Compare cell wal structures of gram positive with gram negative bacteria ?

Danish scientist Hans Christian Gram devised a method to differentiate two types of bacteria
based on the structural differences in their cell walls. In his test, bacteria that retain the crystal
violet dye do so because of a thick layer of peptidoglycan and are called Gram-positive
bacteria. In contrast, Gram-negative bacteria do not retain the violet dye and are colored red or
pink. Compared with Gram-positive bacteria, Gram-negative bacteria are more resistant against
antibodies because of their impenetrable cell wall. These bacteria have a wide variety of
applications ranging from medical treatment to industrial use and Swiss cheese production.

Comparison chart

Gram-negative Bacteria versus Gram-positive Bacteria comparison chart

Gram-negative Bacteria Gram-positive Bacteria

Gram reaction Can be decolourized to accept counter
stain (Safranin or Fuchsine); stain red or
pink, they don't retain the Gram stain
when washed with absolute alcohol and
acetone.
Retain crystal violet dye and stain dark
violet or purple, they remain coloured
blue or purple with gram stain when
washed with absolute alcohol and
water.

Peptidoglycan layer Thin (single-layered) Thick (multilayered)

Teichoic acids Absent Present in many

Periplasmic space present Absent

Outer membrane Present Absent

Lipopolysaccharide High Virtually none

(LPS) content

Lipid and lipoprotein High (due to presence of outer Low (acid-fast bacteria have lipids
content membrane) linked to peptidoglycan)

Flagellar structure 4 rings in basal body 2 rings in basal body

Toxins produced Primarily Endotoxins Primarily Exotoxins

Resistance to physical Low High

disruption

Inhibition by basic Low High

dyes

Susceptibility to Low High

anionic detergents

Resistance to sodium Low High

azide

Resistance to drying Low High

 Gram-negative Bacteria versus Gram-positive Bacteria comparison chart

Gram-negative Bacteria Gram-positive Bacteria

Cell wall composition The cell wall is 70-120 Å (ångström) The cell wall is 100-120 Å thick; single
thick; two layered. Lipid content is 20- layered. Lipid content of the cell wall is
30% (high), Murein content is 10-20% low , whereas Murein content is 70-
(low). 80% (higher).

Mesosome Mesosome is less prominent. Mesosome is more prominent.

Antibiotic Resistance More resistant to antibiotics. More susceptible to antibiotics

which bacteria (gram positive or gram negative) would be more resistant to

antibiotics?
Gram negative bacteria are more resistant to antibiotics because of the following reasons:
 The outer membrane:
The distinctive feature of gram-negative bacteria is the presence of a double membrane surrounding
each bacterial cell. Although all bacteria have an inner cell membrane, gram-negative bacteria have a
unique outer membrane. This outer membrane excludes certain drugs and antibiotics from penetrating
the cell, partially accounting for why gram-negative bacteria are generally more resistant to antibiotics
than are gram-positive bacteria.
Some Examples Of Gram-Negative Bacilli
Bartonella, Brucella, Campylobacter, Enterobacter, Escherichia, Francisella, Hemophilus,
Klebsiella, Morganella, Proteus, Providencia, Pseudomonas, Salmonella, Serratia, Shigella,
Vibrio, and Yersinia.
 The LPS core:
The outer membrane of gram-negative bacteria is rich in a molecule called lipopolysaccharide. If
gram-negative bacteria enter the bloodstream, lipopolysaccharide can trigger a cascade of events,
including high fever and a drop in blood pressure. For this reason, lipopolysaccharide is often
referred to as an endotoxin.
 Mutations & horizontal gene transfer:
Gram-negative bacteria have a great facility for exchanging genetic material (DNA) among strains of
the same species and even among different species.
This means that if a gram-negative bacterium either undergoes a genetic change (mutation) or
acquires genetic material that confers resistance to an antibiotic, the bacterium may later share its
DNA with another strain of bacteria and the second strain can become resistant as well.
Definition of bacterial plasmid ? type of plasmid

A plasmid is an extra-chromosomal element, often a circular DNA. The plasmids we will use in this
class typically have three important elements:

 An origin of replication
 A selectable marker gene (e.g. resistance to ampicillin)
  A cloning site (a place to insert foreign DNAs)

briefly summarize the process of cellular respiration ?

cellular respiration is the enzymatic breakdown of glucose (C6H12O6) in the presence of
oxygen (O2) to produce cellular energy (ATP):

C6H12O6 + 6O2 ® 6 CO2 + 6H2O + 38 ATP

THREE STAGES OF CELLULAR RESPIRATION

1. Glycolysis: (Fig. 18-2)

a ten-step process that occurs in the cytoplasm

converts each molecule of glucose to two molecules of pyruvic acid (a 3-carbon molecule)
an anaerobic process - proceeds whether or not O2 is present ; O2 is not required
net yield of 2 ATP per glucose molecule
net yield of 2 NADH per glucose (NADH is nicotine adenine dinucleotide, a co-enzyme that
serves as a carrier for H+ ions liberated as glucose is oxidized.)
The pyruvic acid diffuses into the inner compartment of the mitochondrion where a transition
reaction (Fig. 18-3) occurs that serves to prepare pyruvic acid for entry into the next stage of
respiration:

(a) pyruvic acid ® acetic acid + CO2 (a waste product of cell metabolism) + NADH+

(b) acetic acid + co-enzyme A ® acetyl CoA

2. Citric Acid or TCA Cycle: (Fig. 18-3)

occurs in the inner mitochondrial matrix

the acetyl group detaches from the co-enzyme A and enters the reaction cycle
an aerobic process; will proceed only in the presence of O2
net yield of 2 ATP per glucose molecule (per 2 acetyl CoA)

net yield of 6 NADH and 2 FADH2 (FAD serves the same purpose as NAD)
in this stage of cellular respiration, the oxidation of glucose to CO2 is completed

3. Electron Transport System:

consists of a series of enzymes on the inner mitochondrial membrane
electrons are released from NADH and from FADH2 and as they are passed along the
series of enzymes, they give up energy which is used to fuel a process called chemiosmosis
by which H+ ions are actively transported across the inner mitochondrial membrane into the
outer mitochondrial compartment. The H+ ions then flow back through special pores in the
membrane, a process that is thought to drive the process of ATP synthesis.
net yield of 34 ATP per glucose molecule
6 H2O are formed when the electrons unite with O2* at the end of electron transport chain. [*
Note: This is the function of oxygen in living organisms!]

when the fermentation process will take place in take place instead of respiration ?
what are the major differences of the two processes?
When the pyruvate is not oxidized, it undergoes the process of fermentation. It is then converted into
the waste products lactate or lactic acid (lactic acid fermentation) and ethanol (ethanol
or alcoholic fermentation).
Describe two methods for microbial growth measurement figure
out the advantage and disadvantages of this method?

Direct Methods of Measurement

Direct Microscopic Count:
4 A specific volume of a bacterial suspension (0.01 ml) is
placed on a microscope slide with a special grid.
4 Stain is added to visualize bacteria.
4 Cells are counted and multiplied by a factor to obtain
concentration.
Advantages:
• No incubation time required.
Disadvantages:
• Cannot always distinguish between live and dead bacteria.
• Motile bacteria are difficult to count.
• Requires a high concentration of bacteria (10 million/ml).

Indirect Methods of Measurement

1. Turbidity:
4 As bacteria multiply in media, it becomes turbid.
4 Use a spectrophotometer to determine % transmission or
absorbance.
4 Multiply by a factor to determine concentration.
Advantages:
• No incubation time required.
Disadvantages:
• Cannot distinguish between live and dead bacteria.
• Requires a high concentration of bacteria (10 to 100 million
cells/ml).
 The differences between continuous culture and batch culture? when continuous culture should
be used instead of batch culture?

Batch vs Continuous Culture

Batch culture technique is used to cultivate Continuous culture technique is used to grow
beneficial microorganisms under limited beneficial microorganisms under optimum
amounts of nutrients in a closed fermenter for level of nutrients in an open system in which
a certain time period. Microbial growth inside nutrients are added continually and waste
the batch culture shows a typical microbial and products are removed at the same rate
growth curve in which four distinct phases can to keep the growth at an exponential phase.
be identified.
Nutrients
Nutrients are supplied once before starting the Nutrients are added many times (at starting
fermentation process. and in between the process).
Type of System
Batch culture is a closed system Continuous culture is an open system.
Termination of Process
The process of the batch culture is stopped The process is not stopped though the
after the product is formed. product is formed. Continuous removal of the
product is done without stopping the process
in continuous culture.
Environmental Conditions
The environmental conditions inside the batch The environmental conditions inside the
culture are not constant. continuous culture are maintained at
constant level.
Microbial Growth
Microbial growth inside the batch culture Microbial growth is maintained at optimum
follows lag, log and stationary phases. level which is an exponential growth stage.
Turnover Rate
Turnover rate is low since the nutrients and Turnover rate is high since the optimum
other conditions are limited inside. levels of nutrients and other conditions are
maintained.
Fermenter Used
A large size fermenter is used for batch A small size fermenter is used for continuous
cultures culture.
Use
Batch culture fermentation is commonly used Continuous culture fermentation is less used
in industries in industries.
Culture Setup
Batch culture setup is easy to make and run. Continuous culture setup is not easy to make
and run.
Contamination
Contaminations are minimum in batch cultures Contamination chance is high in continuous
culture.
Controlling Methods
Control methods are easy and quick. Control methods are complicated and time-
consuming.
Suitability
Batch culture is more suitable for the Continuous culture is more suitable for the
production of secondary metabolites such as production of primary metabolites such as
antibiotics. amino acids and organic acids.

differentiate prokaryotes from eukaryotes?

Prokaryotic Cell Eukaryotic Cell

These are organisms made up of These organisms are made up of

cells that lack a cell nucleus or cells that possess a membrane-
Definition
any membrane-encased bound nucleus as well as
organelles. membrane-bound organelles.

It has a true nucleus, bounded

Nucleus It has no nucleus.
by a double membrane.

DNA arrangement It has a circular loop. It is linear.

Size Small cells ( < 5 µm) Large cells ( < 10 µm)

Cell Always unicellular Mostly multi-cellular

Usually present; chemically When present, chemically

Cell wall
complex in nature simple in nature

It does not contain protein in its It contains proteins in the DNA

Protein
DNA. to form chromatin.
 Ribosome It contains small ribosomes. It contains large ribosomes.

Cytoplasm No cytoskeleton Always have cytoskeleton

Cell division Cell division is by binary fission Cell division is by mitosis

Reproduction is asexual or
Reproduction Reproduction is always asexual
sexual

Huge variety of metabolic

Metabolic pathways Common metabolic pathways
pathways

It is complex in nature and

Consist of two protein building
Flagella consists of multiple
blocks
microtubules

Common with extensive tissue

Multi-cellular forms Rare
formation

They perform functions of

golgi-bodies and mitochondria,
Mesosomes Not present
and also help in separation of
chromosomes.

No carbohydrates and lacks Sterols and carbohydrates are

Plasma membrane
sterols both present

Present as a capsule or slime Present in some cells which lack

Glycocalyx
layer a cell wall

Example Bacteria and Archaea Animal cells and plant cells

Food preservation is to prevent the growth of microorganisms (such as yeasts), or other
microorganisms (although some methods work by introducing benign bacteria or fungi to the food), as
well as slowing the oxidation of fats that cause rancidity. Food preservation may also include
processes that inhibit visual deterioration, such as the enzymatic browningreaction in apples after they
are cut during food preparation.
Protection of foods from microbial spoilage using salt (usually sodium chloride) or sugar (usually
sucrose) has ancient roots and is often referred to as salting, salt curing, corning or sugar curing.
(Pieces of rock salt used for curing are sometimes called corns, hence the name "corned beef.")
Curing may utilize solid forms of salt and sugar or solutions in which salt or sugar is mixed with
water. For instance, brine is the term for salt solutions used in curing or pickling preservation
processes. Examples of foods preserved with salt or sugar include the aforementioned corned beef as
well as bacon, salt pork, sugar-cured ham, fruit preserves, jams and jellies, among others.

describe the step of respiration process

Glycolysis
The first step in aerobic respiration is glycolysis, which literally means the breakdown of glucose.
This process takes place in the cytoplasm, which is a jelly-like substance in your cells. During
glycolysis, molecules of glucose are broken down to yield four molecules of ATP, two three-
carbon molecules called pyruvate and two molecules of nicotinamide adenine dinucleotide, or
NADH. Although four ATP molecules are created here, the net result is only two molecules of
ATP. This is because glycolysis actually uses two ATPs during the first phase of the process to
generate glyceraldehyde-3-phosphate.

Acetyl-CoA
The next step in aerobic respiration is the formation of acetyl-coenzyme A. This occurs in the
mitochondria, which are small energy organelles within your cells. The pyruvate that was created
during glycolysis is converted to a two-carbon acetyl group, which then combines with coenzyme
A to produce acetyl-coA.

Krebs Cycle
The third step in aerobic respiration also takes place in your mitochondria. The acetyl-coA that
was produced from pyruvate combines during the Krebs cycle to produce oxaloacetate, thus
forming citrate. This citrate then undergoes several conversion steps to form the following
compounds, in order: isocitrate, alpha ketoglutarate, succinyl-CoA, succinate, fumarate and
malate. Along the way, one molecule of guanosine triphosphate (GTP), three molecules of NADH
and one molecule of flavin adenine dinucleotide (FADH2), are produced. The GTP is then
converted into a molecule of ATP.

Electron Transport Chain

The final step of aerobic respiration is the electron transport chain, or ETC. This last step uses
the NADH and FADH2 that were created in previous steps to generate ATP. A lot of ATP,
actually -- 34 molecules of ATP to be exact. The ETC accomplishes this large production of ATP
by pumping out the hydrogens from the NADH and FADH2 to the inner membrane of your
mitochondria, thus creating an electrochemical proton (H+) gradient. Chemical energy is
therefore generated, and this energy is used to create energy in the form of ATP via the ATP
synthase enzyme.

how bacteria reproduce ? describe life cycle of bacteria?

Reproduction of Bacteria

Even though they are single-celled microorganisms, bacteria can reproduce prolifically, if the
conditions are right. Like most of the other single-celled organisms, bacteria too undergo
reproduction through binary fission, which is a type of asexual reproduction. In this case,
identical generations are formed, because the daughter cells are identical clone cells only.
Budding is another form of asexual reproduction in bacteria. Even genetic recombination
occurs in different types of bacteria, through transduction, transformation, and conjugation.

Binary Fission

This is a form of asexual reproduction, which is common among bacteria. In this type of
reproduction, a single parent cell divides into two, and forms two daughter cells, which will
be replicas of the parent. In order to undergo binary fission, the bacterial cells must grow to a
fixed size. Once they reach that size, each cell produces a replica of the genetic material,
and form two DNA molecules that attach to the cell membrane in different locations. The cell
membrane starts growing inwards in such a fashion that the two daughter clone cells are
formed with the two DNA molecules. It has been observed that, if the conditions are right,
bacteria can double in number through binary fission, within a short time of about ten
minutes!

Budding

This is another form of asexual reproduction in bacteria. Some types of bacteria reproduce
through budding, which is otherwise known as fragmentation. In this case, the mother cell
forms a bud at one end, and also makes a nucleus for the bud, through the process of
mitosis. The bud grows to the same size as that of the mother cell, that remains constant in
size. The bud separates from the mother, and forms a different organism. Even though the
nucleus of the bud is formed through mitosis, it has been observed that sometimes the
characteristics of the offspring may differ from that of the mother.

Sexual Reproduction

Though sexual reproduction is rare in bacteria, in some cases, genetic recombination is

facilitated through conjugation, transformation, or transduction. One of the reasons is that
bacteria formed through asexual reproduction have the same genetic material, and can get
affected by the same antibiotics. So genetic recombination helps them in creating bacteria
with variations in genetic material. The latter category may be resistant to the particular
antibiotics, or may be adapted to the changing environment.

In case of conjugation, the genetic material will be transferred between bacteria, as one
bacteria connects to the other through a tube called pilus. In case of transformation, DNA is
collected from the remnants of dead bacterial cells. In this case, the bacterial cells get
attached to the DNA of the dead bacteria, and this DNA is transported through the cell
membrane and incorporated to the genetic material of the live bacteria. In case of
transduction, genetic material is transferred through bacteriophages (viruses that attack
bacterial cells). As a bacteriophages attach to the bacterial cell, it inserts its genetic material
into the bacterial cell. This results in the formation of replicated bacteriophages inside the
bacterial cell, which opens up to release the former. The genetic material of this host
bacterial cell can attach to the DNA of any other bacterial cell, that is attacked by these new
bacteriophages.

To summarize, bacteria reproduce both sexually and asexually. Asexual method of

reproduction, especially binary fission, is commonly found in bacteria. Some types of
bacteria may resort to budding too. Sexual reproduction, though rare, happens in bacteria in
some special circumstances.

Bacteria have a number of stages to the life cycle, as detailed below.

Lag Phase – Bacteria do not grow during this stage. However, they do begin to
produce the amino acids needed for division. DNA begins to copy and if the
environment is appropriate for the bacteria then the lag phase may be very short.
Log Phase – During this phase, bacteria multiply rapidly, sometimes exponentially.
The DNA copy within a bacterium moves to the opposite side of the membrane and
then the bacterium pulls apart to produce two identical cells, which then go on to
divide and so on.
Stationary Phase – Bacteria growth slows down, due to the levels of waste increasing
and the shortage of space.
Death Phase – During this phase bacteria are no longer able to reproduce. Death may
occur as rapidly as the growth.

Summary of characteristics of typical bacterial cell structures

Predominant chemical
Function(s)
Structure composition
Flagella
Swimming movement
Protein
Pili
Stabilizes mating bacteria
Sex pilus during DNA transfer by Protein
conjugation
Attachment to surfaces;
Common pili or
protection against Protein
fimbriae
phagotrophic engulfment
Attachment to surfaces;
Capsules protection against
Usually
(includes phagocytic engulfment,
polysaccharide;
"slime layers" occasionally killing or
occasionally
and digestion; reserve of
polypeptide
glycocalyx) nutrients or protection
against desiccation
Cell wall
Prevents osmotic lysis of Peptidoglycan
Gram-positive
cell protoplast and confers (murein) complexed
bacteria
rigidity and shape on cells with teichoic acids
 Peptidoglycan prevents
Peptidoglycan
osmotic lysis and confers
(murein) surrounded
rigidity and shape; outer
Gram-negative by phospholipid
membrane is permeability
bacteria protein-
barrier; associated LPS
lipopolysaccharide
and proteins have various
"outer membrane"
functions
Permeability barrier;
transport of solutes;
Plasma Phospholipid and
energy generation;
membrane protein
location of numerous
enzyme systems
Sites of translation
Ribosomes RNA and protein
(protein synthesis)
Often reserves of Highly variable;
Inclusions nutrients; additional carbohydrate, lipid,
specialized functions protein or inorganic
Chromosome Genetic material of cell DNA
Extrachromosomal genetic
Plasmid DNA
material

Anabolism refers to the process which builds molecules the body

needs; it usually requires energy for completion. Catabolism refers
to the process that breaks down complex molecules into smaller
molecules; it usually releases energy for the organism to use.

Comparison chart
Anabolism versus Catabolism comparison chart

Anabolism Catabolism

Introduction Metabolic process that builds Metabolic process that breaks down large
molecules the body needs. molecules into smaller molecules.

Energy Requires energy Releases energy

Hormones Estrogen, testosterone, insulin, Adrenaline, cortisol, glucagon, cytokines.

growth hormone.

Effects on Anabolic exercises, which are Catabolic exercises are

Exercise often anaerobic in nature, generally usually aerobic and good at burning fat
build muscle mass. and calories.

Example : amino acids becoming polypeptides proteins becoming amino acids, proteins
(proteins), glucose becoming becoming glucose, glycogen becoming
 Anabolism versus Catabolism comparison chart

Anabolism Catabolism
glycogen, fatty acids becoming glucose, or triglycerides becoming fatty
triglycerides. acids.