Universidad Autónoma de Nuevo León

School of chemistry.

Microbiology
Part 2: metabolism and growth

A summary by Carlos Ceron

As taught by Carlos Escárcega González PhD.

Contents
Bioenergetics..............................................................................................................................2
Chemical reactions involved in metabolism..............................................................................2
Glycolysis...................................................................................................................................3
Respiration.................................................................................................................................4

Anaerobic respiration.............................................................................................................5
Bioenergetics
Catabolism is the intracellular process of degrading a compound into smaller and simpler
products (e.g., glucose to co2 and h2o).
Anabolism is involved in the synthesis of more complex compounds (e.g., glucose to
glycogen) and requires energy.
Differences in microbial metabolism can be attributed partly to genetic differences and/or to
differences in their responses to changes in their environment. Even the same species may
produce different products when grown under different nutritional and environmental
conditions.
The energy for living cells is obtained from the catabolism of carbon compounds, mainly
carbohydrates. They are synthesized from co2 and h2o in the presence of light by
photosynthesis. The sun is the ultimate energy source for the life processes on earth.
Energy in biological systems is primarily stored and transferred via adenosine triphosphate
(atp), which contains high-energy phosphate bonds.

Chemical reactions involved in metabolism

Redox reactions fuel cellular metabolism, we define such reactions by the transfer of
electrons among chemical species so that when a species loses electrons it oxidizes and when
it gains electros it reduces, we also define oxidation in a molecule that looses hydrogen atoms
or gains oxygen atoms. Oxidation reduces the potential energy of the molecule.
“the oxidation of glucose can be divided into two phases: the anaerobic and the aerobic. In
the anaerobic or glycolytic phase, glucose is metabolized to lactic acid. In the aerobic or
respiratory phase, pyruvic acid, or its reduction product, lactic acid, is metabolized to carbon
dioxide and water.” (ely, l. O., 1951)
Hydrogen atoms released in biological oxidation–reduction reactions are carried by
nicotinamide adenine dinucleotide (nad+) and nicotinamide adenine dinucleotide phosphate
(nadp+).
Atp formation in respiratory metabolism: the electrons (or h atoms) carried by nadh are
transferred to oxygen (respiratory chain) to produce up to three atp molecules.
Glycolysis
Glycolysis is the metabolic process that serves as the foundation for both aerobic and
anaerobic cellular respiration. In glycolysis, glucose is converted into pyruvate. Glucose is a
six- membered ring molecule found in the blood and is usually a result of the breakdown of
carbohydrates into sugars. It enters cells through specific transporter proteins that move it
from outside the cell into the cell’s cytosol. All the glycolytic enzymes are found in the
cytosol. Glycolysis takes place in cytoplasm, whereas the site for the krebs cycle is the matrix
of mitochondria in eucaryotes. In prokaryotes, these reactions are associated with membrane
bound enzymes.

 Step 1: hexokinase
The first step is the conversion of d-glucose into glucose-6-phosphate. The enzyme
that catalyzes this reaction is hexokinase.
 Step 2: phosphoglucose isomerase
The second reaction of glycolysis is the rearrangement of glucose 6-phosphate (g6p)
into fructose 6-phosphate (f6p) by glucose phosphate isomerase (phosphoglucose
isomerase).
 Step 3: phosphofructokinase
In the third step of glycolysis, fructose-6-phosphate is converted to fructose- 1,6-
bisphosphate (fbp). Similar to the reaction that occurs in step 1 of glycolysis, a second
molecule of atp provides the phosphate group that is added on to the f6p molecule.
The enzyme that catalyzes this reaction is phosphofructokinase (pfk).
 Step 4: aldolase
 The enzyme aldolase splits fructose 1, 6-bisphosphate into two sugars that are isomers
of each other. These two sugars are dihydroxyacetone phosphate (dhap) and
glyceraldehyde 3-phosphate (gap). The enzyme that catalyzes this reaction is aldolase.
 Step 5: triosephosphate isomerase
The enzyme triosephosphate isomerase rapidly inter- converts the molecules
dihydroxyacetone phosphate (dhap) and glyceraldehyde 3-phosphate (gap).
Glyceraldehyde phosphate is removed / used in next step of glycolysis.

 Step 6: glyceraldehyde-3-phosphate dehydrogenase

Glyceraldehyde-3-phosphate dehydrogenase (gapdh) dehydrogenates and adds an
inorganic phosphate to glyceraldehyde 3-phosphate, producing 1,3-
bisphosphoglycerate.
 Step 7: phosphoglycerate kinase
Phosphoglycerate kinase transfers a phosphate group from 1,3-bisphosphoglycerate to
adp to form atp and 3-phosphoglycerate.
 Step 8: phosphoglycerate mutase
The enzyme phosphoglycerol mutase relocates the p from 3- phosphoglycerate from
the 3rd carbon to the 2nd carbon to form 2-phosphoglycerate.
 Step 9: enolase
The enzyme enolase removes a molecule of water from 2-phosphoglycerate to form
phosphoenolpyruvic acid (pep).
 Step 10: pyruvate kinase
The enzyme pyruvate kinase transfers a p from phosphoenolpyruvate (pep) to adp to
form pyruvic acid and atp result in step 10.

Respiration
Respiration may be aerobic or anaerobic, depending on the final electron acceptor. If oxygen
is used as final electron acceptor, the respiration is called aerobic respiration. When other
electron acceptors, such as no3, so4, fe+3, cu+2, and s, are used, the respiration is termed
anaerobic respiration.
Aerobic respiration

Aerobic respiration requires oxygen (O2) in order to create ATP. Although carbohydrates,
fats, and proteins are consumed as reactants, it is the preferred method of pyruvate
breakdown in glycolysis and requires that pyruvate enter the mitochondria in order to be fully
oxidized by the Krebs cycle. The products of this process are carbon dioxide and water, but
the energy transferred is used to break bonds in ADP as the third phosphate group is added to
form ATP (adenosine triphosphate), by substrate-level phosphorylation, NADH and FADH2.
Aerobic metabolism is up to 15 times more efficient than anaerobic metabolism (which yields
2 molecules ATP per 1 molecule glucose) because the double bond in O2 is of higher energy
than other double bonds or pairs of single bonds in other common molecules in the biosphere.
[3] However, some anaerobic organisms, such as methanogens are able to continue with
anaerobic respiration, yielding more ATP by using other inorganic molecules (not oxygen) of
high energy as final electron acceptors in the electron transport chain. They share the initial
pathway of glycolysis but aerobic metabolism continues with the Krebs cycle and oxidative
phosphorylation. The post-glycolytic reactions take place in the mitochondria in eukaryotic
cells, and in the cytoplasm in prokaryotic cells.

Anaerobic respiration

Under anaerobic conditions, pyruvate may be converted to lactic acid, ethanol, or other
products, such as acetone, butanol, and acetic acid. Anaerobic conversion of glucose to the
compounds used to be known as fermentation. Under aerobic conditions, pyruvate is
converted to co2 and nadh through the tca cycle (tricarboxylic acid cycle).
The generation of energy without the electron transport chain is called fermentation
(anaerobic). Other partially oxidized byproducts are acetone, butanol fermentation, propionic
acid, acetic acid (for vinegar), 2,3- butanediol, isopropanol, and glycerol.
Microorganisms like azotomonas, azotococcus, and biejerinckia fix atmospheric nitrogen to
form ammonia. Rhizobium and azospirillum are widely used for agricultural purposes and are
bioprocess products.
Rhizobium and azospirillum are widely used for agricultural purposes and are bioprocess
products.
Growth
The way bacteria reproduce is called binary fission. Binary fission is the primary method of
reproduction of prokaryotic organisms. In protists, binary fission is often differentiated into
types, such as transverse or longitudinal, depending on the axis of cell separation. Regular
transverse fission in some organisms, such as tapeworms and scyphostome polyps, is called
strobilation.
The change in the number of cellular mass by unit of time is called growth speed, the time
taken by the bacteria to double in number during a specified time is known as the generation
time. The generation time tends to vary with different organisms. E. coli divides in every 20
minutes, hence its generation time is 20 minutes, and for Staphylococcus aureus it is 30
minutes.

Bacterial growth curve phases

1) Lag: When a microorganism is introduced into the fresh medium, it takes some time
to adjust with the new environment. The length of the lag phase depends directly on
the previous growth condition of the organism. The organism will start synthesising
the necessary proteins, co-enzymes and vitamins needed for their growth and hence
there will be a subsequent increase in the lag phase.
2) Exponential or Logarithmic (log) phase: During this phase, the microorganisms are in
a rapidly growing and dividing state. The growth medium is exploited at the maximal
rate, the culture reaches the maximum growth rate and the number of bacteria
increases logarithmically and finally the single cell divide into two, which replicate
into four, eight, sixteen, thirty two and so on This will result in a balanced growth.
The generation time tends to vary with different organisms.
3) Stationary phase: As the bacterial population continues to grow, all the nutrients in the
growth medium are used up by the microorganism for their rapid multiplication. This
result in the accumulation of waste materials, toxic metabolites, and inhibitory
compounds such as antibiotics in the medium. This shifts the conditions of the
medium such as pH and temperature, thereby creating an unfavorable environment for
the bacterial growth. The cell number is not increased and thus the growth rate is
stabilized.
4) Decline or Death phase: The depletion of nutrients and the subsequent accumulation
of metabolic waste products and other toxic materials in the media will facilitates the
 bacterium to move on to the Death phase. During this, the bacterium completely loses
its ability to reproduce. Individual bacteria begin to die due to the unfavourable
conditions and the death is rapid and at uniform rate. The number of dead cells
exceeds the number of live cells. Some organisms which can resist this condition can
survive in the environment by producing endospores.

Measurements
The increase in the cell size and cell mass during the development of an organism is termed
as growth. It is the unique characteristics of all organisms. The organism must require certain
basic parameters for their energy generation and cellular biosynthesis. The growth of the
organism is affected by both physical and Nutritional factors. Bacteria are unicellular
organisms. When the bacteria reach a certain size, they divide by binary fission, in which the
one cell divides into two, two into four and continue the process in a geometric fashion. The
bacterium is then known to be in an actively growing phase. The curve thus obtained is a
sigmoid curve and is known as a standard growth curve. The increase in the cell mass of the
organism is measured by using the Spectrophotometer. The Spectrophotometer measures the
turbidity or Optical density which is the measure of the amount of light absorbed by a
bacterial suspension.

Growth control
Sterilization is a method to exterminate microorganisms such as bacteria and virus. It
involves exposure to high temperature and pressure conditions, however if sterilization is not
possible one can limit the microorganism growth by other processes called inhibition.
Physical methods involve high temperatures as this kills most of the microbes including
endospores, this occur at a range of 100°C-130°C. An autoclave is used to sterilize surgical
equipment, laboratory instruments, pharmaceutical items, and other materials. It can sterilize
solids, liquids, hollows, and instruments of various shapes and sizes. Autoclaves vary in size,
shape and functionality. A very basic autoclave is similar to a pressure cooker; both use the
power of steam to kill bacteria, spores and germs resistant to boiling water and powerful
detergents. Autoclave sterilization time as reported by Brock, 2006 can be seen in the
following graph.

Pasteurization
It is mainly used for liquids, the objective of it is not to sterilize, but to inhibit the growth of
microorganism and thus reduce the amount present in the product. In this process liquids are
heated and then cooled rapidly this produces the death of most of the microorganisms without
changing the properties of the liquid. It is commonly used in the food industry.
Radiation is also used to control growth, a controlled amount of electromagnetic radiation is
fired to the substance and this kills most of the microorganisms, this process is preferred by
the medical industry as a way to sterilize tissues, drugs and medical laboratory supplies.

Chemical growth controls

Chemical substances known as bactericide, fungicides, antivirals are used to inhibit growth,
disinfect, or sterilize different objects. Antiseptic and germicides are also used to reduce
microbial growth in living tissue, contrary to disinfect which means to eliminate
microorganisms from inanimate objects.

Metabolic diversity
It is the reason why we are here, learning the different ways that nature has designed to be
able to use the environment, as harsh as it may be, to sustain life gives humanity creative
ways to deal with common known and unsolved problems. Our relationship as a species to
the metabolic diversity of microorganism is ancient, we learned that food and other raw
materials changed when exposed to specific conditions to produce rather interesting and tasty
products.
Know we describe some of the different metabolisms know.

Phototrophic metabolism

Phototrophy is the use of light as a source of energy for growth, more specifically the
conversion of light energy into chemical energy in the form of ATP. Procaryotes that can
convert light energy into chemical energy include the photosynthetic cyanobacteria, the
purple and green bacteria and the "halobacteria" (archaea). The cyanobacteria conduct plant
photosynthesis, called oxygenic photosynthesis; the purple and green bacteria conduct
bacterial photosynthesis or anoxygenic photosynthesis; the extreme halophilic archaea use a
type of nonphotosynthetic photophosphorylation mediated by bacteriorhodopsin to transform
light energy into ATP.
The functional components of the photochemical system are light harvesting pigments, a
membrane electron transport system, and an ATPase enzyme. The photosynthetic electron
transport system of is fundamentally similar to a respiratory ETS, except that there is a low
redox electron acceptor (e.g. ferredoxin) at the top (low redox end) of the electron transport
chain, that is first reduced by the electron displaced from chlorophyll.
Lithotrophy: use of inorganic substances as sources of energy
Photoheterotrophy: use of organic compounds as a carbon source during bacterial
photosynthesis
Anoxygenic photosynthesis: photophosphorylation in the absence of O2
Methanogenesis: an ancient type of archaean metabolism that uses H2 as an energy source
and produces methane
Light-driven nonphotosynthetic photophosphorylation: unique archaean metabolism that
converts light energy into chemical energy
In addition, among autotrophic procaryotes, there are three ways to fix CO2, two of which are
unknown among eucaryotes, the CODH (acetyl CoA pathway) and the reverse TCA cycle.

Energy-Generating Metabolism The term metabolism refers to the sum of the biochemical
reactions required for energy generation AND the use of energy to synthesize cell material
from small molecules in the environment. Hence, metabolism has an energy-generating
component, called catabolism, and an energy-consuming, biosynthetic component, called
anabolism. Catabolic reactions or sequences produce energy as ATP, which can be utilized in
anabolic reactions to build cell material from nutrients in the environment. The relationship
between catabolism and anabolism is illustrated in Figure 1 below.
Final thoughts
The world of the smallest and our world exist thanks to the fact that the individuals living in
them found a way to produce the energy that’s necessary to stay alive using what is present in
their surroundings, however the energy we use is the same, diversity (or progress) happens as
creatures discover new routes to take energy and transform it into a way they can use to grow
and reproduce.
The light we see, the electricity that powers our devices and the fire that makes us warm are
all present inside microscopic living creatures that have learned how to produce them; way
before we build electric generators or even discovered heat or described light, life figured out
how to transform energy, to do so it took atoms and arranged them beautifully to produce
energy by manipulating the fundamental forces of nature we are yet to understand. There is
no doubt life is a miracle, but as science dances with the divine in a never-ending effort to
explain it, even the smallest steps she makes are always giants leaps for mankind.

References:
1. Madigan M.T, Martinko J.M., Stahl D and Clark D.P., Brock Biology of microorganisms,
13th edition, UK, Pearson Benjamin Cummings, 2010.
2. Madigan M.T, Martinko J.M., Dunlap P.V. and Clark D.P., Brock Biología de los
microorganismos, 12a edición, UK, Pearson Education, 2009.
3. Prescott L.M., Harley J.P. and Klein G.A., Microbiología, 3a edición, Madrid, México, Mc
GrawHill-Interamericana, 2009.
4. Tortora G.J., Funke B.R. and Case C.L., Microbiology: An Introduction with Mastering