Graduate Thesis Or Dissertation
 

A filter paper assay for low cellulase activities and the cultivation of Trichoderma reesei on acid whey and sweet whey permeate

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/v692t932r

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  • The traditional filter paper assay for saccharifying cellulase originally described by M. Mandels et al (1976) has been modified to make possible low activity determinations of Trichoderma cellulases. The enzymatic activity appears to decline during a prolonged incubation period if no precautions have been taken. By means of adding bovine serum albumin and potassium chloride as protein stabilizers and sodium azide as an antimicrobial agent filter paper activities in the range from 0.02 to 0.37 (IUPAC assay, 1987) can be estimated by extending the incubation time up to 20 hours. Filter paper activity values obtained by this method may be compared to those obtained by the IUPAC assay by using a conversion factor from 1.4 to 1.7. Acid whey and sweet whey permeate have been investigated as media for growth and metabolite production by Trichoderma reesei QM 9414 using shake flask cultures and spore inocula. In the case of acid whey the mycelial growth after 2 weeks is 13 mg dry weight /ml substrate. The specific growth rate is 0.29/day. The fungus appears to metabolize the whey protein the first 2 weeks. The alkalinity of acid whey rises continuously over a three week period up to a pH of 8.5. In the case of whey permeate the maximal mycelial weight gain is 4.4 mg/ml which appears after 8 days. A rise in net soluble protein level comes after 3-5 days and reaches a maximum value of 0.23 mg/ml after 2 weeks. The pH of whey permeate rises continuously to 7.5 after 3 days and then slowly declines. The net production of cellulases is low on both media. Dilution 1:6 of the acid whey, supplementation with ammonium sulfate and pHadjustments did not enhance the production of cellulases. Acid whey supports a significant growth and sweet whey permeate shows potential for extracellular protein production. A literature review surveys the composition and uses of acid whey, environmental aspects of whey wastes, the fungus Trichoderma reesei, the mode of action of the Trichoderma reesei cellulase system and the structure of cellulose in cotton and wood.
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