In this study, the recombinant endoglucanase was purified by his - tag affinity chromatography with differrent processes, such as using phosphate buffer with or without sodium cloride, pretreatment of samples with ammonium sulphate before supplying into affinity column, using various concentration of imidazole for washing. | TAP CHI SINH HOC 2020 42 1 73 81 DOI 0866-7160 SELECTION OF PURIFICATION CONDITION FOR RECOMBINANT ENDOGLUCANASE ORIGINATED FROM GOAT RUMEN BACTERIA IN Escherichia coli Nguyen Thi Quy1 Nguyen Hong Duong1 Dao Trong Khoa1 Nguyen Khanh Hoang Viet2 Nguyen Khanh Hai3 Truong Nam Hai1 4 Do Thi Huyen1 4 1Institute of Biotechnology VAST Vietnam 2Institute of NewTechnology Academy of Military Science and Technology 3Faculty of Biology VNU Hanoi University of Science 4Graduate University of Science and Technology VAST Vietnam Received 10 October 2019 accepted 2 March 2020 ABSTRACT Cellulase is an important enzyme that plays a role in cleaving β -1 4 glucoside on cellulose to release glucose which is of economic value and can be applied in many different fields. The 1545 bp endoglucanase gene mined from goat rumen s bacterial metagenomic data was expressed in Escherichia coli Rosetta2. In this study the recombinant endoglucanase was purified by his - tag affinity chromatography with differrent processes such as using phosphate buffer with or without sodium cloride pretreatment of samples with ammonium sulphate before supplying into affinity column using various concentration of imidazole for washing. Finally the endoglucanse was sucessfully purified by his-tag affinity column using sodium chloride-free phosphate buffer of which 150 mM and 400 mM imidazole were used for washing and enzyme elution respectively. The resulting enzyme showed its high purity of 99 . CMC plate assay confirmed that although less active than commercial cellulase Sigma the recombinant cellulase hydrolyzed CMC to form a clear zone halo around the well. The purified enzyme is capable of using as material for further analysis. Keywords Escherichia coli Rosetta2 Carboxymethylcellulose CMC clear zone halo protein purification recombinant endoglucanase. Citation Nguyen Thi Quy Nguyen Hong Duong Dao Trong Khoa Nguyen Khanh Hoang Viet Nguyen Khanh Hai Truong Nam Hai Do Thi Huyen 2020. .
