full issue - Association of Biotechnology and Pharmacy

Current Trends in Biotechnology and PharmacyVol. 5 (1) 1043-1053 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1045Serratia liquefaciens (JNMC), Pseudomonasaureginosa (JNMC), Penicillum crysogenum(JNMC), Candida albicans (JNMC-1),Candida albicans (JNMC-2) and 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 and fungi weregrown at 28°C on Sabouraud’s dextrose agar. Allthe cultures were preserved at 4°C and subculturedregularly.Morphological, Physiological and biochemicalcharacteristics: The potent Streptomycesisolates selected after primary screening wascharacterized by morphological, biochemical andphysiological studies (Table III). They were thenobserved for their mycelia structure, and sporearrangements on the mycelia under lightmicroscope at 1000X resolution by using Gramstaining. The observed morphology of the isolateswas compared with the actinomycetesmorphology provided in Bergey’s Manual for thepresumptive identification of 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 of elastin, arbutin, pectin, guanine,tyrosine, esculin and 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 and distilledwater 1000 ml) to which carbon sources wereadded to a final concentration of 1.0% (14). Theplates were incubated at 28°C and growth wasassessed after 7, 14, 21 days using glucose aspositive control and 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 and 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 and distilled water 100ml) to which 0.1% nitrogen sources were added.The plates were incubated at 28°C and growthwas recorded after 7, 14, 21 days using L-asparagine as positive control and a mediumwithout nitrogen source as a negative control (16).Determination of cell wall L, L-diaminopimelicacid was performed by the method of Becker etal., (15). Antibiotic sensitivity against the differentantibiotics (Himedia, Mumbai, India) gentamycin(G 30 ), chloromphenicol (C 10 ), linezolid (LZ 30 ),ciprofloxacin (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 and a recoverable actinomycete groupisolated from the soil. Total fifty sevenStreptomyces isolates were recorded from thesoil of Durg District of Chhattisgarh according todifferences in colony morphology (17) on starchcasein nitrate agar media (Fig. 1). Out of 57isolates, only 19 isolates showed the antimicrobialactivity. Among this 19 isolates, 11 showed bothIsolation and Characterization of Streptomyces sp.