8777799999p999113lecture0706222

BIOLOGY 113 - MICROBIOLOGY
Lecture 7: Microbial Metabolism - Oxidation-reduction, mechanisms of ATP generation,

glycolysis, Krebs cycle
The "trick" in maintaining the energy balance required to carry out cellular metabolism is to control
the chemical energy released during reactions
Many of the reactions of central metabolism are oxidation-reduction reactions, in which electrons
are transferred (Tortora et al., Figure 5.8)
- Oxidation is removal of electrons from a molecule
= In many metabolic reactions, protons (H+) are also removed
= The net effect is to remove a "hydrogen equivalent"
- Reduction is the addition of electrons to a molecule; as in oxidations, protons may also be
involved
- Oxidations are always coupled to reductions; that is, oxidation of one molecule is always
associated with reduction of another
- Two molecules commonly used by living systems as carriers of electrons are NAD+ and
NADP+
= When a molecule is oxidized during metabolism, a pair of electrons (and one proton) may be
transferred to one of these coenzymes to yield NADH or NADPH (Tortora et al. Fig 5.9)
= Similarly, reduction of a molecule during metabolism may be coupled to oxidation of
NAD(P)H to NAD(P)+
- During catabolic metabolism, energy is released during oxidation of nutrients
ATP, formed from phosphorylation of ADP, serves as a convenient carrier of chemical energy;
organisms use three mechanisms of phosphorylation to generate ATP from ADP
- In substrate-level phosphorylation, ATP is generated when a high-energy phosphate group is
directly transferred from another phosphorylated compound
- In oxidative phosphorylation, ATP is generated by a chemiosmotic mechanism (discussed
below) in which electrons obtained from a donor (frequently NADH) are passed through a
controlled series of oxidation-reduction reactions - an electron transport chain
- Photophosphorylation (see Tortora et al., Figure 5.23) is similar to oxidative phosphorylation,
except that the electrons are obtained from light-activated pigments rather than from a donor
molecule
The nature of microbial catabolism can be most readily understood from consideration of metabolic
pathways for oxidation of carbohydrates, especially glucose
- By metabolic pathway, we mean a sequence of enzyme-catalyzed reactions
- Microorganisms obtain energy from glucose using two general processes - respiration and
fermentation (Tortora et al., Figure 5.10)
= In both respiration and fermentation, electrons removed during oxidation of glucose are used
to reduce NAD+
= In respiration, electrons from reduced NADH are transferred through an electron transport
chain to a terminal electron acceptor (usually O2 ), generating ATP by oxidative
phosphorylation
= In fermentation, there is not electron transport chain, and electrons from NADH must be
transferred to another molecule, yielding characteristic fermentation endproducts
Both respiratory and fermentative catabolism of glucose start with glycolysis - oxidation of glucose
to pyruvic acid
- "Glycolysis" translates roughly to "splitting of glucose", in reference to the splitting of the 6-
carbon glucose molecule into two molecules of 3-carbon pyruvic acid
- The most common glycolytic pathway, found in eucaryotes as well as bacteria, is the Embden-
Meyerhof pathway (Tortora et al., Figure 5.11); the steps can be outlined as follows:
= Glucose is phosphorylated, producing glucose-6-phosphate
· In eukaryotes (such as ourselves!), the phosphate is obtained from hydrolysis of ATP
· In bacteria, glucose is phosphorylated upon passage through the membrane and the
phosphate is obtained from hydrolysis of phosphoenolpyruvic acid (PEP) to pyruvic acid
= Glucose-6-phosphate is rearranged (isomerized) to form fructose-6-phosphate
= Fructose-6-phosphate is phosphorylated to fructose-1,6-diphosphate; the phosphate group is
obtained from hydrolysis of ATP
NOTICE THAT, AT THIS POINT, NO OXIDATION HAS OCCURRED
AND NO ENERGY HAS BEEN OBTAINED; IN FACT, WE'VE USED
UP TWO ATP MOLECULES IN THE PROCESS.
= An enzyme catalyzes cleaveage of the 6-carbon fructose-1,6-diphosphate into two 3-carbon
molecules - glyceraldehyde-3-phosphate and dihydroxyacetone phosphate
· For organic chemistry fans (don't laugh; if there weren't any, we wouldn't have Teflon),
this is the reverse of an aldol condensation
· Another enzyme catalyzes the interconversion of dihydroxyacetone phosphate and
glyceraldehyde-3-phosphate
FOR THE PURPOSES OF DISCUSSING GLYCOLYSIS, WE CAN
ASSUME THAT, AT THIS POINT, WE HAVE TWO MOLECULES OF
GLYCERALDEHYDE-3-PHOSPHATE. THIS NEXT STEP IS
CRITICAL FOR OBTAINING A NET ENERGY YIELD FROM
GLYCOLYSIS
= Each molecule of glyceraldehyde-3-phosphate is oxidized to 3-phosphoglyceric acid in a
complex reaction sequence
· Electrons removed from glyceraldehyde-3-phosphate are transferred to NAD+, producing
NADH
· Coupled to the oxidation of glyceraldehyde-3-phosphate, a high energy phosphate bond is
formed, giving the intermediate, 1,3-diphosphoglyceric acid
· The high-energy phosphate is transferred to ADP, producing ATP
THIS IS WHAT WE GET FROM GLYCOLYSIS - 2 MOLECULES OF
NADH AND TWO MOLECULES OF ATP. THE REMAINING
REACTIONS INVOLVE "GETTING BACK" THE 2 ATP INVESTED AT
THE BEGINNING OF THE PATHWAY
= Each molecule of 3-phosphoglycerate is rearranged to 2-phosphoglycerate
= Each molecule of 2-phosphoglycerate is rearranged (and dehydrated) to phosphoenolpyruvic
acid; this creates a high-energy phosphate bond
= Finally, the high-energy phosphate is transferred to ADP, producing ATP and pyruvic acid
- In summary of Embden-Meyerhof glycolysis,
= Glucose has been oxidized to two molecules of pyruvic acid
= Four molecules of ATP have been synthesized; since two ATPs were used to get glycolysis
started, there is a net gain of 2 ATP from glycolysis
= Two molecules of NAD+ have been reduced to NADH; as we shall see, these are the treasure
of respiration and the trash of fermentation
- The Embden-Meyerhof pathway is not the only pathway for oxidation of glucose
= In parallel with glycolysis, many bacteria shunt some of their glucose into the pentose
phosphate pathways, which yield
· five-carbon sugars, such as the ribose used in synthesis of nucleotides
· NADPH, which is "preferred" to NADH in providing electrons to biosynthesis reactions
= The Entner-Deuderoff pathway is another glycolytic pathway used by some (almost always
respiratory) bacteria
If a microorganism possesses respiratory electron transport chains, and if a terminal electron
acceptor such as O2 is available, the pyruvate that is the final product of glycolysis can be further
oxidized to CO2
- The first step of this process is oxidative decarboxylation of pyruvic acid (Tortora et al., Figure
5.12)
= In this reaction, CO2 is released and the remaining acetyl group is transferred to a molecule
of coenzyme A, producing acetyl CoA
= Electrons removed during the reaction are used to reduce NAD+ to NADH
- The acetyl group of acetyl CoA can now enter the Krebs cycle, also known as the tricarboxylic
acid (TCA) cycle or the citric acid cycle (Tortora et al., Figure 5.12)
= The Krebs cycle is a (terrifyingly?) complex set of reactions which, for our present purposes,
can be viewed as a mechanism for stripping electrons from acetyl CoA
· From each molecule of acetyl CoA entering the cycle, three molecules of NAD+ are
reduced to NADH
· In one reaction, a molecule of FAD (flavine adenine dinucleotide), another electron-
carrying coenzyme, is reduced to FADH2
· Two of the oxidation steps are linked to release of CO2
· There is one ATP-producing (GTP-producing in eucaryotes) reaction in the Krebs cycle
= In summary of the Krebs cycle, the acetyl group of acetyl CoA is oxidized to two CO2 , with
electrons being used to produce three molecules of NADH, and one molecule of FADH2 ,
with one molecule of ATP as a "bonus"
So what, at this point, has happened to our molecule of glucose?

- For one thing, it has been thoroughly oxidized to CO2
= It has been oxidized to two molecules of pyruvic acid
= Each molecule of pyruvate has been oxidized to a molecule of acetyl CoA and a molecule of
CO2
= Each molecule of acetyl CoA has been oxidized to two molecules of CO2
- The electrons removed during the oxidations have been transferred to electron-carrying
coenzymes
= Two molecules of NADH were produced during glycolysis
= Two molecules of NADH (one for each molecule of pyruvic acid) were produced during
oxidative decarboxylation of pyruvic acid
= Six molecules of NADH (three for each acetyl CoA) were produced during the Krebs cycle
= Two molecules of FADH2 (one for each acetyl CoA) were produced during the Krebs cycle
- There has been a net production of four ATP molecules (two from glycolysis, two from the
Krebs cycle)

8777799999p999113lecture0706222

Uploaded by

Copyright:

Available Formats

8777799999p999113lecture0706222

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

8777799999p999113lecture0706222

Uploaded by

Copyright:

Available Formats

BIOLOGY 113 - MICROBIOLOGY

Lecture 7: Microbial Metabolism - Oxidation-reduction, mechanisms of ATP generation,

So what, at this point, has happened to our molecule of glucose?

You might also like