بررسي شرايط بهينه توليد آنزيم بتاگلوگوزيداز توسط قارچ Trichoderma reesei )PTCC5142 )

Optimized conditions for production of P-glucosidase enzyme in Trichoderma reesei (PTCCS142)

p-glucosidase, is one of the cellulase enzym es system, hydrolyses cellobiose or cellooligoschrides to gluco se. In this research for production of p-glucosidase enzyme, p-glucosidase activity was measured in the optimum conditions in 30 isolates of Trichoderma sp. In order to obtain the optimum conditions: an isolate of Trichoderma (T. reesei PTCC 5142) was grown in Mandels media containing CMC, Avicel and filter paper as carb on source at pH 4,5,6 and 7. Lactose and cellubiose were added to media as inducer agents . Media were incubated at 25. 29 and 32 oCfor 14 days. Samples were collected in 24h intervals, and enzyme activity was measured . Results showed that the optimum cond itions for hyperproduction of p-glu cosidase were pH=5, CMC as carbon source. lacto se as inducer on seventh day at 29 oc. After screening the -glucosidase activity of all 30 isolates in the optimized conditi ons. it was shown that T. reesei PTCC 5142 had the highest level of -glucosidase specific activity (0.45 U/mg) . For characterization and study of p-Glucosidase gene. CTAB method used for genomic DNA extraction. The expected PCR product with 566 bp was amplified with two specific primers (CPII & CPI2). The amplified fragment was confirmed by restriction pattern .