full issue - Association of Biotechnology and Pharmacy
full issue - Association of Biotechnology and Pharmacy
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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.