Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (1) 1043-1053 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1045Serratia liquefaciens (JNMC), Pseudomonasaureginosa (JNMC), Penicillum crysogenum(JNMC), C<strong>and</strong>ida albicans (JNMC-1),C<strong>and</strong>ida albicans (JNMC-2) <strong>and</strong> from ITRC(Indian Toxicology Research Center, Lucknow)Bacillus cereus (ATCC 10876), Escherichia coli(ATCC 35218). Bacterial cultures were grownat 37°C on nutrient agar medium <strong>and</strong> fungi weregrown at 28°C on Sabouraud’s dextrose agar. Allthe cultures were preserved at 4°C <strong>and</strong> subculturedregularly.Morphological, Physiological <strong>and</strong> biochemicalcharacteristics: The potent Streptomycesisolates selected after primary screening wascharacterized by morphological, biochemical <strong>and</strong>physiological studies (Table III). They were thenobserved for their mycelia structure, <strong>and</strong> sporearrangements on the mycelia under lightmicroscope at 1000X resolution by using Gramstaining. The observed morphology <strong>of</strong> the isolateswas compared with the actinomycetesmorphology provided in Bergey’s Manual for thepresumptive identification <strong>of</strong> the isolates. Thebiochemical tests performed were catalase,lecithinase (egg yolk rection), oxidase, citrateutilization, nitrate reduction, starch hydrolysis,tween-20 hydrolysis (lipolysis), urea hydrolysis,gelatin hydrolysis, ability to produce H 2S,degradation <strong>of</strong> elastin, arbutin, pectin, guanine,tyrosine, esculin <strong>and</strong> xanthine. Carbon utilizationsuch as D-glucose, D-fructose, D-galactose, D-xylose, D-mannitol, D-sorbitol, D-lactose, D-arabinose, D-cellubiose, meso-inositol, dextran,melibiose, melezitose, xylitol, L-rhamnose, L-raffinose was determined on plates containingbasal salt agar media (KH 2PO 42.64 g,K 2HPO 4.3H 2O 5.65 g, (NH4) 2SO 42.64 g, tracesalt solution (CuSO 4.5H 2O 0.64 g, FeSO 4.7H 2O0.11 g, MnCl 2.4H 2O 0.7 g, ZnSO 4.7H 2O 0.15 g,distilled water 100ml) 1ml, agar 20 g <strong>and</strong> distilledwater 1000 ml) to which carbon sources wereadded to a final concentration <strong>of</strong> 1.0% (14). Theplates were incubated at 28°C <strong>and</strong> growth wasassessed after 7, 14, 21 days using glucose aspositive control <strong>and</strong> carbon source free mediumas negative control. The ability to utilize nitrogensources such as L-cystein, L-valine, L-leucine,L-asparagine, L-hydroxyproline, L-serine, L-phenylalanine <strong>and</strong> L-histidine was determined ina basal medium (glucose 1.0 g, MgSO 4.7H 2O0.05 g, FeSO 4.7H 2O 0.001 g, K 2HPO 40.01 g,NaCl 0.05 g, agar 2.0 g <strong>and</strong> distilled water 100ml) to which 0.1% nitrogen sources were added.The plates were incubated at 28°C <strong>and</strong> growthwas recorded after 7, 14, 21 days using L-asparagine as positive control <strong>and</strong> a mediumwithout nitrogen source as a negative control (16).Determination <strong>of</strong> cell wall L, L-diaminopimelicacid was performed by the method <strong>of</strong> Becker etal., (15). Antibiotic sensitivity against the differentantibiotics (Himedia, Mumbai, India) gentamycin(G 30 ), chloromphenicol (C 10 ), linezolid (LZ 30 ),cipr<strong>of</strong>loxacin (CF 5 ), amoxicillin (AM 10 ),methicillin(M 10 ), nalidixic acid (NA 30 ), novobiocin(Nv 5 ), tetracycline (T 10 ), oleondamycin (OL 15 ),streptomycin (S 25 ), vancomycin (VA 5 ),kanamycin (K 30 ), polymixim B (PB 100 ), neomycin(N 30 ), rifamycin (R 5 ), erythromycin (E 15 ),cephalosporin (CR 30 ), penicillin G (P 10 ),amphotericin B (AP 30 ), fluconazole (F 10 ),miconazole (MIC 30 ) were performed using MullerHinton agar medium by paper disc method (16).ResultsThe genus Streptomyces, which is the mostabundant <strong>and</strong> a recoverable actinomycete groupisolated from the soil. Total fifty sevenStreptomyces isolates were recorded from thesoil <strong>of</strong> Durg District <strong>of</strong> Chhattisgarh according todifferences in colony morphology (17) on starchcasein nitrate agar media (Fig. 1). Out <strong>of</strong> 57isolates, only 19 isolates showed the antimicrobialactivity. Among this 19 isolates, 11 showed bothIsolation <strong>and</strong> Characterization <strong>of</strong> Streptomyces sp.
Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (1) 1043-1053 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1046Fig. 1. Growth pattern <strong>of</strong> bioactive Streptomyces isolates on starch casein nitrate agar media fromvarious location <strong>of</strong> Durg.antibacterial <strong>and</strong> antifungal activity while 8exhibited only antibacterial activity. The diameter<strong>of</strong> zone <strong>of</strong> inhibition (in millimeter) showed by theculture filtrate <strong>of</strong> 19 isolates is represented in table(1). In present study results indicated that graycolored isolates were showing activity against themost <strong>of</strong> the test organisms when subjected in tosubmerged culture. The yellow colored isolates(Y8, MS38) <strong>and</strong> grey colored isolates (A8, MS02<strong>and</strong> D15) exhibited highest antibacterial <strong>and</strong>antifungal activity (Fig. 3). Among these isolateY8 (yellow) recorded to show broad spectrumactivity against all gram positive bacteria, gramnegative bacteria <strong>and</strong> fungi. Results <strong>of</strong> antibioticsensitivity test was presented in table-2 showedthat Bacillus cereus (MTCC 1305) was the mostsensitive organism inhibited by the extract <strong>of</strong> 14isolates followed by Bacillus subtilis (MTCC1789), Salmonella typhi (MTCC 531),Escherichia coli (MTCC 1667), Bacillus cereus(ATCC 10876), Staphylococcus aureus (MTCC96), Proteus vulgaris (MTCC 1771), C<strong>and</strong>idaalbicans (MTCC 1637), Klebsiella Pneumoniae(MTCC 2405), Penicillium citrinum (MTCC1751), Staphylococcus epidermis (MTCC 435),Alternaria alternate (MTCC 1779),Escherichia coli (ATCC 35218), Aspergillusniger (MTCC 872), Staphylococcus aureus(JNMC), Serratia liquefaciens (JNMC)Penicillium crysogenum (JNMC), C<strong>and</strong>idaalbicans (JNMC-1), C<strong>and</strong>ida albicans (JNMC-2), Bacillus megaterium (MTCC 1684),Pseudomonas aureginosa (JNMC). Antibioticsensitivity pr<strong>of</strong>ile <strong>of</strong> isolates is shown table-(II)Most <strong>of</strong> the isolates showed resistance to theamoxicillin (AM 10 ), methicillin (M 10 ), nalidixicacid (NA 30 ), cephalosporin (CR 30 ), penicillinG(P 10 ), amphotericin B (AP 30 ), fluconazole (F 10 )<strong>and</strong> showed sensitivity towards the gentamycin(G 30 ), linezolid (LZ 30 ), streptomycin (S 25 ),Singh <strong>and</strong> Rai