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http://dx.doi.org/10.4014/jmb.1510.10092

A Newly Identified Glutaminase-Free L-Asparaginase (L-ASPG86) from the Marine Bacterium Mesoflavibacter zeaxanthinifaciens  

Lee, Su-Jin (Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology)
Lee, Youngdeuk (Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology)
Park, Gun-Hoo (Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology)
Umasuthan, Navaneethaiyer (Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University)
Heo, Soo-Jin (Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology)
Zoysa, Mahanama De (College of Veterinary Medicine, Chungnam National University)
Jung, Won-Kyo (Department of Biomedical Engineering and Centre for Marine-Integrated Biomedical Technology (BK21 Plus), Pukyong National University)
Lee, Dae-Won (Marine Ecosystem and Biological Research Center, Korea Institute of Ocean Science & Technology)
Kim, Hanjun (Marine Ecosystem and Biological Research Center, Korea Institute of Ocean Science & Technology)
Kang, Do-Hyung (Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology)
Oh, Chulhong (Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology)
Publication Information
Journal of Microbiology and Biotechnology / v.26, no.6, 2016 , pp. 1115-1123 More about this Journal
Abstract
L-Asparaginase (E.C. 3.5.1.1) is an enzyme involved in asparagine hydrolysis and has the potential to effect leukemic cells and various other cancer cells. We identified the L-asparaginase gene (L-ASPG86) in the genus Mesoflavibacter, which consists of a 1,035 bp open reading frame encoding 344 amino acids. Following phylogenetic analysis, the deduced amino acid sequence of L-ASPG86 (L-ASPG86) was grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The L-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant L-asparaginase (r-L-ASPG86) showed optimum conditions at 37-40℃, pH 9. Moreover, r-L-ASPG86 did not exhibit glutaminase activity. In the metal ions test, its enzymatic activity was highly improved upon addition of 5 mM manganese (3.97-fold) and magnesium (3.35-fold) compared with the untreated control. The specific activity of r-L-ASPG86 was 687.1 units/mg under optimum conditions (37℃, pH 9, and 5 mM MnSO4).
Keywords
L-Asparaginase; Mesoflavibacter; cloning; expression; manganese; glutaminase-free;
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