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Current Trends in Biotechnology and PharmacyVol. 5 (1) 982-992 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)987Fig. 4. Effect of nitrogen on exoglucanase productionby Aspergillus sp. RCAL-5 under following conditions(pH 4.5, temp. 28 o C, agitation150 rpm , wheatbran 10 gl -1 , incubation time (4 days)).Fig. 6. Effect of temperature on exoglucanase productionby Aspergillus sp. RCAL-5 under followingconditions. (agitation 150 rpm, wheat bran 10 g l -1 ,yeast extract 2.5 g l -1 , pH 4.5, incubation time (4 days)).Fig. 5. Effect of pH on exoglucanase production byAspergillus sp. RCAL-5 under following conditions.(temp. 28 °C, agitation 150 rpm, wheat bran 10 g l -1 ,yeast extract 2.5 g l -1 , incubation time (4 days)).Fig. 7. Effect of agitation on exoglucanase productionby Aspergillus sp. RCAL-5 under following conditions.(temp. 28 o C, wheat bran 10 g l -1 , yeast extract2.5 g l -1 , pH 4.5, incubation time (4 days)).maximum cellulase production (0.775 IU ml -1 )using saw dust as carbon source from A.nigergrown at 28°C.Recently, Deshpande et al. (17) observedmaximum cellulase activity (0.22±0.04 IU ml -1 )at the incubation temperature of 30 o C.Effect of AgitationEffect of different agitation rates rangingfrom 50-250 rpm were studied for maximumFPase (0.81 FPU ml -1 ) production under theconditions (temp. 28 o C, wheat bran 10 g l -1 , yeastextract 2.5 g l -1 , pH 4.5). From the Figure 7, it isChandra Sekhar et al
Current Trends in Biotechnology and PharmacyVol. 5 (1) 982-992 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)988Fig. 8. Enzymatic hydrolysis of control and NaOHpretreated Groundnut shell using the cellulases derivedfrom Aspergillus sp. RCAL-5. The hydrolyticconditions were: substrate and enzyme concentration(1: 15), 50 mM citrate buffer, pH 5.0, 50 °C at 100 rpm.The enzyme cocktail (cellulase 30 FPU, β-glucosidase55 U and xylanase 150 U) was loaded per gram substrate.evident that maximum FPase production wasachieved at 150 rpm followed by a gradualdownfall upon increasing the agitation rates.Agitation has a very important role for theproduction of various metabolites under shakeflask cultivation conditions. Optimum agitation isrequired for the appropriate air supply as well asadequate nutrient availability to the microorganism(18). We observed a maximum of cellulaseproduction (0.81 U ml -1 ) at 150 rpm. Gomes etal. (18) found that shifting of agitation speed from120 rpm to 180 rpm showed 2-fold (2.3 U ml -1 ofCMCase) increment in enzyme production.Our observation for cellulase productionfrom this novel isolate could be fairly comparedwith the previous study of Vyas et al. (19) andDeshpande et al. (17).Cellulase production under optimized set ofconditionsThe cellulolytic enzyme production profileby Aspergillus sp. RCAL-5 under optimizedFig. 9. The time course of growth, sugar utilizationand ethanol production by P. stipitis NCIM 3498 at 30oC using Groundnut shell enzymatic hydrolysate (pH5.5). Initial sugar concentration was 49 g l -1 and thefinal ethanol was 19.4 g l -1 after 84 h.Table 2. Hydrolytic enzymes production profile byAspergillus sp. RCAL-5 under optimized conditions(agitation 150 rpm, temp. 28 o C, wheat bran 10 g l -1 ,yeast extract 2.5 g l -1 , pH 4.5 and after 4 days of incubation).Enzyme Activity (U ml -1 )FPase 0.86CMCase 1.25β – glucosidase 1.68Xylanase 4.57# The data presented are averages of three independentanalysis.conditions is given in Table 2. The organismshowed all the main enzymes required for thedepolymerization of structural polysaccharidesfrom the delignified substrate. Aspergillus sp.RCAL-5 showed maximum cellulase (0.88 FPUml -1 ) after 4 days of incubation, exhibiting higherlevel of CMCase than FPase activity, which is anormal pattern among cellulolytic organisms (20,Fermentation of Enzymatically Saccharified Groundnut
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