137

CHAPTER 6
CONCLUSION

The study was carried out to ascertain the diverse strains of the
Streptomyces from Kolli and Yercaud hills of Eastern Ghats in Tamil Nadu,
southern India. It was identified that a total of four potential strains was
observed to possess the antimicrobial and anticancerous activities and also able
to biosynthesize various nanoparticles such as silver, gold and copper
nanoparticles.
In the present study, four different strains namely Yer11, Yer28 from
Yercaud hills and Kol35, Kol44 strains from Kolli hills were isolated from the
rhizosphere region of medicinal plants. These strains were capable of inhibiting
the cell growth of human pathogenic microorganisms and breast cancer cell
lines to a greater extend.
For the comparison study, a standard strain of Streptomyces
sannanensis MTCC 6285 was procured from the culture collection center,
Chandigarh, India. The number of single colony which was more prominent
and observed to be more on Starch casein nitrate (SCN) agar medium
compared to other media used. Based on the morphological, physiological and
biochemical characterizations, the isolates were identified as Streptomyces
sannanensis.
Molecular characterization studies revealed that the extraction of
DNA from the strains yielded high purity of DNA through STE lysis method

138

compared with that of the other three methods. At the same, the extracted DNA
was subjected to the RAPD analysis using 10 operon primers. Of these, 100%
polymorphic band was obtained using OPA10 followed by OPB09 (95.65%)
and OPB16 (95.45%). Dendrogram analysis using Jaccards similarity
coefficient showed that Kol35 was found to be genetically unique compared to
other strains whereas Yer11, MTCC 6285, Yer28 and Kol44 strains were
observed under a same clade. The protein profile was obtained through SDS
PAGE extraction buffer yielded a higher quantity of protein than the extraction
through TCA and urea protocols tested.
In the case of the antagonistic activity of the isolated strains, Kol35
strain was observed to give prominent activity against the human bacterial and
some of the fungal pathogens. The antibacterial activity was studied by some
reliable methods like conventional streaking, well diffusion and determination
of the Minimum inhibitory concentration (MIC) level. In all the above said
three methods, Kol35 strain possessed the highest antagonistic activity against
S. paratyphi A, E. coliand P. aeuroginosa.
The antifungal and antibacterial activities were screened using the
crude extract of the selected strains of S.sannanensis which showed that all the
strains were observed to inhibit the fungal and bacterial growth, as the
concentration of the crude extract increased the growth of the pathogens was
found to be reduced significantly. Among the different crude extract
concentrations tested for antimicrobial activity, the concentration of 10 mL
and 20 mL were found to be reduced the growth of the fungal and bacterial;
respectively.
The crude extracts containing antibiotics obtained from Yer11,
Yer28, Kol35, Kol44 and MTCC 6285 strains were tested against the human
breast cancerous cell lines such as SKBR3, MCF7, MDA-MB231. The results

139

indicated that the crude extract obtained from the Kol35 strain showed a
significant inhibition of cell growth when compared to that of the other strained
tested. As the study was focused on the biomedical applications, the crude
extract from Kol35 strain was subjected to the purification process through
GC-MS analysis. The results indicated that silane, pyridine, 2,4,6-trimethyl,
amino malonic acid, 4-benzoxazin, Tris methyl and cyclohexydimethoxy
methyl compounds were identified in the crude extract and subsequently
purified for further studies.
After purification of the crude extract, the selected strains of
S.sannanensis were inoculated in the medium containing silver nitrate,
chloroauric acid and copper sulphate or the biosynthesis of the silver, gold and
copper nanoparticles respectively. Each of the nanoparticles was characterized
and confirmed through UV-VIS, FTIR and SEM analysis.
The activity of nanoparticles was verified against certain bacterial
and fungal pathogens including some breast cancerous cell lines which showed
the highest antagonistic activity but superior to antibacterial and antifungal
activities of the crude extract. The anticancerous activity was found to be
maximum with gold nanoparticle followed by the silver and copper
nanoparticles.
In this study, the selected strains of actinomycetes especially
S.sannanensis were found to be of potential candidate for antimicrobial and
anticancerous activities. The search for wide spectrum of antimicrobial
metabolites, especially from actinomycetes, requires a large number of isolates
in order to discover a novel compound of pharmaceutical interest. Thus, it can
be concluded from the results of the present investigation, the actinomycetes
from Yercaud and Kolli hills of Eastern Ghats region are a potential source of
novel antibiotics in biomedical applications.