20BT3998 - Phase1 project (2nd Review)

Isolation, characterization and screening of

marine actinomycetes for the antimicrobial
and anticancer activity
by
Mohanya, K. (PRK20BT1023)

Internal guide: Dr. S. Prathap

Assistant Professor
Department of Biotechnology,
Karunya Institute of technology and sciences, Coimbatore

External guide: Dr. P. Arumugam

Armats Biotek training and Research Institute, Chennai
 Aim
To isolate, characterize and investigate the antimicrobial and anticancer activity of secondary
metabolites produced from the marine actinomycetes.

Objective
• To collect the soil sediment samples from the coastal region of Chennai.
• To isolate the actinomycetes from the samples and perform morphological, physiological,
biochemical and molecular characterization.
• To produce and extract crude secondary metabolites from the isolated marine actinomycetes.
• To perform in vitro antimicrobial, antioxidant and cytotoxic assays.
• To purify and characterize the bioactive compound.
 Introduction
Breast Cancer statistics
• Cancer is the leading cause of deaths worldwide in 2020.
• Of all the cancers, female breast cancer has the highest incidence of 11.7% and fifth leading
cause of cancer mortality of about 6.9% (685,000 deaths) globally.(WHO, 2021)

Signs and symptoms of Breast cancer

Lump formation , Non-lump breast symptoms , Non-breast symptoms (Waks et al, 2017)

Current treatment for breast cancer

1.Surgical treatment 2.Radiation therapy 3.Systemic therapy
Systemic therapy is the use of medication to destroy cancer cells. The types of systemic
therapies used for breast cancer include:
Chemotherapy, Hormonal therapy, Targeted therapy and Immunotherapy (Waks et al, 2017)

Limitations of current treatment

• The drugs developed by synthesis and used in chemotherapy have limitations mainly due to their
toxic effects on non-targeted tissues and consequently furthering human health problems.
• Therefore, there is a demand for alternative treatments, and the naturally-derived anticancer
agents are regarded as the best choice.
Secondary metabolites as anticancer agents
• Secondary metabolites are themselves suitable anticancer agents leading to the development of new clinical drugs
with also new anticancer mechanisms of action.
• Additionally, they are excellent lead compounds and their pharmacological potential is enhanced through structural
modifications, alternative formulations and/or using increasingly effective delivery systems. (Seca et al, 2016)

Actinomycetes
• The actinomycetes comes under phylum “Actinobacteria" and mostly Gram-positive bacteria.
• The colonies often grow as extensive mycelia, like a fungus.
• Actinomycetes strains are usually found in terrestrial soils, but have also been isolated from marine sediments and
marine sponges.
• Based on hyphal and reproductive structures, Actinomycetes are classified into 7 families namely
Streptomycetaceae, Nocardiaceae, Micromonosporaceae, Actinoplanaceae, Dermatophilaceae, Frankiaceae and
Actinomycetaceae.

Marine Actinomycetes as a source of secondary metabolite

• About 200,000– 250,000 metabolites show signs of bioactivity, more than 22,000 of which are produced by
microorganisms.
• Of the 22,500 microbial metabolites, 45% (10,100) are products of actinomycetes fermentation.
• Among filamentous actinomycetes, about 75% (7,600) of metabolites are produced by species of the genus
Streptomyces (Vijayakumar Ramasamy, 2015) and these are richest source of antibiotics and other industrially-
important compounds.
• The metabolites from marine actinomycetes shows high structural variety and exhibit a broad range of activities:
antimicrobial, antiprotozoal, antiparasitic, antitumor, and antimalarial. (Subramani et al, 2012)
 Work Plan
Sample collection and processing

Isolation of marine actinomycetes

Characterization Production
1. Morphological 3. Biochemical Production media
-Colour -Carbon utilization
-Shape -Nitrogen utilization Liquid-liquid extraction
- Spore (SEM) -Enzyme production
Ethyl acetate
4.Molecular
2. Physiological
Genomic DNA • Antioxidant assay (DPPH)
-Melanin
-Pigment • Antimicrobial activity
16s rRNA gene • Anticancer activity
-Organic
amplification
degradation
-NaCl Purification
Sequencing
• Column chromatography
• Characterization ( UV, FTIR, NMR,
NCBI Blast
MS)
 Methodology
Sample collection and processing
• Soil samples were collected from 3 beaches located in Chennai such as Besant
nagar beach, Golden beach, Kovalam beach.
• The samples were collected from 5 locations.

BN- Besant Nagar beach

GB- Golden beach

KB- Kovalam beach

• The samples collected from each beach were dried and pooled together and
taken as one sample.
• The dried samples were used for serial dilution.
BN GB KB
 Isolation of
actinomycetes
The serial diluted samples then plated onto the CSPY-ME agar medium with antibiotics
and incubated for 14 days at 27°C. (Biji, 2003)

Control 10¯¹ A 10¯² A 10¯³ A 10¯⁴ A

(BN01, BN12, (BN03)
BN13)

Stock 10¯¹ B 10¯² B 10¯³ B 10¯⁴ B

(BN05) (BN02, BN14) (BN04)

BN sample
Control 10¯¹ A 10¯² A 10¯³ A 10¯⁴ A
(GB08)

Stock 10¯¹ B 10¯² B 10¯³ B 10¯⁴ B

(GB06) (GB07)

10¯¹ B GB sample
(GB15)
 Control 10¯¹ A 10¯² A 10¯³ A 10¯⁴ A
(KB10, KB11)

Stock 10¯¹ B 10¯² B 10¯³ B 10¯⁴ B

(KB09)
KB sample

The selected 15 colonies were isolated and cultured into CSPY-ME and starch caesin
agar medium.
 Primary cultures of selected
colonies

BN01 BN02 BN03 BN04 BN05

GB06 GB07 GB08 KB09 KB10

KB11 BN12 BN13 BN14 GB15

 Selected 5 cultures
Among 15 colonies, 5 colonies were observed as actinomycetes based on colony
nature and spore formation.

BN01 BN12 BN13

BN14 GB15
 1.Morphological Characterization

Colour
determination
• The color of aerial mycelium and substrate mycelium of the actinomycetes were
observed in 4 different media namely M2, M3, M4 and M5 (Shirling et al, 1966).

M2- Yeast Extract-Malt Extract agar

M3- Oatmeal agar
M4- Inorganic salts-Starch agar
M5- Glycerol-asparagine agar

• The color was observed at the 7 and 14 th day.

 7th day 14th day
M2 M3 M4 M5 M2 M3 M4 M5

Aerial
mycelium

Substrate
mycelium

BN01

Aerial
mycelium

Substrate
mycelium

BN12
 7th day 14th day
M2 M3 M4 M5 M2 M3 M4 M5

Aerial
mycelium

Substrate
mycelium

BN13

Aerial
mycelium

Substrate
mycelium

BN14

Aerial
mycelium

Substrate
mycelium

GB15
Isolate Time Media Aerial Mycelium Substrate Mycelium Pigment Melanin
No.

BN01 7th day M2 Pinkish grey Reddish brown No No

M3 Greyish white Greyish yellow No No
M4 White White No No
M5 Grey Greyish yellow No No
14th day M2 Pinkish grey, White Brown, Yellow No No
M3 Grey Yellowish white No No
M4 Yellowish grey Yellow No No

M5 Yellowish grey Yellowish grey No No

BN12 7th day M2 Grey Yellowish brown No No

M3 Greenish grey Greenish grey No No

M4 Grey Greyish green No No

M5 White, grey Grey No No
14th day M2 Greyish green Yellowish brown, Yellow No No

M3 Greyish green Greyish green No No

M4 Greyish green, greyish Greyish green No No
white

M5 White, greyish green Brown, yellowish green No No

Isolate Time Media Aerial Mycelium Substrate Mycelium Pigment Melanin
No.
BN13 7th day M2 Grey Brown No No
M3 Pinkish grey Yellowish brown No No
M4 White White No No
M5 Yellowish grey, Pinkish grey Yellowish grey No No

14th day M2 Pinkish grey Brown No No

M3 Grey Yellowish grey No No
M4 Yellowish white Yellowish white No No

M5 Pinkish grey, Yellowish grey Yellowish brown, Yellowish No No

grey
BN14 7th day M2 Pinkish grey Yellowish brown No No
M3 White White No No
M4 White, grey White, Yellowish brown No No

M5 White Yellowish white No No

14th day M2 Pinkish grey, whitish grey Brown No No
M3 White, Yellowish white White, Yellowish white No No
M4 Pinkish grey, Yellowish Yellowish brown, Yellowish No No
white white
M5 Yellowish white, Brownish Yellowish white, Greyish No No
yellow yellow
Isolate Time Media Aerial Mycelium Substrate Mycelium Pigment Melanin
No.
GB15 7th day M2 Grey Brown No No
M3 Pinkish grey Yellowish brown No No
M4 White White No No
M5 Yellowish & Pinkish grey Yellowish grey No No

14th day M2 White, green Brown No No

M3 White White No No
M4 Greyish white Yellowish grey No No
M5 Yellowish orange Yellowish orange No No
Microscopic analysis
• The Starch casein agar medium poured onto petriplates.
• The five cultures BN01, BN12, BN13, BN14 and GB15 were streaked onto the
center of the Starch casein agar plate.
• The coverslip was inserted into the streaked plates at 45° angle and incubated for
7 days.

100X 400X 1000X

BN01
100X 400X 1000X
BN12

100X 400X 1000X

BN13
100X 400X 1000X
BN14

100X 400X 1000X

GB15
 2. Physiological characterization
Melanin and pigment
production
• The Peptone – Yeast Extract Iron agar and Tyrosine agar medium was prepared
and poured onto the petriplates.
• The isolates BN01, BN12, BN13, BN14 and GB15 were streaked onto the
plates and incubated for 14 days at room temperature. (Shirling and Gottleib
1966)
M6 M7 M6 M7

Aerial
Mycelium

Substrate
Mycelium

BN01 BN12
 M6 M7 M6 M7

Aerial
Mycelium

Substrate
Mycelium

BN13 BN14

Aerial
Mycelium

Substrate
Mycelium

GB15
Isolate Media Aerial Mycelium Substrate Mycelium Melanin
No.

BN01 M6 Pinkish grey Yellowish brown No

M7 Yellowish grey Yellow No

BN12 M6 Grey Black No

M7 Whitish grey, Grey Greyish yellow No

BN13 M6 Yellowish grey Orange No

M7 Greyish white, Greyish Greyish yellow No

yellow
BN14 M6 Greyish white, Greyish Orange, Yellowish orange No
yellow
M7 Grey, Yellow Yellow, Grey No

GB15 M6 White Yellow No

M7 Yellowish white, Orange Orange, Brown No

Organic
degradation
• The ability of actinomycetes to degrade organic compounds was determined by
using Bennet’s medium for organic compounds Xanthine, Hypoxanthine,
Adenine, Tyrosine, Casein and Aesculin medium for organic compound Aesculin
at pH 7.3.
• The five actinomycetes isolates were streaked onto the agar plates and incubated
for 14 days at room temperature.

NaCl tolerance
• The NaCl tolerance of the actinomycetes was observed by varying the
concentrations of NaCl by 0, 2, 4, 6, 8 and 10% in Starch-Casein agar medium at
pH 7.2.
• The actinomycetes isolates were streaked onto the agar plates and incubated for
14 days at room temperature.
 Production of secondary metabolites
• The 5 marine actinomycetes isolates were inoculated into the ISP-2 broth of each
50ml and incubated for 2 days at shaker incubator at room temparature.
• The spore formation was observed and 5 to 6ml of culture broth was inoculated
into the production media (Medium-14) of each 250 ml broth at pH 7.
• The production medias were incubated at shaker at room temparature for 1 week.
(Biji M, 2003)

BN01 BN12 BN13 BN14 GB15

Production media at the end of 7th

day
Liquid-liquid extraction using separation
funnel
• The production media was centrifuged at 7,000 rpm for 15
mins and supernatant (culture filtrate) was collected.
• The ethyl acetate is used as a solvent for the liquid-liquid
extraction.
• After strong agitation, the upper layer of ethyl acetate along
with metabolite compounds was collected.
• It was condensed to remove ethyl acetate solvent and
remaining liquid with crude metabolite was collected.
• The three different extracts of ethyl acetate, chloroform and
methanol were collected for all the five isolates

Upper layer
 Crude metabolites of marine actinomycetes

Ethyl acetate Chloroform Methanol

BNO1

BN12
 Ethyl acetate Chloroform Methanol

BN13

BN14

GB15
 DPPH Antioxidant assay
• A solution of 0.1mM of DPPH in methanol was prepared, 1 ml of DPPH solution mixed with 1
ml of extracts in methanol solution at different concentrations of 20-100 μg/mL. The positive
control is ascorbic acid.
• The reaction mixture was vortexed and left in dark for 30 mins at room temperature.
• The absorbance of the mixtures was measured by spectrophotometer at 517nm.

% DPPH radical scavenging activity={(A0− A1)/A0}×100

• Then  % of inhibition was plotted against concentration, and from the graph IC50 was
calculated. The experiment was repeated two times at each concentration. (Rahman et al,
2015).

Chloroform Ethyl acetate Methanol

BN01
Chloroform Ethyl acetate Methanol

BN12

BN13
Chloroform Ethyl acetate Methanol

BN14

GB15
 Standard
100.00 95.34 96.30 97.29
86.56
80.00

% of inhibition
60.00 51.57
40.00
20.00 9.05
0.00
5 10 15 20 25 30
Concentration (µg/mL)

BN01- Chloroform BN01- Ethyl acetate BN01- Methanol

100 100 100
88.65 89.81
80 78.36 80
80 69.70

% of inhibition
67.57
% of inhibition

% of inhibition
60 60 55.65 60 50.82
43.71
40 40 40
28.47
20 20 20 13.38
4.43 8.38
0.61 1.57 2.86
0 0 0
20 40 60 80 100 20 40 60 80 100 20 40 60 80 100

Concentration (µg/mL) Concentration (µg/mL) Concentration (µg/mL)

BN12- Chloroform BN12- Ethyl acetate BN12- Methanol

100 97.27 97.75
100 88.52 100 88.77
82.17 83.87 85.77
80 80 69.61 80
% of inhibition
% of inhibittion

% of inhibition
62.51
60 60 57.71 60
44.59
40 40 40
21.77 23.30 24.63 25.85
20.64 20
20 20

0 0 0
20 40 60 80 100 10 20 30 40 50 60 20 40 60 80 100

Concentration (µg/mL) Concentration (µg/mL) Concentration (µg/mL)

 BN13- Chloroform BN13- Ethyl acetate BN13- Methanol
100 100 100

80 80 80

% of inhibition
% of inhibition

% of inhibition
59.4865
60 60 60
47.39 47.3145
39.70 39.29
40 40 32.85 40
25.74 26.763
20 18.63 20 12.397
7.69 20
2.74 4.46 4.80 6.73
0 0 0
20 40 60 80 100 20 40 60 80 100 20 40 60 80 100
Concentration (µg/mL) Concentration (µg/mL) Concentration (µg/mL)

BN14- Chloroform BN14- Ethyl acetate BN14- Methanol

100 100 100
80 80 80
% of inhibition

% of inhibition

% oh inhibition
60 52.56 60 55.82 58.22
60
40.763 44.6095 49.26
40 40
28.6765 40 29.87
18.37 20.76 21.78
20 11.495 20 14.76 19.08
9.84 20
0 0
20 40 60 80 100 20 40 60 80 100 0
20 40 60 80 100
Concentration (µg/mL) Concentration (µg/mL) Concentration (µg/mL)

GB15- Chloroform GB15- Ethyl acetate GB15- Methanol

100 100
100
87.566
80 80 71.989 72.877 75.6705
77.594 82.625 86.48
80
% of inhibition
% of inhibition

% of inhibition
60 63.5465
60 60
40 40 40 31.171
29.009
20 10.668 12.132 13.56 14.5225 20 20
4.8705
0 0 0
20 40 60 80 100 20 40 60 80 100 20 40 60 80 100
Concentration (µg/mL) Concentration (µg/mL) Concentration (µg/mL)
 Samples IC 50 (µg/mL) STDEV
BNO1    
Chloroform 596.91 16.17
Ethyl acetate 35.94 0.25
Methanol 59.03 0.24
BN12    
Chloroform 193.41 0.97
Ethyl acetate 8.66 0.03
Methanol 22.43 0.16
BN13    
Chloroform 650.54 16.93
Ethyl acetate 105.52 0.52
Methanol 84.54 0.37
BN14    
Chloroform 95.13 0.51
Ethyl acetate 229.58 1.79
Methanol 60.9 0.35
GB15    
Chloroform 344.39 7.96
Ethyl acetate 27.78 0.16
Methanol 31.47 0.15
 660 650.54

596.91

560

460
IC50 (µg/mL)

360 344.39

260
229.58
193.41

160
105.52
95.13
84.54
59.03 60.9
60 35.94
22.43 27.78 31.47
8.66

01-EA 01-MET 01-CHL 12-EA 12-MET 12-CHL 13-EA 13-MET 13-CHL 14-EA 14-MET 14-CHL 15-EA 15-MET 15-CHL
-40

Crude Extract Samples

 Antimicrobial assay by Agar well-diffusion method
• The nutrient agar plate for bacterial pathogens and potato dextrose agar plates for fungal pathogen were
prepared and the pathogens were swabbed onto the agar plates.
• The well was prepared by using cork borer (6mm in diameter).
• A volume of 100µL of 10mg/mL of crude extracts were dispensed into each well and allowed to diffuse for
2h and incubated at room temperature for 24h.
• The zone of inhibition around each well was recorded after 24h. (Gebreselema et al, 2013)
• The positive control used for bacterial pathogen was tetracycline and for fungal pathogen was fluconazole.
The negative control used was dimethyl sulfoxide.
• 1-Chloroform, 2-Ethyl acetate, 3-Methanol, 4-Negative Control, 5- Positive Control
• ML- Micrococcus luteus, PA- Pseudomonas aeroginosa, SA- Staphylococcus aureus, EC- Escherichia coli,
CA- Candida albicans
ML PA SA EC CA
1 1 1 1 1

4 5 2 5 2 4 5 2 4 5 2
4 5 2 4
BN01
3 3 3 3 3

1 1 1 1 1

4 2 5 2 5 2
4 5 2 5 4
BN12 5 2 4

3 3 3 3
3
 ML PA SA EC CA

1 1 1 1 1

4 5 2 4 5 2 2 4 5 2
4 5 5 2
BN13 4

3 3
3 3 3

1 1 1 1 1

4 5 2 5 2 2
5 2 4 4 5 5
BN14 4 2 4

3 3 3 3 3

1 1 1 1 1

GB15 4 5 2 4 2 5 2 4 5 2 4 5 2
4

3 3 3 3 3
 ML PA SA EC CA
Samples (mm) (mm) (mm) (mm) (mm)
BN01          
1 0 12 11 13 12
2 18 14 15 16 0
3 10 0 18 17 0
4 0 0 0 0 0
5 33 16 37 30 40
BN12          
1 10 13 0 0 14
2 10 12 15 0 11
3 0 13 0 0 10
4 0 0 0 0 0
5 32 17 33 29 39
BN13          
1 11 11 0 0 14
2 14 14 15 12 0
3 15 12 0 0 0
4 0 0 0 0 0
5 31 17 32 28 38
BN14          
1 12 10 0 10 14
2 14 14 15 20 13
3 0 0 14 16 14
4 0 0 0 0 12
5 28 21 33 35 37
GB15          
1 0 13 13 0 13
2 13 12 27 17 0
3 0 11 0 0 14
4 0 0 0 0 0
5 27 24 33 32 40
 Purification

Thin Layer Chromatography

• The separation of compounds can be analyzed by using thin layer chromatography
technique using suitable solvent fraction.
• The retention factor is used to identify the separated compound by using the
formula:

RF = Distance travelled by Solute / Distance travelled by Solvent

• Since, the crude extract of BN12 has highest anti-oxidant activity, it has been
taken for purification and characterization.

Hexane : Ethyl
acetate

9:1 7:3 5:5

EA:Met
5:5
 Results and Conclusion
• The morphological characterization shows that the color of aerial and substrate
mycelium of actinomycetes varies from white, grey, red, green, pink, orange and
brown.

• The light microscopic analysis showed the shape of mycelium and determination
of spores.
BN01- Spira with branched substrate mycelium
BN12- Rectus. Branched aerial mycelium with short chain spores (3-10). Dense
and branched substrate mycelium with no spores.
BN13- Retinaculum Apertum with branched aerial and substrate mycelium.
Oval shaped short chains of spores.
BN14- Retinaculum Apertum with long chain spores (more than 10). Branched
aerial mycelium and dense substrate mycelium.
GB15- Rectus, branched aerial mycelium with short-chain oval shaped spores
(3-10 spores), dense and unbranched substrate mycelium

• It also showed that all the 5 cultures have not produced melanin pigment.
• The antioxidant assays of 15 crude extracts shows that EA & MET extracts of
isolates BN01, BN12 and BN15 and methanol extract of BN14 have increased
radical scavenging activity. In that, the EA extract of BN12 showed highest
radical scavenging activity of 97.75% at the concentration of 60 µg/mL and has
the half maximal inhibitory concentration (IC50) of 8.66 µg/mL.

• The results of antimicrobial assay showed that the ethyl acetate extract of the
isolate GB15 exhibited a maximum zone of inhibition of 27 mm against S.
aureus and the isolate BN14 showed 20 mm of the zone of inhibition against E.
coli. The isolate BN01 showed a maximum zone of inhibition (18 mm) against
M. luteus. The pathogens P. aeruginosa and C. albicans were moderately
inhibited by all the marine actinomycetes isolates with the average zone of
inhibition of 13 – 14 mm.
 Future works
Characterization
1. Biochemical- Carbon and nitrogen utilization, enzyme production
2. Molecular characterization- Sequencing and phylogenetic analysis

Production
• Bioactive Compound purification- TLC, Column Chromatography
• Characterization- UV, FT-IR, NMR
 References
• Breast Cancer: Types of Treatment | Cancer.Net
• Seca AML, Pinto DCGA. Plant Secondary Metabolites as Anticancer Agents: Successes in
Clinical Trials and Therapeutic Application. Int J Mol Sci. 2018;19(1):263. Published 2018
Jan 16. doi:10.3390/ijms19010263
• Noura El-Ahmady El-Naggar , 2015. Isolation, Screening and Identification of Actinobacteria
with Uricase Activity: Statistical Optimization of Fermentation Conditions for Improved
Production of Uricase by Streptomyces rochei NEAE-25. International Journal of
Pharmacology, 11: 644-658.
• Dholakiya RN, Kumar R, Mishra A, Mody KH and Jha B (2017) Antibacterial and
Antioxidant Activities of Novel Actinobacteria Strain Isolated from Gulf of Khambhat,
Gujarat. Front. Microbiol. 8:2420. doi: 10.3389/fmicb.2017.02420
• Waks AG, Winer EP. Breast Cancer Treatment: A Review. JAMA. 2019 Jan 22;321(3):288-
300. doi: 10.1001/jama.2018.19323. PMID: 30667505.
• Biji, M.. “Marine Actinomycetes as source of antimicrobial compounds and as probiotics and
single cell protein for application in Penaeid Prawn Culture Systems.” (2003).
• Shirling, Elwood B. and David Gottlieb. “Methods for characterization of Streptomyces
species.” International Journal of Systematic and Evolutionary Microbiology 16 (1966): 313-
340.
• Rahman, M.M., Islam, M.B., Biswas, M. et al. In vitro antioxidant and free
radical scavenging activity of different parts of Tabebuia pallida growing in
Bangladesh. BMC Res Notes 8, 621 (2015). https://
doi.org/10.1186/s13104-015-1618-6
• Vijayakumar, Ramasamy. (2015). Actinomycetes are also known as greatest
source of antibiotics.
THANK YOU
 Review of Literature
• Adrienne G et al., 2019
Breast cancer is most common type of cancer among women and it is treated by
means of surgery, radiation therapy, hormone therapy and by medications. The
main limitation of using medication is the side-effects caused during treatment.

• Biji Mathew., 2003

The different pre-treatment conditions of marine soil samples and different medium
for growth of actinomycetes were experimented. CSPY-ME agar medium and
Starch caesin agar medium showed increased growth of actinomycetes species. The
ISP-2 medium and Medium-14 were used for production of secondary metabolites.

• E. B. Shirling and D. Gottlieb, 1966

They Introduced the methods for characterization of Streptomyces species. The
actinomycetes are characterized by means of morphological (color and shape),
physiological (melanin, pigment, NaCl tolerance and organic degradation) and
biochemical methods (carbon and nitrogen utilization).
• Noura El-Ahmady El-Naggar, 2015 & Riddhi et al., 2017
The characterization of actinomycetes at genus and species level is done by
isolation of DNA, amplification of 16s rRNA followed by sequencing and NCBI
blast. The antioxidant activity of compounds is analysed by using DPPH radical
scavenging assay and ferrous ions metal chelating activity

• Jolanta Solecka et al., 2012

They explained on the secondary metabolites obtained from Actinomycetales, their
activities as antibacterial, antitumor, antifungal, antiparasitic and other properties.
The secondary metabolites produced from Streptomyces species includes
tetracyclines, aminoglycosides, macrolides, clavulanic acid, cephamycins,
carbapenems, etc.
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