Photosynthesis: Pathway of Carbon

Fixation
Photosynthesis is the synthesis of organic molecules using the energy of light.
For the sugar glucose (one of the most abundant products of photosynthesis)
the equation is:
6CO
2
+ 12H
2
O -> C
6
H
12
O
6
+ 6H
2
O + 6O
2

A description of the experiments that led to this equation are described
in Discovering the Secrets of Photosynthesis.
Light provides the energy to:
transfer electrons from water to nicotinamide adenine dinucleotide
phosphate (NADP
+
) forming NADPH;
generate ATP.
The details of these processes are described in Photosynthesis: The Role of
Light.
ATP and NADPH provide the energy and electrons to reduce carbon dioxide
(CO
2
) to organic molecules.
The Steps
CO
2
combines with the phosphorylated 5-carbon sugar ribulose
bisphosphate.
This reaction is catalyzed by the enzyme ribulose bisphosphate
carboxylase oxygenase (RUBISCO)(an enzyme which can fairly claim
to be the most abundant protein on earth).
The resulting 6-carbon compound breaks down into two molecules of 3-
phosphoglyceric acid (PGA).
The PGA molecules are further phosphorylated (by ATP) and are
reduced (by NADPH) to form phosphoglyceraldehyde (PGAL).
Phosphoglyceraldehyde serves as the starting material for the synthesis
of glucose and fructose.
Glucose and fructose make the disaccharide sucrose, which travels in
solution to other parts of the plant (e.g., fruit, roots).
Glucose is also the monomer used in the synthesis of
the polysaccharides starch and cellulose.

The graphic shows the steps in the fixation of carbon dioxide during
photosynthesis. All of these reactions occur in the stroma of the chloroplast.
Link to chloroplasts
These steps were worked out by Melvin Calvin and his colleagues at the
University of California and, for this reason, are named the Calvin cycle.
Their Experiment
The experimental apparatus is
shown at the right. After
various intervals of
illumination, a suspension of
unicellular algae is inactivated
and the contents of the cells
extracted. The compounds in
a drop of the extract are then
separated by paper
chromatography.
The identity of each substance
may be determined simply by
comparing its position with
the positions occupied by
known substances under the
same conditions. Or, a
fragment containing the spot
can be cut from the sheet and
chemically analyzed.
To determine which, if any, of
the substances separated on the chromatogram are radioactive, a sheet of X-ray
film is placed next to the chromatogram. If dark spots appear on the film
(because of radiation emitted by the
14
C atoms), their position can be correlated
with the positions of the chemicals in the chromatogram. Using this technique
of autoradiography, Calvin found that
14
C turned up in glucose molecules
within 30 seconds after the start of photosynthesis. When he permitted
photosynthesis to proceed for only 5 seconds, however, the radioactivity was
concentrated in several other, smaller, molecules.



Apparatus used to follow the fate of
14
CO
2
in the dark
reactions. The algal suspension is placed in the
"lollypop", supplied with
14
CO
2
, and illuminated. The
dark reactions are halted by draining the contents of
the lollypop into a flask of hot alcohol. (Courtesy of
Dr. James A. Bassham.)


The dark spots show the radioactive compounds produced after 10 secs (left) and 2 minutes (right) of
photosynthesis by the green alga Scenedesmus. The alga was supplied with carbon dioxide labeled with
14
C, a
radioactive isotope of carbon. At 10 seconds, most of the radioactivity is found in 3-phosphoglyceric acid ("P-
Glyceric"). At 2 minutes, phosphorylated 6-carbon sugars (glucose and fructose) have been synthesized as well
as a number of amino acids. The small rectangle and circle (lower right-hand corners) mark the spots where the
cell extract was applied. (Courtesy of Dr. James A. Bassham.)