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CH 5

Microbes obtain energy through metabolic pathways like catabolism and anabolism. [1] Catabolism breaks down molecules and releases energy as ATP, while anabolism uses this energy to form complex molecules. [2] Aerobic respiration in three stages - glycolysis, the Krebs cycle, and the electron transport chain - generates the most ATP. [3] Enzymes catalyze reactions and help control metabolic pathways.

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51 views

CH 5

Microbes obtain energy through metabolic pathways like catabolism and anabolism. [1] Catabolism breaks down molecules and releases energy as ATP, while anabolism uses this energy to form complex molecules. [2] Aerobic respiration in three stages - glycolysis, the Krebs cycle, and the electron transport chain - generates the most ATP. [3] Enzymes catalyze reactions and help control metabolic pathways.

Uploaded by

arvy
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© © All Rights Reserved
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Download as PPT, PDF, TXT or read online on Scribd
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General Microbiology MICR 130

Chapter 5
Microbial metabolism
Overview
•By combining what we’ve learned about basic chemistry

and anatomy we can consider how microbes obtain


energy.
•Microbes needenergy to sustain metabolic processes,
and to make more microbes.
•Microbes are metabolically diverse. At the species level,
however, they may be fastidious. We distinguish and
classify microbes on the basis of which reactions they
conduct.
Direction of reactions
Metabolic pathway. Sequence of enzyme-catalyzed
reactions in a cell. There are many pathways, but two
directions...
oCatabolic.Enzyme-mediated breakdown of complex
molecules to smaller molecules (degradation,
catabolism). Release energy, e.g., glucoseCO2+H2O
oAnabolic. Enzyme-mediated production of complex
organic molecules from less complex molecules
(biosynthesis, anabolism). Use energy, e.g., proteins
from amino acids.
Net outcome
o Catabolism (breakdown) provides energy in ATP, and
building blocks for anabolism. ATP couples catabolism and
anabolism as it is produced, or its terminal phosphate is
cleaved.

Energy Energy
release use
Catabolism Anabolism
breakdown formation

Fig. 5.1
Reaction fundamentals
 Collision Theory. All molecules are moving. If

colliding molecules react or not depends on, i) how


fast they are traveling, ii) if there is enough activation
energy* to start the reaction, and iii) if they are
properly oriented.
 Reaction rate. Can be enhanced by increasing
[substrate], temperature, pressure, etc. To a point! Or
by adding a catalyst…

*Minimum energy needed for a chemical reaction to occur. Thereafter


it will be self-sustaining.
Enzymes are catalysts
 Biological catalysts that accelerate reactions. Large

proteins (>10kDa MW), with unique shapes and


substrate-specific active site(s).
Their function is shown by -ase, e.g., sucrase hydrolyzes
sucrose to glucose and fructose; lactase hydrolyzes
lactose to glucose and galactose.
Control of enzyme activities
 Temperature and pH
TºC pH
control reaction rate.
Increasing temperature
enhances reaction rates.
Activity falls after an optimum is reached. Activity can
also be controlled by inhibitors (slide #10).

 Reaction rate increases as [substrate]


increases. Active sites on all molecules
become occupied; more substrate does
not accelerate reaction.
Enzyme activities
 Enzymes denature outside their Temp. and pH

range; their tertiary structure is lost as covalent and H-


bonds break.

Active Denatured

Substrate binds to
active site, is
‘transformed’, and the
products are released.
Enzymes
 Reaction may require inorganic co-factor, or organic

coenzyme. Coenzymes may remove e- from substrates


and donate elsewhere later.

To function, the inactive protein on the left needs the co-


factor in the center.
Control of enzyme activities
 Poisons and heavy metals combine with enzymes and
may prevent them functioning. They are inhibitors.
 Different types of inhibitor:
Competitive: Competes with the actual substrate for the
active site (A).
Non-competitive: Binds elsewhere on the enzyme, but by
doing so change the active site’s shape; actual substrate
cannot bind (B). Altered active site

A B

Blocked active site


Control of enzyme activities
 Feedback inhibition
Stops a cell producing too much
product.
Reaction product inhibits an
enzyme early in the pathway.
Shuts down entire production
process.
 Saves substrate or energy for
use elsewhere. Works in vitamin,
amino acid, nucleotide synthesis.
Where does the energy come from?
 e- in bonds in nutrients have energy. This is harnessed

in catabolic (degradative) pathways and stored in ATP.


Breaking unstable bonds in ATP then provides energy to
the cell; enzymes mediate such processes.
 Oxidation-Reduction (redox reaction)
Oxidation is loss of electrons. Often yields energy.
Reduction is gain of electrons. Molecule A gives e- to B.
(Not ox-red!) e- gain
e - e-
A is oxidized as it
donates an e- to B
Fig. 5.9
e- loss
Energy production
 As molecule A is oxidized and molecule B is reduced, 2

protons (H+ ions, or protons) are also removed from A;


A, the organic substrate loses 2e- and 2p+.
The oxidizing agent NAD+ coenzyme (B) is reduced as it
gains 2e- and 1p+; the second p+ is released.
2e-, 2p+
spare H+
(proton)
A B

NAD+
NADH
NAD+ Nicotinamide adenine dinucleotide helps enzymes by accepting H atoms
(or protons)
Energy production
 The now reduced NADH contains more energy than

NAD+. This energy can be used to generate ATP in later


reactions.
 Organic molecules with many hydrogen atoms, e.g.,
glucose (C6H12O6), provide a lot of energy for ATP
production.
C H O

NADH
Respiration
Carbohydrate respiration provides energy as ATP in three
main processes:
• Glycolysis* oxidizes glucose to CO2 through 10 steps to
pyruvic acid, producing NADH and 2 ATP.
• Kreb’scycle† oxidizes acetyl CoA (from pyruvic acid) to
CO2, and production of 2 ATP, NADH, and FADH2. In
bacterial cytoplasm, and in mitochondria in eukaryotes.
• ETS, oxidation of NADH and FADH2 provides electrons
for reactions that provide energy to generate 34 ATP.
*Glycolysis also known as the Embden-Meyerhof pathway
†
Kreb’s cycle also known as the Tricarboxylic acid cycle; glyoxylate in bacteria.
http://library.thinkquest.org/11226/main/c05.htm
Respiration
o Aerobic: Electrons lose energy as they move from NAD

to FAD, and other molecules, ultimately combining with


O and H+ ions (protons) to form water. The energy
derived pumps protons, which are used to produce ATP.
o Anaerobic: Final e- acceptor in many bacteria is not O,
so they occupy niches others cannot, but derive less
energy (and less ATP):
e- acceptor Products
NO3– NO2–, N2 + H2O
SO4– H2 S + H 2 O
CO2 CH4 + H2O (maybe only Archaea)
Metabolic diversity

humans
Testing bacteria physiology
Testing use of single carbon sources detects pathogens;
not all E. coli can kill you!
Durham tube

Mannitol fermentation a) Control. b) Staphylococcus


epidermidis is -ve. c) S. aureus produces lactic acid, no
gas. D) E. coli is +ve, produces gas – presumptive for
coliforms. (Phenol red turns yellow in acid pH.)
Indole test
 Some bacteria use tryptophanase (enzyme) to

hydrolyze tryptophan (amino acid) to indole, pyruvate


and ammonia. Differentiates Enterobacteriaceae species:
Medium contains tryptophan and Kovac’s reagent. p-
DMA in the reagent reacts with indole, if released, to
produce a red color at the top of the
medium.
Distinguishes Enterobacter
Aerogenes(-) and E. coli(+)
.

*p-DMA: p-dimethylaminobenzaldehyde in isoamyl alcohol


‘Modern’ methods…

http://www.jlindquist.net/generalmicro/

http://pages.usherbrooke.ca/biomedias/techniques_microbio.htm
Motility test
 Differentiates species in terms of whether or not they
are motile.
Stab culture into ‘Motility medium’ with inoculating
needle. Medium contains tryptose, sodium chloride,
agar, and a color indicator.
A motile culture grows away from the stab line. A non-
motile one grows only along the stab line. A colored
indicator makes the result easier to judge.
 Presumes the culture can grow in the medium!
Motility test

+ – +
Is it reliable? Is it worth the effort?
Inhibition of an enzyme where the inhibitor does not bind to the
active site but binds to another area of the enzyme is called…?
A. allosterocity
B. plasmolysis
C. competitive inhibition
D. non-competitive inhibition
E. enzymatic disturbance

The three principle stages of cellular respiration are?


A. glycolysis, krebs cycle, electron transport
B. Injury to plasma membrane, injury to proteins, damage to DNA
C. replication, transcription, translation
D. oxidation, reduction, conclusion
E. reduce, reuse, recycle

Which stage of aerobic respiration produces the most ATP?


A. Glycolysis
B. Embden-Meyerhof pathway
C. Krebs cycle
D. Electron transport chain
E. pentose phosphate pathway
Humans, fungi, worms, and most bacteria are…?
A. chemoheterotrophs
B. autotrophs
C. photoheterotrops
D. chemoautotrophs
E. atrophic

Most enzymes are:


A. Effective over a wide range of temperatures compared to the
optimum for the reaction they catalyze.
B. Only effective at temperatures of 0 to 20˚C.
C. Effective at the optimum temperature for the reaction they
catalyze.
D. Are only effective at temperatures of 20 to 40˚C.
E. Are only effective at temperatures of 40 to 80˚C
The removal of electrons (e-) from an atom or molecule is:
A. Oxidation
B. Reduction
C. Competitive inhibition
D. Non-competitive inhibition
E. Feed-back inhibition
Most of a cell’s energy is produced from the oxidation of…?
A. proteins
B. amino acids
C. nucleic acids
D. lipids
E. glucose

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