SCREENING, ISOLATION, AND ANTIBACTERIAL ACTIVITY OF ANTIBIOTIC PRODUCING BACTERIA OBTAINED FROM SAPROPHYTIC SOIL SAMPLES

KAVITHA  R; Dhamodharan N; Dhivya C

doi:10.22159/ajpcr.2017.v10i3.14472

Authors

KAVITHA R SRM University
Dhamodharan N
Dhivya C

DOI:

https://doi.org/10.22159/ajpcr.2017.v10i3.14472

Abstract

ABSTRACT
Objective: The aim of the study was conducted to screen and isolate potential antibiotic producing bacteria from saprophytic soils collected from
Potheri and Nandiambakkam.
Methods: Soil was collected aseptically and subjected to serial dilution. Crowded plate technique was used for the isolation of the colony. Totally
seven isolates were isolated and were screened for their antibacterial activity. The three isolates (S2A, S2B, and S3A) having better zone of inhibition
were selected for morphological, microscopical, and biochemical test to prove their validity. The selected isolates were partially purified. The partially
purified samples further screened for antibacterial activity, minimum inhibitory concentration (MIC) and the isolates, which shown good zone of
inhibition were subjected to 16S rRNA sequencing studies to determine the species.
Results: The isolates screened based on size of the zone formed. Best isolate selected by zone of inhibition was subjected to antibacterial activity,
morphological, microscopical, and biochemical test, partial purification of three isolates and further screened for antibacterial and MIC. The isolate
showed good zone of inhibition compared to others by MIC was selected for 16S rRNA sequencing studies. Genomic DNA extracted from isolate S2B
conforms it belongs to Pseudomonas species which is named as Pseudomonas putida 2435.
Conclusion: The research work revealed that the three isolates showed good antibacterial activity against Gram-positive and Gram-negative bacteria.
The S2B isolate was confirmed to P. putida 2435 by 16S rRNA studies.
Keywords: Isolation of soil microbes, Biochemical characterization, Antimicrobial activity, Minimum inhibitory concentration, 16S rRNA sequencing,
Pseudomonas putida.

Downloads

Download data is not yet available.

Author Biography

KAVITHA R, SRM University

Departemnt of Pharmaceutics, SRM College of Pharmacey, SRM University,

References

REFERENCES

Kaur S, Kaur J, Pankaj PP. Isolation and characterization of antibiotic producing microorganisms from soil samples of a certain area of Punjab region of India. Int J Pharm Clin Res 2014;6(4):312-5.

Singh AP, Mishra S. Isolation and biochemical characterization of antibiotic producing microorganism from waste soil samples of certain industrial areas of India. IOSR J Pharm Biol Sci 2013;5(6):80-9.

Abbas S, Senthilkumar R, Arjunan S. Isolation and molecular characterization of the microorganism producing novel antibiotics from the soil sample. Eur J Exp Biol 2014;4(5):149-55.

Lee YS, Lee HY, Lee CH, Park HS. Isolation of antibiotic-producing bacteria antagonistic to Fusarium oxysporium from sesame-growing soils and evaluation of their antifungal activity. J Microbiol Biotechnol 1995;5(6):46-352.

Chaudhary HS, Yadav J, Shrivastava AR, Singh S, Singh AK, Gopalan N. Antibacterial activity of actinomycetes isolated from different soil samples of Sheopur (A city of central India). J Adv Pharm Technol Res 2013;4(2):118-23.

Dwivedi S, Thakkar R, Upadhyay V, Kumar S. Isolation and characterization of actinomycetes producing antimicrobial substance against human pathogenic bacteria. J Pharm Res 2011;2(4):4066-8.

Kokare C. Identification of morphological characterization of bacteria. Pharm Microbiol Princ Appl 2005;7:4-7.

Pepper IL, Garba CP. Environmental Microbiology. Vol. 2. London: Elsevier Academic Press; 2004. p. 37-49.

Sundararaj T. Motility tests for the isolated microorganism. Taramani, India: Dr. A. L. Mudaliar Post Graduate Institute of Basic Medical Sciences, University of Madras; 1995. p. 52-3.

Nonomura H. The key for classification and identification of 458 species of the Streptomycetes included in ISP. J Ferment Technol 1974;52:107-14.

Krithika S, Chellaram C. Isolation, screening and characterization of chitinase producing bacteria from marine water. Int J Pharm Pharm Sci 2016;8:34-6.

Abirami V, Meenakshi SA, Kanthymathy K, Bharathidasan R, Mahalingam R, Panneerselvam A. Partial purification and characterization of an extracellular protease from Penicillium janthinellum and Neurospora crassa. Eur J Exp Biol 2011;1(3):114-23.

Verma M. Antimicrobial screening of the leaf extracts of Bacopa monnieri (L) Pennell. IJCPR 2016;8:21-3.

Edwards U, Rogall T, BlÃ¶cker H, Emde M, BÃ¶ttger EC. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res 1989;17(19):7843-53.

Castresana J. Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. Mol Biol Evol 2000;17(4):540-52.

Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, Chevenet F, et al. Phylogeny.fr: Robust phylogenetic analysis for the non-specialist. Nucleic Acids Res 2008;36(1):W465-9.

Edgar RC. MUSCLE: Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 2004;32(5):1792-7.

Talavera G, Castresana J. Improvement of phylogenies after removing divergent unambiguously aligned blocks from protein sequence alignments. Syst Biol 2007;56(4):564-77.