Microorganisms as research tools 1

MICROORGANISMS AS RESEARCH TOOLS

Abstract
Microorganisms have been used as a tool to explore fundamental life processes by
researchers. Due to some advantages (e.g. rapid growth, growth manipulation, easy & quick
culture) microbes frequently used as research tools in different fields. Microbes play an
important role in the research of enzyme structure & mode of action, drug invention, cellular
regulatory mechanism, energy metabolism, protein synthesis, structure of viruses. Bacteria
infect host cells, often using toxins, and shield themselves with inbuilt protective
mechanisms.

The team looked at the common bacteria Streptococcus pyogenes to

determine the structures of its toxin and antitoxin. The antitoxin deactivates the toxin by
binding to it. When not bound, the antitoxin changes shape.British bacteriologist Frederick
Griffith discovered that a harmless strain of Streptococcus pneumoniae could be made
virulent after being exposed to heat-killed virulent strains. An efficient and convenient
procedure for transforming bacteria and opened the way for molecular cloning in
biotechnology and research. A biofilm is an aggregate of microorganisms in which cells
adhere to each other and/or to a surface.The engineered bacteria-attacking virus, or phage,
was built using a plug and play library of genes. The same approach could be used to
build viruses custom tailored to target specific bacteria species, the researchers say.
Keywords: advantages, result & rewards of using microorganism as research tools,
Bacterias self defense mechanism, Transformation (genetics),Scientists engineer viruses to
destroy bacteria,Use of bacteria as anticancer agent.

Microorganisms as research tools 2

Any noncellular or unicellular (including colonial) organism, most of

which are too small to be seen with the unaided eye. Microorganisms comprise
bacteria (including cyanobacteria), lichens, microfungi, protozoa, rickettsiae, virinos,
viroids, and viruses, and also some algae; all prokaryotes are included.
The study of microorganisms is called microbiology, a subject that began with Anton
van Leeuwenhoek's discovery of microorganisms in 1675, using a microscope of his
own design.( Pelczar,1990)
Microorganisms have been used as a tool to explore fundamental life processes
because of many advantages(Smith, 2000) :
1. They grow (produce) very rapidly
2. Growth can be manipulated easily by chemical or physical means
3. Microorganisms can be cultured small or vast quantities conveniently and rapidly
4. Their cells can be broken apart & the contents separated into fractions of various
particle sizes
5. Lysed cells can be studied in terms of specific chemical reactions, specific products
and specific structures involved.
Scientists from many disciplines recognized the usefulness of microorganisms
experimental models. Then physicists, chemists and biologist joined with
microbiologists in molecular biology.
The result & rewards from the field of research is spectaculars.( Sharma, 2006) .
The contribution includes :

Microorganisms as research tools 3

1.Elucidation of enzyme structure & mode of action.

2.Celluler regulatory mechanism
3.Energy metabolism
4.Protein synthesis
5.Structure of viruses
Here we discussed about some microbes whcich were used as a resechal model or tool
in experiments:
Bacterias self defense mechanism might work against fighting infections
Bacteria infect host cells, often using toxins, and shield themselves with inbuilt
protective mechanisms. These pathways could become their Achilles heel,
according to researchers from Washington University in a study published on Feb. 9
in the journal Structure.
The team looked at the common bacteria Streptococcus pyogenes to determine the
structures of its toxin and antitoxin. The antitoxin deactivates the toxin by binding to
it. When not bound, the antitoxin changes shape.
Thats the Achilles heel that we would like to exploit, said Dr. Thomas Ellenberger,
head of the universitys Department of Biochemistry and Molecular Biophysics, in a
release. A drug that would stabilize the inactive form of the immunity factor would
liberate the toxin in the bacteria.
Experts say that the bacterium protects itself from the secreted toxic by keeping it in
an inactive form, by counter producing its antidote. When toxin complexes with the

Microorganisms as research tools 4

antitoxin molecule (immunity factor), it remains inactive and is unable to cause any
harm to the host or the bacterial cell itself.

Figure 1 :Microalgae under the electron microscope.

The bacterias toxin is called Streptococcus pyogenes beta-NAD+ glycohydrolase, or
SPN. Cells store the coenzyme NAD+ as part of their metabolism, and the toxin
works by draining these stores. The bacterias energy supply is protected by the
antitoxin, the immunity factor for SPN (IFS), which blocks the toxins access to
NAD+.
The most important aspect of the structure is that it tells us a lot about how the
antitoxin blocks the toxin activity and spares the bacterium, said Ellenberger.
Understanding how these bacteria cause disease in humans is important in drug
design.
There is a war going on between bacteria and their hosts, said coauthor Dr. Craig
Smith, a postdoctoral researcher at the university, in the press release. Bacteria
secrete toxins and we have ways to counterattack through our immune systems and

Microorganisms as research tools 5

with the help of antibiotics. But, as bacteria develop antibiotic resistance, we need to
develop new generations of antibiotics.
Antibiotics work in different ways; some block bacterial cell wall synthesis, while
others interfere with DNA synthesis or even inhibit bacterial metabolism.
As of yet, there are no classes of drugs that attack the protective antitoxin
mechanisms of bacteria.
Obviously they could evolve resistance once you target the antitoxin, Ellenberger
said. But this would be a new target. Understanding structures is a keystone of drug
design.

( Arshdeep 2011)
Transformation (genetics)

Transformation was first demonstrated in 1928 by British

bacteriologist Frederick Griffith. Griffith discovered that a harmless
strain of Streptococcus pneumoniae could be made virulent after
being exposed to heat-killed virulent strains. Griffith hypothesized
that some "transforming principle" from the heat-killed strain was
responsible for making the harmless strain virulent. In 1944 this
"transforming principle" was identified as being genetic by Oswald
Avery, Colin MacLeod, and Maclyn McCarty. They isolated DNA from
a virulent strain of S. pneumoniae and using just this DNA were
able to make a harmless strain virulent. They called this uptake and
incorporation of DNA by bacteria "transformation" See AveryMacLeod-McCarty experiment.

Microorganisms as research tools 6

The results of Avery et al.'s experiments were at first sceptically

received by the scientific community and it was not until the
development of genetic markers and the discovery of other
methods of genetic transfer (conjugation in 1947 and transduction
in 1953) by Joshua Lederberg that Avery's experiments were
accepted. Transformation did not become routine procedure in
laboratories until 1972 when Stanley Cohen, Annie Chang and
Leslie successfully transformed Escherichia coli by treating the
bacteria with calcium chloride. This created an efficient and
convenient procedure for transforming bacteria and opened the
way for molecular cloning in biotechnology and research.
Transformation using electroporation was developed in the late
1980s thus increasing the efficiency and number of bacterial
strains that could be transformed. Transformation of animal and
plant cells was also investigated with the first transgenic mouse
being created by injecting a gene for a rat growth hormone into a
mouse embryo in 1982. In 1907 a bacterium that caused plant
tumors, Agrobacterium tumefaciens, was discovered and in the
early 1970s the tumor inducing agent was found to be a DNA
plasmid called the Ti plasmid. By removing the genes in the
plasmid that caused the cancer and adding in novel genes
researchers were able to infect plants with A. tumefaciens and let
the bacteria insert their chosen DNA into the genomes of the

Microorganisms as research tools 7

plants. Not all plant cells are susceptible to infection by A.

tumefaciens so other methods were developed including
electroporation and micro-injection. Particle bombardment was
made possible with the invention of the Biolistic Particle Delivery
System (gene gun) by John Sanford in 1990. (wikipedia, 2011)
Scientists engineer viruses to destroy bacteria
Synthetic creation attacked biofilms that can form on teeth, in crevices.
A biofilm is an aggregate of microorganisms in which cells adhere to each other
and/or to a surface. These adherent cells are frequently embedded within a selfproduced matrix of extracellular polymeric substance (EPS). Biofilm EPS, which is
also referred to as slime (although not everything described as slime is a biofilm), is
a polymeric conglomeration generally composed of extracellular DNA, proteins, and
polysaccharides. Biofilms may form on living or non-living surfaces and can be
prevalent in natural, industrial and hospital settings.

Microorganisms as research tools 8

Figure 2: Staphylococcus aureus biofilm

The engineered bacteria-attacking virus, or phage, was built using a plug and
play library of genes. The same approach could be used to build viruses custom
tailored to target specific bacteria species, the researchers say.
The library could contain different phages that target different species or strains of
bacteria, each constructed using related design principles to express different
enzymes, said study leader James Collins, a biomedical engineer at Boston
University. (Jain KK 2011)
Use of bacteria as anticancer agent
Historically, bacteria were used as oncolytic agents for malignant
brain tumours. Advances in bacteriology and molecular biology
have widened the scope of bacterial approaches to cancer therapy
and various possibilities include the use of bacteria as sensitising
agents for chemotherapy, as delivery agents for anticancer drugs,
and as vectors for gene therapy. Bacterial toxins can be used for
tumour destruction and cancer vaccines can be based
immunotoxins of bacterial origin. The most promising approaches
are the use of genetically modified bacteria for selective
destruction of tumours, and bacterial gene-directed enzyme
prodrug therapy.

Microorganisms as research tools 9

Figure 3 : Salmonella
Knowledge gained from study of bacterial genomes forms an
important basis of use of bacteria as anticancer agents. TAPET (Tumour
Amplified Protein Expression Therapy) uses a genetically altered strain
of Salmonella as a bacterial vector, or vehicle, for preferentially
delivering anticancer drugs to solid tumours. Verotoxin 1 (VT1) of
Escherichia coli has been used for ex vivo purging of human bone
marrow of cancer cells before autologous bone marrow transplant.E.
coli genes and enzymes have become part of well-known prodrug
approaches to cancer in which inert prodrugs can be converted in vivo
to highly active species. IL-4 fused with Pseudomonas exotoxin has
been administered directly into malignant brain tumours and binds
with high affinity to IL-4 receptors, which do not exist on normal brain
cells, thus destroying a major part of the tumour without harming the

Microorganisms as research tools 10

normal brain tissue. It is in Phase I/II clinical trials in patients with

glioblastoma. No ideal anticancer agent of bacterial origin that is
applicable to all types of cancers has been discovered yet. The most
promising approach to malignant brain tumours appears to be the use
of genetically engineered bacteria that destroy the tumour selectively
while sparing the normal brain tissue. (Ker Than 2007)
References
Arshdeep Sarao,Epoch Times 2011, Bacterias Self Defense Mechanism Might
Work Against It.
A.Smith (2000), Oxford Dictionary of Biochemistry and Molecular Biology,Revised
edition (February 24, 2000) , Oxford University Press, USA
Jain KK ,PharmaBiotech, Blsiring 7, CH-4057 Basel, Switzerland.
Ker Than ,Live Science Magazine 2007
Pelczar, M.J, Chan, E.C.S & N. R. Krieg. Microbiology- Concepts and Applications
(International Edition), .McGraw- Hill Inc. New Delhi
S. Sharma,,General microbiology,Microbial World, History and Development
of Microbiology, Scope of Microbiology,(Revised 12-Dec-2006)
Wikipedia , 23 February 2011
From : http://en.wikipedia.org/wiki/Transformation_%28genetics%29

Microorganisms as research tools 11