CHAPTER 6

PHOTOSYNTHESIS:
CARBON METABOLISME
 A summary of
photosynthesis
Consist of two reactions:
 Photochemical reactions
Occur in thylakoids and grana, produce
O2, ATP, and reduced NADP+ (or NADPH).
 Biochemical reactions
Known as Calvin cycle. Occur in stroma,
uses the ATP and reduced NADP+ to reduce
CO2 to carbohydrate. The byproducts ADP,
Pi, and NADP+ are returned from the Calvin
cycle to the photochemical reactions
 6.1 Photosynthetic Carbon
Reduction (PCR) Cycle @ Calvin
cycle
 Pathway which all organisms
(photosynthetic eukaryotic)
incorporate CO2 into carbohydrate;
known as carbon fixation @
Photosynthetic Carbon Reduction (PCR)
cycle
 Referred as Calvin Cycle

 Consume ATP and NADPH produced by

photosynthetic electron transport
 PCR cycle divided into 3 primary
stage :
(i) Carboxylation fixes CO2 in the
presence of 5-C acceptor
molecule; ribulose bisphosphate
(RuBP); and converts into 2
molecules of 3-C acid
(ii) Reduction consume the ATP and
NADPH produced by photosynthetic
electron transport to convert the
3-C acid to tiose phosphate
(iii) Regeneration consume
(i) Carboxylation
 Calvin’s study by using radiolabeled
CO2; 14CO2

 Radioactivity was found in a 3-Carbon

acid (3-phosphoglycerate); 3-PGA

 3-PGA first stable product of

photosynthesis

 PCR cycle referred as C3 cycle

 Acceptor molecule ribulose-1,5-

biphosphate (RuBP)
 Carboxylation reaction; which
CO2 added to RuBP forming six-
carbon intermediate which is
transient & unstable

 six-carbon intermediate which

is transient & unstable remain
bound to enzyme and hydrolized
to 2-molecules 3-PGA

 Carboxylation reaction
Catalyzed by enzyme ribulose-
1,5-biphosphate carboxylase-
oxygenase (Rubisco)
 Carboxylation Reaction of the PCR cycle

O CH20-P CH20-P
CH20-P
C-COH HCOH
C=O
+ *CO2 HO C=O CO2-
HCOH
HCOH
HCOH CO2-
CH2OH-P
CH2OH-P
HCOH
CH20-P
Ribulose-1,5- Six-carbon
biphosphate intermediate 3-phosphoglycerate
(RuBP) (3-PGA)

* Ribulose-1,5-biphosphate carboxylase-oxygenase
(Rubisco)
 Energy from light reaction of
photosynthesis is required at 2
points. i.e.:
i. Reduction of 3-PGA
ii. Regeneration of RuBP
acceptor molecule
(ii) Reduction of 3-PGA

 For chloroplast continue take

up CO2
 Involved 2 conditions :
i. Production of 3-PGA
continually removed
ii. Maintain an adequate
supply of RuBP
 Both require energy ATP and
NADPH
 3-PGA is removed by reduction
 Reduction 3-PGA to G3P

ATP ADP NADPH NADP+

CH20-P CH20-P
CH20-P
HCOH HCOH
HCOH
CO2- CH + Pi
C
O O O

P
3-PGA Glycelaraldehyde-3-
1,3-biphosphoglycerate
phosphate(G3P)
 2-step reaction:
3-PGA phosphorylated to
1,3bisphosphoglycerate
1,3-bisphosphoglycerate reduced
to glyceraldehyde -3-phosphate
 Both ATP and NADPH required in 2
step reaction
 Triose sugar-phosphate; G3P
available for export to cytoplasm
(iii) Regeneration of RuBP

 Continuing supply of acceptor

molecule; RUBP
 Accomplished by series of
reaction involving 4-, 5-, 6-,
and 7- carbon sugars
 Reactions include the
condensation of 6-C fructose
phosphate with triose-phosphate
to forms 5-C and 4-C sugar
 Another triose join with 4-C
 7-C sugar combined with 3 rd triose-
phosphate 2 molecules 5-C sugar
formed
 5-C sugars can be isomerized to form
Ribulose-5-phosphate (Ru5P)
 Ru5P phosphorylated to regenerate
required ribulose-1,5-biphosphate
 Net effect of reaction recycle
carbon from 5 out of 6 G3P molecules
to regenerate 3 RuBP molecules
 Sum = 3 RuBP + 3 CO2 3 RuBP
+ G3P
 6 turns of cycle would regenerate 6
molecules RuBP and one additional
hexose sugar as net product
 6.1.2 Energy input in PCR
cycle
 3 turns of cycle ⇨ uptake of 3
molecules CO2 ⇨ total 6
molecules NADPH and 9 molecules
ATP required
 Reduction of each molecule of
CO2 requires 2 molecules NADPH
and 3 molecules ATP ratio of
ATP/NADPH of 3/2 @ 1.5
 Each NADPH stores 2 electrons
total of 4 electrons required
to fix each molecule CO2
6.2 Activity and regulation of
PCR Cycle
 Rate of carbon reduction is partly
dependent on the availability of an
adequate pool of acceptor molecules;
CO2 and RuBP

 PCR cycle can utilize fixed carbon to

increase the pool size through
autocatalytic regeneration of RuBP
 Normally, extra carbon taken through
PCR cycle accumulated as starch @
exported from chloroplast
 However, PCR cycle has potential to
 Amount of acceptor quickly
built up within chloroplast to
support rapid photosynthesis @
increase the rate of
photosynthesis
 Time required to built up the
levels of PCR cycle
intermediates in transition
from dark to night; called
photosynthesis induction time
6.2.2 Regulation of Rubisco
activity
 Rapidly to zero when light is
turn off
 Slowly when light is once again
turned on
 Rubisco activity is regulated
indirectly by light
 Involves complex interaction
between Mg 2+ fluxes across the
thylakoid, CO2 activation,
chloroplast pH changes and
 Light –driven electron
transport leads to proton net
movement into lumen of
thylakoid
 Proton across thylakoid
membrane generates a proton
gradient = pH 3.0 and increase
pH of the stroma (pH 5.0 in the
dark to pH 8.0 in the light)
 Light bring increase in the
free Mg 2+ of the stroma
 Mg 2+
moves out of the lumen to
 CO2 reacts with amino group
forming carbamate
 Carbamate requires the release
of 2 proton ⇨ consequently,
increasing pH
 Mg 2+ then coordinated to
carbamate to form carbamate- Mg
2+
complex ⇨ active form of the
enzyme
6.2.3 Regulation of other PCR
enzymes
 eg: ferredoxin/thioredoxin for
light-driven enzyme activation
in chloroplast

 PSI drives the ferredoxin

reduction which in turn reduces
thioredoxin
 Thioredoxin reduces the
appropriate disulphide bond
(-S-S-) state to sulfhydryl
(-SH-SH-) state
6.3 Photorespiration
 Photorespiration ⇨ process involves
reoxidation of products assimilated
in photosynthesis
 Involves activities of 3 different
organelles:
i. chloroplast
ii. peroxisome
iii. Mitochondria
 Photorespiration consume
oxygen, release carbon dioxide
 Because CO2 is evolved, results in
net loss of carbon from cell
 Plants that incorporate carbon
through PCR @ Calvin cycle known
as C3 plants; cause the 1st
product to incorporate CO2 in 3C
acid PGA
(3-PGA)

 C3 plants have built-in

metabolic inefficiency in
photosynthetic process
 Acquire CO2 from atmosphere
through stomata
 when hot dry weather stomata
close to reduce water loss
 3 CO2

6 PGA
Rubisco
3 RuBP 6 ATP
3 ADP
Calvin 6 ADP
3 ATP Cycle 6 1,3-
biphosphoglycerate
5 G3P 6 NADPH

6 NADP+ 6Pi
6 G3P

1 G3P

glucose
 H 2O CO2

Light
Chloroplast
NADP+

Photosynthe ADP
Calvin Cycle
tic electron
transport
ATP

NADPH

O2 CH2O
(Glucose)
 6.3.1 Rubisco catalyzes
fixation of both
CO2 and O2
 Glycolate (2-C) metabolism related to
photorespiration
 Enzyme involved in process located in
3 organelles
 Bifunctional nature of Rubisco
⇨Rubisco catalyzes an in
carboxylation reaction oxygenase
reaction ⇨ Ribulose 1-5-biphosphate
carboxylase-oxygenase

RuBP + O2 3 PGA +
phosphoglycolate (2C)
P-glycolate CO2 +
recovery of remaining
carbon of PCR

 Known as C2 glycolate cycle @

PCO(Photosyntheitic carbon
oxidation) cycle
 Begins with RuBP oxidation to
3-PGA and P-Glycolate
 Glycolate exported from chloroplast
and diffuses to peroxisome
 Glycolate oxidized to glyoxylate and
hidrogen peroxide
 Peroxide broken down by catalase
 Glyoxylate undergoes transamination
reaction form amino acid glycine
 Glycine transferred to mitochondria
⇨ 2 mol. glycine (4-C) converted to
1 mol. serine (3-C) + 1 mol. CO2
(released)
 Serine leaves mitocondria ⇨returning
to peroxisome; amino (-NH2) group
given up in transmination
 The product (Hydroxy
6.4 C4 Syndrome
 C-4 plants:
⇨ First product is 4-carbon acid
oxaloacetate(OAA)
⇨ exhibit specific anatomical,
physiological and
biochemical characteristic
constitute C4 syndrome
 C-4 Leaves anatomy :
⇨ presence of 2 distinct
photosynthesis tissues;
between vascular bundles
mesophyll cells called
 Under condition of high fluence rates
and high temperature (30-40ºC),
photosynthesis rates of C4 species
may 2 -3 times greater than C3
species
 Maintain active photosynthesis under
of water stress ⇨ lead stomatal
closure
 CO2 concentrated in bundle sheath
cells
 Hatch & Slack proposed a cyclic
mechanism ⇨ carbon incorporated into
C4 acid ⇨ β –carboxyl carbon
transferred as CO2 to PCR cycle
C4 photosynthetic carbon
assimilation cycle
 6.4.1 C4 photosynthetic carbon
assimilation cycle
 Carboxylation phosphoenol pyruvate
(PEP) using HCO3- by PEPcase
 Oxaloacetate (OAA) formed is
unstable
 OAA reduced to malate or
transaminated to aspartate
 Transported from mesophyll cell into
bundle sheath cell
 C4-Acid undergoes decarboxylation,
CO2 formed reduce to triose sugars
via PCR cycle
 C3-acid (pyruvate or alanine)
transported back into mesophyll cell
 C4 Syndrome Ecological
Significance
Comparison of C4 and C3 plants
(Refer to Table 5.4 page 112;
Textbook)

 Photosynthesis of C4 is not
inhibited by oxygen
 Photorespiraton is absent of C4
plants or the process suppressed
 C4 plants have a low CO2
compensation point(range of 0 to 5
ul/l CO2 ) ; While for C3 plants,
range of 20 to 100 ul/l CO2
 CO2 compensation point :
ambient CO2 concentration at which
 High level of CO2 in bundle sheath
cell outcompeting O2 for binding
with Rubisco
 Adaptations of C4 leaves ensure CO2
that might escape bundle-sheath cell
is trapped and reassimilated by
PEPcase in mesophyll cells before
escape from leaf ⇨ thus, C4 leaves
efficient in CO2 absorbers, trap and
recirculate CO2 produced in leaf
 C4 plants have higher temperature
optimum (30-45oC) than C3 plants
(20-25oC)
 High temperature (between 40 oC and
50 oC) ⇨ rate of photosynthesis
decrease to greater extent than rate
 C4 plants can maintain
photosynthesis when stomata
partially closed to conserve water;
while C3 plants , moderate water
stress will close stomata

 Transpiration ratio of C4 plants is

low (range of 200 to 350), while C3
plants higher ( range of 500 to
1000)

 Does not saturate, even in full

sunlight; while light saturation of
C3 occur about 25 % of ful,l
Crassulacean Acid
Metabolism
(CAM)
 Studied most extensively in Family
Crassulaceae
 Now found in 23 different flowering
plants (Cactaceae, Euphobiaceae)
fern (Polypodiaceae) primitive plant
(Welwitschia)
 Survive in extremely dry or
xerophytic habitats
 Succulent plants ⇨ characteristics
by thick, fleshy leaves, cells
contain large water-filled vacuoles)

 Involve Carboxylation
phosphoenol pyruvate (PEP) @
enzyme PEP carboxylase
 Immediate product OAA reduce to
malate which is store in
vacuole
 During daylight hours
⇨ malate retrieved from vacuole
⇨ decarboxylated
⇨ CO2 diffuses into chloroplast
and converted to triose
Comparison between CAM and C4
syndrome
 CAM similar to C4
⇨ use PEPcase to form C4 acids
from PEP and bicarbonate
⇨ Acids decarboxylated to provide
CO2 for PCR cycle

 Significant differences
⇨ C4 requires a special anatomy
by which C4 carboxylation spatially
separated from C3 PCR cycle.
In CAM both occur in same cell
but separate time
⇨ CAM no close cycle of carbon
intermediate as in C4 plants. PEP
 Significant of CAM
 Represents a adaptation to dry habitat
 Growing in shallow & sandy soil with
little available water
 CO2 uptake at night (during minimum
evaporative water loss)
 Daylight hours
⇨ stomata closed to reduce water loss
⇨ photosynthesis can proceed using
the reservoir of stored CO2
 Daily carbon assimilation by CAM
plants about ½ of C3 plants, 1/3 of C4
plants
 CO2 uptake by CAM plants continue
under water stress conditions which in