tr ca
cc nhm chc nng m cn th hin tnh a dng ca cc phn t
enzyme. Tnh a dng ca nhiu enzyme khc nhau c pht hin
cc c th sng khc nhau t ngi, ng vt, thc vt n vi sinh
vt.
Nhng enzyme xc tc cng mt phn ng ha hc v c cng tnh c
hiu c cht nhng c ngun gc khc nhau nn th hin nhiu tnh
cht khc nhau.
Vd: amylase ca dch nc bt v dch ty ca ngi th ging nhau,
nhng chng khc vi amylase thu c t ty ln v ha tan, v
pH thch hp v mt s tnh cht khc.
23