Biological Oxidation

Introduction
ATP
Oxidative Phosphorylation
Oxidation not producing
ATP

1
 Introduction
Biological Oxidation
The process in which nutrient substances
such as saccharides, lipids and proteins
are oxidized into water and carbon dioxide
with simultaneous production of energy.

2
Nutrient + O2  H2O + CO2 + ATP +
energy heat

3
 Characteristics of Biological Oxidation

Biological Oxidation Burning

Temperature 37℃ high
Condition neutral dry
Catalyst enzyme no
Velocity of energy slow fast
released
Form of energy ATP heat
released
Way of producing organic acid decarboxylation Oxygen directly
CO2and H2O produce CO2; wide addition of combine with
water and dehydrogenation , carbon(hydrogen)
the hydrogen combine with , produce
oxygen in an indirect way. CO2(H2O).
4
 Forms of Oxidoreduction reactions in
Biological Oxidation
Oxidation
Loss of electrons
Dehydrogenation [O
]
Addation of oxygen -e / -H / +O
Reduction +e / +H / -O
Gain of electrons
Addition of hydrogen [R]
Removal of oxygen

5
 General course of Biological Oxidation

glucogen triglyceride protei

n
glucose fatty acid + amino acids
Glycerol

AcetylCoA

TAC
CO2 ADP+P AT
i P
2H respiratory chain H2
6
 Section 1. ATP
NH2
N
N

O- O- O- N N
-
O P O ～P O ～P CH2 O
O O O H H
H H
OH OH

Adenosine Triphosphate
ATP + H2O ADP + Pi + 30.5 kJ/mol
7
activation effect

8
 ATP is the main form of energy
utilization and store in body and the
center of energy conversion.

H2O + CO2 ATP

有机物 生物大分子 主动运输 肌肉收缩 信息传递

氧化 合成

O2 ADP + Pi

9
 High-energy compound
分类及举例 释放能量
（ pH7.0,25℃)
UTP 、 CTP 、 GTP 30.5 kJ/mol
1,3- 二磷酸甘油酸、 61.9 kJ/mol
磷酸烯醇式丙酮酸

磷酸肌酸 43.9 kJ/mol

乙酰 CoA 、琥珀酰 CoA 31.4 kJ/mol
、脂酰 CoA

10
Transform 　 between high-energy compounds

nucleoside
GDP diphosphate kinase GTP
ATP + UDP ADP + UTP

CDP CTP
adenylate kinase
ADP + ADP  ATP +
AMP

11
creatine phosphate– store form of ATP
in brain and muscle.

12
 Section 2. oxidative
phosphorylation

Ways producing ATP:

substrate level phosphorylation
oxidative phosphorylation

substrate level phosphorylation

--formation of ATP by the way of
straight transfer high-energy substrate
energy to ADP.

13
1,3- 二磷酸甘油酸 + ADP 3- 磷酸甘油酸 + ATP

磷酸烯醇式丙酮酸 + ADP 丙酮酸 + ATP

PK
琥珀酰 CoA + H3PO4 + GDP 琥珀酸 + CoA-SH +
GTP

ADP

ATP
14
I respiratory chain （呼吸链 )
a oxidoreduction system which consists of a
series of enzyme, coenzyme aligning in
mitochondrial inner membrane, function as
linksystem transferer of hydrogen and electron.

15
Mitochondria

Respiratory chain 16
 electron transfer in respiratory chain

Cyt 内外膜间隙侧
e c

e- -
e Q
I e- e- - 线粒体内
Ⅱ Ⅲ Ⅳ 膜

NADH+ 延胡素 H2 基质侧

H
NAD 酸 1/2O2+2 O
+ 琥珀酸 H
+
+

17
 succinic acid oxidation
respiratory chain

NADH oxidation
respiratory chain

18
 Component of respiratory chain

Complex name Number of Prosthetic

peptide chain group
ComplexⅠ NADH- CoQ 42 FMN, Fe-S
reductase
ComplexⅡ Succinate-CoQ 4 FAD, Fe-S
reductase
ComplexⅢ Q-Cyt C 11 Fe-S, iron
reductase protoporphyrin
,
Complex Cyt C oxidase 13 Cu, iron
Ⅳ protoporphyrin
,
 Co-Q and Cyt 19
 (1) complexⅠ—
NADH-Q reducase
iron-sulphur
protein,
Flavoprotein with
FMN, 42 peptide
chains, 850 kD.

Bind and oxidize

NADH, transfer
electrons to Q,
release 4H+ to
interspace of inner
and outer
membrane.
ADH FMN ， Fe-S CoQ 20
 Structure of NAD+ and
NADP+
NH2
O
C NH2 N
N
+
N O- O- N N
CH2 O P O P O CH2
O O
H H O O H H
H H H
OH OH OH OR

R=H:NAD+;
+ R=H PO 3:NADP +
NAD :nicotinamide adenine 2dinucleotide,CoI
NADP+:nicotinamide adenine dinucleotide
21
phosphate,CoII
22
黄素单核甘酸
23
 ironsulfur protein

Fe-S
Fe2+ Fe3+ + e

Fe2S2, Fe4S4
Fe4S4
24
Ubiquinone,Q
(Coenzyme Q,CoQ) 人体中： CoQ10

 quinones contain a polyisoprene

poly side
chain.
 liposolubility ， make it move in
mitochondrial inner membrane easily.
 the only one electron carrier without
protein in respiratory chain.


25
 2H+

复 FM Fe- N- Q QH
合 N S 2 2
体 2e
-
Ⅰ

子传 2H
的递 NAD+
+
NADH+H
过电
+

程
NADH+ FMN Fe2+ Q
H
+
NAD + FMNH Fe3+ QH
2 2
NADH FM Fe- Q
26
N S
 (2) complexⅡ-
Succinate-CoQ reductase

ccinic acid → FAD→ Fe-S→ Q 。
i.e. succinate
dehydrogenase,
consists at list 4
peptides. Contain
one FAD, two
ironsulfur protein and
one Cyt b560.
Transfer electron
from succinic acid to
Q, do not release H+
to the interspace.
27
cytochrome,Cyt 细胞色素
A 、 structure: colourant protein containing iron
porphyrin.
B 、 typing: Cyta: Cytaa3
Cytb: Cytb562 、 Cytb566 、 Cytb560
Cytc: Cytc 、 c1
C 、 difference:
① different side chain of iron porphyrin.
② Different linkage form of iron porphyrin with the
protein.

CytFe3+ + e 
CytFe2+
28
 多聚异戊二烯长链

甲酰基

29
 Difference between Cyt a and Cyt b, Cyt
c.
αband
Linkage with
　 prothetic group color wavelen
protein
gth
Non-covalent
Cytb heme red 560nm
bonding

Cytc red 550nm Bind with – SH

heme
of Cys

Non-covalent
Cyta heme A green 600nm
bonding 　

30
 (3) Complex Ⅲ i.e. cyt c reducase,
Q-cytc reducase consists of 11 peptide
chains different,
existing as a dimer.
every monomer
contains two cyt b
(b562, b566), one cyt
c1 and a iron sulphur
protein.
Catalyze electron
transfer from Q to cyt
c. every two
electrons’s transfering
lead four proton
pumped to the
QH2 b566 b562 Fe- Cyt intermembrance
S c1 space.
c 31
 Cyt c

Complex 32
 Electron transfering process in
complexⅢ
first time oxidation of QH2 secondary time oxidation of QH2
Cyt Cyt
Cytc c 2H+ Cytc c 2H+
Fe- Fe-
bL
1
e -
bL
1
e -
e- QH S Q e- QH S Q
bH Q 
Q 2 bH 
Q QH2

2H

33
(4) complex Ⅳ
Cyt c oxidase
Dimer. Every monomer
consists of 13 peptide
chains different, as 3
subunits: I include 2
heme(a,a3),a cuproprotein
(CuB);Ⅱinclude a dikaryon
center formed by two
copper ion(CuA);Ⅲ not
clear
Cu2+ + e  Cu+
CuA a  a3  O2
CuB 34
 NADH
氧
化
链
呼
吸

NADH FMN(Fe-
S) Q b c1 c aa3 O2

琥
珀
酸
氧
链化
呼
吸
succinic acid FAD(Fe-S) Q b c1 c aa3 O2 35
 overall reaction

呼吸链
NADH + H + +
NAD+ +
o 1/2O2 H2
r 呼吸链
FADH2 + O
FAD +
1/2O2 H2
furthe O
r
呼吸链
2H + H2
1/ O
DH2 or NADH 2O
2
36
呼吸链中电子传递体的排列顺序的确定

 利用脱氧胆酸处理线粒体
内膜、分离出呼吸链的 4
种复合物，辅酶 Q 和细
胞色素 C 及 ATP 合酶。
 根据标准氧还电位 E0’ 的
高低 排列
 根据电子传递体氧化还原
态时的吸收光谱变化进行
检测
 利用阻断剂研究分析
 四种复合物的电子传递再
造实验

37
II 、 oxidative phosphorylation

oxidative phosphorylation refer to the ATP

producing form which the reaction ADP change
into ATP couple with respiratory chain oxidation.
It is main form of ATP producing in body.

39
 Why does the reaction: ADP + Pi  ATP + H2O
request a couple?

The product
state(ATP+H2O) is higher

energy
energy level than
ATP+H2
reactant state(ADP + Pi).
So, ADP change into O
ATP isn’t spontaneous
30.5kJ/mo
process. Energy
l
obtaining is requested for
the process.
ADP +
P
energy level diagram
i
40
Whichsegment in respiratory chain can
produce enugh energy for ADP
phosphorylation ？
What’s P/O ratio ？ What’s its
meaning ？
--- number of moles of ATP produced as
consuming a mole of oxygen atom in a reaction, i.e. the
number of moles of phosphor cosumed when consume a
mole of oxygen atom in the reaction.
 What’s the way of ADP
phosphorylation coupled with
respiratory chain oxidation ？
氧化磷酸化偶联机制有： 化学物质偶联学说、
构象偶联学说及 化学渗透学说。目前公认度较大的是
化学渗透学说。

Chemiosmotic hypothesis
 the energy of respiratory chain
oxidation change into proton
gradient across the inner
membrane.
 the proton gradient drive ATP-
synase produce ATP. 1961~1978
43
 In electron
transfer process,
respiratory chain
put proton to
intermembrance of
inner and outer
membrance, result
in proton
concentration
different of the two
side of the inner
membrance.

44
 conceptual diagram of Chemiosmotic hypothesis

H+ H+ H+ H+
+ + + + + + + + + + + + + + + + + +
+
e-

延胡素 H2
- - - - - - - - - - - - - -
NADH+ 琥珀酸 酸 O
- 1/2O2+2
HNAD
+ + H
+

ADP+P AT
i
45
 ATP
syna
Consists of se
hydrophobic
F0(a1b2c912)and
hydrophilic
F1(33).
When proton go
straight through
a,push c loop
turning , and as a
result, spur the F1
turning. 46
Position of ATP
synase in
mitochondria

47
 work principle of ATP

Three conformations of  subunit: 松 L, 紧 T, 放

O
Procedure 松开
of ATP H+ 结好 H+ 脱水 H+
production: （O  （L  （T  （O
） ） ） ）
48
 Ⅲ. Factors affecting
oxidation
phosphorylation

 Inhibitors
 Regulation by ADP
 Thyroid hormone
 Mitchondrial DNA mutation

49
 (1)Inhibitors

 Inhibitors of respiratory chain

block electron transfer of respiratory chain.
 Uncoupler
destroy the coupling of oxidation with
phosphorylation, like uncoupling protein, 2,4-
dinitrophenol.
 Inhibitors of oxidative phosphorylation 　
restrain the proton return to matrix side in ATP
synase, like oligomycin

50
Blocking sites of inhibitor of
respiratory chain CO 、 CN-
抗霉素Ａ
、 N3- 及Ｈ
二巯基丙醇
２Ｓ

  

异藤酮
粉蝶霉素Ａ
异戊巴比妥 51
 Machanism of
uncoupling
Ｈ＋ Ｈ＋ Ｈ＋ 解偶联蛋
白

热
Ｈ＋
Ｈ＋ ADP+P ATP+H2
i O
 inhibitory action of oligomycin

stop proton
flow from
F0 proton 寡霉素
channel.

53
(2) regulation by
ADP

main regulation
factor ： ADP/ATP
ratio
Respiratory control ratio

ADP + Pi  ATP + H2O 离体线粒体实验 , 过量底物存在

时 , 加入 ADP 后的耗氧速率与
仅有底物时的耗氧速率之比

54
(3). Thyroid
Hormone
Thyroid Hormone
甲状腺激素

Na+-K+ ATP enzyme

ATP degradation

ADP/ATP

Oxidative phosphorylation
55
 (4) Mitchondrial DNA mutation
function of mitochondrium Mitochondrium diseases
 all of the 13 peptides (7 peptides in
NADH dehydrogenase, 1 in Cytc
reducase, 3 in Cyt c oxydase,2 in
Maternally
ATP synase coded) by
inherited
Mitochondrium join in oxidative
diseases
phosphorylation.
Mutation affect oxidative (heritage
neurosis,
phosphorylation, ATP production
heritage
decrease.
Naked cyclic duble helix DNA, lack diabetes and
deafness)
of defend system and restoration
Aging related
system.
Symptoms are dependent on the
degree of mutation and the different 56
IV 、 Mitochondria Entry and Exit of
Molecules

Mitochondrial porin,
the major protein of
the outer
mitochondrial
membrane, allows
molecules less
than 10 kD to pass

Inner membrance
were controled by
differnet
transporter. 57
58
ation of NADH in cytosol

 α-glycerophosphate shuttle
(α- 磷酸甘油穿梭 )

 malate-asparate shuttle
( 苹果酸 - 天冬氨酸穿梭 )

59
 Comparison of the two ways of NADH
oxidation
氧化途 主要存 主要承 胞液中 线粒体 被完全 完全氧
径 在的组 担酶 主要承 内主要 氧化时 化时产
织 担酶的 承担酶 经过的 生的
辅基 的辅基 呼吸链 ATP 量

- 磷酸 骨骼肌 α- 磷酸 NAD+ FAD 琥珀酸 2ATP

甘油穿 、神经 甘油脱 氧化呼
梭 细胞 氢酶 吸链

苹果酸 肝、心 苹果酸 NAD+ NAD+ NADH 3ATP

穿梭 肌组织 脱氢酶 氧化呼
吸链

60
  -glycerol phosphate shuttle

呼吸链

NADH+H FADH
+
 - 磷酸甘 2
油脱氢酶  - 磷酸甘
油脱氢酶
NAD
+ FA
D
malate-asparate shuttle

3ATP

62
 Section 3. other oxidation
system
Ⅰ. aerobic dehydrogenase and oxydase

hydrogen prosthetic produc example

acceptor group t
不需氧脱 辅酶 NADH 脱氢酶
氢酶

需氧脱氢 O2 FMN(FAD) H2O2 氨基酸氧化酶、

酶 单胺氧化酶、
黄嘌呤氧化酶
氧化酶 O2 含 Cu H2O 细胞色素 c 氧化酶
、酚氧化酶 、抗
坏血酸氧化酶
63
 H
D- 氨基酸氧化酶
R C COOH + O 2 + H 2 O R C COOH + H 2 O 2 + NH 3

NH 2 O

O O
C C
HO C O C
O 抗坏血酸氧化酶 O +
O + 1/2 2 O C H2O
HO C
HC HC
HO CH HO CH
CH2OH CH2OH

L-抗坏血酸 脱氢抗坏血酸
64
 Ⅱ. Erzymes in peroxisome
过氧化酶体中的酶类
(1).catalase
(1). 过氧化氢酶
• catalytic reaction: one molecule H2O2 offer
electron;another molecule H2O2 accept
electron.
2H22O
2H  2H
O22  OO
2H22O O22

• prosthetic group: 4 heme

• Function: wide distribution, wipe out toxical
H2O2

65
. peroxidase （过氧化物酶）
 Catalytic reaction:catalyze H2O2 straight
oxidize phenols and amines

H O22 AH
H22O 2H
AH22  OA
2H22O A

 prosthetic group ： 1 heme

Protect body. glutathione peroxidase

H O22 GSH
H22O GSH2H OGSSG
2H22O GSSG
Clinical diachorema occult blood test:
过氧化物酶
联苯胺 + H 2 O 2 联苯胺蓝 + H 2 O
66
 Ⅲ . superoxide dimutase,
（ SOD, 超氧化物歧化
酶）
呼吸链电子传递过程中产生超氧
离子 (O2-.)
• O2-. H2O2 + .OH
• 损伤生物膜、生成脂褐素

SOD
+ 2H +
H2O2 + O2

H2O +
过氧化氢酶
基含 Cu 、 Zn( 胞液 )
辅基含
O
( 线粒体 ) 。 2
67
 Ⅳ. Oxidases in
microsome
(1). monooxygenase （加单氧酶）
Catalytic
reactio+
RH+NADPH+H
n: +O2
ROH+NADP +
+H2Ooxidase( 混合功能氧化酶 )
mixedfunction
or Hydroxylase( 羟化酶 ).

composition: NADPH-Cytc reducase, flavoprotein(FAD),

ironsulfur protein(Fe2S2) 、 CytP450 。

function ： hydroxylation 羟化。胆汁酸、胆固醇

的生成；药物、毒物的转化；肾上腺皮
质、类固醇激素的生物合成。
68
mechanism

RH.P450.Fe3+

RH

H2O

ROH

69
(2). dioxygenases ( 双加氧酶 )

incorporate both oxygen

atoms into the substrate.

色氨酸吡咯酶 O2

70
11. Except iron, Cyt aa3 contain ( ) ion.

A Zn
B Mg
C Cu
D Mn
E K

71
12. Which one can be inhibited by CO in
respiratory chain ?
A FAD
B FMN
C Fe-S
D Cyt aa3
E Cyt c

72
13. Which one is uncoupler?

A CO
B piericidin A
C KCN
D 2,4-dinitrophenol
2,4-
E H2 S

73
14. The right electron tansferation sequence is ( )

A b→c→c1→aa3→O2
B c1→c→b→aa3→O2
C c→c1→b→aa3→O2
D c→b→c1→aa3→O2
E b→c1→c→aa3→O2

74
19. Which make Fe-S as prosthetic group
in the respiratory chain?

A Complex Ⅰ
B Complex Ⅱ
C Complex Ⅲ
D Complex Ⅳ
E Cyt c

75
20. Where does the phosphorylation couple
with the oxidation and can produce ATP?

A NADH→CoQ
B CoQ→Cyt b
C CoQ→Cyt c
D FADH2→CoQ
E Cyt aa3→O2

76
Energy of reaction

G < 0 spontaneous G1
G = 0 　 equilibrium
G2
G > 0 　 non-spontaneous
G = G2-G1

G0’= - 2.303 R T log Keg = - n F E0’

77