HCMC March 2004

MOT SO KHAI NIEM

ong hoc phan ng nghien cu g?
1. ong hoc nghien cu s phu thuoc gia toc o phan ng vao nong o c
chat, vao ieu kien tien hanh phan ng;
2. ong hoc
Toc o phan ng chnh la ai lng bieu dien s bien thien ve lng cua mot c
chat nao o theo thi gian trong mot n v the tch thiet b. Khi phan ng la ong
the no c xac nh theo:
t Vd
dn
R
i
i
=
oi vi phan ng xay ra tren be mat tiep xuc pha (phan ng d the) no c xac
nh cho 1 n v dien tch tiep xuc pha, co ngha:
t Sd
dn
R
i
i
=
hay mot n v khoi lng vi sinh vat:
Neu phan ng co the tch khong oi, toc o phan ng c xac nh cho mot n
v the tch phan ng, khi o tng ng la:
t d
dS
R =
hay:
t Xd
dS
R =
t Gd
dn
R
i
=
HCMC March 2004
MOT SO KHAI NIEM
ong hoc phan ng sinh hoc xay ra nh the nao?
Moi sinh vat e duy tr c hoat ong cua mnh la do trong c the cua chung luon
xay ra hang tram, hang nghn phan ng hoa hoc, mot cach nhp nhang, nhanh, nhay.
Bnh thng cac phan ng nay (nhat la phan ng hu c) khi xay can co xuc tac,
nhiet o cao, ap suat ln. Nhng trong c the sinh vat chung xay ra ngay nhiet
o thng, ap suat kh quyen va pH gan nh trung tnh. o la nh xuc tac sinh hoc
ac hieu cua enzim trong c the sinh vat.
C che xuc tac cua enzim la rat phc tap, en nay van cha co mot giai thch hoan
hao ve c che nay. Ngi ta ch co the giai thch gan ung phan ng xuc tac
enzim bang thuyet toc o au.
Theo o, muon cho phan ng xuc tac bi enzim E xay ra th c chat S can c gan
vao trung tam hoat ong cua phan t enzim, tao thanh phc enzim c chat ES. Trung
tam hoat ong cua phan t enzim co nhng nhom hoa hoc ac biet trc tiep tham
gia vao c che xuc tac.
HCMC March 2004
MOT SO KHAI NIEM
ong hoc phan ng sinh hoc xay ra nh the nao?
Di tac dung cua enzim, hoat tnh cua c chat tang len ro ret, nen ch can mot
nang lng hoat hoa nho hn nhieu cung u lam cho phan ng xay ra tc th
chuyen hoa c chat thanh san pham.
Viec giam nang lng hoat hoa trong phan ng xuc tac bi enzim, so vi phan ng
hoa hoc thong thng co the thay ro qua v du cua phan ng thuy phan ng
sacazo bang H
+
va bang enzim o amilaza:
C
12
H
22
O
11
+ H
2
O C
6
H
12
O
6
+ C
6
H
12
O
6

Hang so toc o cua phan ng c bieu dien theo phng trnh Arrhenius di dang:
Trong o: R = 1,98 cal/mol hang so kh; T nhiet o tuyet oi, K; E nang lng
hoat hoa, cal/mol; B hang so bieu th tan so va cham cua cac phan t va s t
nh hng ac hieu cua cac phan t va cham. 37
0
C, phan ng tren xuc tac
bang enzim amilaza co E
E
= 8.000 10.000 cal/mol; con E
H
+
= 25.600 cal/mol.
RT
E
B k = ln
HCMC March 2004
MOT SO KHAI NIEM
ong hoc phan ng sinh hoc xay ra nh the nao?
C
12
H
22
O
11
+ H
2
O C
6
H
12
O
6
+ C
6
H
12
O
6

Thay E
E
= 8.000 10.000 cal/mol; con E
H
+
= 25.600 cal/mol.vao phng trnh (*) ta co:
RT
E
B k = ln
(*)
= 28.67
( ) 8 , 613
17600
37 273 . 98 , 1
8000 25600
ln
25600
ln
8000
ln
=
+

=
=
+
+ +
H
E
H H
E E
k
k
RT
B k
RT
B k
Suy ra:
=
+
H
E
k
k
2,826.10
12

Ro rang, hang so toc o phan ng xuc tac bi enzim ln hn hang nghn ty lan so
vi xuc tac thong thng (bang ion H
+
) cung mot nhiet o.

HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
Trong c the sinh vat luon xay ra hang tram, hang ngan phan ng hoa hoc mot cach
nhp nhang, lien quan chat che vi nhau. e duy tr s song va phat trien cua moi
sinh vat sinh. Nang lng can thiet cho hoat ong song cua chung cung c tao ra t
nhng phan ng nay. Khong nhng vay cac phan ng nay xay ra trong te bao sinh
vat ngay nhiet o thng, pH gan nh trung tnh, vi nong o thap (cac phan
ng nay bnh thng co the xay ra ch khi co xuc tac, nhiet o cao, ap suat ln
). o la nh xuc tac cua enzim (men) co trong te bao sinh vat.
2. Cac tnh chat cua enzim
1. Cau tao enzim
3. Cac yeu to anh hng en hoat o cua enzim Cach goi ten
4. Cach goi ten
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
1. Cau tao enzim:
Enzim co the hoa tan trong nc, trong dung dch muoi loang, trong cac dung moi hu
c co cc, nhng khong tan trong cac dung moi khong phan cc. Dung dch enzim co
tnh chat cua dung dch keo a nc, trong nc cac ion, nhom ion hay nhom phan cc
cua no se ket hp vi phan t nc (lng cc) tao thnah lp vo hydrat.

Enzim la nhng chat hu c co ban chat protein (phan ln co cau truc bac 4), phan
t lng ln t 20.000 (nho nhat la Ribonucleaza 12.700) en 1.000.000 dalton.
Giong nh phan t protein, enzim khong ben di tac dung cua nhiet, nhiet o cao
enzim b bien tnh va mat kha nang xuc tac cua mnh, mc o giam hoat tnh xuc
tac tng ng vi mc o bien tnh cua protein. Ngoai ra cac yeu to gay bien tnh
cua protein (axit, kiem, hay muoi kim loai nang ) cung lam mat kha nang hoat ong
cua enzim.

HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
1. Cau tao enzim:
Ngi ta a thu c tinh the enzim: ureaza t au tng (Summer 1026), pepxin
(Northrop 1930) va tripxin (Kunitz 1931) , ieu o cho thay enzim co ban chat
protein.
Enzim co tnh lng tnh, co the ton tai di dang anion, cation hay trung hoa ve ien.
Enzim co cau tao t cac axit amin va cac chat khac (khong phai protein) lien ket vi
nhau qua moi lien ket peptit. Neu ch gom cac axit amin th goi la enzim mot cau t
(protein n gian), con lai la enzim phc tap (hai cau t hay thanh phan). Trong enzim
phc tap phan protein goi la feron hoac apoenzim. Phan khong phai protein goi la
nhom ngoai agon, hay coenzim. Neu tach rieng apoenzim (bang mang ban tham) th
coenzim co the ton tai oc lap.
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
1. Cau tao enzim:
Trong nhieu enzim (mot va hai cau t) con co cha ca nguyen t kim loai, va chnh
ion kim loai tham d vao hoat ong xuc tac cua enzim.
Chnh apoenzim quyet nh tnh ac hieu cao cua enzim va lam tang hoat tnh xuc tac
cua coenzim. Phan agon quyet nh kieu phan ng ma enzim xuc tac, trc tiep tham
gia vao phan ng va tang o ben cua apoenzim oi vi cac yeu to gay bien tnh.
Trung tam hoat ong cua enzim la phan ket hp ac hieu vi c chat, cau tao cua
tam hoat hoa rat phc tap va cha c nghien cu ky. Trong enzim mot cau t,
tam hoat ong la to hp cac nhom nh chc cua axit amin khong tham gia tao thanh
truc chnh cua si polypeptit, nh: SH cua xystein, OH cua xerin Trong enzim hai cau
t thng gom nhom ngoai (vitamin, ion kim loai ) va cac nhom nh chc cua axit
amin phan apoenzim.
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
1. Cau tao enzim:
2. Cac tnh chat cua enzim.:
Trung tam hoat ong co cau hnh khong gian tng ng vi cau truc cua c chat nh
o va cha khoa. Cau truc nay c hnh thanh trong qua trnh tiep xuc vi c chat.
2.1 o xuc tac:
Enzim la chat xuc tac sinh hoc, no co ay u cac tnh chat nh moi chat xuc tac
khac. Nhng co hoat o xuc tac manh hn nhieu, nh: 1 mol Fe
3+
xuc tac phan ly c
10
6
mol H
2
O
2
/ph; Mot phan t catalaza co mot nguyen t Fe xuc tac phan ly 5.10
6
mol
H
2
O
2
/ph; 1 phan t | amilaza sau mot giay, co the phan giai 4.000 lien ket glucosit
trong phan t tinh bot;

2.2 Tnh ac hieu cua enzim:
ac hieu phan ng: Moi enzim ch co the xuc tac cho mot kieu phan ng chuyen
hoa mot chat nhat nh. V du: phan ng oxy hoa kh, thuy phan, chuyen v
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
2. Cac tnh chat cua enzim.:
2.1 o xuc tac:
2.2 Tnh ac hieu cua enzim:
ac hieu c chat: Moi enzim ch co kha nang xuc tac chuyen hoa mot hay mot so
loai c chat (ket hp vao tam hoat hoa) ma thoi.
ac hieu tuyet oi: Ch tac dung vi mot loai c chat nhat nh, khong tac dung
vi c chat khac, nh: ureaza ch tac dung vi ure, thuy phan no: H
2
N CO NH
2
+
H
2
O CO
2
+2NH
3
; vi hydroxyure gan giong ure ch tac dung vi toc o rat thap.
ac hieu tng oi: Tac dung len mot kieu lien ket hoa hoc nhat nh trong phan
t c chat ma khong phu thuoc vao cau tao cua phan tham gia tao thanh moi lien
ket o. V nh: lipaza thuy phan tat ca cac moi lien ket este, aminopeptidaza co the
thuy phan nhieu peptit.
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
2. Cac tnh chat cua enzim.:
2.1 o xuc tac:
2.2 Tnh ac hieu cua enzim:
ac hieu nhom: Tac dung len mot kieu lien ket hoa hoc khi mot trong hai thanh
phan tao lien ket co cau tao xac nh.
ac hieu quang hoc: Tac dung vi mot trong hai ong phan quang hoc cua cac chat.
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
3. Cac yeu to anh hng en hoat o cua enzim:
1. Cac chat km ham:
Hoat o cua enzim co the b thay oi di tac dung cua mot so chat co ban
chat hoa hoc rat khac nhau. Chat lam giam hoat o nhng khong b chuyen hoa bi
enzim goi la chat km ham hay chat c che (I). o co the la cac ion, phan t vo,
hu c ke ca protein. Chung tham gia vao qua trnh kiem tra, ieu chnh san xuat.
Cac chat lam bien tnh protein la nhng chat km ham khong ac hieu cua
enzim.
Nhieu chat khong lam bien tnh protein enzim nhng van lam giam hoat o theo
c che khac.
Cac chat co the lam km ham thuan nghch hay khong thuan nghch, o la do
phan ng thuan nghch tao phc enzim chat c che xay ra theo chieu nao.
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
3. Cac yeu to anh hng en hoat o cua enzim:
1. Cac chat km ham:
Chat km ham canh tranh: la nhng chat km ham thuan nghch, co cau truc tng
t c chat, nen de ket hp vao tam hoat ong cua enzim, chiem cho cua c chat.
V du: trong len men glyxerin th axetaldehyt (CH
3
CH=O) la chat km ham canh tranh
cua enzim etanoldehydrogenaza. Co cau tao kha giong aldehyt glyxerinic (CH
2
OH CHOH
CH=O) la c chat cua enzim.

HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
3. Cac yeu to anh hng en hoat o cua enzim:
2. Nhiet o:
Moi Enzim thch nghi trong mot ieu kien nhat nh cua moi trng (ve nhiet o,
pH ). Trong cac phan ng enzim, nhiet o co tac dung lam tang toc o phan ng
len toc o cc ai. o la do khi tang nhiet o hoat o xuc tac cua enzim tang nh
giam nang lng hoat hoa. Nh qua trnh hieu kh khi tang 1
0
C hoat o tang gap oi.
Noi chung khi tang 1
0
C, hoat o tang khoang 7,2 %. Nhng khi nhiet o tang len
nhiet o ti han (phan ln cac enzim khoang 60 70
0
C) th hoat tnh cua chung b
mat. Moi enzim eu co mot nhiet o thch hp nhat cho hoat ong xuc tac ma
o phan ng enzim at toc o cc ai. Noi chung chung gan nhiet o c the, nh
ong vat nhiet o o khoang 40
0
C. Nhng cung co loai len en 70
0
C (nh o -
amilaza cac sinh vat chu nhiet o, hay trong thc vat, tham ch gan nhiet o soi
(trong VSV cac suoi nc nong).

HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
3. Cac yeu to anh hng en hoat o cua enzim:
2. Nhiet o:
nhiet o thap, hoat o cua enzim giam i, 0
0
C hoat o cua cac enzim
khong ang ke. Khi tang nhiet o th hoat o phuc hoi tr lai, nen ta co the bao
ve cac enzim nhiet o thap (di 20
0
C).
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
3. Cac yeu to anh hng en hoat o cua enzim:
2. o pH:
Enzim rat nhay cam vi pH cua moi trng, nen pH co tac dung rat ln oi
vi toc o phan ng enzim. Moi enzim co mot gia tr pH thch hp cho s hoat ong
cua no, o toc o pgan ng ln nhat, gia tr khac toc o eu giam. Gia
tr thch hp cua pH con phu thuoc vao loai va nong o c chat, nhiet o, noi chung
no cung thay oi theo cac thong so nay.
Nhieu enzim co pH thch hp vung trung tnh, nhng cung co loai kha thap
(pepsin pH = 1,5), cung co loai khac cao (photphataza pH = 9 10)
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
Ten c chat ac hieu ten kieu phan ng xuc tac uoi aza
V du: ureaza: cacbamit amidohydrolaza
4. Cach goi ten:
Khi co hai s chuyen hoa th co them phan th hai trong ngoac
V du: L axitamin oxydoreductaza (deamin).
Theo tnh ac hieu phan ng chia thanh 6 lp:
- Oxydoreductaza: xuc tac cho phan ng oxy hoa khu;
- Transpheraza: cho phan ng chuyen v;
- Hydrolaza: phan ng thuy phan;
- Liaza: Phan cat khong can nc, loai nc tao thanh lien ket oi, hay ket hp
phan t nc vao lien ket oi;
- Izomeraza: ong phan hoa;
- Ligaza: Tong hp co s dung lien ket giau nang lng ATP,
HCMC March 2004
MOT SO KHAI NIEM
Enzim Xuc tac sinh hoc la g?
Moi lp c chia lam nhieu to, trong mot to co nhieu nhom, nen trc ten enzim
co 4 con so: so au ch lp, th hai ch to, th ba ch nhom va cuoi cung ch
enzim.
4. Cach goi ten:
- V du: 1.1.1.1 alcoholdehydrogenaza la enzim cha Zn, co coenzim NAD
+
, xuc tac cho
phan ng:
Chang han: CH
3
CH
2
OH + NAD
+
CH
3
CHO

+ NADH + H
+


5. Hoat o enzim:
n v hoat o: n v enzim quoc te (UI) la lng enzim xuc tac lam chuyen hoa 1
micromol c chat sau mot phut ieu kien tieu chuan: 1 UI = 1 mol S (10
6
mol)/ph.
1 katal (Kat) la lng enzim co the lam xuc tac 1 mol c chat sau 1 giay ieu kien
tieu chuan: 1 Kat = 1 mol c chat/s
Hoat o rieng cua mot che pham enzim la so n v UI hay Katal ng vi 1 ml hay 1
mg protein cua che pham.
HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
ong hoc phan ng xuc tac enzim (men) la phc tap, en nay cha co mot c che
nao giai thch ro rang. Ma ch co the giai thch gan ung khi dung thuyet nong
o dng va thuyet phc chat trung gian nh Michaelis Menten a tng lam
Theo Michaelis Menten th trc het enzim E ket hp vi c chat S tao nen phc
trung gian gia enzim c chat ES mot cach thuan nghch. Hang so toc ong phan
ng thuan trong giai oan nay la k
1
, phan ng nghc la k
1
. Trong phan ng phan
giai phc ES e tao thanh san pham P va giai phong enzim t do ban au E vi
hang so toc o k
2
.
Qua trnh o c mo ta bang s o sau:
E + S ES P + E
k
1
k
2

k
1

HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
S tao phc ES nhanh, phc nay khong ben se b phan ly thanh san pham P, giai
phong enzim t do E. Giai oan sau co toc o cham, quyet nh toc o chung cua
qua trnh.
E + S ES P + E
k
1
k
2

k
1

ong hoc tao san pham theo Michaelis - Menten
Theo Michaelis Menten toc o tao san pham cua phan ng, phu thuoc vao nong
o phc ES. Toc o chuyen hoa phc ES c xac nh theo:
Theo nguyen ly Bodenstein th phc ES la hp chat trung gian khong ben, nen co toc
o chuyen hoa bang 0, tc la:
| |
| || | | | | | ES k ES k S E k
d
ES d
R
ES 2 1 1
= =

t
| |
| || | | | | | 0
2 1 1
= = =

ES k ES k S E k
d
ES d
R
ES
t
HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
E + S ES P + E
k
1
k
2

k
1

ong hoc tao san pham theo Michaelis - Menten
Goi E
0
la tong nong o enzim, th:
Ta co toc o tao san pham se la:
| | | | | | ES E E =
0
| |
| | ES k
d
ES d
R
2
= =
t
| |
| || | | | | | 0
2 1 1
= = =

ES k ES k S E k
d
ES d
R
ES
t
Thay vao va bien oi ta co:
| | | | ( )| | | | | | | || | | | ( )| | ES k k S k S E k ES k ES k S ES E k
2 1 1 0 1 2 1 0 1
0 + + = =

Ket qua nhan c:
| |
| || |
| |
2 1 1
0 1
k k S k
S E k
ES
+ +
=

| || |
| |
| || |
| |
m
K S
S E
k
k k
S
S E
+
=
+
+

0
1
2 1
0
=
Cho nen toc o tao san pham la:
| | | || |
| |
| |
| | S K
S
R
S K
S E
k
d
ES d
R
m
MAX
m
+
=
+
= =
0
2
t
Trong o:
| |
0 2
E k R
MAX
=
toc o tao san pham cc ai v khi o toan
bo tam hoat hoa cua cac enzim eu a tham gia phan ng [ES] = [E
0
] (nong o c
chat rat ln so vi enzim).
HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
E + S ES P + E
k
1
k
2

k
1

ong hoc tao san pham theo Michaelis - Menten
| |
| || | | | | | 0
2 1 1
= = =

ES k ES k S E k
d
ES d
R
ES
t
Phng trnh tren goi la phng trnh Michaelis Menten.
la hang so MichaelisMenten, ac trung cho moi enzim.
| | | || |
| |
| |
| | S K
S
R
S K
S E
k
d
ES d
R
m
MAX
m
+
=
+
= =
0
2
t
Co the viet theo mot so dang khac:
Hay:
Dang nghch ao:
1
2 1
k
k k
K
m
+
=

| |
|
.
|

\
|
= 1
R
R
S K
MAX
m
| | | | S
K
R
R
Hay
S
K
R
R
m
MAX
m
MAX
+
=
+
=
1
1
1
| |
m MAX
m
K S R
K
R
1 1 1
+ =
Ro rang, khi
| |
| | S K
S
K
th
R
R
m
m
MAX
= =
+
=
2
1
1
1
2
HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
E + S ES P + E
k
1
k
2

k
1

ong hoc tao san pham theo Michaelis - Menten
| |
| || | | | | | 0
2 1 1
= = =

ES k ES k S E k
d
ES d
R
ES
t
Mot so dang khac cua phng trnh
Michaelis Menten:
| | | || |
| |
| |
| | S K
S
R
S K
S E
k
d
ES d
R
m
MAX
m
+
=
+
= =
0
2
t
| | | | S
K
R
R
Hay
S
K
R
R
m
MAX
m
MAX
+
=
+
=
1
1
1
| |
m MAX
m
K S R
K
R
1 1 1
+ =
Nh vay, hang so K
m
chnh la nong o c chat (mol/l) u lam cho toc o phan ng
at en mot na toc o cc ai R
MAX
. e at toc o cc ai thng phai co
nong o c chat ln vao khoang 100 lam K
m
.
| |
| | S K
S
K
th
R
R
m
m
MAX
= =
+
=
2
1
1
1
2
HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
E + S ES P + E
k
1
k
2

k
1

ong hoc tao san pham theo Michaelis - Menten
K
m
the hien ai lc cua enzim oi vi c chat, K
m
cang ln th ai lc cang nho va
ngc lai. K
m
cua moi enzim oi vi tng c chat khac nhau la khac nhau.
Tr so cua K
m
, phan ln phan ng enzim nam trong khoang 10
2
10
5
mol/l.
HCMC March 2004
ONG HOC PHAN NG XUC TAC MEN
E + S ES P + E
k
1
k
2

k
1

ong hoc tao san pham theo Michaelis - Menten
Nh vay, phng trnh Michaelis Menten mo ta quan he gia toc o phan ng vao
nong o c chat va hoat o cua enzim.
Toc o phan ng ay co the hieu la:
Toc o tieu thu c chat S: lng c chat b tieu thu trong mot n v thi gian (h) do
1 g vi sinh vat tieu thu:
| |
t Xd
S d
R =
Toc o s dung oxy (trong qua trnh hieu kh): lng oxy do mot 1 g VSV s dung trong
1h:
Toc o tao kh metan (san pham trong len men k kh): lng metan tao thanh t 1 g
VSV trong mot n v thi gian;
Cac ai lng K
m
va R
MAX
co the xac nh bang nhieu phng phap khac nhau, nhng
hay s dung nhat la dung thc nghiem ket hp vi phng trnh nghch ao.
| |
t Xd
O d
R
2
=
HCMC March 2004
ONG HOC TAO SINH KHOI MONOD
Tng t nh Michaelis Menten, e mo ta toc o sinh trng cua vi sinh vat (sinh
khoi) Monod a a ra phng trnh sau:
Trong o, toc o tao sinh khoi la s gia tang sinh khoi trong mot n v thi gian
(ngay hay gi) t mot n v sinh khoi (X) goi la toc o tao sinh khoi rieng [1/ngay,
hay 1/gi):
Toc o tao sinh khoi cc ai, at c khi [S] >> K
S
; K
S
hang so Monod, o la nong
o c chat khi toc o tao sinh khoi bang
| |
| | S K
S
S
MAX
+
=
t

d X
dX
=
2
MAX

Hang so Monod K
S
phu thuoc vao chung sinh vat va c chat, c xac nh bang thc
nghiem trong bang
HCMC March 2004
ONG HOC TAO SINH KHOI MONOD
Khi lay nghch ao phng trnh Monod ta cung co cac dang khac nhau:
| |
MAX MAX
S
S
K

1 1 1
+ =
| | | |
| | S
K
hay
K S S
S
MAX MAX
S
MAX
+
= + =
1
1

HCMC March 2004
ONG HOC MONOD M RONG
Tren c s mo hnh Monod, e mo ta toc o sinh trng cua vi sinh vat (sinh khoi)
ngi ta con dung mo hnh cai tien sau:
Phng trnh Monod co the viet di dang:
S K
S
Xd
dX
S
m
+
= =
t

Hay:
Goi:
la he so chuyen hoa, va coi no la khong oi th:
t d dX
XS
S K
m
S
=
+
dS
dX
Y =
( ) S S Y X X + =
0 0
Thay vao ta co:

(
(

|
|
.
|

\
|
+ +

=
+
= =
} }
0
0
0
ln 1 ln
1 1
0
X
X
X
YK
X X
YS
X
YK
X
dX
S
S K
d
m
S
m m
S
m
S
S
S
m

t t
t
( ) | |
( ) | | S S Y X
S S Y X d
S
S K
X
dX
S
S K
d
S
m
S
m
+
+ +
=
+
=
0 0
0 0
1 1

t
Lay tch phan t S
0
en S:

Neu at:
0
1
ln
S
S
x =
m m
S m
X
YK X
a

+
=
1
X X
YS
x
m

=
0
2
ln
m m
S
X
YK
a

=
2
Th phng trnh tren co dang:
2 2 1 1
x a x a + = t
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
Co nhieu yeu to anh hng en toc o chuyen hoa c chat, trong o quan trong
nhat la anh hng cua:
Nong o c chat;
Nhiet o;
o pH;
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
1. Nong o c chat:
Toc o tieu thu c chat co the viet di dang:
Nen toc o tieu thu c chat bi VSV se la:
| | | |
| | S K
S
R
Xd
S d
R
m
MAX
+
= =
t
| | | |
| | S K
s
XR
d
S d
m
MAX
+
=
t
Khi lng VSV khong oi (X = const) th:
- Neu nong o c chat rat nho ([S] << K
m
), ta co K
m
+ [S] ~ K
m
nen:
| | | |
| |
| |
| |
m
MAX
H H
m
MAX
m
MAX
K
R
K S XK
K
S
XR
S K
S
XR
d
S d
= = ~
+
= :
t
- Bieu thc tren cho thay toc o tieu thu c chat ty le vi nong o c chat bac 1,
goi la mien phan ng bac 1
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
1. Nong o c chat:
- Khi ([S] << K
m
), ta co K
m
+ [S] ~ K
m
nen:
| | | |
| |
| |
| |
m
MAX
H H
m
MAX
m
MAX
K
R
K S XK
K
S
XR
S K
S
XR
d
S d
= = ~
+
= :
t
- Khi bien oi va tch phan ta co:
| |
| | S
S
XK
R
m
MAX
0
ln
1
t
=
- Ro rang, toc o cc ai ban au nho hn K
m
nhieu (S
0
nong o c chat tai thi
iem au: t = 0);
- Neu nong o c chat rat ln ([S] >> K
m
), ta co K
m
+ [S] ~ [S] nen:
| | | |
| |
| |
| |
MAX MAX
m
MAX
XR
S
S
XR
S K
S
XR
d
S d
= ~
+
=
t
ay toc o phan ng khong phu thuoc vao nong o c chat (phan ng bac 0,
oan c)
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
2. Anh hng cua nhiet o:
Moi loai VSV thch nghi trong mot ieu kien nhat nh cua moi trng (ve nhiet o,
pH ). Trong cac phan ng enzim, nhiet o co tac dung lam tang toc o phan
ng len toc o cc ai. Cung giong nh cac phan ng hoa hoc, khi nhiet o tang
len 10
0
C th toc o phan ng tang len khoang 2 lan, co ngha la he so nhiet o
cua phan ng khoang bang 2. o la do khi tang nhiet o hoat o xuc tac cua
enzim tang nh giam nang lng hoat hoa. Nh qua trnh hieu kh khi tang 1
0
C hoat
o tang gap oi. Noi chung khi tang 1
0
C, hoat o tang khoang 7,2 %. Nhng khi
nhiet o tang len nhiet o ti han (phan ln cac enzim khoang 60 70
0
C) th hoat
tnh cua chung b mat. ieu o co the giai thch bang tnh ac hieu xuc tac protein
cua enzim.
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
2. Anh hng cua nhiet o:
Moi enzim eu co mot nhiet o thch hp nhat cho hoat ong xuc tac ma o
phan ng enzim at toc o cc ai. Noi chung chung gan nhiet o c the, nh
ong vat nhiet o o khoang 40
0
C. Nhng cung co loai len en 70
0
C (nh o -
amilaza cac sinh vat chu nhiet o, hay trong thc vat, tham ch gan nhiet o soi
(trong VSV cac suoi nc nong).
nhiet o thap, hoat o cua enzim giam i, 0
0
C hoat o cua cac enzim khong
ang ke. Khi tang nhiet o th hoat o phuc hoi tr lai, nen ta co the bao ve cac
enzim nhiet o thap (di 20
0
C).
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
2. Anh hng cua nhiet o:
Trong phan ng enzim neu cho rang toc o phan ng ty le thuan vi hang so toc
o th ta co quan he:
Nh vay, tren toa o ln R :
RT
E
B R
E
MAX
= ln
T
1
quan he tren co dang ng thang
vi he so goc
R
E
E

HCMC March 2004

CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
2. Anh hng cua nhiet o:
Ngi ta co the bieu dien anh hng
cua nhiet o len toc o phan ng di
dang:
Trong o u = 1,05 1,1 la he so nhiet o
cua phan ng, thng lay bang 1,072.
( ) 15
15

=
t
t
R R u
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
2. Anh hng cua nhiet o:
Ngi ta co the bieu dien anh hng
cua nhiet o len toc o phan ng di
dang:
Trong o u = 1,05 1,1 la he so nhiet o
cua phan ng, thng lay bang 1,072.
( ) 15
15

=
t
t
R R u
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
3. Anh hng cua pH
Enzim rat nhay cam vi pH cua moi trng, nen pH co tac dung rat ln oi vi
toc o phan ng enzim. Moi enzim co mot gia tr pH thch hp cho s hoat ong cua
no, o toc o pgan ng ln nhat, gia tr khac toc o eu giam. Gia tr
thch hp cua pH con phu thuoc vao loai va nong o c chat, nhiet o, noi chung
no cung thay oi theo cac thong so nay.
Nhieu enzim co pH thch hp vung trung tnh, nhng cung co loai kha thap (pepsin
pH = 1,5), cung co loai khac cao (photphataza pH = 9 10)
.
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
3. Anh hng cua pH
Nhieu enzim co pH thch hp vung trung tnh, nhng cung co loai kha thap (pepsin
pH = 1,5), cung co loai khac cao (photphataza pH = 9 10)
.
HCMC March 2004
CAC YEU TO ANH HNG EN TOC O
CHUYEN HOA C CHAT VA TOC O TAO SINH
KHOI
4. Anh hng cua chat hoat hoa, c che va ion kim loai:
Chat hoat hoa se lam cho enzim t kem hoat ong tr nen hoat ong hn, hay t
khong hoat ong tr thanh hoat ong. Co nhieu loai khac nhau, qua v du: Cl
-
hoat
hoa enzim a amilaza; glution cho proteaza ;
Khi mot chat ket hp vi enzim lam cho no kem hoat ong, co the do bien tnh phan
t enzim, nhom hoat ong hay giam ai lc cua no vi c chat. Co nhieu loai chat
c che, tuyet oi, canh tranh
V enzim co ban chat protein, nen anh hng cua ion kim loai len hoat ong cua
chung cc ky phc tap. Nhieu loai enzim chu anh hng ro ret cua nong o va
ban chat cua ion kim loai. Cac ion kim loai co anh hng len tam hoat hoa, len
nhom chc hay chung tac dung vi enzim lam can tr viec tao phc ES, lam giam
ai lc vi c chat.
HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
e xac nh ca he so K
m
R
MAX
cua phng trnh Michaelis Menten (2.6) hay K
S
va
m
Max
cua phng trnh Monod (2.11), co nhieu phng phap. Thong thng ngi ta
tien hanh thc nghiem trong thiet b khuay tron ly tng hoat ong lien tuc (xem 3.)
Thc hien nuoi cay thuan chung, moi trng tinh khiet (mot loai c chat tinh). Gi
co nh thanh phan moi trng, thay oi lu lng dong vao sao cho vi moi lu
lng qua trnh at trang thai on nh.
Trc het cay vi sinh vat vao moi trng, khuay tron eu (at che o ly tng).
Thay oi lu lng, khi a at trang thai on nh (thng sau thi gian kha lau) th o
nong o c chat S [mg/l] va mat o VSV X au ra [mg/l].
HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
Khi at che o on nh th ta co:
Vi:
| | | | | | ( ) | | | | | | | |
R
X
S S
Q
V
X
S S
VX
S S Q
Xd
S d
=

=
t t
0 0 0
Q
V
= t
Do dong vao la moi trng tinh khiet nen [S
0
] = 0, coi khong co vi sinh vat dong
vao X
0
= 0, th can bang vat chat oi vi sinh khoi se la:
thi gian lu trung bnh cua cac phan t trong thiet b.

t t
= = =
1
VX
QX
Xd
dX
Nh vay khi the tch phan ng khong oi (V = const), th ng vi lu lng Q thay oi,
thi gian lu trung bnh thay oi theo, o cung co cac gia tr [S
0
], [S], va X tng ng.
Theo so lieu o c se xac nh toc o tao sinh khoi rieng
t

1
=
[1/h, hay 1/ngay] theo (2.21), va toc o tieu thu c chat | | | |
t X
S S
R

=
0
HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
Ket qua tnh toan c bieu dien theo o th (phng trnh Michaelis Menten) va o
th (phng trnh Monod). ong thi theo phng trnh nghch ao cua chung ta co
quan he:
la tuyen tnh.
Xay dng hai o th nay, t ay ta xac nh c tung o goc ng vi
he so goc tng ng la
| |
|
|
.
|

\
|
=
S
f
R
1 1
| |
|
|
.
|

\
|
=
S
g
1 1

S m
K K
1 1
va
MAX
S
MAA
S
K
R
K

va
T o xac nh K
m
, K
S
, R
MAX
va
MAX
.
Ta cung co the viet (2.6) di dang:
| |
| |
MAX
m
MAX
R
K
S R R
S
+ =
1 1
Bieu thc tren cung la c s e tuyen tnh hoa so lieu thc nghiem e xac nh
cac he so K
m
va R
MAX

HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
HCMC March 2004
XAC NH CAC HANG SO K
M
, R
MAX,
K
S
VA
M
MAX
BANG THC NGHIEM
| |
MAX MAX
S
S
K

1 1 1
+ =
| | | |
| | S
K
hay
K S S
S
MAX MAX
S
MAX
+
= + =
1
1

HCMC March 2004
ONG HOC PHAN NG ENZIM VI
NHIEU C CHAT
Phan ln cac enzim xuc tac cho nhng phan ng t hai hay nhieu
c chat, nh phan ng len men giam, phan ng x ly nc thai ... Cac
phan ng nay co ong hoc phc tap hn nhieu so vi phan ng mot
c chat.
Xet phan ng enzim E vi hai c chat S
1
va S
2
, th enzim se xuc tac
chuyen mot nhom ac hieu t c chat nay sang c chat khac.
Co the bieu dien nh sau:
Q P S S
E
+ +
2 1
Phan ng loai nay phc tap v co the tao ra nhieu phc hai thanh
phan gia enzim c chat: ES
1
, ES
2
, EP, EQ; hay phc ba thanh phan: ES
1
S
2
,
EPQ, ES
1
Q, EPS
2
Viec xac nh va R
MAX
cho nhng he hai c chat cung
c tien hanh tng t nh phan ng vi mot c chat.
HCMC March 2004
ONG HOC PHAN NG ENZIM VI
NHIEU C CHAT
Khi gi nong o mot c chat (gia s S
1
) mc bao hoa va
khong oi, tien hanh thay oi S
2
e xac nh .
Tiep theo lai gi S
2
mc bao hoa e xac nh va R
MAX
.
Nh vay, phan ng vi hai c chat co the a ve dang phan ng
mot c chat, khi mot trong chung c gi mc bao hoa khong oi.
Noi chung, cac phan ng vi hai c chat xay ra di hai dang: phan
ng hai c chat mot bc hoac hai bc. Co the phan biet hai dang nay
bang phan tch ong hoc ket hp vi thc nghiem. Neu nh san pham
cuoi cung cua phan ng c che phan ng tong bang c che canh tranh
vi c chat dan ng, th o la phan ng hai c chat co th t.
HCMC March 2004
ONG HOC PHAN NG ENZIM VI
NHIEU C CHAT
Phan ng hai c chat mot bc
Khi ca hai c chat S
1
va S
2
cung ong thi co mat trung tam
hoat ong cua enzim e tao ra mot phc ba thanh phan th phan ng
mi xay ra. Chung co the xay ra mot cach ngau nhien hay co th t.
Phan ng hai c chat mot bc ngau nhien
Xay ra khi bat ky mot trong hai co chat eu co the ket hp trc
vi enzim e tao phc ba thanh phan theo mot trong hai dang:
2 1 2 1 1 1
S ES S ES ES S E + +

2 1 1 2 2 2
S ES S ES ES S E + +

La ong kha nang
HCMC March 2004
ONG HOC PHAN NG ENZIM VI
NHIEU C CHAT
Phan ng hai c chat mot bc co th t
Co mot c chat goi la c chat dan ng phai ket hp trc vi
enzim, sau o mi co the tac dung vi c chat th hai e tao phc ba
thanh phan
Phng trnh tong quat cho phan ng hai c chat co th t vi S
1
la
c chat dan ng mo ta nh sau:
Hay dang nghch ao (e xac nh K
m
S
va R
MAX

2 1 2 1 1 1
S ES S ES ES S E + +

o sau
| || | | | | |
1
2 1 2 1
2 1 2 1
+ + +
=
S
K
S
K
S S
K K
R
R
S
m
S
m
S
m
S
m
MAX
| | | | | |
|
|
.
|

\
|
+ +
|
|
.
|

\
|
+ =
2 1 2
2 2 1
1
1
1 1 1 1
S
K
R S S
K K
K
R R
S
m
MAX
S
m
S
m
S
m
MAX
HCMC March 2004
ONG HOC PHAN NG ENZIM VI
NHIEU C CHAT
Phan ng hai c chat hai bc
Mot c chat S
1
phai c gan vao enzim E tao phc ES
1
roi giai
phong ra mot san pham P va E
*
. Sau o khi c chat th hai S
2
ket hp
vi enzim E
*
tao phc E
*
S
2
th giai phong ra san pham th hai Q va E.
Trong giai oan au mot nhom chc cua c chat th nhat chuyen cho
enzim. Nhom chc nay se chuyen cho c chat th hai trong giai oan sau.
Phng trnh bieu dien toc o phan ng thay oi theo cac c chat
nh sau:
Dang nghch ao
| | | |
2 1
2 1
1
S
K
S
K
R
R
S
m
S
m
MAX
+ +
=
| | | |
|
|
.
|

\
|
+ + =
2 1
2 1
1
1 1 1
S
K
R S R
K
R
S
m
MAX MAX
S
m
HCMC March 2004
ONG HOC PHAN NG ENZIM VI
NHIEU C CHAT
Phan ng hai c chat hai bc
.