Metabolism

Part I: Fermentations Part II: Respiration

Learning objectives are to gain an appreciation of:

Catabolism and anabolism ATP Generation and energy conservation Fermentation

Importance of Metabolism
Industrial
Yogurt, cheese Bread, wine, beer

Medical/Health
Strain identification Digestion

Environmental
Cycling of elements Pollutant transformation

Images: (1) www.bact.wisc.edu (2) en.wikipedia.org

Metabolism: the bigger picture

What is metabolism?
Chemical reactions that occur in living organisms in order to maintain life.

What does maintain life mean?

Growth and division Maintaining cellular structures Sense/respond to environment

Two parts of metabolism:

nutrients + C-source

Cell components

chemicals or light

Two parts of metabolism

Anabolism- synthesis of complex molecules from simpler ones during which energy is added as input Catabolism- the breakdown of larger, more complex molecules into smaller, simpler ones, during which energy is released, trapped, and made available for work

Anabolism and Growth

Appropriate nutrients:
Carbon source N, S, P sources Trace metals

Appropriate environmental conditions:

pH Oxygen Temperature. Light

Carbons Sources
Heterotroph Organic Compounds

Autotroph CO2

Anabolism (biosynthesis)

Energy Sources:
Chemotrophs Chemical compounds Organotrophs Organic compounds, i.e., glucose, succinate Litotrophs Inorganic compounds, S, Fe2+,CO2, H2, CH4 Phototrophs Light

Catabolism

ATP, pmf

ATP and Energy

Adenosine triphosphate two of the phosphate bonds are high energy bonds breaking bond to remove phosphate releases energy

Figure 5.6

The Role of ATP in Metabolism

Reactions in which the terminal phosphate of ATP is removed results in a:

Exergonic breakdown of ATP can be coupled with:

Energy conserving reactions are used to catalyze the formation of ATP from ADP and Pi, and thus to restore the energy balance of the cell

ATP is produced by three processes:

_________________ phosphorylation
Two reactions are coupled to make ATP.

__________________ phosphorylation
Carried out by: Requires:

Photophosphorylation

Chemical-based ATP synthesis:

Substrate level phosphorylation Respiration-linked phosphorylation

ATP synthesis by substrate-level phosphorylation

Phosphorylated intermediates are generated in: Glycolysis (Embden-Meyhof pathway) Tricarboxylic Acid Cycle (TCA) Fermentation Finally, the Pi is transferred from a high energy phosphorylated intermediate to ADP by a kinase

ADP obtains phosphate from metabolic intermediate

molecule which has a high energy bond

ATP is formed

Embden Meyerhof Pathway a.k.a. glycolysis

Major pathway for: Major pathway of: Widespread method of: The end result is the release of a small amount of energy conserved as ATP through: And fermentation end products.

Glycolysis and Fermentation

Glycolysis is an anoxic process It is divided into two major stages Nets two ATPs and two NADHs End product of glycolysis is:

The fate of this metabolite varies:

Energy Input

Reductions

ATP by SLP

Glucose + 6O2

6CO2 + 6H2O

G= -2830 kJ/mol

Glycolysis and NADH

Glycolysis also generates NADH But, cells need: Fermentation can regenerate:
This can be done through: Substrates can be reduced by NADH

Example:
Lactic acid fermentation

NADH

Fermentation
ATP production by substrate level phosphorylation

What is fermentation?
Widespread method of anaerobic metabolism The end result is the release of a small amount of energy
conserved as ATP through substrate level phosphorylation

Incomplete oxidation of substrates Need to have a fermentation balance

Oxidation-reduction state of products equal the substrates NAD+ recycled

Fermentation end products are generally secreted

Why is fermentation important?

Ecologically important for decomposition of organic material in anaerobic environments Byproducts are usually energy rich and used by other microbes as energy/carbon sources Useful in food industry Digestion

Lots of fermentable carbon sources

Substrate level P

Lactic Acid Fermentation: I

Carried out by several groups of bacteria
Lactobacillus and Lactococcus

Gram + No cytochromes Anaerobes Only use sugars

Lactic Acid Fermentation

Fermentations have to maintain redox balance.

Key Reaction

Lactic acid bacteria and dairy products

These bacteria are used to to make cheese and yogurt from milk. Carbon/energy source in milk is lactose Lactose is hydrolyzed Causes pH to decrease Milk proteins coagulate
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QuickTime and a TIFF (Uncompressed) decompressor are needed to see this picture.

http://web.mit.edu/esgbio/www/lm/s ugars/lactose.gif http://www.reluctantgourmet.com/images/cheese.jpg

What happens to the lactate?

Transported out by lactate symporter
Takes a proton out with it.

Lactate is still a good carbon/energy source.

Glucose  2 Lactate G= -196 kJ/mol Made 2 ATPs = +63 kJ/mol -133 kJ/mol left in lactate Wasted energy!

Lactic acid bacteria are wasteful or are they?

They live in organic rich environments without oxygen They generate a proton gradient so more ATP can be made They make acid which inhibits competitors They can grow in low iron environments

Substrates other than glucose

Monosaccharides
Fructose, mannose, galactose get converted to glucose-6-phosphate or fructose-6-phosphate

Disaccharides--get cleaved in monosaccharides by specific enzymes

Lactose => galactose and glucose Maltose => 2 glucoses Sucrose => glucose and fructose

Large complex polysaccharides

Starch, cellulose: found in plant material Glycogen: found in animals Bacterium needs specific enzyme to break down the sugar chains into monomers
The enzymes are often secreted.

Some Complex Polysaccharides

Cellulose
Aerobic decomposition: myxobacteria cytophaga sporocytophaga Fermented by: some clostridia glucanases

Starch
Amylases in: Bacillus acidocaldarius Streptococcus bovis Bacteroides amylophilus

Polysaccharides and Catabolic Enzymes

In many cases the sugar monomers are ultimately metabolized either by glycolysis or another pathway to generate pyruvate.

More energy can be conserved from glucose by oxidizing it to CO2

Fermentations products are not fully oxidized

G G

glucose >> 2 lactate + 2H+

= -198 kJ/mol

glucose + 6O2 >> 6CO2 + 6H2O = -2830 kJ/mol

Next Lecture
The maximum energy stored in glucose can be conserved only when its complete oxidation is coupled to the reduction of an external electron accepting substrate.

Cells do this by: Oxidative phosphorylation The tricarboxylic acid cycle

Study Questions
1. How is ATP produced when organisms grow fermentatively? Does it matter what type of fermentation is occurring? 2. Why is pyruvate reduced in fermentations? To what can it be reduced? What do the fermentation products have in common? 3. Do fermentations produce a lot of ATP? Why not? What types of environments might fermentative bacteria (such as lactic acid bacteria) do well in? 4. What are the roles of ATP and NAD+ in glycolysis? 5. Compare and contrast the metabolism of lactose or maltose with that of glucose? 6. Can molecules such as cellulose enter gram + cells? Gram cells? Why/why not? If not, how are they consumed?