Home > Archives > IJSRST173473 IJSRST-Library

Production, Purification and Advance Applications of L- Asparaginase (Review)

Authors(4) :-Sadia Sanawer, Sikander Ali, Tahira Mohsin, Ammara Nasir

L-Asparaginase, an enzyme (E.C. that is responsible for breakdown of L-asparagine into aspartic acid and ammonia, has significant applications in the therapeutics and food technology. Presently L-Asparaginase is mostly produced by using microbial sources including bacteria, fungi and actinomycetes. In therapeutics L-Asparaginase is used as antileukemic agent to treat acute lymphoblastic leukemia (ALL), a malignancy mostly present in children. In food industry this enzyme is used to diminish the quantity of acrylamide that is present in fried and baked starchy foods and carcinogenic in nature. L-Asparaginase done deamination of asparagine thus reduced acrylamide concentration. Another application is its use in biosensors. Both at industrial scale and at laboratory level production of L-Asparaginase can be done by submerged fermentation (SmF) or by solid-state fermentation (SSF). Comparison of both methods revealed that SSF is preferable over SmF because it is inexpensive, environment-friendly and it provides high yield of L-Asparaginase enzyme. SSF method consumes both industrial and agricultural wastes as their substrate. The contamination is significantly decreased in SSF because of presence of low moisture content. Purification of enzyme can be done by using different techniques i.e. precipitation of sample, dialysis and various chromatographic techniques are used in different combinations. A wise combination of steps increase the purification and yield
Sadia Sanawer, Sikander Ali, Tahira Mohsin, Ammara Nasir
E.C., ALL, SSF, L-Asparaginase enzyme, pH
  1. Amena, S., N. Vishalakshi,M.Prabhaka,A. Dayanand and K.Lingappa.2010. Production purification and characterization of L-asparaginase from Streptomyces gulbargensis.BJM., 41: 173-178.
  2. Arima, K.,T.Sakamoto,C.Araki and G.Tamura.1972. Production of extracellular L-Asparaginases by microorganisms. Agric Biol Chem., 36(3): 356-361.
  3. Avramis,I.,S. Sencer,P. Antonia,P.Sather,B.Bostrom, L.Cohen, A.Ettinger, L.Ettinger, J.Franklin,P.Gaynon, J.Hilden, B.Lange, F.Majlessipour, P.Mathew, M.Needle, J.Neglia, G.Reaman and J.Holcenberg.2002. A randomized comparison of native Escherichia coli asparaginase and polyethylene glycol conjugated asparaginase for treatment of children with newly diagnosed standard-risk acute lymphoblastic leukemia: a Children's Cancer Group study. Blood., 99 (6):1986?1994.
  4. Basha,S.,R.Rekha,M.Komala and S.Ruby. 2009. Production of extracellular antileukaemic enzyme L-asparaginase from marine actinomycetes by solid- state and submerged fermentation: Purification and characterisation. TJPR., 8: 353-360.
  5. Broome,D.1961. Evidence that the L-asparaginase activity of guinea pig serum is responsible for its antilymphoma effects. Nature., 191(4793): 1114-1115.
  6. Broome, D. 1963. Evidence that the L-Asparaginase of guinea pig serum is responsible for its antilymphoma effects I. Properties of the L-Asparaginase of guinea pig serum in relation to those of the antilymphoma substance. J. Exp. Med., 118(1): 99-120.
  7. Ciesarova, Z., E.Kiss and Boegl. 2006. Impact of L-Asparaginase on acrylamide content in potato products. J FOOD NUTR RES., 45(4): 141-146.
  8. De-Angeli,C.,F.Pocchiari,S.Russi, A.Tonolo,E. Zurita and E.Ciaranfi. 1970. Effect of L-asparaginase from Aspergillus terreus on ascites sarcoma in the rat. Nature., 225(7): 549-550.
  9. Dhanam,G. and S.Kannan.2013. L-asparaginase: types perspectives and applications. Adv Bio Technol., 13: 1-5.
  10. Dharmaraj, S.2011. Study of L-Asparaginase production by Streptomyces noursei MTCC 10469, isolated from marine sponge Callyspongia diffusa.IJB., 9(2): 102-108.
  11. Dhevagi, P. and E.Poorani. 2006. Isolation and characterization of L-asparaginase from marine actinomycetes. IJBT., 5: 514-520.
  12. El-Bessoumy, A.,M. Sarhan and J.Mansour.2004. Production, isolation, and purification of L-Asparaginase from Pseudomonas aeruginosa 50071 using solid-state fermentation. IJBMB., 37(4): 387-393.
  13. Ferrara, A., N.Severino, J.Mansure,S.Martins,M.Oliveire and C.Siani.2006. Asparaginase production by a recombinant Pichia pastoris strain harbouring Saccharomyces cerevisiae ASP3 gene. Enzyme Microb Technol., 39(7): 1457-1463.
  14. Friedman, M. 2003. Chemistry biochemistry and safety of acrylamide. J. Agric. Food Chem., 51: 4504-4526.
  15. Gervais, D.,N.Allison,A.Jennings,S.Jones and T.Marks. 2013. Validation of a 30-yearold process for the manufacture of L-asparaginase from Erwinia chrysanthemi. Bioprocess Biosyst Eng., 36: 453-60.
  16. Gokmen, V. and K.Palazoglu.2008. Acrylamide formation in foods during thermal processing with a focus on frying. Food Bioprocess Tech., 1(168): 35-42.
  17. Golunski, S.,V.Astolfi,N.Carniel,D.Oliveira,M.Di Luccio,A.Mazutti and H.Treichel.2011. Ethanol precipitation and ultrafiltration of inulinases from Kluyveromyces marxianus. Sep Purif Technol., 78: 261-265.
  18. Grslund,S.,P.Nordlund,J.Weigelt,J.Bray,O.Gileadi,S.Knapp,U.Oppermann,C.Arrowsmith, R.Hui,J.Ming and W.Park. 2008. Protein production and purification. Nat Methods., 5: 135-146.
  19. Gulati, R.,K.Saxena and R.Gupta.1997. A rapid plate assay for screening L-Asparaginase producing micro-organisms. Lett Appl Microbiol., 24(1): 23-26.
  20. Hendriksen,V.,B.Kornbrust,R.Ostergaard and M.Stringer.2009. Evaluating the potential for enzymatic acrylamide mitigation in a range of food products using an asparaginase from Aspergillus oryzae. J. Agri Food Chem., 57(10): 4168-4176.
  21. Hill, M.,J.Roberts,E.Loeb,A.Khan,A.MacLellan and W.Hill. 1967. LAsparaginase therapy for leukemia and other malignant neoplasms. Remission in human leukemia. JAMA., 202(9): 882-888.
  22. Imada, A.,S.Igarasi,K.Nakahama and M.Isono.1973. Asparaginase and glutaminase activities of micro-organisms.J Gen Microbiol., 76(1): 85-99.
  23. Jain, R.,U. Zaidi, Y.Verma and P.Saxena.2012. L-asparaginase: a promising enzyme for treatment of ALL. People's J Sci Res., 5: 29-35.
  24. Kidd, G. 1953. 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(6): 565-582.
  25. Killander, D.,A. Dohlwitz,L.Engstedt,S.Franzen,G.Gahrton,B.Gullbring,G.Holm,A. Holmgren,S.H-glund,A,Killander and D.Lockner. 1976. Hypersensitive reactions and antibody formation during L-asparaginase treatment of children and adults with acute leukemia. Cancer., 37 (1): 220-228.
  26. Kumar, S., V.Dasu and K.Pakshirajan.2011. Purification and characterization of glutaminase-free L-Asparaginase from Pectobacterium carotovorum MTCC 1428. Bioresour Technol., 102(2): 2077-2082.
  27. Kumar, S. and K.Sobha.2012. L-Asparaginase from microbes: A comprehensive review. Adv. Biores., 3(4): 137-157.
  28. Kumar,M. and K.Manonmani. 2013. Purification, characterization and kinetic properties of extracellular L-asparaginase produced by Cladosporium sp. World J Microbiol Biotechnol., 29: 577-587.
  29. Kumar, K.,M. Kataria and N. Verma. 2013. Plant asparaginase-based asparagine biosensor for leukemia. Artif Cells Nanomed Biotechnol., 41: 184-188.
  30. Lam ,H.,M. Kavoosi and A.Haynes . 2004. Affinity-enhanced protein partitioning in decyl b-D-glucopyranoside two-phase aqueous micellar systems. Biotechnol Bioeng., 89: 381-392.
  31. Lee, S.,T. Ross,E.Gustafson,H.Wroble and M.Muschik.1986. Large-scale recovery and purification of L-Asparaginase from Eriwinia carotovora. Appl Biochem Biotech., 12: 229-247.
  32. Lingnert,H.,S.Grivas, M.Jagerstad, K.Skog, M.Tornqvist and P.Aman. 2002. Acrylamide in food mechanisms of formation and influencing factors during heating of foods. Scand J Food Nutr., 46: 159-172.
  33. Loureiro,B.,S. Borges, F.Andrade,G.Tone and S.Said. 2012. Purification and biochemical characterization of native and pegylated form of L-asparaginase from Aspergillus terreus and evaluation of its antiproliferative activity. Adv Microbiol., 2: 138-45.
  34. Mashburn,T. and C.Wriston.1964. Tumor inhibitory effect of L-Asparaginase from Escherichia coli. Arch Biochem and Biophy., 105: 450-452.
  35. Michalska, K. and M.Jaskolski. 2006. Structural aspects of L-Asparaginases, their friends and relations. Acta Biochimica Polonica., 53(4): 627.
  36. Mishra, A. 2006. Production of L-Asparaginase, an anticancer agent, from Aspergillus niger using agricultural waste in solid state fermentation. Appl Biochem Biotech., 135(1): 33-42.
  37. Mohan, S. and K.Manonmani.2013. Purification, characterization and kinetic properties of extracellular L-Asparaginase produced by Cladosporium sp. IUMS., 29(4): 577-587.
  38. Mohapatra,R., K.Sani and C. Banerjee.1995. Characterization of L-asparaginase from Bacillus sp. isolated from an intertidal marine alga (Sargassum sp.). Lett Appl Microbiol., 21(6): 380-383.
  39. Moorthy, V.,A. Ramalingam,A.Sumantha and T.Shankaranaya.2010. Production, purification and characterisation of extracellular L-Asparaginase from a soil isolate of Bacillus sp. Afr. J. Microbiol. Re., 4(18): 1862-1867.
  40. Patro, R. and N. Gupta. 2012. Extraction, purification and characterization of L-As paraginase from Penicillium sp. by submerged fermentation. IJBM Res., 3(3): 30-34.
  41. Pedreschi, F.,K. Kaack and K. Granby. 2008. The effect of asparaginase on acrylamide formation in French fries. Food Chem., 109: 386-392.
  42. Pritsa, A. and A. Kyriakidis. 2001. L-Asparaginase of Thermus thermophilus: Purification, properties and identification of essential amino acids for its catalytic activity. Mol Cell Biochem., 216: 93-101.
  43. Qin ,M. and F. Zhao. 2003. L-asparaginase release from Escherichia coli cells with aqueous two-phase micellar systems. Appl Biochem Biotechnol., 110: 11-21.
  44. Sahu,K.,K.Sivakumar,E.Poorani,T.Thangaradjou,L.Kannan.2007. Studies on L-asparaginase enzyme of actinomycetes isolated from estuarine fishes. J Environ Biol., 28:465-474.
  45. Salzer,L.,L. Asselin,V.Plourde,T.Corn and S.Hunger.2014. Development of asparaginase Erwinia chrysanthemi for the treatment of acute lymphoblastic leukemia. Ann. N. Y. Acad. Sci.,1329: 81-92.
  46. Sarquis, M.,M. Oliveira,S.Santos and L.Costa.2004. Production of L-Asparaginase by filamentous fungi. Mem Inst Oswaldo Cruz., 99(5): 489-492.
  47. Shrivastava, A.,A.Khan,A.Shrivastav, K.Jain and K.Singhal.2012. Kinetic studies of L-Asparaginase from Penicillium digitatum. Prepara Biochem Biotech., 42(6): 574-581.
  48. Sinha, R.,R. Singh and K.Jha.2013. L-Asparaginase: present and future prospective. IJIRSET., 2: 7031-7051.
  49. Soares,L., M.Guimares,B.Polakiewicz,N.Pitombo and J.Neto.2002. Effects of polyethylene glycol attachment on physicochemical and biological stability of E. coli l-asparaginase. Int J Pharm., 237: 163-70.
  50. Souza,L., P.Ventura,M.Soares,A.Coutinho and A.Lima.2015. Lipase purification using ionic liquids as adjuvants in aqueous two-phase systems. Green Chem., 17: 3026-34.
  51. Sudhir,P.,R.Dave, K.Trivedi and B. Subramanian. 2012. Production and amplification of an L-Asparaginase gene from actinomycete isolate Streptomyces ABR2. Ann Microbiol., 62(4): 1609-1614.
  52. Szymanska, B., U.Wilczynska-Kalak,H.Kang,L. Liem,H.Carol and B.Lock.2012. Pharmacokinetic modelling of an induction regimen for in vivo combined testing of novel drugs against pediatric acute lymphoblastic leukemia xenografts. PloS One., 7(3): 33894-33994.
  53. Takahashi,H., J. Inoue,K .Sakaguchi, M. Takagi, S. Mizutani and J. Inazawa.2017. Autophagy is required for cell survival under L-asparaginase-induced metabolic stress in acute lymphoblastic leukemia cells.Oncogene.,59:1-2.
  54. Theantana, T., D.Hyde and S. Lumyong. 2009. Asparaginase production by endophytic fungi from Thai medicinal plants: cytotoxicity properties. Int J Integr Biol., 7: 1-8.
  55. Uppuluri,B.,R. Dasari, V.Sajja,S. Jacob and D.Reddy. 2013. Optimization of L-Asparaginase production by Isolated Aspergillus niger C4 from sesame (black) oil cake under SSF using Box Behnken design in column bioreactor.INT J CHEM REACT ENG., 11(1): 1-7.
  56. Van, H. 2011. Asparaginase revisited. Leuk Lymphoma., 52(2): 168-178.
  57. Verma, N.,M.Bansal and S.Kumar.2012. Whole cell based miniaturized fiber optic biosensor to monitor L-asparagine. J.Appl Sci Res., 3: 809-814.
  58. Warangkar,C. and N.Khobragade. 2010. Purification, characterization, and effect of thiol compounds on activity of the Erwinia carotovora L-asparaginase. Enzyme Res., 1: 1-10.
  59. Watanabe,I., N.Miyata, A.Ando, R.Shiroma, K.Tokuyasua and T.Nakamura.2002. Biotechnological advantages of laboratory-scale solid-state fermentation with fungi. Biochem Eng Jour., 93(1): 90-95.
  60. Zeidan, A.,S.Wang and M.Wetzler.2009. Pegasparaginase: where do we stand? Expert Opin Biol Ther., 9 (1): 111-119.
  61. Zia,A.,R.Bashir,I.Ahmed and T.Iftikhar.2013. Production of L-Asparaginase from Aspergillus niger using agro wastes by-products in submerged fermentation process. Jurnal Teknologi., 62(2): 47-51.
  62. Zyzak,V.,R. Sanders, M.Stojanovic, H.Tallmadge, L. Eberhart ,K.Ewald, C.Gruber, R.Morsch, A.Strothers, P.Rizzi and D.Villagran. 2003. Acrylamide formation mechanism in heated foods. J Agric Food Chem., 51(16):4782-4787.
Publication Details
  Published in : Volume 3 | Issue 4 | May-June 2017
  Date of Publication : 2017-06-30
License:  This work is licensed under a Creative Commons Attribution 4.0 International License.
Page(s) : 341-351
Manuscript Number : IJSRST173473
Publisher : Technoscience Academy
PRINT ISSN : 2395-6011
ONLINE ISSN : 2395-602X
Cite This Article :
Sadia Sanawer, Sikander Ali, Tahira Mohsin, Ammara Nasir, "Production, Purification and Advance Applications of L- Asparaginase (Review) ", International Journal of Scientific Research in Science and Technology(IJSRST), Print ISSN : 2395-6011, Online ISSN : 2395-602X, Volume 3, Issue 4, pp.341-351 , May-June-2017
URL : http://ijsrst.com/IJSRST173473