Generic placeholder image

Current Biotechnology

Editor-in-Chief

ISSN (Print): 2211-5501
ISSN (Online): 2211-551X

Research Article

Bioprocess Optimization for Enhanced Production of L-asparaginase via Two Model-Based Experimental Designs by Alkaliphilic Streptomyces fradiae NEAE-82

Author(s): Hoda M. Soliman, Noura El-A. El-Naggar* and Sara M. El-Ewasy

Volume 9, Issue 1, 2020

Page: [23 - 37] Pages: 15

DOI: 10.2174/2211550109666200206100445

Abstract

Background: L-asparaginase is one of the most widely used chemotherapeutic agents for the treatment of a variety of lymphoproliferative disorders and particularly acute lymphoblastic leukemia. Due to increased applications of L-asparaginase in several industrial fields including food processing and medical fields, its production needs to be increased to several folds.

Objectives: The aim was (i) to identify the significant factors which affect L-asparaginase production by Streptomyces fradiae NEAE-82 and (ii) to achieve higher production of L-asparaginase.

Methods: Sixteen assigned factors and three dummy factors were screened using Plackett-Burman experimental design to determine the most important factors for the production of L-asparaginase by Streptomyces fradiae NEAE-82.

Results: L-asparagine was determined to be the most significant positive independent factor (P-value 0.0092) affecting L-asparaginase production by Streptomyces fradiae NEAE-82 followed by pH and NaCl with significant P-values of 0.0133 and 0.0272; respectively. These factors were further optimized by Box-Behnken experimental design. The optimized fermentation conditions, which resulted in the maximum L-asparagine activity of 53.572 UmL-1 are g/L: dextrose 4, L-asparagine 15, KNO3 2, MgSO4.7H2O 0.5, K2HPO4 1, FeSO4.7H2O 0.02, NaCl 0.2, ZnSO4 0.01 and inoculum size 2 %, v/v for 7 days incubation at temperature 37°C, agitation speed 100 rpm, pH 8.5.

Conclusion: A total of 3.41-fold increase in the production of L-asparaginase was achieved in the medium after statistical improvement (53.572 UmL-1) as compared to the unoptimized basal medium used prior to the application of Plackett-Burman (15.704 UmL-1).

Keywords: L-asparaginase production, Streptomyces fradiae NEAE-82, Plackett-Burman, Box-Behnken designs, lymphoblastic leukemia, lymphocytes.

Graphical Abstract
[1]
Kwan JM, Fialho AM, Kundu M, et al. Bacterial proteins as potential drugs in the treatment of leukemia. Leuk Res 2009; 33(10): 1392-9.
[http://dx.doi.org/10.1016/j.leukres.2009.01.024] [PMID: 19250673]
[2]
Jain R, Zaidi K, Verma Y, Saxena P. L-asparaginase: A promising enzyme for treatment of acute lymphoblastic leukemia. People’s J Sci Res 2012; 5(1): 29-35.
[3]
Verma N, Kumar K, Kaur G, Anand S. L-asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol 2007; 27(1): 45-62.
[http://dx.doi.org/10.1080/07388550601173926] [PMID: 17364689]
[4]
Kumar K, Verma N. The various sources and application of L-asparaginase. Asian J Biochem Pharm Res 2012; 2: 197-205.
[5]
Savitri AN, Azmi W. Microbial L-asparaginase: A potent antitumour enzyme. Int J Biotechnol 2003; 2: 184-94.
[6]
Keating MJ, Holmes R, Lerner S, Ho DH. L-asparaginase and PEG asparaginase--past, present, and future. Leuk Lymphoma 1993; 10: 153-7.
[http://dx.doi.org/10.3109/10428199309149129] [PMID: 8481665]
[7]
Duval M, Suciu S, Ferster A, et al. 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 2002; 99(8): 2734-9.
[http://dx.doi.org/10.1182/blood.V99.8.2734] [PMID: 11929760]
[8]
Reynolds D, Taylor J. The fungal holomorph: A consideration of mitotic, meiotic and pleomorphic speciation Wallingford, UK. Wallingford, UK: CAB International 1993.
[9]
Thomas X, Cannas G, Chelghoum Y, Gougounon A. [Therapeutic alternatives to native L-asparaginase in the treatment of adult acute lymphoblastic leukemia]. Bull Cancer 2010; 97(9): 1105-17.
[PMID: 20693115]
[10]
Pedreschi F, Kaack K, Granby K. The effect of asparaginase on acrylamide formation in French fries. Food Chem 2008; 109(2): 386-92.
[http://dx.doi.org/10.1016/j.foodchem.2007.12.057] [PMID: 26003362]
[11]
Maysa E, Amira M, Gamal E, Sanaa T, Sayed E. Production, immobilization and anti-tumor activity of L-asparaginase of Bacillus sp. R36. J Am Sci 2010; 6: 157-65.
[12]
Hendriksen HV, Kornbrust BA, Østergaard PR, Stringer MA. Evaluating the potential for enzymatic acrylamide mitigation in a range of food products using an asparaginase from Aspergillus oryzae. J Agric Food Chem 2009; 57(10): 4168-76.
[http://dx.doi.org/10.1021/jf900174q] [PMID: 19388639]
[13]
Patro KKR, Satpathy S, Gupta N. Evaluation of some fungi for Lasparaginase production. Ind J Fund Appl Life Sci 2011; 1: 219-21.
[14]
Deshpande N, Choubey P, Agashe M. Studies on optimization of growth parameters for L-asparaginase production by Streptomyces ginsengisoli. ScientificWorldJournal 2014; 2014895167
[http://dx.doi.org/10.1155/2014/895167] [PMID: 24616652]
[15]
El-Naggar NE, Abdelwahed NA. Optimization of process parameters for the production of alkali-tolerant carboxymethyl cellulase by newly isolated Streptomyces sp. strain NEAE-D. Afr J Biotechnol 2012; 11: 1185.
[16]
El-Naggar NE. Extracellular production of the oncolytic enzyme, L-asparaginase, by newly isolated Streptomyces sp. strain NEAE-95 as potential microbial cell factories: optimization of culture conditions using response surface methodology. Curr Pharm Biotechnol 2015; 16(2): 162-78.
[http://dx.doi.org/10.2174/1389201015666141113123910] [PMID: 25395212]
[17]
El-Naggar NE. Isolation, screening and identification of actinobacteria with uricase activity: Statistical optimization of fermentation conditions for improved production of uricase by Streptomyces rochei NEAE-25. Int J Pharmacol 2015; 11: 644-58.
[http://dx.doi.org/10.3923/ijp.2015.644.658]
[18]
Narayana KJ, Kumar KG, Vijayalakshmi M. L-asparaginase production by Streptomyces albidoflavus. Indian J Microbiol 2008; 48(3): 331-6.
[http://dx.doi.org/10.1007/s12088-008-0018-1] [PMID: 23100730]
[19]
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(1): 173-8.
[http://dx.doi.org/10.1590/S1517-83822010000100025] [PMID: 24031478]
[20]
Hymavathi M, Sathish T, Subba Rao Ch, Prakasham RS. Enhancement of L-asparaginase production by isolated Bacillus circulans (MTCC 8574) using response surface methodology. Appl Biochem Biotechnol 2009; 159(1): 191-8.
[http://dx.doi.org/10.1007/s12010-008-8438-2] [PMID: 19052920]
[21]
Coman G, Bahrim G. Optimization of xylanase production by Streptomyces sp. P12-137 using response surface methodology and central composite design. Ann Microbiol 2011; 61(4): 773-9.
[http://dx.doi.org/10.1007/s13213-010-0195-0] [PMID: 22131948]
[22]
El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM. Optimization of culture conditions for production of the antileukemic glutaminase free L-asparaginase by newly isolated Streptomyces olivaceus NEAE-119 using response surface methodology. BioMed Res Int 2015c; 2015: 627031
[http://dx.doi.org/10.1155/2015/627031] [PMID: 26180806]
[23]
Selamat SN, Halmi MIEB, Abdullah SRS, Idris M, Hasan HA, Anuar N. Optimization of lead (Pb) bioaccumulation in Melastoma malabathricum L by response surface methodology (RSM). Rend Lincei Sci Fis Nat 2018; 29: 43-51.
[http://dx.doi.org/10.1007/s12210-017-0656-5]
[24]
El-Naggar NE, Deraz SF, Soliman HM, El-Deeb NM, El-Ewasy SM. Purification, characterization, cytotoxicity and anticancer activities of L-asparaginase, anti-colon cancer protein, from the newly isolated alkaliphilic Streptomyces fradiae NEAE-82. Sci Rep 2016; 6: 32926.
[http://dx.doi.org/10.1038/srep32926] [PMID: 27605431]
[25]
De jong PJ. L-asparaginase production by Streptomyces griseus. Appl microbial 1972; 23: 1163-4.
[26]
Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening Lasparaginase producing micro-organisms. Lett Appl Microbiol 1997; 24(1): 23-6.
[http://dx.doi.org/10.1046/j.1472-765X.1997.00331.x] [PMID: 9024001]
[27]
Saxena RK, Sinha U. L-asparaginase and glutaminase activities in the culture filtrates of Aspergillus nidulans. Curr Sci 1981; 50: 218-9.
[28]
Wriston JC Jr, Yellin TO. L-asparaginase: a review. Adv Enzymol Relat Areas Mol Biol 1973; 39: 185-248.
[PMID: 4583638]
[29]
Plackett RL, Burman JP. The design of optimum multifactorial experiments. Biometrika 1946; 33: 305-25.
[http://dx.doi.org/10.1093/biomet/33.4.305]
[30]
Krishnan S, Prapulla S, Rajalakshmi D, Misra M, Karanth N. Screening and selection of media components for lactic acid production using Plackett–Burman design. Biopro Eng 1998; 19: 61-5.
[http://dx.doi.org/10.1007/PL00009003]
[31]
Box GE, Behnken DW. Some new three level designs for the study of quantitative variables. Technometrics 1960; 2: 455-75.
[http://dx.doi.org/10.1080/00401706.1960.10489912]
[32]
El-Naggar NE, Moawad H, El-Shweihy NM, El-Ewasy SM, Elsehemy IA, Abdelwahed NAM. Process development for scale-up production of a therapeutic L-asparaginase by Streptomyces brollosae NEAE-115 from shake flasks to bioreactor. Sci Rep 2019; 9(1): 13571.
[http://dx.doi.org/10.1038/s41598-019-49709-6] [PMID: 31537817]
[33]
El-Naggar NE, El-Ewasy SM, El-Shweihy NM. Microbial L-asparaginase as a potential therapeutic agent for the treatment of acute lymphoblastic leukemia: The pros and cons. Int J Pharmacol 2014; 10: 182-99.
[http://dx.doi.org/10.3923/ijp.2014.182.199]
[34]
Li Y, Horsman M, Wang B, Wu N, Lan CQ. Effects of nitrogen sources on cell growth and lipid accumulation of green alga Neochloris oleoabundans. Appl Microbiol Biotechnol 2008; 81(4): 629-36.
[http://dx.doi.org/10.1007/s00253-008-1681-1] [PMID: 18795284]
[35]
Kaushik R, Saran S, Isar J, Saxena R. Statistical optimization of medium components and growth conditions by response surface methodology to enhance lipase production by Aspergillus carneus. J Mol Catal, B Enzym 2006; 40: 121-6.
[http://dx.doi.org/10.1016/j.molcatb.2006.02.019]
[36]
El-Naggar NE, Moawad H, Nayera AM. Optimization of fermentation conditions for enhancing extracellular production of L-asparaginase, an anti-leukemic agent, by newly isolated Streptomyces brollosae NEAE-115 using solid state fermentation. Ann Microbiol 2017; 67: 1-15.
[http://dx.doi.org/10.1007/s13213-016-1231-5]
[37]
Sarquis MIdM Oliveira EMM, Santos AS, Costa, GLd. Production of L-asparaginase by filamentous fungi. Mem Inst Oswaldo Cruz 2004; 99: 489-92.
[http://dx.doi.org/10.1590/S0074-02762004000500005]
[38]
Sreenivasulu V, Jayaveera K, Rao PM. Optimization of process parameters for the production of L-asparaginase from an isolated fungus. Res J Pharmaco Phytochem 2009; 1: 30-4.
[39]
Sudhir AP, Dave BR, Trivedi KA, Subramanian RB. Production and amplification of an l-asparaginase gene from actinomycete isolate Streptomyces ABR2 Ann microbial 2012; 62: 1609-14.
[40]
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 bioproc Eng 2011; 16: 50-8.
[41]
Mostafa SA. Activity of L-asparaginase in cells of Streptomyces karnatakensis. Zentralbl Bakteriol Naturwiss 1979; 134(4): 343-51.
[http://dx.doi.org/10.1016/S0323-6056(79)80007-5] [PMID: 44600]
[42]
Pattnaik S, Kabi R, Ram KJ, Bhanot K. L-Asparaginase activity in Aeromonas sp. isolated from freshwater mussel. Indian J Exp Biol 2000; 38: 1143-6.
[43]
Jain P, Pundir R. Effect of fermentation medium, pH and temperature variations on antibacterial soil fungal metabolite production. Agric Technol Thail 2011; 7: 247-69.
[44]
Farag AM, Hassan SW, Beltagy EA, El-Shenawy MA. Optimization of production of anti-tumor l-asparaginase by free and immobilized marine Aspergillus terreus. The Egypt J Aqu Res 2015; 41(4): 295-302.
[http://dx.doi.org/10.1016/j.ejar.2015.10.002]
[45]
Sahu MK, Sivakumar K, Poorani E, Thangaradjou T, Kannan L. Studies on L-asparaginase enzyme of actinomycetes isolated from estuarine fishes. J Environ Biol 2007; 28(2)(Suppl.): 465-74.
[PMID: 17929767]
[46]
Mathew A, Dhevendaran K, Georgekutty M, Natarajan P. L-asparaginase activity in antagonistic Streptomycetes associated with clam Villorita cyprinoides (Hanley). Indian J Geo-Mar Sci 1994; 23: 204-8.
[47]
Monica T, Lincoln L, Niyonzima F, Sunil S. Isolation, purification and characterization of fungal extracellular L-asparaginase from Mucor Hiemalis. J Biocatal Biotransf 2013; 9: 12-4.
[48]
Kavitha A, Vijayalakshmi M. A study on L-asparaginase of Nocardia levis MK-VL_113. ScientificWorldJournal 2012; 2012160434
[http://dx.doi.org/10.1100/2012/160434] [PMID: 22619604]
[49]
Geckil H, Gencer S. Production of L-asparaginase in Enterobacter aerogenes expressing Vitreoscilla hemoglobin for efficient oxygen uptake. Appl Microbiol Biotechnol 2004; 63(6): 691-7.
[http://dx.doi.org/10.1007/s00253-003-1482-5] [PMID: 14593509]
[50]
Nei M, Kumar S. Molecular evolution and phylogenetics. New York: Oxford University Press 2000.
[51]
Fisher SH, Wray LV. Bacillus subtilis 168 contains two differentially regulated genes encoding L-asparaginase. J bacterial 2002; 184: 2148-54.
[52]
Michalska K, Jaskolski M. Structural aspects of L-asparaginases, their friends and relations. Acta Biochim Pol 2006; 53(4): 627-40.
[http://dx.doi.org/10.18388/abp.2006_3291] [PMID: 17143335]
[53]
Eisenberg H, Mevarech M, Zaccai G. Biochemical, structural, and molecular genetic aspects of halophilism. Adv Protein Chem 1992; 43: 1-62.
[http://dx.doi.org/10.1016/S0065-3233(08)60553-7] [PMID: 1442321]
[54]
Kumar MS, Selvam K. Singaravel. Statistical assessment of medium components by factorial design and surface methodology of L-asparaginase production by isolated Streptomyces radiopugnans MS1 in submerged fermentation using Tapioca Effluent. Asian J Appl Sci 2012; 5: 252-65.
[http://dx.doi.org/10.3923/ajaps.2012.252.265]
[55]
Shakambari G, Sumi BM, Ashokkumar B, Palanivelu P, Varalakshmi P. Industrial effluent as a substrate for glutaminase free. Lasparaginase production from Pseudomonas plecoglossicida strain RS1; media optimization, enzyme purification and its characterization RSC Adv 2015; 5: 48729-38.
[56]
Mangamuri U, Vijayalakshmi M, Ganduri VSRK, Babu S, Poda RS. Extracellular L-asparaginase from Streptomyces labedae VSM-6: Isolation, production and optimization of culture conditions using RSM. Pharmacogn J 2017; 9(6): 931-40.
[http://dx.doi.org/10.5530/pj.2017.6.146]
[57]
Weisberg S. Applied Linear Regression. 2nd ed. New York, USA: John Wiley and Sons 1985.
[58]
Box GEP, Hunter WG, Hunter JS. Statistics for Experiments. New York, USA: John Wiley and Sons Inc. 1978; pp. 291-334.

© 2024 Bentham Science Publishers | Privacy Policy