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

이영덕|정원교|허수진|이대원|김한준|강도형|오철홍|이수진|박건후|디조이사마하나마|Navaneethaiyer Umasuthan
한국미생물·생명공학회 (The Korean Society For Microbiology And Biotechnology)|2016|Journal of Microbiology and Biotechnology|제26권 제6호|pp. 1115-1123|DOI 10.4014/jmb.1510.10092

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expressionmanganesecloningʟ-AsparaginaseMesoflavibacterglutaminase-free
주제분류생물학

초록

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 Lasparaginase 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 37-40oC, 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-LASPG86 was 687.1 units/mg under optimum conditions (37°C, pH 9, and 5 mM MnSO4).

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 Lasparaginase 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 37-40oC, 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-LASPG86 was 687.1 units/mg under optimum conditions (37°C, pH 9, and 5 mM MnSO4).

참고문헌 (49)

  1. Abakumova O, "Antitumor activity of L-asparaginase from Yersinia pseudotuberculosis", Biomed. Khim, 54, pp.712~719 (2008)
  2. Aghaiypour K, "Structural basis for the activity and substrate specificity of Erwinia chrysanthemi L-asparaginase", Biochemistry, 40, pp.5655~5664 (2001)
  3. Angélica Moreno-Enríquez, "Biochemical Characterization of Recombinant L-Asparaginase (AnsA) from Rhizobium etli, a Member of an Increasing Rhizobial-Type Family of L-Asparaginases", Journal of Microbiology and Biotechnology, 22(3), pp.292~300 (2012), 한국미생물·생명공학회
  4. Arnold K, "The SWISSMODEL workspace: a web-based environment for protein structure homology modelling", Bioinformatics, 22, pp.195~201 (2006)
  5. Bansal S, "Structural stability and functional analysis of L-asparaginase from Pyrococcus furiosus", Biochemistry (Moscow), 75, pp.375~381 (2010)
  6. Basha NS, "Production of extracellular anti-leukaemic enzyme L-asparaginase from marine actinomycetes by solid-state and submerged fermentation:purification and characterisation", Trop. J. Pharm. Res, 8, pp.353~360 (2009)
  7. Broome J, "Evidence that the L-asparaginase activity of guinea pig serum is responsible for its antilymphoma effects", Nature (Lond.), 191, pp.1114~1115 (1961)
  8. Broome J, "Studies on the mechanism of tumor inhibition by L-asparaginase. Effects of the enzyme on asparagine levels in the blood, normal tissues, and 6C3HED lymphomas of mice: differences in asparagine formation and utilization in asparaginase-sensitive and -resistant lymphoma cells", J. Exp. Med, 127, pp.1055~1072 (1968)
  9. Campbell H, "Two Lasparaginases from Escherichia coli B. Their separation, purification, and antitumor activity", Biochemistry, 6, pp.721~730 (1967)
  10. DeLano WL, "The PyMOL user’s manual" (2002), DeLano Scientific
  11. Duval M, "Comparison of Escherichia coli-asparaginase with Erwinia-asparaginase in the treatment of childhood lymphoid malignancies: results of a randomized European Organisation for Research and Treatment of Cancer-Children's Leukemia Group phase 3 trial", Blood, 99, pp.2734~2739 (2002)
  12. Ashraf A. El-Bessoumy, "Production, Isolation, and Purification of L-Asparaginase from Pseudomonas Aeruginosa 50071 Using Solid-state Fermentation", BMB reports, 37(4), pp.387~393 (2004), 한국생화학분자생물학회
  13. Elshafei AM, "Purification, characterization and antitumor activity of L-asparaginase from Penicillium brevicompactum NRC 829", Br. Microbiol. Res. J, 2, pp.158~174 (2012)
  14. Ghasemi Y, "An optimized medium for screening of L-asparaginase production by Escherichia coli", Am. J. Biochem. Biotechnol, 4, pp.422~ (2008)
  15. 한상원, "Biochemical Characterization of L-Asparaginase in NaCl-Tolerant Staphylococcus sp. OJ82 Isolated from Fermented Seafood", Journal of Microbiology and Biotechnology, 24(8), pp.1096~1104 (2014), 한국미생물·생명공학회[DOI]
  16. Hong SJ, "Cloning, expression, and characterization of thermophilic L-asparaginase from Thermococcus kodakarensis KOD1", J. Basic Microbiol, 54, pp.500~508 (2014)
  17. Imada A, "Asparaginase and glutaminase activities of micro-organisms", J. Gen. Microbiol, 76, pp.85~99 (1973)
  18. Joner PE, "Purification and properties of L-asparaginase A from Acinetobacter calcoaceticus", Biochim. Biophys. Acta, 327, pp.146~156 (1973)
  19. Kamble V, "Purification of L-asparaginase from a bacteria Erwinia carotovora and effect of a dihydropyrimidine derivative on some of its kinetic parameters", Ind. J. Biochem. Biophys, 43, pp.391~ (2006)
  20. Khushoo A, "Extracellular expression and single step purification of recombinant Escherichia coli L-asparaginase II", Protein Expr. Purif, 38, pp.29~36 (2004)
  21. Kidd JG, "Regression of transplanted lymphomas induced in vivo by means of normal guinea pig serum. I. Course of transplanted cancers of various kinds in mice and rats given guinea pig serum, horse serum, or rabbit serum", J. Exp. Med, 98, pp.565~582 (1953)
  22. Kornbrust BA, "Asparaginase - an enzyme for acrylamide reduction in food products", Enzymes Food Technol, 2, pp.59~87 (2010)
  23. Kotzia GA, "L-Asparaginase from Erwinia chrysanthemi 3937: cloning, expression and characterization", J. Biotechnol, 127, pp.657~669 (2007)
  24. Kozak M, "A comparison between the crystal and solution structures of Escherichia coli asparaginase II", Acta Biochim. Pol, 49, pp.509~514 (2002)
  25. Krasotkina J, "One-step purification and kinetic properties of the recombinant L-asparaginase from Erwinia carotovora", Biotechnol. Appl. Biochem, 39, pp.215~221 (2004)
  26. Kumar S, "Purification and characterization of glutaminase-free L-asparaginase from Pectobacterium carotovorum MTCC 1428", Bioresour. Technol, 102, pp.2077~2082 (2011)
  27. Law AS, "Purification and properties of Bacillus coagulans L-asparaginase", Arch. Biochem. Biophys, 147, pp.744~752 (1971)
  28. Lee MB, "L-Asparaginase activity in human and animal sera", Nature, 217, pp.758~759 (1968)
  29. Mahajan RV, "Efficient production of L-asparaginase from Bacillus licheniformis with low-glutaminase activity: optimization, scale up and acrylamide degradation studies", Bioresour. Technol, 125, pp.11~16 (2012)
  30. Manna S, "Purification, characterization and antitumor activity of Lasparaginase isolated from Pseudomonas stutzeri MB-405", Curr. Microbiol, 30, pp.291~298 (1995)
  31. Mercado L, "Escherichia coli L-asparaginase induces phosphorylation of endogenous polypeptides in human immune cells", Sangre, 44, pp.438~442 (1999)
  32. Mottram DS, "Food chemistry: acrylamide is formed in the Maillard reaction", Nature, 419, pp.448~449 (2002)
  33. Muharram M, "Recombinant expression, purification of L-asparaginase-II from thermotolerant E. coli strain and evaluation of its antiproliferative activity", Afr. J. Microbiol. Res, 8, pp.1610~1619 (2014)
  34. Oh C, "Whole-genome sequence of the xylanase-producing Mesoflavibacter zeaxanthinifaciens strain S86", J. Bacteriol, 193, pp.5557~ (2011)
  35. Onishi Y, "Effective treatment for suppression of acrylamide formation in fried potato chips using Lasparaginase from Bacillus subtilis", 3 Biotech, 5, pp.783~789 (2015)
  36. Oza VP, "Withania somnifera (Ashwagandha): a novel source of Lasparaginase", J. Integr. Plant Biol, 51, pp.201~206 (2009)
  37. Paul JH, "Regulation of L-asparaginase in a Chlamydomonas species in response to ambient concentrations of combined nitrogen", J. Bacteriol, 147, pp.9~12 (1981)
  38. Peterson R, "L-Asparaginase production by various bacteria", Appl. Microbiol, 17, pp.929~ (1969)
  39. Pokrovskaya M, "Cloning, expression and characterization of the recombinant Yersinia pseudotuberculosis L-asparaginase", Protein Expr. Purif, 82, pp.150~154 (2012)
  40. Sarquis MIdM, "Production of L-asparaginase by filamentous fungi", Memorias do Instituto Oswaldo Cruz, 99, pp.489~492 (2004)
  41. Savitri NA, "Microbial L-asparaginase: a potent antitumour enzyme", Ind. J. Biotechnol, 2, pp.184~194 (2003)
  42. Streeter JG, "Asparaginase and asparagine transaminase in soybean leaves and root nodules", Plant Physiol, 60, pp.235~239 (1977)
  43. Tamura K, "MEGA6: molecular evolutionary genetics analysis version 6.0", Mol. Biol. Evol, 30, pp.2725~2729 (2013)
  44. Tareke E, "Analysis of acrylamide, a carcinogen formed in heated foodstuffs", J. Agric. Food Chem, 50, pp.4998~5006 (2002)
  45. Thompson JD, "CLUSTAL W:improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice", Nucleic Acids Res, 22, pp.4673~4680 (1994)
  46. Warangkar SC, "Purification, characterization, and effect of thiol compounds on activity of the Erwinia carotovora L-asparaginase", Enzyme Res, 2010, pp.165878~ (2010)
  47. Yao M, "Structure of the type I L-asparaginase from the hyperthermophilic archaeon Pyrococcus horikoshii at 2.16 resolution", Acta Crystallogr. D Biol. Crystallogr, 61, pp.294~301 (2005)
  48. Yun M-K, "Crystal structure and allosteric regulation of the cytoplasmic Escherichia coli L-asparaginase I", J. Mol. Biol, 369, pp.794~811 (2007)
  49. Zuo S, "Recent research progress on microbial L-asparaginases", Appl. Microbiol. Biotechnol, 99, pp.1069~1079 (2015)

관련 논문AI추천

Anticancer Activity of Extremely Effective Recombinant L-Asparaginase from Burkholderia pseudomalleiAl-Awthan Yahya S., Alnour Tarig M., Darwesh Doaa B., Darwish Ahmed B., Elfaki Imadeldin, Habib Salem A., Youssef Magdy M.한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2022 · 제32권 제5호Biochemical Characterization of L-Asparaginase in NaCl-Tolerant Staphylococcus sp. OJ82 Isolated from Fermented Seafood박우준, 정재준, 한상원한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2014 · 제24권 제8호Secretory Expression of Escherichia coli L-Asparaginase in Corynebacterium glutamicumKim Geun-Joong, Lee Jin-Young, Yoo Ho-Seok, Yoo Su-Kyoung한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2025 · 제35권 Expression and Characterization of a Novel Nitrilase from Hyperthermophilic Bacterium Thermotoga maritima MSB8Huayou Chen, Jinru Jia, Rui Tian, Shengli Yang, Tianxi Zhang, Zhi Chen, Zhong Ni한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2015 · 제25권 제10호Characterization of a Glycoside Hydrolase Family 50 Thermostable β-agarase AgrA from Marine Bacteria Agarivorans sp. AG17오철홍, 이숙경, 이영득, 이제희, 황일손, NIKAPITIYACHAMILANI LALANTHI한국수산과학회 · Fisheries and Aquatic Sciences2010 · 제13권 제1호Characterization of a Recombinant Thermostable Arylsulfatase from Deep-Sea Bacterium Flammeovirga pacificaChao Gao, Min Jin, Runying Zeng, Zhiwei Yi한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2015 · 제25권 제11호

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