ENHANCED ENZYMATIC ACTIVITY OF STREPTOMYCES GRISEOPLANUS L-ASPARGINASE VIA ITS INCORPORATION IN AN OIL-BASED NANOCARRIER
Keywords:L-asparginase, Nanoemulsion, Streptomyces griseoplanus, Immobilization, Enzymatic activity
Objective: L-asparaginase (L-asp) is a vital enzyme used as a therapeutic agent in combination with other drugs in the treatment of acute lymphoma, melanosarcoma and lymphocytic leukemia. Immobilization of enzymes through loading on nanoemulsion (NE) results in some advantages such as enhancing their stability and increasing their resistance to proteases. Aim of the present study is to formulate L-asp loaded nanoemulsion to enhance its efficiency and thermal stability.
Methods: Nanoemulsion loaded with L-asp crude extract (specific activity 13.23U/mg protein) was prepared employing oleic acid as oil, tween 20/tween 80 as surfactants and propylene glycol (PG) as co-surfactant. L-asp loaded NE underwent several thermodynamic stability studies and the optimized formulae were further examined for their biochemical properties and thermal stability.
Results The developed formulations were spherical in shape and their sizes were in the nanometric dimensions with negatively charged zeta potential values. Upon comparing the enzyme activity of L-asp loaded NE employing tween 20 (F1) or tween80 (F4) at different concentrations, the results revealed that F4 NE showed higher enzymatic activity [323 U/ml] compared to F1 NE [197 U/ml] at the same concentration. The nanosized immobilized L-asp was more stable in the pH range from 8 to 8.5 as compared to free L-asp. The immobilized enzyme preserved about 59.11% of its residual activity at 50 °C; while free L-asp preserved about 33.84%.
Conclusion: In the view of these results, NE composed of oleic acid, tween 80 and PG represents a promising dosage form for enhancing the activity and stability of Streptomyces griseoplanus L-asp.
Verma N, Kumar K, Kaur G, Anand S. L-asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol 2007;27:45-62.
Amena S, Vishalakshi N, Prabhakar M, Dayanand A, Lingappa K. Production, purification and characterization of L-asparaginase from streptomyces gulbargensis. Braz J Microbiol 2010;41:173-8.
Stecher A, De Deus PM, Polikarpov I, Abrahao Neto J. Stability of L-asparaginase: an enzyme used in leukemia treatment. Pharm Acta Helv 1999;74:1-9.
Reynolds D, Taylor J. The fungal holomorph: mitotic meiotic and pleomorphic speciation. CAB Int Wallingford UK 1993;4:293-301.
Klibanov AM. Immobilized enzymes and cells as practical catalysts. J Sci 1983;219:722-7.
Desai N. Challenges in the development of nanoparticle-based therapeutics. AAPS J 2012;14:282-95.
El-Ridy MS, Yehia SA, Mohsen AM, El-Awdan SA, Darwish AB. Formulation of niosomal gel for enhanced transdermal lornoxicam delivery: in vitro and in vivo evaluation. Curr Drug Delivery 2018;15:122-33.
El-Ridy MS, Badawi AA, Safar MM, Mohsen AM. Niosomes as a novel pharmaceutical formulation encapsulating the hepatoprotective drug silymarin. Int J Pharm Pharm Sci 2012;4:549-59.
Asfour MH, Mohsen AM. Formulation and evaluation of pH-sensitive rutin nanospheres against colon carcinoma using HCT-116 cell line. J Adv Res 2018;9:17-26.
Abousamra MM, Mohsen AM. Solid lipid nanoparticles and nanostructured lipid carriers of tolnaftate: design, optimization and in vitro evaluation. Int J Pharm Pharm Sci 2015;8:380-5.
Shah P, Bhalodia D, Shelat P. Nanoemulsion: a pharmaceutical review. Sys Rev Pharm 2010;1:24.
Baker Jr JR, Wright DC, Hayes MM, Hamouda T, Brisker J. Methods of inactivating bacteria including bacterial spores. Google Patents; 2000.
Velikov KP, Pelan E. Colloidal delivery systems for micronutrients and nutraceuticals. Soft Matter 2008;4:1964-80.
Aboofazeli R. Nanometric-scaled emulsions (Nanoemulsions). Iran J Pharm Res 2010;9:325-6.
Bouchemal K, Briançon S, Perrier E, Fessi H. Nano-emulsion formulation using spontaneous emulsification: solvent, oil and surfactant optimization. Int J Pharm 2004;280:241-51.
El-Hadi AA, Ahmed HM, Hamzawy RA. Optimization of culture conditions for production of L-asparaginase by a novel isolated streptomyces strain. Egypt Pharm J 2019;18:111-22.
El-Refai HA, Shafei MS, Mostafa H, El-Refai A-MH, El-Beih FM, Easa SM, 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.
Khamna S, Yokota A, Lumyong S. L-asparaginase production by actinomycetes isolated from some Thai medicinal plant rhizosphere soils. Int J Integr Biol 2009;6:22-6.
Ghosh V, Mukherjee A, Chandrasekaran N. Eugenol-loaded antimicrobial nanoemulsion preserves fruit juice against microbial spoilage. Colloids Surf B 2014;114:392-7.
Grigoriev DO, Miller R. Mono-and multilayer covered drops as carriers. Curr Opin Colloid Interface Sci 2009;14:48-59.
Azeem A, Rizwan M, Ahmad FJ, Iqbal Z, Khar RK, Aqil M, et al. Nanoemulsion components screening and selection: a technical note. AAPS PharmSciTech 2009;10:69-76.
Lawrence MJ, Rees GD. Microemulsion-based media as novel drug delivery systems. Adv Drug Delivery Rev 2000;45:89-121.
Ahmed M, Ramadan W, Rambhu D, Shakeel F. Potential of nanoemulsions for intravenous delivery of rifampicin. Die Pharmazie 2008;63:806-11.
Han D, Rhee JS. Characteristics of lipase‐catalyzed hydrolysis of olive oil in AOT–isooctane reversed micelles. Biotechnol Bioeng 1986;28:1250-5.
Meghavarnam A, Janakiraman S. Purification and characterization of therapeutic enzyme L-asparaginase from a tropical soil fungal isolate Fusarium culmorum ASP-87. J Anesth Crit Care Open Access 2015;2:00064.
Kassem AA, Mohsen AM, Ahmed RS, Essam TM. Self-nanoemulsifying drug delivery system (SNEDDS) with enhanced solubilization of nystatin for treatment of oral candidiasis: design, optimization, in vitro and in vivo evaluation. J Mol Liq 2016;218:219-32.
Kanuganti S, Jukanti R, Veerareddy PR, Bandari S. Paliperidone-loaded self-emulsifying drug delivery systems (SEDDS) for improved oral delivery. J Disper Sci Technol 2012;33:506-15.
Parveen R, Baboota S, Ali J, Ahuja A, Vasudev SS, Ahmad SJIjop. Oil based nanocarrier for improved oral delivery of silymarin: in vitro and in vivo studies. Int J Pharm 2011;413:245-53.
Shakeel F, Haq N, Alanazi FK, Alsarra IA. Polymeric solid self-nanoemulsifying drug delivery system of glibenclamide using coffee husk as a low-cost biosorbent. Powder Technol 2014;256:352-60.
Zhao Y, Wang C, Chow AH, Ren K, Gong T, Zhang Z, et al. Self-nanoemulsifying drug delivery system (SNEDDS) for oral delivery of zedoary essential oil: formulation and bioavailability studies. Int J Pharm 2010;383:170-7.
Agrawal AG, Kumar A, Gide PS. Formulation of solid self-nanoemulsifying drug delivery systems using N-methyl pyrrolidone as cosolvent. J Drug Dev Ind Pharm 2015;41:594-604.
Schmidts T, Dobler D, Nissing C, Runkel F. Influence of hydrophilic surfactants on the properties of multiple W/O/W emulsions. J Colloid Interface Sci 2009;338:184-92.
Balakumar K, Raghavan CV, Abdu S. Self nanoemulsifying drug delivery system (SNEDDS) of rosuvastatin calcium: design, formulation, bioavailability and pharmacokinetic evaluation. Colloids Surf B 2013;112:337-43.
Tabandeh MR, Aminlari M. Synthesis, physicochemical and immunological properties of oxidized inulin–l-asparaginase bioconjugate. J Biotechnol 2009;141:189-95.
Dias FFG, Ruiz ALTG, Della Torre A, Sato HH. Purification, characterization and antiproliferative activity of L-asparaginase from aspergillus oryzae CCT 3940 with no glutaminase activity. Asian Pac J Trop Biomed 2016;6:785-94.
Elshafei AM, Hassan MM, Abd M, Abouzeid E, Mahmoud DA, Elghonemy DH. Purification, characterization and antitumor activity of L-asparaginase from penicillium brevicompactum 2 NRC 829. Br Microbiol Res 2011;2:158-74.
El-Sabbagh SM, El-Batanony NH, Salem TA. L-Asparaginase produced by streptomyces strain isolated from Egyptian soil: purification, characterization and evaluation of its anti-tumor. Afr J Microbiol Res 2013;7:5677-86.
Muharram M, Abulhamd A, Salem-Bekhet MMJAjomr. Recombinant expression, purification of L-asparaginase-II from thermotolerant E. Coli strain and evaluation of its antiproliferative activity. Afr J Microbiol Res 2014;8:1610-9.
Kumar NM, Manonmani H. Purification, characterization and kinetic properties of extracellular l-asparaginase produced by cladosporium sp. World J Microbiol Biotechnol 2013;29:577-87.
Dange V, Peshwe S. Production, purification and characterization of fungal L-asparaginase. Bionano Frontier 2011;4:162-7.
Kumar S, Kikon K, Upadhyay A, Kanwar SS, Gupta R. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. Protein Expr Purif 2005;41:38-44.
Huang L, Liu Y, Sun Y, Yan Q, Jiang Z. 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.
Hiol A, Jonzo MD, Rugani N, Druet D, Sarda L, Comeau LC. Purification and characterization of an extracellular lipase from a thermophilic Rhizopus oryzae strain isolated from palm fruit. Enzyme Microb Technol 2000;26:421-30.
Nguyen TTH, Nguyen CT, Le Nguyen TS, Do TT. Optimization, purification and characterization of recombinant L-asparaginase II in Escherichia coli. Afr J Biotechnol 2016;15:1681-91.
Asker MM, Mohamed SF, El-sayed OH. Characterization of Immobilized L-asparaginase produced from pseudomonas aeruginosa 50071 by solid-state fermentation. Biosci Biotech Res Asia 2012;9:569-76.