Microorganisms as research tools 1

MICROORGANISMS AS RESEARCH TOOLS

Submitted to:
Abu Taiab Md !amaluddin
Senior Lecturer
"e#artment o$ %harmac&
East 'est (ni)ersit&
Submitted b&:
Md Sa*ib Hossain
East 'est (ni)ersit&
+asic Microbiolog& %HRM,-,

Microorganisms as research tools 2
Abstract
Microorganisms ha)e been used as a tool to e.#lore $undamental li$e #rocesses b&
researchers "ue to some ad)antages /eg ra#id gro0th1 gro0th mani#ulation1 eas& 2 3uic4
culture5 microbes $re3uentl& used as research tools in di$$erent $ields Microbes #la& an
im#ortant role in the research o$ en6&me structure 2 mode o$ action1 drug in)ention1 cellular
regulator& mechanism1 energ& metabolism1 #rotein s&nthesis1 structure o$ )iruses +acteria
in$ect host cells1 o$ten using to.ins1 and shield themsel)es 0ith inbuilt #rotecti)e
mechanisms The team loo4ed at the common bacteria Stre#tococcus #&ogenes to
determine the structures o$ its to.in and antito.in The antito.in deacti)ates the to.in b&
binding to it 'hen not bound1 the antito.in changes sha#e+ritish bacteriologist 7rederic4
Gri$$ith disco)ered that a harmless strain o$ Stre#tococcus #neumoniae could be made
)irulent a$ter being e.#osed to heat84illed )irulent strains An e$$icient and con)enient
#rocedure $or trans$orming bacteria and o#ened the 0a& $or molecular cloning in
biotechnolog& and research A bio$ilm is an aggregate o$ microorganisms in 0hich cells
adhere to each other and9or to a sur$aceThe engineered bacteria8attac4ing )irus1 or :#hage1;
0as built using a :#lug and #la&; librar& o$ genes The same a##roach could be used to
build )iruses custom tailored to target s#eci$ic bacteria s#ecies1 the researchers sa&
Keywords: ad)antages1 result 2 re0ards o$ using microorganism as research tools1
+acteria<s sel$ de$ense mechanism1 Trans$ormation /genetics51Scientists engineer )iruses to
destro& bacteria1(se o$ bacteria as anticancer agent
An& noncellular or unicellular /including colonial5 organism1 most o$
0hich are too small to be seen 0ith the unaided e&e Microorganisms com#rise

Microorganisms as research tools 3
bacteria /including c&anobacteria51 lichens1 micro$ungi1 #roto6oa1 ric4ettsiae1 )irinos1
)iroids1 and )iruses1 and also some algae= all #ro4ar&otes are included
The stud& o$ microorganisms is called microbiolog&1 a sub*ect that began 0ith Anton
)an Leeu0enhoe4>s disco)er& o$ microorganisms in ?@AB1 using a microsco#e o$ his
o0n design/

%elc6ar1?CC-5
Microorganisms ha)e been used as a tool to e.#lore $undamental li$e #rocesses
because o$ man& ad)antages/Smith1 ,---5

:
? The& gro0 /#roduce5 )er& ra#idl&
, Gro0th can be mani#ulated easil& b& chemical or #h&sical means
D Microorganisms can be cultured small or )ast 3uantities con)enientl& and ra#idl&
E Their cells can be bro4en a#art 2 the contents se#arated into $ractions o$ )arious
#article si6es
B L&sed cells can be studied in terms o$ s#eci$ic chemical reactions1 s#eci$ic #roducts
and s#eci$ic structures in)ol)ed
Scientists $rom man& disci#lines recogni6ed the use$ulness o$ microorganisms
e.#erimental models Then #h&sicists1 chemists and biologist *oined 0ith
microbiologists in molecular biolog&
The result 2 re0ards $rom the $ield o$ research is s#ectaculars/ Sharma1 ,--@5
The contribution includes :
?Elucidation o$ en6&me structure 2 mode o$ action

Microorganisms as research tools 4
,Celluler regulator& mechanism
DEnerg& metabolism
E%rotein s&nthesis
BStructure o$ )iruses
Here 0e discussed about some microbes 0hcich 0ere used as a resechal model or tool
in e.#eriments:
+acteria<s sel$ de$ense mechanism might 0or4 against $ighting in$ections
+acteria in$ect host cells1 o$ten using to.ins1 and shield themsel)es 0ith inbuilt
#rotecti)e mechanisms These #ath0a&s could become their :Achilles< heel1;
according to researchers $rom 'ashington (ni)ersit& in a stud& #ublished on 7eb C
in the *ournal Structure
The team loo4ed at the common bacteria Stre#tococcus #&ogenes to determine the
structures o$ its to.in and antito.in The antito.in deacti)ates the to.in b& binding to
it 'hen not bound1 the antito.in changes sha#e
:That<s the Achilles< heel that 0e 0ould li4e to e.#loit1; said "r Thomas Ellenberger1
head o$ the uni)ersit&<s "e#artment o$ +iochemistr& and Molecular +io#h&sics1 in a
release :A drug that 0ould stabili6e the inacti)e $orm o$ the immunit& $actor 0ould
liberate the to.in in the bacteria;
E.#erts sa& that the bacterium #rotects itsel$ $rom the secreted to.ic b& 4ee#ing it in
an inacti)e $orm1 b& counter #roducing its antidote 'hen to.in com#le.es 0ith the

Microorganisms as research tools 5
antito.in molecule /immunit& $actor51 it remains inacti)e and is unable to cause an&
harm to the host or the bacterial cell itsel$
Figure 1 :Microalgae under the electron microscope.
The bacteria<s to.in is called Stre#tococcus #&ogenes beta8NA"F gl&coh&drolase1 or
S%N Cells store the coen6&me NA"F as #art o$ their metabolism1 and the to.in
0or4s b& draining these stores The bacteria<s energ& su##l& is #rotected b& the
antito.in1 the immunit& $actor $or S%N /I7S51 0hich bloc4s the to.in<s access to
NA"F
:The most im#ortant as#ect o$ the structure is that it tells us a lot about ho0 the
antito.in bloc4s the to.in acti)it& and s#ares the bacterium1; said Ellenberger
(nderstanding ho0 these bacteria cause disease in humans is im#ortant in drug
design
:There is a 0ar going on bet0een bacteria and their hosts1; said coauthor "r Craig
Smith1 a #ostdoctoral researcher at the uni)ersit&1 in the #ress release :+acteria
secrete to.ins and 0e ha)e 0a&s to counterattac4 through our immune s&stems and

Microorganisms as research tools 6
0ith the hel# o$ antibiotics +ut1 as bacteria de)elo# antibiotic resistance1 0e need to
de)elo# ne0 generations o$ antibiotics;
Antibiotics 0or4 in di$$erent 0a&s= some bloc4 bacterial cell 0all s&nthesis1 0hile
others inter$ere 0ith "NA s&nthesis or e)en inhibit bacterial metabolism
As o$ &et1 there are no classes o$ drugs that attac4 the #rotecti)e antito.in
mechanisms o$ bacteria
:Ob)iousl& the& could e)ol)e resistance once &ou target the antito.in1; Ellenberger
said :+ut this 0ould be a ne0 target (nderstanding structures is a 4e&stone o$ drug
design; / Arshdee# ,-??5
Trans$ormation /genetics5
Trans$ormation 0as $irst demonstrated in ?C,G b& +ritish bacteriologist 7rederic4
Gri$$ith Gri$$ith disco)ered that a harmless strain o$ Stre#tococcus #neumoniae
could be made )irulent a$ter being e.#osed to heat84illed )irulent strains Gri$$ith
h&#othesi6ed that some Htrans$orming #rinci#leH $rom the heat84illed strain 0as
res#onsible $or ma4ing the harmless strain )irulent In ?CEE this Htrans$orming
#rinci#leH 0as identi$ied as being genetic b& Os0ald A)er&1 Colin MacLeod1 and
Macl&n McCart& The& isolated "NA $rom a )irulent strain o$ S #neumoniae and
using *ust this "NA 0ere able to ma4e a harmless strain )irulent The& called this
u#ta4e and incor#oration o$ "NA b& bacteria Htrans$ormationH See A)er&8MacLeod8
McCart& e.#eriment
The results o$ A)er& et al>s e.#eriments 0ere at $irst sce#ticall& recei)ed b& the
scienti$ic communit& and it 0as not until the de)elo#ment o$ genetic mar4ers and the

Microorganisms as research tools 7
disco)er& o$ other methods o$ genetic trans$er /con*ugation in ?CEA and transduction
in ?CBD5 b& !oshua Lederberg that A)er&>s e.#eriments 0ere acce#ted
Trans$ormation did not become routine #rocedure in laboratories until ?CA, 0hen
Stanle& Cohen1 Annie Chang and Leslie success$ull& trans$ormed Escherichia coli b&
treating the bacteria 0ith calcium chloride This created an e$$icient and con)enient
#rocedure $or trans$orming bacteria and o#ened the 0a& $or molecular cloning in
biotechnolog& and research
Trans$ormation using electro#oration 0as de)elo#ed in the late ?CG-s thus
increasing the e$$icienc& and number o$ bacterial strains that could be trans$ormed
Trans$ormation o$ animal and #lant cells 0as also in)estigated 0ith the $irst
transgenic mouse being created b& in*ecting a gene $or a rat gro0th hormone into a
mouse embr&o in ?CG, In ?C-A a bacterium that caused #lant tumors1
Agrobacterium tume$aciens1 0as disco)ered and in the earl& ?CA-s the tumor
inducing agent 0as $ound to be a "NA #lasmid called the Ti #lasmid +& remo)ing
the genes in the #lasmid that caused the cancer and adding in no)el genes
researchers 0ere able to in$ect #lants 0ith A tume$aciens and let the bacteria insert
their chosen "NA into the genomes o$ the #lants Not all #lant cells are susce#tible
to in$ection b& A tume$aciens so other methods 0ere de)elo#ed including
electro#oration and micro8in*ection %article bombardment 0as made #ossible 0ith
the in)ention o$ the +iolistic %article "eli)er& S&stem /gene gun5 b& !ohn San$ord in
?CC-

/0i4i#edia1 ,-??5
Scientists engineer )iruses to destro& bacteria
S&nthetic creation attac4ed bio$ilms that can $orm on teeth1 in cre)ices

Microorganisms as research tools 8
A bio$ilm is an aggregate o$ microorganisms in 0hich cells adhere to each other
and9or to a sur$ace These adherent cells are $re3uentl& embedded 0ithin a sel$8
#roduced matri. o$ e.tracellular #ol&meric substance /E%S5 +io$ilm E%S1 0hich is
also re$erred to as slime /although not e)er&thing described as slime is a bio$ilm51 is
a #ol&meric conglomeration generall& com#osed o$ e.tracellular "NA1 #roteins1 and
#ol&saccharides +io$ilms ma& $orm on li)ing or non8li)ing sur$aces and can be
#re)alent in natural1 industrial and hos#ital settings
Figure 2: Staphylococcus aureus biofilm
The engineered bacteria8attac4ing )irus1 or :#hage1; 0as built using a :#lug and
#la&; librar& o$ genes The same a##roach could be used to build )iruses custom
tailored to target s#eci$ic bacteria s#ecies1 the researchers sa&
:The librar& could contain di$$erent #hages that target di$$erent s#ecies or strains o$
bacteria1 each constructed using related design #rinci#les to e.#ress di$$erent

Microorganisms as research tools 9
en6&mes1; said stud& leader !ames Collins1 a biomedical engineer at +oston
(ni)ersit& /!ain II ,-??5
(se o$ bacteria as anticancer agent
Historicall&1 bacteria 0ere used as oncol&tic agents $or malignant brain tumours
Ad)ances in bacteriolog& and molecular biolog& ha)e 0idened the sco#e o$ bacterial
a##roaches to cancer thera#& and )arious #ossibilities include the use o$ bacteria as
sensitising agents $or chemothera#&1 as deli)er& agents $or anticancer drugs1 and as
)ectors $or gene thera#& +acterial to.ins can be used $or tumour destruction and
cancer )accines can be based immunoto.ins o$ bacterial origin The most #romising
a##roaches are the use o$ geneticall& modi$ied bacteria $or selecti)e destruction o$
tumours1 and bacterial gene8directed en6&me #rodrug thera#&
Figure 3 : Salmonella

Microorganisms as research tools 10
Ino0ledge gained $rom stud& o$ bacterial genomes $orms an im#ortant basis o$ use
o$ bacteria as anticancer agents TA%ET /Tumour Am#li$ied %rotein E.#ression Thera#&5
uses a geneticall& altered strain o$ Salmonella as a bacterial )ector1 or )ehicle1 $or
#re$erentiall& deli)ering anticancer drugs to solid tumours Jeroto.in ? /JT?5 o$
Escherichia coli has been used $or e. )i)o #urging o$ human bone marro0 o$ cancer cells
be$ore autologous bone marro0 trans#lantE coli genes and en6&mes ha)e become #art
o$ 0ell84no0n #rodrug a##roaches to cancer in 0hich inert #rodrugs can be con)erted in
)i)o to highl& acti)e s#ecies IL8E $used 0ith %seudomonas e.oto.in has been
administered directl& into malignant brain tumours and binds 0ith high a$$init& to IL8E
rece#tors1 0hich do not e.ist on normal brain cells1 thus destro&ing a ma*or #art o$ the
tumour 0ithout harming the normal brain tissue It is in %hase I9II clinical trials in
#atients 0ith glioblastoma No ideal anticancer agent o$ bacterial origin that is a##licable
to all t&#es o$ cancers has been disco)ered &et The most #romising a##roach to
malignant brain tumours a##ears to be the use o$ geneticall& engineered bacteria that
destro& the tumour selecti)el& 0hile s#aring the normal brain tissue /Ier Than ,--A5
Re$erences
Arshdee# Sarao1E#och Times ,-??1 +acteria<s Sel$ "e$ense Mechanism Might
'or4 Against It
ASmith /,---51 O.$ord "ictionar& o$ +iochemistr& and Molecular +iolog&1Re)ised
edition /7ebruar& ,E1 ,---5 1 O.$ord (ni)ersit& %ress1 (SA
!ain II 1%harma+iotech1 +lKsiring A1 CH8E-BA +asel1 S0it6erland
Ier Than 1Li)e Science Maga6ine ,--A

Microorganisms as research tools 11
%elc6ar1 M!1 Chan1 ECS 2 N R Irieg Microbiolog&8 Conce#ts and A##lications
/International Edition51 McGra08 Hill Inc Ne0 "elhi
S Sharma11General microbiolog&1Microbial 'orld1 Histor& and "e)elo#ment
o$ Microbiolog&1 Sco#e o$ Microbiolog&1/Re)ised ?,8"ec8,--@5
'i4i#edia 1 ,D 7ebruar& ,-??
7rom : htt#:99en0i4i#ediaorg90i4i9Trans$ormationLM,GgeneticsM,C