102

Chng 5
Trao i cht vi sinh vt
I. Cc con ng phn gii hexose
1. Con ng Embden - Meyerhof - Parnas (EMP hay ng phn)
1.1 C ch (hnh 5.1)
Glucose
(1)
Glucose - 6 P
(2)
Fructose - 6P
ATP
(3) ADP
Fructose - 1,6 DP
(5) (4)

(AlPG) Glyceraldehyd - 3P Dioxy acetone P (DAP)

H3PO4 NAD
(6)
NADH2
a. 1,3 diphosphoglyceric (a. 1,3DPG)
ADP
(7)
ATP
a.3 P - Glyceric (A3PG)
(8)
a.2P- Glyceric (A2PG)
(9)
Phosphoenolpyruvic (PEP)
(10) ADP
A TP

Trong iu kin c hoc khng c O2, glucose u c chuyn

a. Pyruvic
thnh pyruvate qua 10 phn ng, trong 3 phn ng 1, 3 v 10 l phn
Hnh
ng mt chiu, cn cc 5.1:
phn Con
ng ng
khc EMPng thun nghch. Tr cht
l phn
 103

u v cht cui, tt c cc cht trung gian trong chu trnh u dng

phosphoryl ha. Gc phosphoryl mang 3 chc nng l gip cho cht trung
gian mang in tch m cao khng th thot ra ngoi, l v tr gn enzyme
v l v tr cung cp lin kt cao nng.
Phng trnh chung:
Glucose 2 pyruvate + 2ATP + 2 NADH2
ATP c to thnh t 2 phn ng 7 v 10. S to thnh ATP 2
phn ng trn gi l phosphoryl ha mc c cht v phosphate cao
nng trc l mt phn ca phn t cht trao i (1,3 di - P - glycerate
v PEP), khi c mt ca ADP th cht trao i s chuyn gc P cho ADP
to thnh ATP.
C th chia con ng EMP thnh 4 giai on:
1. Hot ha glucose to fructose - 1,6 diphosphate (c th xy ra
nhiu monosaccharide khc).
2. B i phn t fructose - 1,6 diphosphate to AlPG v DAP.
3. xy ha AlPG thnh A2PG.
4. Chuyn ha A2PG to acid pyruvic (hnh 5.1).
1.2. ngha ca con ng EMP
- Qu trnh ng phn cung cp cho t bo 6 trong s 12 tin cht
dng tng hp cc n v kin trc: glucose - 6-P, fructose - 6-P, 3-P-
glyceraldehyd, 3P - glycerate, PEP, pyruvate.
- Cung cp nng lng cho hot ng sng ca t bo tuy hiu sut
khng cao (khong 10%), nhng y l con ng nguyn thy nht gp
hu ht sinh vt.
2. Con ng pentosophosphate xy ho (PP, HMP)
Do mt t bin no (thiu enzyme) m mt s vi sinh vt c th
chuyn ha glucose thnh pyruvate theo con ng PP (hnh 5.2).
T glucose qua qu trnh decarboxyl ha to ribuloso - 5P v CO2.
Cc phn ng tip theo l s chuyn ha gia pentoso- 5P v hexoso - 6P.
ngha ca chu trnh: Xt v mt nng lng, t 1 phn t glucose
khi phn gii to 35 ATP nhng qua con ng ny to riboso - 5P tham
gia vo qu trnh sinh tng hp acid nucleic, to cc loi ng C4, C7...
tham gia vo qu trnh sinh tng hp acid amin. Con ng ny to
NADPH2 cn thit cho cc phn ng sinh tng hp trong t bo. y l
con ng phn gii ng thng gp vi khun thuc ging
Pseudomonas.
 104

Phng trnh tng qut:

Glucose + 6O2 6CO2 + 6H2O + 35ATP

Glucose

Glucose

Ribulose
Glucose

Fructos Frutose Xilulose

Ribose
Erytrose
Xilulose

Xilulose
Erytros
Ribose
Fructos Fructos
Xilulose

Hnh 5.2 Con ng pentosophosphate

3. Con ng KDPG (2keto - 3deoxy - 6Pgluconate)

Phng trnh tng qut nh sau:
Glucose 2 Pyruvate + ATP + NADH2 + NADPH2
Con ng KDPG gip cho nhiu vi khun s dng gluconate v ch
gp mt s vi khun. E. coli v nhiu loi ca Clostridium chuyn ha
glucose qua con ng EMP nhng phn gii gluconate qua con ng
KDPG. Qua cc con ng phn gii glucose, pyruvate c to thnh.
Nu trong iu kin hiu kh cc vi sinh vt s xy ho pyruvate to
CO2 v nc. Nu iu kin k kh cc vi sinh vt s tin hnh qu trnh
ln men hoc h hp k kh (hnh 5.3).
 105

Glucose

Glucose -6-
h h

6 - phosphoglucono-18-lactose

6 - phosphogluconate

2-keto-3 deoxy-6-phosphogluconate

Glyceraldehyd pyruvate

1,3 Phosphoenol-pyruvate

3-phosphoglycerat 2-phosphoglycerate

Hnh 5.3 Con ng KDPG

4. xy ho pyruvate
Pyruvate chim v tr trung tm trong trao i cht trung gian v l
tin cht ca nhiu sn phm. Nhiu vi sinh vt xy ha pyruvate thnh
acetyl - CoA ch yu qua 3 phn ng sau (hnh 5.4):
 106

Pyruvate +CoA+NAD+ Acetyl - CoA + NADH2 + CO2 (1)

Pyruvate + CoA + 2Fd Acetyl - CoA + 2FdH + CO2 (2)
Pyruvate + CoA + NAD+ Acetyl - CoA + formate (3)
(Formate - Fd - ferredoxin)

Pyruvate-
carboxylase

Vng tiasolium
Pyruvate
ca thiamin
pyrophosphate
Enzyme

Acid lipoic

Dihydrolipoid-
transacetylase

Enzyme Dihydrolipoid
-
dehydrogenas

Enzyme Enzyme
Acid acetyl-lipoic Acid dihydrolipoic

Hnh 5.4 Qu trnh xy ha pyruvate

Phn ng (1) do phc h pyruvate - dehydrogenase (PDH) xc tc.

PDH gp hu ht vi sinh vt hiu kh (khng gp vi khun k kh bt
buc). PDH gm 3 enzyme, ngoi CoA v NAD+ cn cn TPP (thiamine -
pyrophosphate) v acid lipoic. Phn ng (2) xc tc bi pyruvate - Fd -
oxidoreductase, enzyme ny gp nhiu vi khun k kh nh Clostridium.
Phn ng (3) do pyruvate - formate - liase xc tc, enzyme ny gp nhiu
vi khun k kh tit acid formic (thuc h Enterobacteriaceae) v vi
khun quang nng.
 107

II. Chu trnh tricarboxylic (Krebs)

Trong qu trnh xy ha hon ton, pyruvate s c xy ho trit
thng qua chu trnh Krebs (chu trnh acid tricarboxylic = ATC).
C ch c trnh by hnh 5.5.

Hnh 5.5 Chu trnh Krebs (theo Lehninger)

Trong chui phn ng vng ca chu trnh ATC, acid pyruvic b phn
hu trit to CO2, coezyme dng kh v cc hp cht trung gian. Cc
coezyme thng qua chui h hp vn chuyn H v electron n cho O2
phn t to nc ng thi gii phng nng lng ATP.
 108

ngha ca chu trnh: Ngoi chc nng xy ha tn cng, chu trnh

ATC cn cung cp cho t bo 3 tin cht l oxalacetate (tng hp
aspactate), - ketoglutarate v succinyl - CoA (tng hp nhn hem). V
nng lng, chu trnh cung cp 1ATP mc phosphoryl ha c cht
nhng sn sinh NADH2 i vo chui vn chuyn electron to ATP.
Trong iu kin k kh chu trnh TCA khng c chc nng sinh nng
lng nhng cng phi cung cp cho t bo 3 tin cht trn. T bo phi
s dng phn ln pyruvate ln men thu nng lng, cn phn nh
chuyn thnh acetyl - CoA nh pyruvate - ferredoxin - oxidoreductase.
Phn ln cc vi sinh vt hiu kh xy ha cht dinh dng hu c
thnh CO2 v H2O (v carbon trong phn t CO2 t mc xy ha cao
nht nn ngi ta gi l qu trnh xy ha hon ton). Mt s vi sinh vt
c kh nng xy ha c cht trong iu kin hiu kh nhng li sinh ra cc
sn phm cha c xy ha hon ton (ketoacid, acid gluconic, acid
malic, acid fumaric...). Nhiu qu trnh xy ha khng hon ton c
ngha quan trng trong cng nghip ln men v chng sn sinh ra nhiu
sn phm c gi tr.
III. Chui h hp (respiratory chain) v phosphororyl ha xy
ha (oxidative phosphorylation)
Khc vi vi khun k kh, vi khun hiu kh c th tng hp ATP mnh m
hn nhiu nh c chui h hp v ATP - synthetase. Hai h thng ny nm
mng t bo Prokaryota hoc mng trong ti th ca t bo Eukaryota. Nng
lng thot ra c chuyn thnh ATP, mt phn nh gii phng ra dng
nhit. Cc thnh phn ca chui nm trong lp lipid kp, gm nhiu enzyme vn
chuyn electron v hydro, cc coenzyme v nhm thm, cc dehydrogenase v
h thng vn chuyn. Cc thnh phn quan trng nht tham gia vo xy ha
hydro l :
- Flavoprotein l cc enzyme cha nhm thm l FMN hoc FAD - vn
chuyn hydro.
- Protein Fe-S vn chuyn electron, cha nguyn t Fe, va lin kt vi S
ca cysteine va lin kt vi S ca H2S. Cysteine l mt phn ca chui
polypeptide, c th coi trung tm Fe - S l nhm thm ca chui polypeptide.
Cc protein Fe - S cng tham gia vo s c nh N, kh
sulphite, kh nitrite, quang hp...
- Quinol: nm mng trong ca ti th, vi khun G- l ubiquinol
(CoQ), vi khun G+ l naphtoquinol v lc lp l plastoquinol. Quinol,
c bit l CoQ c c tnh a lipid do lp vo lp lipid kp ca mng,
vn chuyn hydro hoc enzyme.
 109

- Cytochrome: vn chuyn electron, khng vn chuyn hydrogen.

Cytochrome nhn electron t quinol, trong qu trnh vn chuyn mt s
lng H+ tng ng vi s lng electron b tch vo mi trng (hnh
5.6).

Cyt Cyt Cyt Cyt

Hnh 5.6 S chui h hp

Trong qu trnh vn chuyn 2[H] t NADH2 n O2 c 3 bc

chuyn electron lin quan n to thnh ATP v c biu th bng h s
P/O (mol ATP/mol nguyn t O). H s P/O = 3 khi 2[H] c chuyn
qua NAD+ v P/O = 2 khi 2[H] t succinate qua FAD n O2. Do 1mol
glucose khi phn gii qua chu trnh EMP v TCA vi tt c [H] tch ra
c chuyn vo chui h hp to thnh nc s to 38 ATP. S tnh
ton trn ng vi ti th v mt s vi khun. Tuy nhin, mt s vi khun
do ch c 2 v tr phosphoryl ha nn s h hp hiu kh glucose ch cho
26 ATP (nh E. coli khng c NAD, Proteus vulgaris khng c
cytochrome C).
C ch phosphoryl ho xy ho (s to thnh ATP trong chui h
hp) c gii thch theo thuyt ho thm thu ca Mitchell (1979). Theo
thuyt ny, mng t bo vi khun v mng trong ti th khng thm cc ion
k c H+ v OH- v km dn in. S sp xp cc protein ca mng (cc
thnh phn vn chuyn electron, permease, ATP - sinthetase...) khin
mng mang tnh cht khng i xng. S nh hng khng gian ca cc
phn t enzyme dn n trao i cht c c tnh vect. Chui h hp bao
gm mt dy cc cht vn chuyn hidro v electron sp xp lun phin
trong mng. Do s xy ho c cht m pha trong mng c s tiu th H+
v pha mng ngoi gii phng H+. Trong s xy ho NADH2, 6H+
 110

c vn chuyn ra pha ngoi qua 3 nm. Vic vn chuyn proton do h

hp lm xut hin mt gradien in ho gia 2 pha ca mng. Th proton
ny s l ng lc ca s phosphoryl ho ngha l s to thnh ATP. Qu
trnh c xc tc bi ATP - sinthetase, enzyme ny chuyn nng lng
pht ra t dng electron thnh lin kt esterphosphate cao nng ca ATP.
ATP - sinthetase gp cc mng tham gia vo s chuyn ho nng lng
l mng ti th, lc lp v mng t bo vi khun. C l, dng H+ t ngoi
i qua rnh ca ATP - sinthetase tr vo bn trong ti th hoc vi khun,
nng lng thot ra c dng cho phn ng phosphoryl ho to ATP.
IV. S vn chuyn cht dinh dng vo t bo
tn ti, sinh trng v pht trin, t bo vi sinh vt phi thng
xuyn trao i vt cht v nng lng vi mi trng bn ngoi. Mt mt
chng nhn cc cht dinh dng cn thit t mi trng, mt khc chng
thi ra cc sn phm trao i cht. Gia mi trng xung quanh v mi
trng bn trong t bo tn ti mt hng ro thm thu, hng ro ny
chnh l mng t bo cht.
T bo nhn v thi cc cht mt cch chn lc. S xm nhp ca
nc v cc cht ha tan qua mng t bo l mt qu trnh ng hc; t
bo sng khng bao gi trng thi cn bng vi cc cht ca mi trng
chung quanh. S vn chuyn cc cht qua mng t bo vi sinh vt tun
theo mt trong hai c ch: khuch tn n gin (hay cn gi l vn chuyn
th ng hay vn chuyn xui dng) v vn chuyn ch ng (vn chuyn
ngc dng).
Theo c ch khuch tn th ng cc phn t i qua mng nh s
chnh lch nng (trong trng hp cc cht khng in phn) hay
chnh lch in th (trong trng hp cc ion) hai pha ca mng. Hng
lot nghin cu khng nh, tr nc ra rt t hp cht c th c vn
chuyn qua mng theo c ch ni trn.
Tri li, a s cc cht ha tan qua mng do c ch vn chuyn c
bit. Nhng phn t vn chuyn sp xp trong mng lin kt vi cc phn
t cht ha tan v chuyn chng vo b mt bn trong ca mng, t y
cc phn t cht ha tan c chuyn vo t bo cht. Kiu khuch tn
ny c gi l khuch tn xc tin. Cc phn t protein vn chuyn
(carrier protein) ni trn gi l protein thm permease.
S vn chuyn cc cht nh permease c th l th ng (khng cn
nng lng ca t bo) hoc ch ng (cn nng lng). Theo c ch vn
chuyn th ng, cht ha tan lin kt thun nghch vo mt v tr c bit
trn phn t permease nm bn trong mng (c th cc l ca mng).
Phc hp cht ha tan - permease c vn chuyn theo hai pha ca
 111

mng nh s chnh lch nng mt cht no , ngha l s vn

chuyn din ra theo kiu xui dng. S vn chuyn th ng nh
permease c chng minh mt s vi sinh vt.
Tuy nhin, vi sinh vt c kh nng tch ly mt s cht vi nng
cao hn nhiu so vi nng bn ngoi. Chng hn, nng K+ bn trong
mt s t bo vi sinh vt c th ln hn nng bn ngoi hng ngn ln.
m bo trung ha in, t bo cng ng thi thi ra bn ngoi cc
ion H+ hoc Na+. Thm vo , ngi ta pht hin thy cc mng vi
khun c hot tnh ca ATP - ase l enzyme c lin quan n vic vn
chuyn cc cht.
R rng, trong t bo vi sinh vt ngoi c ch vn chuyn th ng
cn tn ti c ch vn chuyn ch ng nh permease. S vn chuyn ny
c tin hnh ngc vi gradien nng ngha l theo kiu ngc
dng, nng lng tiu th c th do ATP cung cp.
Cho ti nay ngi ta phn lp c hng lot protein vn chuyn
trong cc loi vi sinh vt. Ging nh enzyme, chng c tnh c hiu c
cht khc nhau. Mt s c tnh c hiu gn nh tuyt i. Chng hn,
permease ca galactose E.coli ch vn chuyn galactose. Cc permease
ca cc ng v acid amin khc th hin tnh c hiu yu hn i vi
cc cht ha tan.
iu ng ch l trong vi khun s vn chuyn ch ng ca hu
ht cc loi ng ph thuc vo qu trnh phosphoryl ha v h thng
enzyme phosphotransferase. H thng ny bao gm hai enzyme EI, EII v
mt protein vn chuyn bn nhit (Hpr: heat stable carried protein) c khi
lng phn t thp. Cc thnh phn protein ca h thng ny c
thun khit v phn ng din ra theo hai bc.
Trc ht EI chuyn phosphate t phosphoenolpyruvate (PEP) n
Hpr:
Hpr + PEP Hpr - P + Pyruvate
Sau EII chuyn phosphate t Hpr - P n C6 ca ng n.
EI l chung cho nhiu loi ng nhng EII li c trng cho mi
loi ng. Ngha l mt t bin no nh hng n vic tng hp EI
s dn n mt kh nng vn chuyn nhiu loi ng. Tri li vi t
bin nh th i vi EII ch nh hng n s vn chuyn ca mt loi
ng (hnh 5.7).
 112

Mng Trong
Ngoi

EII Glucose- 6P
Glucose
Hpr
PEP

EII glucose Hpr-P pyruvic

Hnh 5.7 S vn chuyn ng qua mng t bo vi sinh vt

V. Trao i cht v nng lng

1. Cc ngun nng lng vi sinh vt
Ty theo bn cht ca ngun nng lng s cp, c hai con ng
c bn cung cp cho vi sinh vt: con ng quang dng v ha
dng. Trong c hai trng hp, s thu nhn nng lng ca t bo c
lin h vi s vn chuyn electron qua mng, do to ra mt gradient
electron proton t phn ny i vi phn kia ca mng t bo cht. Chnh
gradien nng ion l ngun gc ca lc bm proton (s khc bit in
tch) c th c dng vo nhng mc ch khc nhau nh vn chuyn
tch cc, sn xut cc cht giu nng lng, chuyn ng v.v.. .
Trong th gii vi sinh vt c s khc bit rt ln v kh nng s
dng cc ngun nng lng. Mt s vi khun hiu kh cng ging nh
ng vt c kh nng tng hp ATP cho mnh t ng b phn gii theo
con ng ng phn (glycolyse) v chu trnh Krebs nh chui h hp
trn mng t bo cht (tng t nh chui h hp mng trong ca ty
th).
Cn nhng vi sinh vt k kh li thu nng lng cho hot ng sng
ca mnh nh qu trnh ln men ca cc loi ng hoc nh mt loi
chui vn chuyn electron trong c s dng cc hp cht khng phi l
xy phn t lm cht nhn electron cui cng trong chui vn chuyn.
y, chui vn chuyn cng khu tr trn mng t bo cht tng t nh
chui vn chuyn nm trong cc ty th. i vi cc c th quang dng
th li khc, chng c th thu nhn nng lng ca nh sng.
 113

ng v quan im nng lng sinh hc, th ty th, lc lp v t bo

vi khun c nhiu nt tng ng. Tt c cc vi khun c trn mng t bo
cht mt h enzyme ATP - synthetase tng t nh ATP - synthetase ca
cc ty th v lc lp. Phc hp protein vn chuyn qua mng m bo
bin i dng proton thnh nng lng trong mi lin kt phosphate
dng ATP.
1.1.Cc c th quang dng (phototroph) v quang tng hp
(photosynthesis)
Hnh thi ca c th quang dng v quang tng hp gia vi khun,
to, thc vt c s khc bit r rng. Mt khc, cu to cc c quan quang
hp, sc t quang tng hp, cng khc bit (xem chng II). i vi thc
vt, cht cho electron l H2O v qu trnh quang hp gii phng xy phn
t. i vi vi khun quang hp khng thi xy v cht cho electron l
nhng hp cht v c (nh H2S, S) hoc hp cht hu c (isopropanol)
(xem thm chng VII).
1.2. Cc c th ha dng v xy ha sinh hc
Phn ln vi khun khng c c quan v sc t quang hp. Chng ly
nng lng hu ch cho cc qu trnh tng hp ca t bo nh gii phng
nng lng trong cc phn ng ha hc.
S trao i cht ca cc c th ho dng ni chung bao gm 3 giai
on:
- Phn ct cc i phn t ngoi t bo.
- Phn gii cc phn t b to ra cc cht trao i trung gian
(pyruvate, acetyl - CoA) v to nng lng hu ch (ATP).
- Phn gii trit cc cht trao i trung gian thnh CO2 v H2O
vi vic to ra lng ln ATP. l nhng phn ng xy ha c cht hu
c v v c.
Qu trnh xy ha mt c cht A c th xem l mt chui phn ng
m trong c cht A mt i cc eletron ca mnh. C cht A gi l cht
cho electron s bin thnh mt sn phm c xy ha. Tng t, mt
hp cht B khc gi l cht nhn electron s bin thnh sn phm kh.
Trong phn ln trng hp, mt hp cht A l mt cht hu c c
th xem l cht c hydro ha, nh vy th qu trnh xy ha v thc
cht l mt qu trnh kh hydro (dehydrogenation), trong khi qu trnh
kh l qu trnh hydro ha (hydrogenation), t hp hai qu trnh ny l
qu trnh xy ha kh (oxidoreduction).
Qu trnh xy ho:
 114

- 2H+
AH2 (cht cho hydro) A (sn phm xy ho) + Nng lng
- 2e
Qu trnh kh:
+ 2H+
B (cht nhn hydro) BH2 (sn phm kh)
+ 2e
Kt qu:
AH2 + B A + BH2 + Nng lng
Trong chui phn ng, vic vn chuyn hydro hay electron t c
cht sang cht nhn c thc hin nh hng lot enzyme, cc enzyme
ny to thnh chui vn chuyn in t.
Cc cht xc tc thch ng cao vi qu trnh xy ho c cht l
nhng dehydrogenase. Nhng enzyme ny c trng cho mt loi c cht
v t hp vi cc coenzyme m n ng vai tr l cht nhn hydro t c
cht. l cc dn xut ca flavin (FMN, FAD) hoc ca pyridine (NAD,
NADP) v nhiu hp cht khc. cc c th hiu kh, s vn chuyn cc
electron gia coenzyme ca dehydrogenase v xy phn t c thc hin
nh cc hp cht trung gian ca cytochrome.
2. Cc kiu h hp
Thng thng khi cht nhn electron cui cng l xy phn t th
gi l qu trnh h hp hiu kh v vi sinh vt thuc dng ny gi l vi sinh
vt hiu kh (aerobes). Khi cht nhn electron cui cng l mt c cht
khc khng phi l xy t do, th ngi ta gi l qu trnh ln men hoc c
th l qu trnh h hp k kh v vi sinh vt thc hin cc qu trnh ny gi
l cc c th k kh (anaerobes).
Trong s cc c th k kh, cht nhn electron c th l cht hu c
hoc cht v c. Nu cht nhn electron l cht hu c ngi ta gi l ln
men, nu cht nhn l electron l cht v c ngi ta gi l h hp k kh
(anaerobic respiration).
H hp ca mt c th hiu kh l mt qu trnh vn chuyn electron
v proton qua mng vi xy phn t lm cht nhn electron cui cng, cn
h hp k kh dng ch mt qu trnh vn chuyn qua mng ca cc
electron v proton n cht nhn electron cui cng l mt cht khc xy
phn t, nh nitrate trong trng hp h hp nitrate (nitrate respiration),
fumarate nh h hp fumarate (fumarate respiration ) v.v...
 115

Rt nhiu vi sinh vt c nhiu chui vn chuyn electron c th cng

hot ng, hoc c hot ng trong nhng iu kin nui cy nht nh.
V d nh E. coli, vi khun ng rut ny c chui vn chuyn
electron vi xy phn t l cht nhn electron cui cng (chui hot ng
nh c th hiu kh ) v chui h hp nitrate hot ng k kh trong
mi trng nitrate, mt chui h hp fumarate, hot ng k kh trn
mi trng cha c cht hu c m vi khun ny c th ln men.
H hp c c trng trc ht l ni khu tr ca chui vn
chuyn electron mng t bo cht i vi cc c th nhn s v mng
trong ca ty th i vi cc c th nhn chun. H hp cn c trng
bn cht ca cht nhn electron cui cng l xy phn t. S xy ho c
cht v ng thi s kh ca xy c thc hin nh mt chui phn ng
enzyme, trong c s tham gia ca dehydrogenase v cc coenzyme lin
kt vi n.
C ch thng thy h hp l con ng ca cc cytochrome gin
tip. Ngy nay ngi ta hon ton bit r thnh phn v c ch hot ng
ca cc cytochrome ny trong t bo nhn chun v trong mt s c th
nhn s.
Bn cnh qu trnh phosphoryl xy ho c cht, cn tn ti nhng
con ng khc, c bit l con ng xy ho trc tip nh cc enzyme
vn chuyn cc electron t c cht n xy vi s to thnh H2O2 .
FADH2 + O2 FAD- oxydase FAD + H2O2
Hp cht ny rt c v cn phi c phn gii nh catalase,
peroxydase hoc superoxyd dismutase trnh cho t bo khi b cht.
H2O2 catalase H2O + 1/2 O2
+ -
H2O2 + 2H + 2e peroxydase 2H2O

Nhng con ng ny to rt t hoc khng to nng lng hu ch.

Mt vi khun khng c enzyme trn, nhng con ng phn gii trn buc
chng phi sng k kh bt buc, bi v xy phn t l tc nhn gy c
vi chng. C mt s vi khun chu c hiu kh nh Streptococcus,
chng khng c catalase, nhng c cc enzyme flavin (NAD - oxydase v
NAD - peroxydase) cho php chng thc hin s cn bng trong h hp
mng t bo cht. Tu thuc vo s lng v lng cc enzyme c th
phn gii H2O2 m quyt nh tnh hiu kh hay k kh cc vi khun.
Ln men ch s c mt ca chui vn chuyn electron nm trong
t bo cht, khng c s t ng i qua ca dng electron hoc proton
phn bn ny v phn bn kia ca mng t bo (mt dng vn chuyn th
 116

cp c thit lp nu cn thit, nh nng lng ca ATP). V phng

din trao i nng lng, y cng l mt qu trnh xy ha kh, nhng
cht nhn electron cui cng l cc hp cht hu c, ch khng phi l
cc phn t xy. Qu trnh ny cng gii phng nng lng nhng t hn
nhiu so vi qu trnh h hp. Cc hp cht cho electron (AH2) v cht
nhn electron (B) u l cc hp cht hu c. Vic vn chuyn electron
c thc hin nh NAD. y c s khc bit r rt so vi h hp v
bn cht ca cht vn chuyn electron v sn phm cui cng...
C ch vn chuyn electron trong qu trnh ln men c tm tt
nh sau:
AH2 NAD+ BH2

A NADH2 B
3. Nghin cu s trao i nng lng
S khc bit v cc phn ng xy ha kh cc nhm vi sinh vt
khc nhau l mt tiu chun dng nh loi chng, mt s nh nghin
cu xut nhng phng php v mi trng c th nh loi nhanh
chng v n gin cc nhm vi sinh vt.
3.1 Nghin cu kiu h hp
xc nh kiu h hp ca mt loi vi khun, trong phng th
nghim ngi ta s dng mi trng VF (nc tht - gan) ch c v
a vo ng nghim, ri cy ging vo su, da vo kh nng pht trin
trong cc ng nghim ta bit c l loi h hp g.
3.2 Dng phn ng oxydase
Phn ng ny cho php s c mt ca chui h hp ang hot ng
vi cytochrome C mng nh TMPD (tetra - methyl - paraphenylene -
diamine). Phn ng ny c bit hiu qu i vi vi khun G-, phn ng
dng tnh vi cc vi khun hiu kh bt buc (tr Pseudomonas
maltophyla v Acinatobacter), phn ng m tnh i vi cc vi khun k
kh khng bt buc tr cc Vibrio v Aeromonas. c bit i vi cc vi
khun ng rut l c phn ng m tnh mc d vi khun ny c chui
h hp vi cc cytochrome.
3.3 Th catalase
Enzyme catalase thng c cc vi khun G-, tr cc ging
Streptococcus v Lactobacilus. Khi c mt H2O2, mt vi git huyn ph
vi khun c catalase dng s lm si bt gii phng xy phn t. Nu:
 117

- Si nhiu bt: l huyn ph vi khun hiu kh

- Si t bt: vi khun hiu - k kh
- Khng c bt: vi khun k kh
3.4 Phn ng nitrate reductase
S kh nitrate c th c nghin cu trn mi trng c nitrate
(mi trng lng hoc c) pht hin s c mt ca nitrite (nh thuc
th Griess), c th dn n 2 kt qu theo 2 c ch khc nhau:
- Kh nitrate ng ha c xc tc bi nitrate reductase B to
nitrite, cht ny c th c chuyn ha tip thnh mui ammon l ngun
nit d c c th s dng.
- Kh nitrate d ha hay h hp nitrate di nh hng ca nitrate
reductase A v cc enzyme khc, cc nitrate s c kh hon ton thnh
N2O v N2 s c gii phng ngoi t bo. H hp nitrate l loi h hp
k kh.
3.5 Nghin cu kiu trao i cht
Nghin cu ny c thc hin nh s dng glucose lm c cht,
loi ng c dng ph bin nht nhiu loi vi khun. Th nghim
cn nui cy 2 iu kin hiu kh v k kh. Sau khi nui t m vi
nhit thch hp, c th c 4 trng hp xy ra:
- Vi khun ln men khi c ging pht trin trong canh trng v lm
acid ha trong 2 ng nghim.
- Vi khun xy ha khi n ch lm acid ha trong ng nghim iu
kin hiu kh.
- Vi khun bt hot khi khng thy acid ha trong c 2 ng nghim.
- Vi khun kim ha khi thy mi trng b kim ho trong iu
kin hiu kh, iu ny c th thy khi dng peptone.
3.6 Kh cc sn phm lu hunh
Th nghim kh cc sn phm xy ha ca lu hunh (sulfate,
sulfite, thiosulfate v.v...) c th thy r nh to ra hydro c lu hunh
ha (H2S...) pht hin di dng sulfua kim loi b en (sulfua st hay
sulfua ch...).
3.7 o mc h hp
H hp k (Respirometre) Warburg l dng c u tin dng xc
nh mc s dng xy ca t bo sng. N bao gm mt bnh cch
thy chu nhit mu cn nghin cu. Mu ny c t trong mt bnh
cha ni vi mt p k, bnh cha mu v p k to thnh mt nhnh c
 118

c nh chc chn trn ng c mc nc di ng, s di chuyn y l do

s giao ng ca mu nghin cu gy nn.
Bnh cha mu c ni vi mt nhnh ca bnh cong ca p k v
c iu chnh c mt dung tch kh nht nh phn trn ca mu
nghin cu. loi b nh hng ca CO2 sinh ra trong qu trnh h hp,
ngi ta t mt ng nh gia bnh ng mu nghin cu, trong , cha
KOH 10% hp th CO2 do qu trnh h hp sinh ra. S thay i p sut
trong p k s c tnh chuyn nh dung tch xy tiu th.
Vi dng c h hp k ny khng nhng dng o s tiu th xy
phn t m cn c th nghin cu cc phn ng enzyme khc na, trong
s thay i kh xut hin do kh carbonic, kh amin ha.
4. S tch tr v s dng nng lng
4.1. Cc mi lin kt giu nng lng
T bo sng tch tr nng lng vo cc hp cht v chng c th
c gii phng d dng khi t bo cn. Phn ng ha hc ph bin l
thy phn mi lin kt esterphosphate. Hp cht c t bo s dng
thun li v thng xuyn nht l ATP. Ngoi ra cn c cc hp cht giu
nng lng khc nh cc nucleotide UTP, GTP, CTP, TTP; cc
acylphosphate, v d nh 1-3 diphosphoglycerate; cc enolphosphate nh
PEP; cc acyl - thio - ester nh CoA - SH.
CH2

R- C-OP
V d: PEP (G = - 53,5 Kj.mole -1)
+ Cc acyl - thioester: O

R-C-SP V d: Coenzyme A
4.2. Ngun gc hp cht cao nng
ATP c th c tng hp bng hai c ch:
- Qu trnh phosphoryl ha c cht xy ra trong t bo cht.
- Qu trnh phosphoryl ha lin h vi gradien electron v proton
xy ra mng t bo cht, y ngi ta thy c qu trnh phosphoryl
xy ha v phosphoryl quang ha (photophosphorylation).
Trong c hai trng hp qu trnh tng hp ATP ph thuc vo
lng ADP v phosphate v c.
 119

+ Qu trnh phosphoryl ha c cht: trong qu trnh ny, mt

phosphate v c c lin kt (nh xc tc bi enzyme) vi cc sn phm
ca s phn gii. Hp cht xy ha l cht mang mt mi lin kt
phosphate giu nng lng v c th c chuyn n ADP. Kiu hnh
thnh ATP ny thng thy s phn gii glucose theo con ng EMP
(giai on xy ha 3 - phosphoglyceraldehyd thnh acid 1,3-
diphosphoglyceric v s bin i hp cht ny thnh acid pyruvic).
+ Qu trnh phosphoryl ha cng lin h vi gradien proton. C khi
vn chuyn electron trong qu trnh h hp (hiu kh hoc k kh) cng
nh trong quang hp to ra mt gradien proton quan trng v n l
nng lng d tr (c th c so vi mt t in c np sn), nng
lng d tr ny c th c thu hi bng cch li cc proton mt bn
mng m ngi ta thng so snh nh mt ng c sn sinh ATP.
5. S trao i carbohydrate
5.1. S phn gii carbohydrate
Carbohydrate nu dng hp cht c trng lng phn t ln phi
c thu phn thnh cc dng ng n c th i vo cc con ng
phn gii ng cung cp nng lng cn thit cho hot ng sng ca
c th hoc to ra cc hp cht trung gian cn cho s chuyn ho trao i
cht.
5.2. S tng hp carbohydrate
Tng t nh cc thc vt bc cao, vi sinh vt c kh nng tng hp
cc oligo - v polysaccharide. Lng cc oligo - v polysaccharide ni bo
t ti 60% khi lng kh ca t bo, cn polysaccharide ngoi bo c
th vt nhiu ln khi lng ca vi sinh vt. Thnh t bo cng cha mt
lng ln polysaccharide.
Tt c cc oligo v polysaccharide c tng hp bng cch ko
di chui saccharide c trc nh vic thm n v monosaccharide (X).
n v monosaccharide ny tham gia vo phn ng dng nucleotide -
monosaccharide c hot ha, thng l dn xut ca uridine -
diphosphate (UDP.X) nhng i khi cng vi cc nucleotide purine v
pirimidine khc. S tng hp din ra theo phn ng chung nh sau:
...X.X.X.X.X.X + UDP - X = ...X.X.X.X.X.X.X. + UDP
(n - nhnh) (n + 1 - nhnh)
Trong trng hp polysaccharide bao gm hai loi monosaccharide
lin tip (X v Y) phn ng chung xy ra qua hai bc:
Bc 1: ...X.Y.X.Y.X.Y + UDP - X = ... X.Y.X.Y.X.Y.X. + UDP
 120

Bc 2: ...X.Y.X.Y.X.Y.X + UDP - Y = ... X.Y.X.Y.X.Y.X.Y + UDP

Con ng tng hp polysaccharide phn nhnh ta cn bit rt t.
Trong dch chit t bo ca mt s loi nm si c cha enzyme kitine -
sinthetase xc tc phn ng chuyn cc nhnh N - acetylglucosamine t
UDP - acetylglucosamine n phn t nhn. Mt cch tng t vic tng
hp mannan ca thnh t bo nm men cng bao gm vic chuyn cc
nhnh mannose t GDP - mannose n cht nhn. Trong vic tng hp
tinh bt Chlorella v glycogen Arthrobacter c s tham gia ca ADP -
glucose.
- Tng hp glycogen: Khi c iu kin ngoi cnh hn ch s pht
trin ca vi khun th chng c th tch ly glycogen nh h enzyme trng
hp cc glucose. Glycogen tch ly di dng cc ht trong cht ngyn
sinh m chng ta c th pht hin bng cch nhum mu vi iode (hnh
5.8).

Hnh 5.8 S tng hp glycogen

- Tng hp levan v dextran: Cc levan (poly - 2,6-fructose) v

cc dextran (poly -1,6 glucose) vi s phn nhnh - 1,3 hoc -1,4
c tng hp t saccharose theo phn ng sau:
n saccharose n glucose + (fructose)n
n saccharose (glucose)n + n fructose
S hnh thnh nhng hp cht ny chnh l nguyn nhn ca hin
tng nhy nht khi nui cy mt s loi vi khun trn mi trng giu
saccharose. Cc hp cht polysaccharide cng vi hp cht nit s hnh
thnh nn glycopeptide ca thnh t bo.
6. S trao i protein
6.1. S phn gii protein
Khc vi ln men, c cht ca qu trnh thi ra l protein. y l
mt trong nhng thnh phn quan trng ca xc b ng vt, thc vt v
vi sinh vt. S phn gii cc hp cht hu c cha nit c ngha to ln
 121

i vi nng nghip v i vi vng tun hon vt cht trong t nhin.

Ngi ta gi qu trnh phn gii ny l qu trnh ammon ha
(ammonification).
Mun phn gii protein, cng nh cc hp cht cao phn t khc,
trc ht vi sinh vt phi tit ra protease ngoi bo v chuyn ha protein
thnh cc hp cht c phn t nh hn (polypeptide, olygopeptide). Cc
cht ny tip tc c phn hy thnh acid amin nh cc peptidase ngoi
bo, hoc c xm nhp ngay vo t bo vi sinh vt sau mi chuyn
ha thnh acid amin. Mt phn cc acid amin ny c vi sinh vt s dng
trong tng hp protein, mt phn khc c phn gii theo nhiu con
ng khc nhau to NH3, CO2 v nhiu sn phm trung gian khc.
Nhng vi sinh vt khng c kh nng sn sinh cc enzyme phn gii
protein ngoi bo s khng ng ha c cc loi protein thin nhin.
Chng ch c th s dng c cc sn phm thy phn ca protein.
Rt nhiu loi vi sinh vt khc nhau tham gia vo qu trnh ammon
ha trong t nhin. ng ch l cc vi sinh vt sau:
- Vi khun: Bacillus mycoides, B. subtilis, Proteus vulgais,
Pseudomonas aeruginosa, E. coli...
- X khun v nm: Streptomyces griseus, S. fradiae, Aspergillus
niger, A. awamori, Penicillium camemberti, Mucor spp., ...
Trong c th vi sinh vt, cc acid amin thng c chuyn ha nh
qu trnh kh amin, qu trnh kh carboxyl hoc ng thi va kh amin
va kh carboxyl.
Khi phn gii cc acid amin ty theo tng loi phn ng s to cc
sn phm trung gian, y l tin cht cho cc phn ng sinh tng hp hoc
cn thit cho mt s qu trnh trao i cht, nh s to thnh acid fumaric
t acid aspactic:
HOOC- CH2- CH (NH2) - COOH HOOC-CH=CH-COOH+ NH3
hay to - ketoglutaric t acid glutamic, acid acetic t glycine, acid
succinic t aspactic...). Trong qu trnh kh amin thy phn c s lin kt
vi cc ion H+ v OH- nh s to thnh acid lactic t alanine:
CH3-CH(NH2)-COOH + H2O CH3-CHOH-COOH + NH3
 122

Protein Polypeptide
Olygopeptide Amino acid
Enzyme phn gii Peptidase
protein ngoi bo

Cc Amino acid ni bo

Kh amin v Kh amin Chuyn amin v Trc tip dng

phn gii mch carbon phn gii carbon trong qu
trnh sinh
tng hp protein
Nhiu sn phm sinh ra trong qu trnh phn gii acid amin (ru,
acid hu c ...) s c li cun vo qu trnh phn gii to nng lng.
Nhiu loi amin do vi khun sn sinh ra trong c th ngi v ng
vt c tnh c, nh s tch ly histamine c th gy chng co git mch
mu, cc hp cht diamine nh acmatine, putrecine gp nhiu trong tht c
i thiu u c tnh c.
Mt s vi khun c kh nng chuyn ha tryptophan thnh 2 cht c
mi hi l indol v skatol. Trong qu trnh phn gii cc acid amin cha
lu hunh (methionine, cysteine, cystine) s to ra H2S lm cho phn hoc
tht c c mi hi.

NH2

HOOC- CH- (NH2)-CH2-SH + 2H2O H2S + NH3 + CH3COOH + HCOOH

(cysteine)

Khi qu trnh thi ra xy ra trong iu kin thong kh th qu trnh

xy ha c tin hnh n cng, khi hu ht carbon c chuyn
thnh CO2. Nu qu trnh ny din ra trong iu kin k kh th c s tch
ly nhiu sn phm trung gian ni trn.
Ngoi kh nng phn gii protein, vi sinh vt cn c kh nng phn
gii mt s cht m khc nh urea, acid uric... S phn gii cc cht ny
cng to ra cc sn phm trung gian nh acid formic, acid acetic, CO2,
 123

NH3..., cc cht ny c th tham gia trc tip vo qu trnh trao i cht v

trao i nng lng ca t bo.
Urea l cht hu c ch yu trong nc tiu ca ngi v ng.
Trong urea c cha 47% nit nhng nu khng c vi sinh vt phn gii
thnh NH3 th ngun nit ln lao ny cng tr thnh v ch i vi mi
thc vt.
Rt nhiu vi khun, x khun, nm si sng trong t c kh nng
phn gii mnh m urea. ng ch l cc loi nh Planosarcina urea,
Micrococcus urea, Bacillus pasteurii, Proteus vulgaris... Qu trnh phn
gii urea xy ra rt n gin vi s xc tc ca urease:
H2N- CO- NH2 + 2H2O (NH4)2CO3
(NH4)2 CO3 t bn vng do d dng phn gii tip:
(NH4)2CO3 2NH3 + CO2 + H2O
NH3 sinh ra do s phn gii hp cht hu c thng c xy ha
thnh NO2- v tip tc xy ha n NO3-. Phn m ((NH4)2SO4 hoc
NH4Cl) sau khi bn rung cng nhanh chng chuyn thnh NO3-. Qu
trnh chuyn ho ny gi l qu trnh nitrate ha. Qu trnh ny bao gm 2
giai on:
- Qu trnh nitrite ha c biu th bng phng trnh phn ng
sau:
2NH3 + 3O2 2HNO2 + 2H2O + Q
Nng lng sinh ra trong phn ng ny cng c vi khun nitrite
s dng ng ha CO2 ca khng kh. Thc ra qu trnh ny tri qua
nhiu phn ng trung gian v to nhiu sn phm trung gian, trong sn
phm trung gian quan trng nht l hydroxilamin (NH2OH). Enzyme xc
tc cho vic chuyn hydro l cc enzyme ca qu trnh h hp hiu kh. Vi
khun nitrite l bn t dng bt buc vi cc chi i din l
Nitrosomonas, Nitrosococcus, Nitrosospira v Nitrosocystis.
- Qu trnh nitrate ha c biu th bng phng trnh sau:
NO2- + O2 NO3- + Q

Cng nh cc vi khun t dng ha nng khc, nng lng sinh

trong phn ng xy ho NO2- c s dng ng ha CO2 khng kh.
Vi khun nitrate ha in hnh l cc loi thuc ging Nitrobacter. y l
nhng t bo hnh bu dc, kch thc khong 0,8-1,0m, thuc loi G-,
khng to bo t, khun lc nh b v trong sut. Ngoi ra cn c mt s
ging khc nh Nitrococcus, Nitrospira...
 124

6.2. S tng hp protein

Ba qu trnh ch yu trong vic bin i v truyn t thng tin di
truyn trong c th l:
- S nhn i DNA to ra 2 phn DNA ging nhau v ging vi
phn t DNA m.
- Qu trnh sao m theo thng tin di truyn t DNA c sao
sang RNA, sau c chuyn n ribosome.
- Qu trnh dch m cho php chuyn dch thng tin di truyn thnh
cc axit amin cu trc nn protein. Cng ngh di truyn cho php gii m
trt t cc nucleotide thnh trt t cc axit amin c xc lp (h. 5.9).

Hnh 5.9 S sinh tng hp

axit amin (theo Lehninger)
 125

Phn ln cc vi sinh vt c kh nng tng hp 20 loi axit amin.

Con ng tng hp axit amin vi sinh vt c nghin cu kh t m.
Nguyn liu dng cho qu trnh tng hp l nhng sn phm trung gian
nh pyruvate, - ketoglutarate, oxaloacetate, fumarate...
Trong phn ln trng hp, nhm amin c a vo giai on
cui cng ca sinh tng hp nh s chuyn amin, mt s acid amin c
hnh thnh nh hng lot bin i ca cc acid amin khc, trong trng
hp khng cn s chuyn amin. Nhiu loi L - acid amin c sinh
tng hp bng con ng ln men nh vi sinh vt nh lysine, glutamic,
alanine, glycine...
Hin nay nhiu loi acid amin t nhin l sn phm ca cng ngh
sinh hc bng cch thy phn protein, tng hp ha hc hoc sinh tng
hp nh vi sinh vt. Trong nhiu trng hp ngi ta phi hp gia tng
hp ha hc v sinh tng hp, phng php ny c nhiu u im l cho
cc dng L - acid amin l dng thng gp trong c th.
7. S trao i lipid
7.1. S phn gii lipid
Lipid (ester phc tp ca glycerine v acid bo) v cc cht sp
(ester phc tp ca acid bo v ru bc mt) c nhiu vi sinh vt dng
lm ngun thc n v nng lng. So vi cc c cht khc th y l loi
c cht c ng ha vi tc chm, qu trnh ny c thc hin bi
nhiu nhm vi sinh vt khc nhau nh vi khun, x khun, vi nm
Bc u tin ca qu trnh ng ha lipid l s phn gii thnh
glycerine (hoc ru n nguyn t) v cc acid bo. Qu trnh ny c
xc tc nh lipase ni bo hoc ngoi bo.
Sau khi c phosphoryl ha, glycerine s c chuyn ha theo
con ng EMP, cn cc acid bo i vo chu trnh - xy ha (hnh
5.10).
Nh vy, c qua mt vng xy ha han ton chui carbon ca phn
t acid bo li mt i 2C. C tip tc nh vy th ton b chui carbon s
c chuyn ha thnh acetyl - CoA, cht ny c chuyn ha nh chu
trnh TCA hoc chu trnh acid glyoxilic. Mt s nm mc thuc cc chi
nh Penicillium v Aspergillus c th xy ha acid bo mt cch khng
trit v tch ly li trong mi trng metylketone to mi kh chu (gp
trong du m b nhim mc).
 126

Hnh 5.10: Chu trnh - xy ha axit bo

Cu hi n tp ca chng 5
1. Cc hnh thc vn chuyn cc cht qua mng?
2. C ch ca qu trnh sinh nng lng ca h hp hiu kh v k
kh?
3. Cc kiu h hp vi sinh vt?
4. Cc con ng phn gii hexose?
5. Cc kiu trao i cht vi sinh vt?
6. C ch v ngha ca chu trnh acid tricarboxylic (ATC)?
7. S phn gii cc hp cht protein?
8. Vai tr ca vi sinh vt trong vng tun hon nit?
9. Vai tr ca vi sinh vt trong tun hon carbon?