title: A Newly Identified Glutaminase-Free ʟ-Asparaginase (ʟ-ASPG86) from the Marine Bacterium Mesoflavibacter zeaxanthinifaciens.
abstract: ʟ-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 Lasparaginase gene (ʟ-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 ʟ-ASPG86 (ʟ-ASPG86) was grouped as a type I asparaginase with respective homologs in Escherichia coli and Yersinia pseudotuberculosis. The ʟ-ASPG86 gene was cloned into the pET-16b vector to express the respective protein in E. coli BL21 (DE3) cells. Recombinant ʟ-asparaginase (r-ʟ-ASPG86) showed optimum conditions at 37-40oC, pH 9. Moreover, r-ʟ-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-LASPG86 was 687.1 units/mg under optimum conditions (37°C, pH 9, and 5 mM MnSO4).
authors.affiliation: Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology, Jeju Special Self-Governing Province 63349, Republic of Korea.
authors.forename: Su-Jin
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology, Jeju Special Self-Governing Province 63349, Republic of Korea.
authors.forename: Youngdeuk
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology, Jeju Special Self-Governing Province 63349, Republic of Korea.
authors.forename: Gun-Hoo
authors.initials: GH
authors.lastname: Park
authors.affiliation: Department of Marine Life Sciences, School of Marine Biomedical Sciences, Jeju National University, Jeju Special Self-Governing Province 63243, Republic of Korea.
authors.forename: Navaneethaiyer
authors.initials: N
authors.lastname: Umasuthan
authors.affiliation: Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology, Jeju Special Self-Governing Province 63349, Republic of Korea.
authors.forename: Soo-Jin
authors.initials: SJ
authors.lastname: Heo
authors.affiliation: College of Veterinary Medicine, Chungnam National University, Daejeon 34134, Republic of Korea.
authors.forename: Mahanama
authors.initials: M
authors.lastname: De Zoysa
authors.affiliation: Department of Biomedical Engineering and Centre for Marine-Integrated Biomedical Technology (BK21 Plus), Pukyong National University, Busan 48513, Republic of Korea.
authors.forename: Won-Kyo
authors.initials: WK
authors.lastname: Jung
authors.affiliation: Marine Ecosystem and Biological Research Center, Korea Institute of Ocean Science & Technology, Ansan 15627, Republic of Korea.
authors.forename: Dae-Won
authors.initials: DW
authors.lastname: Lee
authors.affiliation: Marine Ecosystem and Biological Research Center, Korea Institute of Ocean Science & Technology, Ansan 15627, Republic of Korea.
authors.forename: Hanjun
authors.initials: H
authors.lastname: Kim
authors.affiliation: Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology, Jeju Special Self-Governing Province 63349, Republic of Korea.
authors.forename: Do-Hyung
authors.initials: DH
authors.lastname: Kang
authors.affiliation: Jeju International Marine Science Research & Education Center, Korea Institute of Ocean Science & Technology, Jeju Special Self-Governing Province 63349, Republic of Korea.
authors.forename: Chulhong
authors.initials: C
authors.lastname: Oh
mesh_terms: D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000302:isolation & purification; D001215:Asparaginase / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001216:Asparagine / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D041965:Flavobacteriaceae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D005972:Glutaminase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008958:Models, Molecular; D010802:Phylogeny; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012623:Seawater / Q000382:microbiology*; D013379:Substrate Specificity; D015011:Yersinia pseudotuberculosis / Q000201:enzymology / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D011994:Recombinant Proteins; D008345:Manganese; D001216:Asparagine; D001215:Asparaginase; D005972:Glutaminase; D008274:Magnesium
keywords: Mesoflavibacter; cloning; expression; glutaminase-free; manganese; ʟ-Asparaginase