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Production and characterization of thermostable cellulases from Streptomyces transformant T3-1
Authors:Jang  Hung-Der  Chen   Kuo-Shu
Affiliation:(1) Department of Food Science, Yuanpei University of Science and Technology, Hsinchu, 300, Taiwan; Tel.:;(2) Department of Medical Technology, Yuanpei University of Science and Technology, Hsinchu, 300, Taiwan
Abstract:A total of 26 thermophilic isolates, selected from a compost of agricultural waste, which was mostly composed of vegetable, corncob and rice straw, were cultivated at 50 °C for further studies of thermostable cellulase production. The thermostable cellulase gene from the chromosomal DNA of actinomycetes isolate no. 10 was shotgun-cloned and transformed into Streptomyces sp. IAF 10-164. A transformant, T3-1, was found to be a good strain for the production of thermostable cellulases. Cultivation of T3-1 in modified Mandels–Reese broth containing 1% carboxymethylcellulose (CMC)-sodium salt and the optimal condition for microbial growth were studied. Batch cultivation in a flask revealed that CMCase and Avicelase production reached the maximum between the third to fifth day, whereas maximum beta-glucosidase production occurred on the ninth day. Microbial biomass increased from the first day to the fifth day and then decreased. The crude enzyme had the highest activity at 50 °C and at pH 6.5. The enzyme was shown to be a thermostable cellulase whose activities were stable at 50 °C for more than 7 days.
Keywords:Compost  gene cloning  Streptomyces  thermostable cellulase  transformants
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