Problems (source: http://web.virginia.edu/Heidi/chapter1/problems.

htm)

1. The nutritional requirements oI Escherichia coli cells are Iar simpler than those oI humans, yet the
macromolecules Iound in bacteria are about as complex as those oI animals. Since bacteria can make all
their essential biomolecules while subsisting on a simpler diet, do you think bacteria may have more
biosynthetic capacity and hence more metabolic complexity than animals? Organize your thoughts on this
question, pro and con, into a rational argument.
. ithout consulting chapter Iigures, sketch the characteristic prokaryotic and eukaryotic cell types and
label their pertinent organelle and membrane systems.
3. Escherichia coli cells are about 2 mm (microns) long and 0.8 mm in diameter.
a. How many E coli cells laid end to end would Iit across the diameter oI a pin head? (Assume a pinhead
diameter oI 0.5 mm.)
-. hat is the volume oI an E. coli cell? (Assume it is a cylinder, with the volume oI a cylinder given
by V p r
2
h, where p 3.14.)
.. hat is the surIace area oI an E. coli cell? hat is the surIace-to-volume ratio oI an E coli cell?
d. Glucose, a major energy-yielding nutrient, is present in bacterial cells at a concentration oI about 1
mM. How many glucose molecules are contained in a typical E. coli cell? (Recall that Avogadro`s
number 6.023 x 10
23
.)
e. A number oI regulatory proteins are present in E. coli at only one or two molecules per cell. II we
assume that an E. coli cell contains just one molecule oI a particular protein, what is the molar
concentration oI this protein in the cell?
1. An E. coli cell contains about 15,000 ribosomes, which carry out protein synthesis. Assuming
ribosomes are spherical and have a diameter oI 20 nm (nanometers), what Iraction oI the E. .4 cell
volume is occupied by ribosomes?
. The E. coli chromosome is a single DNA molecule whose mass is about 3 x 10
9
daltons. This
macromolecule is actually a linear array oI nucleotide pairs. The average molecular weight oI a nucleotide
pair is 660, and each pair imparts 0.34 nm to the length oI the DNA molecule. hat is the total length oI
the E coli chromosome? How does this length compare with the overall dimensions oI an E. coli cell?
How many nucleotide pairs does this DNA contain? The average E. coli protein is a linear chain oI 360
amino acids. II three nucleotide pairs in a gene encode one amino acid in a protein, how many diIIerent
proteins can the E. coli chromosome encode? (The answer to this question is a reasonable approximation
oI the maximum number oI diIIerent kinds oI proteins that can be expected in bacteria.)
. Assume that mitochondria are cylinders 1.5 mm in length and 0.6 mm in diameter.
a. hat is the volume oI a single mitochondrion?
-. Oxaloacetate is an intermediate in the citric acid cycle, an
important metabolic pathway localized in the mitochondria oI eukaryotic cells. The concentration oI
oxaloacetate in mitochondria is about 0.03 mM. How many molecules oI oxaloacetate are in a single
mitochondrion?
5. Assume that liver cells are cuboidal in shape, 20 mm on a side.
a. How many liver cells laid end to end would Iit across the diameter oI a pin head? (Assume a pinhead
diameter oI 0.5 mm.)
-. hat is the volume oI a liver cell? (Assume it is a cube.)
.. hat is the surIace area oI a liver cell? hat is the surIace-to-volume ratio oI a liver cell? How does
this compare to the surIace-to-volume ratio oI an E. coli cell (compare this answer to that oI problem 3c)?
hat problems must cells with low surIace-to-volume ratios conIront that do not occur in cells with high
surIace-to-volume ratios?
d. A human liver cell contains two sets oI 23 chromosomes, each set being roughly equivalent in
inIormation content. The total mass oI DNA contained in these 46 enormous DNA molecules is 4 x
10
12
daltons. Since each nucleotide pair contributes 660 daltons to the mass oI DNA and 0.34 nm to the
length oI DNA, what is the total number oI nucleotide pairs and the complete length oI the DNA in a liver
cell? How does this length compare with the overall dimensions oI a liver cell? The maximal inIormation
in each set oI liver cell chromosomes should be related to the number oI nucleotide pairs in the
chromosome set`s DNA. This number can be obtained by dividing the total number oI nucleotide pairs
calculated above by 2. hat is this value? II this inIormation is expressed in proteins that average 400
amino acids in length and three nucleotide pairs encode one amino acid in a protein, how many diIIerent
kinds oI proteins might a liver cell be able to produce? (In reality, liver cells express at most about 30,000
diIIerent proteins. Thus, a large discrepancy exists between the theoretical inIormation content oI DNA in
liver cells and the amount oI inIormation actually expressed.)
6. Biomolecules interact with one another through molecular surIaces that are structurally
complementary. How can various proteins interact with molecules as diIIerent as simple ions,
hydrophobic lipids, polar but uncharged carbohydrates, and even nucleic acids?
7. hat structural Ieatures allow biological polymers to be inIormational macromolecules? Is it possible
Ior polysaccharides to be inIormational macromolecules?
8. hy is it important that weak Iorces, not strong Iorces, mediate biomolecular recognition?
9. hy does the central role oI weak Iorces in biomolecular interactions restrict living systems to a
narrow range oI environmental conditions?
10. Describe what is meant by the phrase 'cells are steady-state systems.



Source hLLp//wwwLexLbookofbacLerlologyneL/growLh_3hLml
The Growth of Bacterial Populations (page 3)

(This chapter has 4 pages)

2011 Kenneth Todar, PhD

The Bacterial Growth Curve
In the laboratory, under favorable conditions, a growing bacterial population doubles at regular intervals.
Growth is by geometric progression: 1, 2, 4, 8, etc. or 2
0
, 2
1
, 2
2
, 2
3
.........2
n
(where n = the number of
generations). This is calledexponential growth. In reality, exponential growth is only part of the bacterial
life cycle, and not representative of the normal pattern of growth of bacteria in Nature.
hen a fresh medium is inoculated with a given number of cells, and the population growth is monitored
over a period of time, plotting the data will yield atypical bacterial growth curve (Figure 3 below).

Figure 3. The typical bacterial growth curve. When bacteria are grown in a closed system (also
called a batch culture), like a test tube, the population of cells almost always exhibits these
growth dynamics: cells initially adjust to the new medium (lag phase) until they can start
dividing regularly by the process of binary fission (exponential phase). When their growth
becomes limited, the cells stop dividing (stationary phase), until eventually they show loss of
viability (death phase). Note the parameters of the x and y axes. Growth is expressed as change
in the number viable cells vs time. Generation times are calculated during the exponential phase
of growth. Time measurements are in hours for bacteria with short generation times.
Four characteristic phases of the growth cycle are recognized.
1. Lag Phase. Immediately after inoculation of the cells into fresh medium, the population remains
temporarily unchanged. Although there is no apparent cell division occurring, the cells may be growing in
volume or mass, synthesizing enzymes, proteins, RNA, etc., and increasing in metabolic activity.
The length of the lag phase is apparently dependent on a wide variety of factors including the size of the
inoculum; time necessary to recover from physical damage or shock in the transfer; time required for
synthesis of essential coenzymes or division factors; and time required for synthesis of new (inducible)
enzymes that are necessary to metabolize the substrates present in the medium.
2. Exponential (log) Phase. The exponential phase of growth is a pattern of balanced growth wherein all
the cells are dividing regularly by binary fission, and are growing by geometric progression. The cells divide
at a constant rate depending upon the composition of the growth medium and the conditions of incubation.
The rate of exponential growth of a bacterial culture is expressed asgeneration time, also the doubling
time of the bacterial population. Generation time (G) is defined as the time (t) per generation (n = number
of generations). Hence, G=t/n is the equation from which calculations of generation time (below) derive.
3. Stationary Phase. Exponential growth cannot be continued forever in abatch culture (e.g. a closed
system such as a test tube or flask). Population growth is limited by one of three factors: 1. exhaustion of
available nutrients; 2. accumulation of inhibitory metabolites or end products; 3. exhaustion of space, in this
case called a lack of "biological space".
During the stationary phase, if viable cells are being counted, it cannot be determined whether some cells
are dying and an equal number of cells are dividing, or the population of cells has simply stopped growing
and dividing. The stationary phase, like the lag phase, is not necessarily a period of quiescence. Bacteria
that produce secondary metabolites, such as antibiotics, do so during the stationary phase of the growth
cycle (Secondary metabolites are defined as metabolites produced after the active stage of growth). It is
during the stationary phase that spore-forming bacteria have to induce or unmask the activity of dozens of
genes that may be involved in sporulation process.
4. Death Phase. If incubation continues after the population reaches stationary phase, a death phase
follows, in which the viable cell population declines. (Note, if counting by turbidimetric measurements or
microscopic counts, the death phase cannot be observed.). During the death phase, the number of viable
cells decreases geometrically (exponentially), essentially the reverse of growth during the log phase.
Growth Rate and Generation Time
As mentioned above, bacterial growth rates during the phase of exponential growth, under standard
nutritional conditions (culture medium, temperature, pH, etc.), define the bacterium's generation time.
Generation times for bacteria vary from about 12 minutes to 24 hours or more. The generation time for E.
coli in the laboratory is 15-20 minutes, but in the intestinal tract, the coliform's generation time is estimated
to be 12-24 hours. For most known bacteria that can be cultured, generation times range from about 15
minutes to 1 hour. Symbionts such as Rhizobium tend to have longer generation times. Many lithotrophs,
such as the nitrifying bacteria, also have long generation times. Some bacteria that are pathogens, such
as Mycobacterium tuberculosis and Treponema pallidum, have especially long generation times, and this is
thought to be an advantage in their virulence. Generation times for a few bacteria are are shown in Table 2.
Table 2. Generation times for some common bacteria under optimal conditions of growth.
a.9erium Medium Genera9ion Time (minu9es)
Escherichia coli Glucose-salts 17
Bacillus megaterium Sucrose-salts 25
Streptococcus lactis Milk 26
Streptococcus lactis Lactose broth 48
Staphylococcus aureus Heart inIusion broth 27-30
Lactobacillus acidophilus Milk 66-87
Rhi:obium faponicum Mannitol-salts-yeast extract 344-461
Mycobacterium tuberculosis Synthetic 792-932
Treponema pallidum Rabbit testes 1980
Calculation of Generation Time
hen growing exponentially by binary fission, the increase in a bacterial population is by geometric
progression. If we start with one cell, when it divides, there are 2 cells in the first generation, 4 cells in the
second generation, 8 cells in the third generation, and so on. The generation time is the time interval
required for the cells (or population) to divide.
G (generation time) = (time, in minutes or hours)/n(number of generations)
G = t/n
t = time interval in hours or minutes
B = number of bacteria at the beginning of a time interval
b = number of bacteria at the end of the time interval
n = number of generations (number of times the cell population doubles during the time interval)
b = B x 2
n
(This equation is an expression of growth by binary fission)
Solve for n:
logb = logB + nlog2
n = logb - logB
log2
n = logb - logB
.301
n = 3.3 logb/B
G = t/n
Solve for G
G = t
3.3 log b/B


Example: What is the generation time of a bacterial population that increases from 10,000 cells
to 10,000,000 cells in four hours of growth?


G = t_____
3.3 log b/B
G = 240 minutes
3.3 log 10
7
/10
4

G = 240 minutes
3.3 x 3
G = 24 minutes