Author name:
عذراء حرجان محسن الدحيدحاوي
Supervisor name:
فاطمة عبد الحسين التميمي | محسن هاشم رسن
University location:
Najaf
Abstract:
هدفت الدراسة الحالية الى تقييم قابلية بعض انزيمات السليليز الفطري على تخمير بقايا النباتات على انتاج الايثانول من بعض الفطريات المحلية المعزولة من 50 نموذج من التربة والذي تضمن Aspergillus oryzae , A. niger, A. terreus , A.flavus , A.fumigatus, A.parasiti | The present study was conducted to proceed a Bioethanol production using some of the waste fermentation plant by cellulosic hydrolysis enzyme produced by some local fungi isolated from 50 samples from soil which were including Aspergillus oryzae, A.niger, A.terreus , A.flavus , A.fumigatus, A.parasiticus, A.nidulans, Penicilliuum chrysogenum, Trichoderma longi, Rhizopus stolonifer, Cladosporium spp, Mucor indicus, Trichothecium spp. The prominent isolates were A. oryzae, A.niger, A. terreus which were chosen for further studies and screening to producing cellulase enzymes the fungal isolates revealed variation values of clear zone It’s (5.1, 5, 5, 4.8) cm to A. oryzae, A.niger, A. terreus , A.flavus respectively. The results revealed that the suitable carbon source to the cellulase activity was induced in different raw plant substrate, the highest was produced when using Corn cobs, Rice husk and Reed reached to (1.72, 2, 1.26)IU/ ml respectively when using A. oryzae and (1.24, 1.17, 1.89) IU/ ml when using A.niger, and (1.31, 1.19, 1.93)IU/ ml when using A. terreus. but using the cellulose powder and CMC, Avical sole carbon source total cellulase given least activity compare with raw plant substrate.on the other hand, using Mandels - Weber medium activity of enzyme production by A. oryzae, A.niger, A. terreus recorded (6.51, 4.14, 5.61) U/ ml respectively when using Corn cobs and (5.79, 4.03, 5.24) U/ ml and (5.55, 3.97, 5.19) U/ ml when using Rice husk and Reed by different fungi. The results refers that , the best chemical pretreatment results were appear when using NaOH at 100Co in 30min on Corn cobs, the total cellulase activity were (1.63, 1, 1.21) U / ml when using fungal isolated to degradation after treatment, while using hot water at100Co without NaOH in 15 min to pretreatment Rice husk activity were (2.16, 2.07, 2.12) U / ml but Reed are appear high activity when pretreatment with NaOH at 100C0 in 15min activity It’s (2.28, 1.65, 2.09) U / ml. The results revealed that, The optimized conditions of the enzymes were different in this study that give activities of Fpase, CMCase and ? - glucosidases to fungus A. oryzae compared to the rest of the fungi using Corn cobs as carbon source, it reached to(2.21, 2.30, 46.72) IU / ml respectively when pH6, and temperature tested for these enzymes recorded (1.64, 1.61, 30.86) IU\ ml respectively at a temperature of 30 C° and when using the concentration of substrate gave activity (2.4o, 2.48, 42.34) IU / ml of 6 % from Corn cobs and recorded when using nitrogen source concentration at 6 % (1.92, 1.89) IU\ ml in KNO3 and (NH4)2SO4 to Fpase and CMCase and the effectiveness of ? - glucosidase at the same nitrogen source (43.54, 50) IU / ml. on the other hand, showed enzymes Fpase, CMCase and ? - glucosidase high activities to ? - glucosidase, CMCase to fungus A. oryzae compared to the rest of the fungi using Reed as carbon source reached (2.31, 2.21, 46.15) IU/ml respectively at pH 6, and at different temperatures were (1.66, 30.94) IU/ml to Fpase and ? - glucosidase at 30 C°, but CMCase gave the highest efficacy (1.60) IU \ml at 25 C° and decreased effectiveness to (2.60, 2.57, 49.69) IU / ml at 6 % of the carbon source recorded effectiveness of the enzyme Fpase (1.19, 1.09) IU / ml when using (NH4)2PO4 and (NH4)2SO4 at 6 % as a nitrogen source of from either enzymatically CMCase and ? - glucosidase was effective at same nitrogen source (1.29, 2) and (35.19, 40.41) IU / ml respectively.The results showed when you use a Rice husk recorded higher effective enzymatic Fpase and CMCase and ? - glucosidases of fungus A. oryzae was (2.32, 39.27) IU /ml for Fpase and ? - glucosidases at pH 6 but CMCase gave the highest efficacy at 5 pH as recorded (2.04) IU / ml at temperatures 30 C°, reached to (1.64, 1.58, 29.13) IU / ml respectively for Fpase and CMCase and ? - glucosidases at a temperature of 30 C°, and when using 6% of the substrate was effective (2.47, 2.19, 44.86) IU / ml, activity recorded when using (NH4)2PO4 and (NH4)2SO4 at 6 % as a nitrogen source of (1.1) IU / ml to FPase on either enzymatically CMCase and ? - glucosidase was effective when use same a nitrogen source (1.18, 1.83) and (31.92, 36.91) IU/ml, respectively. In addition, The results refers when precipitation by ammonium sulfate specific activity were (38.69, 1.25, 5767.5) IU of the enzyme with 0.012 mg protein and then, Elution protein of crude enzyme solution of A.oryzae from DEAE - cellulose column was shown to the protein was separated in (33) fractions.It was found that only the fractions (F - 8) contained cellulase activity, while fraction (F - 30) eluted in column containing buffer and different concentrations of NaCl then using gel filtration and gave three peak of enzymes alone and Characterize enzymes purified FPase activity was found at 5.5 pH which (0.91) U and temperature was having activity at 25 and 30 C°, were (0.14, 0.15) U and CMCase found that the enzyme exhibited maximum activity at 25 C° was (0.33) U but 30 C° for ? - glucosidase was (38.22) U and the molecular weight of the protein was found to be about (38, 52 and 49) kD for (FPase, CMCase, ? - glucosidase) respectively. Finally, estimated ethanol density (0.80 - 0.91) g /ml and ethanol concentration (60 - 97) % but using chromic acid test of purified ethanol the complex enzyme with Reed, complex enzyme with Corn cobs recoded high value of ethanol which were (0.98, 1.72, 1.87, 1.89 ) % and (1.09, 1.83, 1.92, 1.99) % respectively but complex enzyme with Rice husk were given (1.01, 1.12, 1.48, 1.79) % in 4, 5, 6, and 7 days respectively on the other hand, when using the A.oryzae and A.terreus with Reed gave high ethanol value from other fungi with substrate in this study, and observed Rice husk in all results was recorded less absorbance at 350 nm and determination of ethanol concentration by gas chromatography mass used ethanol concentration 99 % as standard, also we showed the ethanol when using enzyme pure with substrate in which were (93.7, 97.1, 88) % ethanol to enzyme with Corn cobs and Reed and Rice husk respectively
