kasem, M., Fayed, A., Eldemery, S., Okba, A., Ibrahim, A. (2016). PURIFICATION, CHARACTERIZATION AND SEQUENCING OF AMYLASE A FROM ALKALIPHLIC BACILLUS LICHENIFORMIS STRAIN-MK7. Research Journal of Applied Biotechnology, 2(Special issue (2) for the first International Conference of Genetic Engineering and Biotechnology), 11-20. doi: 10.21608/rjab.2016.59670
Mohamed kasem; Aysam M. Fayed; Samah Eldemery; Adel Okba; Atef Ibrahim. "PURIFICATION, CHARACTERIZATION AND SEQUENCING OF AMYLASE A FROM ALKALIPHLIC BACILLUS LICHENIFORMIS STRAIN-MK7". Research Journal of Applied Biotechnology, 2, Special issue (2) for the first International Conference of Genetic Engineering and Biotechnology, 2016, 11-20. doi: 10.21608/rjab.2016.59670
kasem, M., Fayed, A., Eldemery, S., Okba, A., Ibrahim, A. (2016). 'PURIFICATION, CHARACTERIZATION AND SEQUENCING OF AMYLASE A FROM ALKALIPHLIC BACILLUS LICHENIFORMIS STRAIN-MK7', Research Journal of Applied Biotechnology, 2(Special issue (2) for the first International Conference of Genetic Engineering and Biotechnology), pp. 11-20. doi: 10.21608/rjab.2016.59670
kasem, M., Fayed, A., Eldemery, S., Okba, A., Ibrahim, A. PURIFICATION, CHARACTERIZATION AND SEQUENCING OF AMYLASE A FROM ALKALIPHLIC BACILLUS LICHENIFORMIS STRAIN-MK7. Research Journal of Applied Biotechnology, 2016; 2(Special issue (2) for the first International Conference of Genetic Engineering and Biotechnology): 11-20. doi: 10.21608/rjab.2016.59670
PURIFICATION, CHARACTERIZATION AND SEQUENCING OF AMYLASE A FROM ALKALIPHLIC BACILLUS LICHENIFORMIS STRAIN-MK7
1Microbial biotechnology dept., Genetic Engineering and Biotechnology Research Institute, University of Sadat City (USC), Egypt
2Molecular biology dept., Genetic Engineering and Biotechnology Research Institute, University of Sadat City (USC), Egypt
3MicrobialBiotechnology Dept., Genetic Engineering & Biotechnology Research Institute (GEBRI), University of Sadat City (USC), Sadat City, Egypt
Abstract
Two degenerative primers were designated from the closest Bacillus, Bacillus licheniformis strain DSM13 (ATCC 14580) which has complete genome to get amylase gene from our Bacillus licheniformis strain MK7 (KP322016) by PCR technique. We got about 1500 bp PCR, the sequence analysis revealed that the alkaliphilic cellulase belongs to glycosyl hydrolase family 12A which has 100 % identity to glycoside hydrolase from Bacillus sp. (WP009329478) and has 99 % identity to family 12 xyloglucanase from Bacillus licheniformis (2JEMIA). The nucleotide sequence of the alkaliphilic cellulase gene (cel12A) was deposited to the GenBank under accession number (KP322015). The cel12A gene from B. licheniformis strain MK7 has open reading frame of 1500 bp that codes for 449 amino acid which has a molecular weight of 55KDa protein. The enzyme protein was further purified using ammonium sulphate precipitation and chromatography. The characterization of amylase protein showed that the enzyme has an optimum temperature at (37oC), optimum pH (9.0) and Some metals increase the enzyme activity such as CoCl2 +, K+ and EDTA , but others decrease the activity such as Ag +, Zn2 + and urea ions.
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