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Recent Patents on Biotechnology


ISSN (Print): 1872-2083
ISSN (Online): 2212-4012

Review Article

Isolation, Purification, Characterisation and Application of L-ASNase: A Review

Author(s): Tania Paul*, Abhijit Mondal and Tarun Kanti Bandyopadhyay

Volume 13, Issue 1, 2019

Page: [33 - 44] Pages: 12

DOI: 10.2174/1872208312666181012150407

Price: $65


Background: L-ASNase (L-asparagine aminohydrolase EC is used for the conversion of L-asparagine to L-aspartic acid and ammonia and also it was found as an agent of chemotherapeutic property according to recent patents. It is known as an anti-cancer agent and recently it has received an immense attention. Various microorganisms have the ability to secrete the L-ASNase. It is famous world-wide as anti-tumor medicine for acute lymphoblastic leukemia and lymphosarcoma. L-ASNase helps in deamination of Asparagine and Glutamine.

Source: L-ASNase mainly found in two bacterial sources; Escherichia coli and Erwinia carotovora. Isolation from plants: Endophytes were also a great source of L-ASNase. It was isolated from four types of plants named as; C. citratus, O. diffusa, M. koengii, and also P. bleo.

Applications: L-ASNase is used as a potential anti-tumor medicine. It plays a very much essential role for the growth of tumor cells. Tumor cells require a lot of asparagine for their growth. But ASNase converts to aspartate and ammonia from asparagine. So the tumor cell does not proliferate and fails to survive. The L-ASNase is used as the medicine for the major type of cancer like acute lymphocytic leukemia (ALL), brain. It also used as a medicine for central nervous system (CNS) tumors, and also for neuroblastoma. Two types of L-ASNase have been found.

Conclusion: L-ASNase becomes a powerful anti-tumor medicine and researchers should develop a potent strain of asparaginase which can produce asparaginase in the industrial level. It is also used in the pharmaceutical industry and food industry on a broader scale.

Keywords: L-ASNase, enzyme, isolation, purification, microorganisms, anti-tumor medicine.

Graphical Abstract
Mashburn LT, Wriston JC Jr. Tumor inhibitory effect of L-asparaginase from Escherichia coli. Arch Biochem Biophys 1964; 105: 450-3.
Shrivastava A, Khan AA, Khurshid M, et al. Recent developments in l-asparaginase discovery and its potential as anticancer agent. Crit Rev Oncol Hematol 2016; 100: 1-10.
Verma N, Kumar K, Kaur G, et al. L-asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol 2007; 27: 45-62.
Saeed H, Ali H, Soudan H, et al. Molecular cloning, structural modeling and production of recombinant Aspergillus terreus l. asparaginase in Escherichia coli. Int J Biol Macromol 2018; 106: 1041-51.
Schalk AM, Nguyen H-A, Rigouin C, et al. Identification and structural analysis of an L-asparaginase enzyme from guinea pig with putative tumor cell killing properties. J Biol Chem 2014; 289: 33175-86.
Aguayo A, Cortes J, Thomas D, et al. Combination therapy with methotrexate, vincristine, polyethylene-glycol conjugated‐asparaginase, and prednisone in the treatment of patients with refractory or recurrent acute lymphoblastic leukemia. Cancer 1999; 86: 1203-9.
Wakaizumi M, Yamamoto H, Fujimoto N, et al. Acrylamide degradation by filamentous fungi used in food and beverage industries. J Biosci Bioeng 2009; 108: 391-3.
Ou S, Shi J, Huang C, et al. Effect of antioxidants on elimination and formation of acrylamide in model reaction systems. J Hazard Mater 2010; 182: 863-8.
Curtis TY, Muttucumaru N, Shewry PR, et al. Effects of genotype and environment on free amino acid levels in wheat grain: implications for acrylamide formation during processing. J Agric Food Chem 2009; 57: 1013-21.
Halford NG, Muttucumaru N, Curtis TY, et al. Genetic and agronomic approaches to decreasing acrylamide precursors in crop plants. Food Addit Contam 2007; 24: 26-36.
Ciesarova Z, Kukurova K, Bednáriková A, et al. Improvement of cereal product safety by enzymatic way of acrylamide mitigation. Czech J Food Sci 2009; 27: S96-8.
Sharma SK, Sehgal N, Kumar A. Biomolecules for development of biosensors and their applications. Curr Appl Phys 2003; 3: 307-16.
Jha SK, Pasrija D, Sinha RK, et al. Microbial L-asparaginase: a review on current scenario and future prospects. Int J Pharm Sci Res 2012; 3: 3076.
Bansal S, Gnaneswari D, Mishra P, et al. Structural stability and functional analysis of L-asparaginase from Pyrococcus furiosus. Biochemistry (Mosc) 2010; 75: 375-81.
Moorthy V, Ramalingam A, Sumantha A, et al. Production, purification and characterisation of extracellular L-asparaginase from a soil isolate of Bacillus sp. Afr J Microbiol Res 2010; 4: 1862-7.
Pritsa AA, Kyriakidis DA. L-asparaginase of Thermus thermophilus: Purification, properties and identificaation of essential amino acids for its catalytic activity. Mol Cell Biochem 2001; 216: 93-101.
Borkotaky B, Bezbaruah R. Production and properties of asparaginase from a new Erwinia sp. Folia Microbiol 2002; 47: 473-6.
Paglla U, Rao C, Rajulapati SB. Studies on L-asparaginase production by using Staphylococcus Capitis. J Chem Biol Phys Sci (JCBPS) 2012; 3: 201.
Agarwal A, Kumar S, Veeranki V. Effect of chemical and physical parameters on the production of l-asparaginase from a newly isolated Serratia marcescens SK-07. Lett Appl Microbiol 2011; 52: 307-13.
Sun Y, Su J, Liu G, et al. Advances of blood cell-based drug delivery systems. Eur J Pharm Sci 2017; 96: 115-28.
He H, Ye J, Wang Y, et al. Cell-penetrating peptides meditated encapsulation of protein therapeutics into intact red blood cells and its application. J Control Release 2014; 176: 123-32.
Baskar G, Renganathan S. Application of latin square design for the evaluation and screening of supplementary nitrogen source for L-asparaginase production by Aspergillus terreus MTCC 1782. Indian J Sci Technol 2009; 2: 50-4.
Baskar G, Renganathan S. Statistical screening of process variables for the production of L-asparaginase from cornflour by Aspergillus terreus MTCC 1782 in submerged fermentation. Indian J Sci Technol 2009; 2: 45-8.
Baskar G, Sriharini C, Sripriya R, et al. Statistical screening of supplementary nitrogen source for enhanced production of L-Asparaginase by Aspergillus terreus 1782. Chem Biochem Eng Q 2010; 24: 467-72.
Gurunathan B, Sahadevan R. Design of experiments and artificial neural network linked genetic algorithm for modeling and optimization of L-asparaginase production by Aspergillus terreus MTCC 1782. Biotechnol Bioprocess Eng 2011; 16: 50-8.
Farag AM, Hassan SW, Beltagy EA, et al. Optimization of production of anti-tumor l-asparaginase by free and immobilized marine Aspergillus terreus. Egypt J Aquat Res 2015; 41: 295-302.
Dias FFG, de Castro RJS, Ohara A, et al. Simplex centroid mixture design to improve l-asparaginase production in solid-state fermentation using agroindustrial wastes. Biocatal Agric Biotechnol 2015; 4: 528-34.
Uppuluri KB, Dasari RKV, Sajja V, et al. 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 2013; 11: 103-9.
Dias FF, Sato HH. Sequential optimization strategy for maximum l-asparaginase production from Aspergillus oryzae CCT 3940. Biocatal Agric Biotechnol 2016; 6: 33-9.
Dutta S, Ghosh S, Pramanik S. L-asparaginase and L-glutaminase from Aspergillus fumigatus WL002: Production and some physicochemical properties. Appl Biochem Microbiol 2015; 51: 425-31.
Patro KR, Basak UC, Mohapatra AK, et al. Development of new medium composition for enhanced production of L-asparaginase by Aspergillus flavus. J Environ Biol 2014; 35: 295.
Kumar NM, Manonmani H. Purification, characterization and kinetic properties of extracellular l-asparaginase produced by Cladosporium sp. World J Biol Psychiatry 2013; 29: 577-87.
Kumar NM, Ramasamy R, Manonmani H. Production and optimization of l-asparaginase from Clados-porium sp. using agricultural residues in solid state fermentation. Ind Crops Prod 2013; 43: 150-8.
Eisele N, Linke D, Bitzer K, et al. The first characterized asparaginase from a basidiomycete, Flammulina velutipes. Bioresour Technol 2011; 102: 3316-21.
Heba A, El-Shafei MS, Mostafa H, et al. Statistical optimization of anti-leukemic enzyme l-asparaginase production by Penicillium cyclopium. Curr Trends Biotechnol Pharm 2014; 130-8.
Shafei MS, El-Refai HA, Mostafa H, et al. Purification, characterization and kinetic properties of Penicillium cyclopium L-asparaginase: Impact of lasparaginase on acrylamide content in potato products and its cytotoxic activity. Curr Trends Biotechnol Pharm 2015; 9: 132-40.
Huang L, Liu Y, Sun Y, et al. Biochemical characterization of a novel L-Asparaginase with low glutaminase activity from Rhizomucor miehei and its application in food safety and leukemia treatment. Appl Environ Microbiol 2014; 80: 1561-9.
Strobel G. Genetic diversity of microbial endophytes and their biotechnical applications Genomics Applications for the Developing World. Springer: Germany 2012.
Patil MP, Patil RH, Maheshwari VL. A novel and sensitive agar plug assay for screening of asparaginase-producing endophytic fungi from aegle marmelos. Acta Biol Szeged 2012; 52: 175-7.
Mhatre A, Narwankar R, Rawat A, et al. Characterization of endophytic fungi from medicinal plants for application in therapeutic enzyme extraction. Current Perspectives in Sustainable Environment Management. Indian Institute of Environment Management: Navi Mumbai 2017.
Harper JK, Arif AM, Ford EJ, et al. Pestacin: a 1, 3-dihydro isobenzofuran from Pestalotiopsis microspora possessing antioxidant and antimycotic activities. Tetrahedron 2003; 59: 2471-6.
MP Gutierrez R. MN Gonzalez A, M Ramirez A. Compounds derived from endophytes: a review of phytochemistry and pharmacology. Curr Med Chem 2012; 19: 2992-3030.
Cassady JM, Chan KK, Floss HG, et al. Recent developments in the maytansinoid antitumor agents. Chem Pharm Bull 2004; 52: 1-26.
Zhao J, Li C, Wang W, et al. Hypocrea lixii, novel endophytic fungi producing anticancer agent cajanol, isolated from pigeon pea (Cajanus cajan [L.] Millsp.). J Appl Microbiol 2013; 115: 102-13.
Sim K, Sri Nurestri A, Norhanom A. Phenolic content and antioxidant activity of crude and fractionated extracts of Pereskia bleo (Kunth) DC.(Cactaceae). Afr J Pharm Pharmacol 2010; 4: 193-201.
Yun-Cheung K, Kam-Hung N, Pui-Hay BP, et al. Sources of the anti-implantation alkaloid yueh-chukene in the genus Murraya. J Ethnopharmacol 1986; 15: 195-200.
Ting AS, Meon S, Kadir J, et al. Endophytic microorganisms as potential growth promoters of banana. BioControl 2008; 53: 541-53.
Theantana T, Hyde KD, Lumyong S. Asparaginase production by endophytic fungi from Thai medicinal plants: cytoxicity properties. Int J Integr Biol 2009; 7: 1-8.
Tan M, Sulaiman S, Najimuddin N, et al. Methanolic extract of Pereskia bleo (Kunth) DC.(Cactaceae) induces apoptosis in breast carcinoma, T47-D cell line. J Ethnopharmacol 2005; 96: 287-94.
Diaz PL, Hennell JR, Sucher NJ. Genomic DNA extraction and barcoding of endophytic fungi Plant DNA Fingerprinting and Barcoding. Springer: Germany 2012.
Nakahama K, Imada A, Igarasi S, et al. Formation of L-asparaginase by Fusarium species. Microbiol 1973; 75: 269-73.
Chow Y, Ting AS. Endophytic L-asparaginase-producing fungi from plants associated with anticancer properties. J Adv Res 2015; 6: 869-76.
Sati S, Belwal M. Aquatic hyphomycetes as endophytes of riparian plant roots. Mycologia 2005; 97: 45-9.
Rodríguez JA, Astudillo L, Schmeda-Hirschmann G. Oleanolic acid promotes healing of acetic acid-induced chronic gastric lesions in rats. Pharmacol Res 2003; 48: 291-4.
Krishnamurthy Y, Hemalatha T. Isolation of endophytic fungi from some grasses. J Mycol Pl Path 2003; 33: 305-6.
Baskar G, Renganathan S. Statistical and evolutionary optimisation of operating conditions for enhanced production of fungal l-asparaginase. Chem Pap 2011; 65: 798-804.
Baskar G, Renganathan S. Optimization of L‐asparaginase production by Aspergillus terreus MTCC 1782 using response surface methodology and artificial neural network‐linked genetic algorithm. Asia-Pac J Chem Eng 2012; 7: 212-20.
Baskar G, Sathya SRKLJ, Jinnah RB, et al. Statistical optimization of polysaccharide production by submerged cultivation of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (W.Curt.: Fr.) P. Karst. MTCC 1039 (Aphyllophoromycetideae). Int J Med Mushrooms 2011; 13: 41-9.
Gurunathan B, Sahadevan R. Optimization of culture conditions and bench-scale production of L-asparaginase by submerged fermentation of Aspergillus terreus MTCC 1782. J Microbiol Biotechnol 2012; 22: 923-9.
Imada A, Igarasi S, Nakahama K, et al. Asparaginase and glutaminase activities of micro-organisms. Microbiol 1973; 76: 85-99.
Broome J. Evidence that the L-asparaginase activity of guinea pig serum is responsible for its antilymphoma effects. Nature 1961; 191: 1114.
Lopes AM. Oliveira-Nascimento Ld, Ribeiro A, et al. Therapeutic l-asparaginase: upstream, downstream and beyond. Crit Rev Biotechnol 2017; 37: 82-99.
Yun M-K, Nourse A, White SW, et al. Crystal structure and allosteric regulation of the cytoplasmic Escherichia coli L-asparaginase I. J Mol Biol 2007; 369: 794-811.
Pieters R, Hunger SP, Boos J, et al. L-asparaginase treatment in acute lymphoblastic leukemia. Cancer 2011; 117: 238-49.
Evans WE, Tsiatis A, Rivera G, et al. Anaphylactoid reactions to Escherichia coli and Erwinia asparaginase in children with leukemia and lymphoma. Cancer 1982; 49: 1378-83.
Keating MJ, Holmes R, Lerner S, et al. L-asparaginase and PEG asparaginase-past, present, and future. Leukemia Lymphoma 1993; 10: 153-7.
Zhang Y. Silk fibroin nanoparticles fixed with enzyme and their production. WO2007112679A1, 2007.
Abribat T. PEGylated L-asparaginase. US20160060 613A1, 2018.
Golestaneh D, Varshosaz J. Enhancement in bio-logical activity of L-asparginase by its conjugation on silica nanoparticles. Recent Pat Nanotechnol 2018; 12: 70-82.
Dinu CZ, Dordick JS, Kane RS, et al. Enzyme-based nanoscale decontaminating composites. US201200 21486, 2012.
Godfrin Y. Test for predicting neutralization of asparaginase activity. US8852880B2, 2014.

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