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GJESR RESEARCH PAPER VOL. 2 [ISSUE 10] NOVEMBER, 2015

ISSN:- 2349283X

ISOLATION AND CHARACTERIZATION OF

ANTIBACTERIAL
COMPOUNDS PRODUCING MICROBES FROM
SOIL AND WATER SAMPLES
1. Zainab Siddiqui, Graduation student 2.Saima Azmi, PhD scholar
Department of biotechnology
Integral university, Lucknow
227105
Abstract Actinomycetes are one of the most attractive
sources of antibiotic.In the present studies total of four
Actinomycetes strains were isolated from two different
places in Lucknow, U.P, India. Isolated strains were
identified for their activity but only two isolated show good
resultsthey were Neisseria mucosa and Streptococcus
equisimilis. Isolation of bacterial strain was done by serial
dilution method. Primary screening of the culture was done
by sub-culturing through simple streaking and quadrant
streaking. Identification of bacteriawas done with help of
Bergeys manual and antibiotic sensitivity test was
performing by agar will diffusion method. The pathogens
were used E.coli, S.aureus, and P.aeruginosa against
intracellular and extracellular antibacterial compounds
which were isolated from the microbes. Best results were
obtained for Neisseria mucosa against E.coli. Further
Growth Kinetics study was done to know the log phase
because in log phase culture will produce maximum
secondary which works like antibacterial compounds. The
present study is carried out by isolation of anti-bacterial
compound producing microbes from the soil sample and
characterization as well as production for this culture to get
maximum secondary metabolites

1. INTRODUCTION
The actinomycetes are gram positive, high G+C
(>55%) organisms that tend to grow slowly as
branching filaments. Many actinomycetes will grow
on the common bacteriological media used in the
laboratory, such as nutrient agar, trypticase soy agar,
blood agar, and even brain-heart infusion agar.
Actinomycetes encompass a wide range of bacteria.
They can be terrestrial or aquatic.Actinobacteria is
one of the dominant phyla of the bacteria. Analysis of
glutamine synthetase sequence has been suggested
for phylogenetic analysis of Actinobacteria.
Actinomycetes are best known for their ability to
produce antibiotics and are gram positive bacteria
which comprise a group of branching unicellular
microorganisms. They produce branchingmycelium

which may be of two kinds viz., substrate mycelium

andaerial mycelium. Among actinomycetes, the
streptomycetes
are
the
dominant.
The
nonstreptomycetes are called rare actinomycetes,
comprising approximately 100 genera .Actinobacteria
are well known as secondary metabolites producers
and hence of high pharmacological and commercial
interest. In 1940 Selman Waksman discovered that
the soil bacteria he was studying made actinomycin, a
discovery for which he received a Noble Prize. Since
then, hundreds of naturally occurring antibiotics have
been discovered in this terrestrial microorganism,
especially from the genus Streptomyces.
Antibiotics are compounds produced by
microorganisms that are able to inhibit the growth of
other microorganisms. Antibiotics are medicinal
products that have an anti-bacterial effect, they either
kill bacteria in the system or keep away them from
reproducing, allowing the infected body to heal by
producing its own defenses and overcome the
infection. Screening of antibiotics has been widely
performed for about last 50 years and new antibiotics
are still being found. In screening of new antibiotics,
new approaches are required and following three
factors must be considered i.e. detection of antibiotic
producing microorganisms, selection of producing
microorganisms and cultivation methods.
These are closely related to each other, and
their efficient combination is essential for successful
screening of an antibiotic interest
2. SOURCE OF ANTIBIOTICS
Soil are cheap source of antibiotics, many
bacteria are present in soil. Many antibiotics produce
in microorganisms. People have been using herbal
antibiotics traditionally for centuries to combat
microbes that cause illnesses. Recently, scientists
have studied and reported results supporting the use
of these antimicrobial plants. Herbal antibiotics are
milder than their pharmaceutical counterparts, which

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makes using them for minor illnesses a logical

option. According to the Centers for Disease Control
and Prevention, the overuse of pharmaceutical
antibiotics causes microbes to mutate and grow
stronger. As a result, some experts say it makes sense
to use herbal treatments in place of pharmaceutical
antibiotics when appropriate.

4.

Reishi mushrooms medicinally for centuries, Reishi

Mushroom has antibacterial properties, Tea Tree Oil
can effectively treat and prevent fungal infections,
Goldenseal have been traditionally used goldenseal to
treat skin disorders, digestive problems and eye
infections, Garlic can effectively treat and prevent the
common cold., Neem is a multi-purpose Ayurvedic
herb that is especially effective against skin diseases,
Myrrh has been used by herbalists for centuries and is
even mentioned in the Bible. It is antiseptic,
antibiotic and antiviral. It can be taken internally,
used as a gargle or used as a wash for wounds;
Echinacea is one of the most popular herbal remedies
for colds, flues and other bacterial infections. It has
been used by herbalists as a blood purifier and to
treat a number of infections.

5.

6.

3. DIFFERENT TYPES OF ANTIBIOTICS

Of the 100 plus antibiotics substances produced
naturally or synthetically,very few have been proven
safe and effective. The commonly used antibiotic
types are:
1.

2.

3.

Penicillin:Penicillin is a group of antibiotics

derived from Penicillium fungi. It is an
antibiotic which destroys the cell walls of
the bacteria, while they are in the process of
reproduction. They include penicillin G,
procaine penicillin, benzathine penicillin,
and penicillin V. Penicillin antibiotics are
historically significant because they are the
first drugs that were effective against many
previously serious diseases, such as syphilis,
and infections caused by staphylococci and
streptococci .
Cephalosporin:
Cephalosporins
are
bactericidal and have the same mode of
action as other beta-lactam antibiotics (such
as penicillins) but are less susceptible to
penicillinases. Cephalosporins disrupt the
synthesis of the peptidoglycan layer of
bacterial cell walls. The Cephalosporin is a
class of -lactam antibiotics originally
derived from the fungus Acremonium, which
was
previously
known
as
"Cephalosporium".
Aminoglycosides: Thistype of antibiotics
hinders protein formation of bacteria

ISSN:- 2349283X

invading cells. This antibiotic encompasses

gentamicin, streptomycin and neomycin. As
aminoglycosides are effective in inhibiting
protein production in invading bacterial
cells, they are administered to treat typhus.
Macrolides: Macrolides are protein
synthesis inhibitors. The mechanism of
action of macrolides is inhibition of bacterial
protein biosynthesis, and they are thought to
do this by preventing peptidyltransferase
from adding the peptidyl attached to tRNA
to the next amino acid (similarly to
chloramphenicol) as well as inhibiting
ribosomal translocation.Another potential
mechanism is premature dissociation of the
peptidyl-tRNA from the ribosome.
Sulfonamides: The sulfa-related group of
antibiotics, which are used to treat bacterial
infections and some fungal infections. The
sulfonamide family includes sulfadiazine,
sulfamethizole,
sulfamethoxazole
,
sulfasalazine , sulfisoxazole, and various
high-strength combinations of three
sulfonamides. Sulfa drugs kill bacteria and
fungi by interfering with cell metabolism.
Tetracyclines: Tetracycline is a broadspectrum polyketideantibiotic produced by
the Streptomycesgenus of Actinobacteria,
indicated for use against many bacterial
infections. It is a protein synthesis inhibitor.
It is used to treat various eye infections.

A simplified enrichment method for the highly

selective isolation of the zoosporic actinomycetes
Actinoplanes spp. From soil. The method consists of
baiting the species with Pinus pollen grains,
desiccating (300C, 2 h) the baits bearing sporangia in
dried soil particles with the aid of silica gel and
following the spore liberation upon immersion in
water. Portions of the liquid enriched with zoospores
are plated out on humic acid-vitamin (HV) agar
supplemented with nalidixic acid at a concentration
of 10 g/ml. The desiccation stage had enabled the
almost complete elimination of associated bacteria
from colonized baits while allowing the
Actinoplanessporangia to survive and still possess the
ability to release many spores. A total of four
different soil samples from fields of corn, peach,
vegetable and paddy rice were examined. The pollenbaiting and drying method consistently resulted in the
highly selective isolation of Actinoplanes spp. which
accounted for over 83% of the total number of
microorganisms recovered on HV agar containing
nalidixic acid [1].
An actinomycete, designed strain KS3-5, was
from a soil sample collected from Kaohsiung,

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Taiwan, ROC. This organism is capable of producing

a series of antibiotics that strongly inhibit the growth
of gram-positive and gram-negative bacteria and
yeast like fungi. The spore morphology and cell wall
chemotype suggest that strain KS3-5 is a
Streptomycete. Further cultural and physiological
characterization and the DNA homology suggest that
strain KS3-5 is identical to Stretomycestoxytricini [2].
The presence and types of antibiotic-producing
bacteria, fungi and actinomycetes using nutrient agar,
potato Dextrose agar and starch casein nitrate agar
respectively as culture media was determined in
university of lagos from the compost soil. A variety
of bacteria were isolated and these included
Staphylococcus aureus, B. subtilis, B. pumilis, B.
lactesporus, B. megaterium, B. pulvifaciens, B.
licheniformis, Streptococus spp., Corynebacterium
spp. and E. coli. The fungal isolates encountered
were Aspergillusniger, A. flarus, T. viridae, P.
chrysogenum, P. pinofylum and Absida spp., while
the following actinomycetes were identified:
Norcadia spp., Micromonospora spp., Streptomyces
scabies, S. reticuli and S. hygroscopicus.
When these organisms were screened for
antibiosis, the following species were found to be
antibiotic producers: B. licheniformis, B. subtilis,
Penicilliumchrysogenum, Streptomyces reticuli, S.
hygroscopicus and Micromonospora spp. The fungus
Penicilliumchrysogenum had the highest rate of
antibiotic production with an inhibitory zone width of
17mm while Trichodermaviridae produced toxins
lytic to other fungal hyphae[3].
Bacillus subtilis and Bacillus pumiluswere
isolated from soil and screened for the production of
antibiotics by plate assay and then cultured in shake
flask fermentation at 300C for further studies.
Identification of antibiotics was done by paper
chromatography. Bacitracin was found to be
produced by both the strains against Micrococcus
luteus(ATCC#10240),whereas;Staphylococcusaureus
(ATCC# 6538) proved to be resistant to Bacitracin
produced by Bacillus pumilus. The maximum
production of Bacitracin from B. subtilis and B.
pumilus against Staphylococcus aureus and
Micrococcus luteus at different pH (6-9), incubation
time (0-144 hours) and glucose concentration (1-5%)
was checked by agar diffusion assay as detected by
the size of zones of inhibition. Maximum zones of
inhibition were observed at pH 8, 5% glucose and
after 24 hours of incubation at 30oC against
Staphylococcus aureusand Micrococcus luteus [4].
Isolation of different microorganisms from
some soils' rhizosphere in Al-Madinah AlMunawwarah, viz. corn (Zea mays), datepalm
(Phoenix dactylifera), catnip (Menthapiperita),
sunflower
(Helianthus),
balessan

ISSN:- 2349283X

(Amyrisgileadensis),
nabk-tree
(ZiziphusSpinaChristi Willd)and basil (Marrubiumvulgare) was
carried out. All microbial isolated were then screened
for their antagonistic activity against the most
resistant eight target bacteria isolated from caesarean
section site infections (E.coli, Klebsiellaspp.,
Pseudomonas spp., Proteus spp., Citrobacterspp.,
Acinetobacterspp.,
methicillin
resistant
Staphylococcus aureus MRSA, and coagulase
negative Staphylococcus). Among the total 86 fungal
and bacterial isolates, only 15 of them (17.44%) were
capable of biosynthesizing antimicrobial metabolites.
One of the actinomycetes that was obtained from
catnip rhizosphere, Al-Ouayna district in Al-Madinah
Al-Munawwarah, found to exhibit the highest
antimicrobial activity and it matched with
Streptomyces ramulosusin the morphological,
physiological and biochemical characters. Thus, it
was given the suggested name Streptomyces
ramulosus, A-MM-24[5].
4 .MATERIAL AND METHODS
Soil sample (C1, C2, C3, C4) was collected
from garden soil of different places of Indira Nagar
and Aliganj area of Lucknow city. Actinomyceties
Agar ,Nutrient Agar medium, Nutrient Broth,
Production media, different Pathogens(Pseudomonas
, E.coli , S.aureus) , Antibiotics(Tetracycline),
Crystal violet, Grams iodine, Ethanol, Saffranine ,
Malacnite Green were used for the purpose of study.

4.1Isolation
of
antibiotic
actinomycetes from soil

producing

Serial dilution agar plating method or viable

count method is one of the most commonly used
procedures for the isolation and enrichment of the
most prevalent micro-organism such as fungi,
bacteria. This method is based on the principle that
when sample containing the micro-organism is
cultured each viable micro-organism will develop
into a colony; this method use is number of reduced
bacterial colonies in order to get pure cultures. The
numbers of micro-organism are diluted because the
soil sample which was collected had a population of
micro-organism. The numbers of colonies are same
as the number of organism contain in the sample.
Microbes from soil were isolated by serial dilution
method and mixed culture was obtained by spreading
as shown in figure 1. Colony morphology of culture
from sample C1, C2, C3 and C4 is also depicted in
table 1.

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3.1 PRIMARY SCREENING

After incubated overnight four types of
colonies were present in the plates, Zone of inhibition
around the colonies was observed. Colonies showing
zones of inhibition were selected for streaking and
secondary screening.

A. Sub culturing
Pure culture is usually derived from a mixed
culture (containing many species) by methods that
separate the individual cells so that, when they
multiply, each will form an individually distinct
colony, which may then be used to establish new
cultures with the assurance that only one type of
organism will be present. Pure cultures may be more
easily isolated if the growth medium of the original
mixed culture favours the growth of one organism to
the exclusion of others. 60 ml nutrient agar was
prepared and poured in to four sterile petriplates.
Simple or zigzag streaking of four cultures was done
on petriplates with the help of inoculation loop. All
Figure 1- Mixed colonies in spread plate

the plates were incubated at 370C for overnight. After

Table 2- Colony morphology of culture from sample

zigzag streaking quadrant streaking is done.

C1, C2, C3 and C4.

Colony
morpholog
y

Culture
C1

Culture
C2

Cultur
e C3

Cultur
e C4

Shape

Circular

Circular

Circul
ar

Circul
ar

Margin

Entire

Entire

Entire

Entire

Opacity

Transluce
nt

Transluce
nt

Opaqu
e

Opaqu
e

Pigmentati
on

White

Off White

Off
White

Off
White

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GJESR RESEARCH PAPER VOL. 2 [ISSUE 10] NOVEMBER, 2015

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C4
Fig 2: Pure colonies obtained through simple
streaking

C2
Quadrant streaking is done when colonies
are not clearly identified. For this, 30 ml nutrient agar
was prepared and poured in to two sterile petri plates.
Quadrant or four ways streaking of the two cultures
was done on petriplates with the help of inoculation
loop. Both the plates were incubated at 370C for
overnight. C3 and C4 pure colonies obtained through
Quadrant Streaking. The cultures C1, C2, C3 and C4
obtain through primary screening were purified by
zigzag or simple streaking was shown in fig. 2. C3

C3

and C4 cultures obtained after quadrant streaking is

shown in fig.3

C3

C4
Fig 3:- Pure colonies obtained through Quadrant
streaking
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E.coli

B. STAINING AND BIO-CHEMICAL

CHARACTERIZATION OF SAMPLE
Gram staining was given by Hans Christian
Gram. It is a method of differentiating bacterial species
into two large groups i.e. Gram-positive and Gramnegative. It is based on the chemical and physical
properties of their cell wall. Primarily, it detects
peptidoglycan, which is present in a thick layer in Gram
positive bacteria. A gram positive results in a purple/blue
color while Gram negative results in a pink/red color.
Gram-positive bacteria have a thick mesh-like cell wall
made of peptidoglycan (50-90% of cell envelope), which
are stained purple by crystal violet, whereas Gramnegative bacteria have a thinner layer (10% of cell
envelope), which are stained pink by the counter-stain.a
primary stain (crystal violet) to a heat-fixed smear of a
bacterial culture. After grams staining slide were
observed under microscope and some colonies (C1, C2,
C3) get pink colour and some get purple colour (C4). The
colonies showing pink colour are gram negative cocci
and colony which shows purple colour are gram positive
cocci bacteria.

S.aureus

3.2 SECONDARY SCREENING

A. Antibiogram Analysis
Activity of biological compounds agent
pathogen an important task of the clinical microbiology
laboratory is the performance of antimicrobial
susceptibility testing of significant bacterial isolates. The
goals of testing are to detect possible drug resistance in
common pathogens and to assure susceptibility to drugs
of choice for particular infections. Antibiogram of
purified cultures C1, C2, C3 and C4 was performed
against various pathogens (E.coli, P.aeruginosa and
S.aureus). There were zones of inhibition obtained in
each culture. Each cultures show positive result against
pathogensas shown in fig 4 and table 2.

P.aeruginosa
Fig 4: Antibiogram analysis of cultures against various
pathogens.
Table 3 Antibiogram of Isolated cultures against various
pathogens.
S.No. Pathogen C1
C2
C3
C4
Dia(mm) Dia(mm) Dia (mm) Dia (mm)
1

E.coli

10

10

S.aureus

10

Pseud
omona
s

10

10.5

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B. Endospore Test
We observe pink color cocci under microscope, which
confirm that these are non-endospore forming bacteria.
C. Catalase Test
In this test bubble formation was not observed. By this
we confirmed that the bacteria are from group
Streptococcus.
D.6.5% NaCl Test
No growth was observed in 6.5% NaCl. Therefore,
further glycerol test was done to confirm the species of
Streptococcus.
E. Acid from Glycerol Test
After 72 hrs appearance of yellow halo surrounding the
streak was appeared showing that these gram positive
bacteria are producing acid, which confirmed
Streptococcus equisimilis species as shown in fig 5.

Blank

Fig 5:- Gram positive bacteria are producing acid.

F. Of Glucose Test

Colour change

The oxidative-fermentative (OF) test was

developed by Hugh and Leifson. This test determines
whether an organism metabolizes a given
carbohydrate (usually glucose) by oxidation or by
fermentation.
The power of breakdown of carbohydrates is
possessed by a large number of bacteria, fungi and
yeasts. After performing this test blue colour of the
solution changes to yellow due to glucose
fermentation and confirm Neisseria group, the of
glucose test results are shown in fig 6.

Fig 6:- glucose fermentation

G. Nitrate Test:
This test is done to confirm the species of
Neisseria. In this test the red colour of solution was
observed which shows that nitrate was reduced and
confirmed Neisseria mucosa species. Result is shown in
figure 7.

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S.No. Pathogen

Intracellular Extracellular Intra + Extra

Diameter(m Diameter(m Diameter(mm
m)
m)
)

E.coli

32.5

27.5

S.aureus

14.5

32.5

29

Pseudom
onas

14.5

29

29

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3.3ANTIBIOTIC SENSITIVITY TEST

A. Antibiogram of Intracellular and Extracellular
Antibiotics Extracts
Antibiogram analysis of intra-extracellular from
our sample C3 and C4 was performed against various
pathogens. The results can be seen in table 3 and fig 8
below For Neisseria. Antibiogram for of intracellular and
extracellular antibiotic extract from S.equisimilis is
shown in fig 9 and table 4.

For Neisseria

Table 3- Antibiogram analysis of intra-extracellular

Antibiotic extracts from N.mucosa.

Initial colour

E.coli

Final colour showing nitrate reduction

Fig 7: Nitrate reduction.

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monas

P.aeruginosa
S.Aureus
S.aureusP.aeruginosa

S.aureus
Fig 8: Antibiogram of intracellular and extracellular
antibiotic extract from N.mucosa
For S.equisimilis
Table5.Antibiogram analysis of
antibiotic extract from S.equisimilis

intra-extracellular

E.coli

S.No. Pathogen Intracellular Extracellular Intra + Extra

Diameter(
mm)

Diameter(
mm)

Diameter(m
m)

E.coli

27.5

22

S.aureus

18

12.5

Pseudo

19

20
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0.5

Decline

P.aeruginosa

Fig 9:- Antibiogram for of intracellular and extracellular

antibiotic extract from S.equisimilis

Growth kinetics of S.equisimilis and

N.mucosa is graphically represented for better
understanding in figure.10&11 along with table 6&7
showing optical density at different intervals of the
day and stages of them.
Growth is an orderly increase in the quantity
of cellular constituents. It depends upon the ability of
the cell to form new protoplasm from nutrients
available in the environment. In most bacteria,
growth involves increase in cell mass and number of
ribosome, duplication of the bacterial chromosome,
synthesis of new cell wall and plasma membrane,
partitioning of the two chromosomes, septum
formation, and cell division.
1.2

3.4GROWTH KINETICS OF CULTURE

Growth kinetics process is applied to
determine the time period at which the culture show
its optimum activity (stationary phase). Growth of
any microbes occurs in different stages which are
indicated by growth curve. Growth shows different
four stages Lag, Log, stationary and decline phase.

O.D

0.8
0.6
O.D

0.4
0.2

Table 6- Growth Kinetics of S. equisimilis

DAYS O.D OF THE CULTURE GROWTH PHASE

0
0.77

Lag

1.03

Log

1.05

Log

10

No. of days

Fig 10: Graph of Growth Kinetics of S. equisimilis

Table 7- Growth Kinetics of N. mucosa

DAYS O.D OF THE CULTURE GROWTH PHASE

1.03

Stationary

0.24

Lag

0.57

Log

0.95

Log

0.94

Stationary

1
1

Stationary

2
0.89

Decline

3
0.84

Decline
4

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0.93

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Stationary

5
0.87

Decline

0.83

Decline

S.no

Pathoge
n

Tetracy
cl-ine
(mm)

Intracell
ular
(mm)

E.coli

41

24

P.aerugi
nosa

26

29.5

22

S.aureus

35

22

14.3

Intraextra
mixture
(mm)
16.5

7
For determining growth kinetics of the
bacteria we have to prepare 50 ml NB in two conical
flasks then it is Autoclaved and cool to ensure the
impurity autoclave is the method for steam
sterilization generally used for media sterilization.
After that Inoculatation of the first culture in first
flask and second culture in another flask is done.
Incubate the media in shaker incubator and do check
the O.D day by day for 4-5 days at 600 nm. Finally
Plot the graph of growth kinetics.
1.2
1

O.D

0.8
0.6
O.D

0.4

E.coli
0.2
0
0

10

No. of days
Fig 11: Graph of Growth Kinetics of N.mucosa
3.5 ANTIBIOGRAM ANALYSIS
Antibiogram analysis of intracellular and extravellular
antimicrobial extract agsinst various pathogens is sown in
table 8 and figure 12.
By Antibiogram analysis of intra and extra
cellular against various pathogens and by comparing with
tetracycline, intracellular componentshows better result
than tetracycline against P.aeruginosa. Intracellular give
29.5mm of zone of inhibition and tetracycline give
26mm.
Table 8- Antibiogram analysis of intra and extra
cellular antimicrobial extract against various
pathogens.
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5. REFERENCES

S.aureus
Fig 12: Antibiogram analysis of culture sample against
various pathogens.

1. Masayuki Hayakawa, Takayuki Kajiura and

Hideo Nomomura, 1991, new methods for the
highly selective isolation of Streptosporangium
and Dactylosporangium from soil, Journal of
Fermentation and Bioengineering, 72(5):327-333.
2. Rong-Yang Wu and Ming-Ho Chen, 1995,
Identification of theStreptomyces strain KS3-5.
Bot Bull Acad Sin, 36:201-205.
3. Adeleye IA, Eruba S, Ezeani CJ2004, Isolation
and characterization of antibiotic producing
microorganisms in composted Nigerian soil. J
Environ Biol.; 25(3):313-6.
4. MuhammadAwais2007, Isolation, identification
and optimization of bacitracin produced by
Bacillussp., Pak. J. Bot., 39(4): 1303-1312.
5. Abo-Shadi
Al-RahmanMahaAbd2010,
Antimicrobial Agent Producing Microbes from
some Soils' Rhizosphere in Al-Madinah AlMunawwarah, KSA,Journal of American Science,
6(10):915-925.

4. CONCLUSION AND FUTURE EXPECTS

Screening of antibiotics has been widely performed
for about last 50 years and new antibiotics are still
being found. In screening of new antibiotics, new
approaches are required and following three factors
must be considered i.e. detection of antibiotic
producing microorganisms, selection of producing
microorganisms and cultivation methods. Finally it
can be concluded that the Actinomycetes are the best
source for antibiotic isolation. In this project by
Antibiogram analysis intracellular component shows
better result than tetracycline against P.aeruginosa.
Intracellular give 29.5mm of zone of inhibition and
tetracycline give 26mm.
As the antibiotics are secondary metabolites,
they are synthesized in trace amounts. Moreover the
synthesis of antibiotic is regulated by tight metabolic
and genetic regulation. Therefore it is the task to the
biotechnologists to modify the wild type strain and to
provide cultural conditions to improve the
productivity of antibiotics. Improvement of the
microbial strain offers the greatest opportunity for
cost reduction without significant capital investment.
The problem of the bacterial resistance to antibiotics
had evolved and new compounds or derived from the
known antibiotics had to be found to replace existing
ones. Water and air microbes can also be used for the
isolation, characterizing, purification and production
of antimicrobial compound.

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