The nutritional and cultural conditions in shake flask culture for improved production of L-Asparaginase from endophytic fungus Fusarium sp. LCJ324: A sequential statistical method
DOI:
https://doi.org/10.6092/issn.2531-7342/16067Keywords:
Modified Czapek Dox broth, L-Asparagine, One Factor at a Time, Response Surface Methodology, Submerged fermentationAbstract
L-Asparaginase has a greater demand due to its potential application in the food industry as well as in the pharmaceutical industry. To meet this demand, optimization of nutritional as well as physical conditions is critical for scaling up the production of L-Asparaginase. This study aims to achieve enhanced L-Asparaginase production, a therapeutic enzyme from the endophytic fungus Fusarium sp. LCJ324 isolated from Glycosmis mauritiana (Lam.) Tanaka belonging to the Rutaceae family. Fusarium sp. LCJ324 exhibited L-Asparaginase activity of 8.75 ± 0.55 U mL-1 in modified Czapek Dox broth, the highest among all the seven different media tested by submerged fermentation. Different sources and concentrations of carbon, nitrogen and amino acid inducers (nutritional factors) as well as pH and temperature (physical parameters) were optimized to achieve enhanced L-Asparaginase production. Maximum L-Asparaginase activity of 19.94 ± 0.35 U mL-1 was obtained at 30º C with dextrose (3 g L-1), ammonium sulphate (30 g L-1), and L-Asparagine (20 g L-1) at pH 6. Response Surface Methodology was employed for statistical optimization of minimum and maximum levels of the selected parameters. The levels of parameters suggested by the response surface methodology for maximum production of L-Asparaginase were similar to the conventional optimization. A 2.29 fold increase in L-Asparaginase production was obtained through conventional and statistical optimization.
References
Abdollahzadeh R, Pazhang M, Najavand S, Fallahzadeh-Mamaghani V, Amani-Ghadim, AR (2020) Screening of pectinase-producing bacteria from farmlands and optimization of enzyme production from selected strain by RSM. Folia microbiologica 65(4):705‒719. https://doi.org/10.1007/s12223-020-00776-7
Ali SS, Rai V, Soni K, Kulshrestha P (1994) A fungal L-asparaginase with potential antitumor activity. Indian Journal of Microbiology 34:73‒73.
Alrumman SA, Mostafa YS, Al-Izran KA, Alfaifi MY, Taha TH, Elbehairi SE (2019) Production and anticancer activity of an L-Asparaginase from Bacillus licheniformis isolated from the Red Sea, Saudi Arabia. Scientific reports 9(1):1‒14. https://doi.org/10.1038/s41598-019-40512-x
As-Suhbani AE, Bhosale H (2020) Optimizing l-asparaginase production by Fusarium solani clr-36: a statistical approach using response surface methodology. Journal of Advanced Scientific Research 11(Suppl. 1):365‒374.
Balbool BA, Abdel-Azeem AM (2020) Diversity of the culturable endophytic fungi producing L-asparaginase in arid Sinai, Egypt. Italian Journal of Mycology 49:8‒24. https://doi.org/10.6092/issn.2531-7342/10063
Bano M, Sivaramakrishnan VM (1980) Preparation and properties of L-Asparaginase from green chillies (Capsicum annum L.). Journal of Biosciences 2(4):291‒297. https://doi.org/10.1007/BF02716861
Chandrasekhar PA (2012) Isolation, purification and characterization of Asparaginase from Aspergillus species. International Journal of Research in Chemistry and Environment (IJRCE) 2(1):38‒43.
Costa-Silva TA, Camacho-Córdova DI, Agamez-Montalvo GS, Parizotto LA, Sánchez-Moguel I, Pessoa-Jr A (2019) Optimization of culture conditions and bench-scale production of anticancer enzyme L-Asparaginase by submerged fermentation from Aspergillus terreus CCT 7693. Preparative Biochemistry and Biotechnology 49(1):95‒104. https://doi.org/10.1080/10826068.2018.1536990
Couto SR, Sanromán MA (2006) Application of solid-state fermentation to food industry—a review. Journal of Food Engineering 76(3):291‒302. https://doi.org/10.1016/j.jfoodeng.2005.05.022
Da Rocha WRV, Costa‐Silva TA, Agamez‐Montalvo GS, Feitosa VA, Machado SEF, de Souza Lima GM, Alves HDS (2019) Screening and optimizing fermentation production of l‐asparaginase by Aspergillus terreus strain S‐18 isolated from the Brazilian Caatinga Biome. Journal of applied microbiology 126(5):1426‒1437. https://doi.org/10.1111/jam.14221
Ekpenyong M, Asitok A, Antigha R, Ogarekpe N, Ekong U, Asuquo M, Antai S (2021) Bioprocess optimization of nutritional parameters for enhanced anti-leukemic L-Asparaginase production by Aspergillus candidus UCCM 00117: a sequential statistical approach. International Journal of Peptide Research and Therapeutics 27(2):1501‒1527. https://doi.org/10.1007/s10989-021-10188-x
El-Gendy MMAA, Awad MF, El-Shenawy FS, El-Bondkly AMA (2021) Production, purification, characterization, antioxidant and antiproliferative activities of extracellular L-Asparaginase produced by Fusarium equiseti AHMF4. Saudi Journal of Biological Sciences 28(4):2540‒2548. https://doi.org/10.1016/j.sjbs.2021.01.058
El-Hadi AA, El-Refai HA, Shafei MS, Zaki R, Mostafa H (2017) Statistical optimization of L-asparaginase production by using Fusarium solani. Egyptian Pharmaceutical Journal 16(1):16‒23. https://doi.org/10.4103/1687-4315.205825
El-Naggar NEA, Deraz SF, El-Ewasy SM, Suddek GM (2018) Purification, characterization and immunogenicity assessment of glutaminase free L-Asparaginase from Streptomyces brollosae NEAE-115. BMC Pharmacology and Toxicology 19(1):1‒15. https://doi.org/10.1186/s40360-018-0242-1
El-Naggar NE, El-Ewasy SM, El-Shweihy NM (2014) Microbial L-asparaginase as a potential therapeutic agent for the treatment of acute lymphoblastic leukemia: the pros and cons. International Journal of Pharmacology 10(4):182‒199. https://doi.org/10.3923/ijp.2014
Elshafei AM, El-Ghonemy DH (2015) Screening and media optimization for enhancing L-asparaginase production, an anticancer agent, from different filamentous fungi in solid state fermentation. British Biotechnology Journal 9(3):1‒15. https://doi.org/10.9734/BBJ/2015/19728
Farag AM, Hassan SW, Beltagy EA, El-Shenawy MA (2015) Optimization of production of anti-tumor L-Asparaginase by free and immobilized marine Aspergillus terreus. The Egyptian Journal of Aquatic Research 41(4):295‒302. https://doi.org/10.1016/j.ejar.2015.10.002
Fazeli N, Alimadadi N, Nasr S (2021) Screening and optimization of process parameters for the production of L-Asparaginase by indigenous fungal-type strains. Iranian Journal of Science and Technology, Transactions A: Science 45(2):409‒416. https://doi.org/10.1007/s40995-020-01056-2
Hamed M, Osman AA, Ateş M (2021) Statistical optimization of L-asparaginase production by Cladosporium tenuissimum. Egyptian Pharmaceutical Journal 20(1):51. https://doi.org/10.4103/epj.epj_47_20
Hatamzadeh S, Rahnama K, Nasrollahnejad S, Fotouhifar KB, Hemmati K, White JF, Taliei F (2020) Isolation and identification of L-Asparaginase-producing endophytic fungi from the Asteraceae family plant species of Iran PeerJ 8: e8309. https://doi.org/10.7717/peerj.8309
Hosamani R, Kaliwal BB (2011) Isolation, molecular identification and optimization of fermentation parameters for the production of L-Asparaginase, an anticancer agent by Fusarium equiseti. International Journal of Microbiology Research 3(2):108. https://doi.org/10.9735/0975-5276.3.2.108-119
Imada A, Igarasi S, Nakahama K, Isono M (1973) Asparaginase and glutaminase activities of micro-organisms. Microbiology 76(1):85‒99. https://doi.org/10.1099/00221287-76-1-85
Jenila AV, Gnanadoss JJ (2018) Formulation of a suitable medium and its optimization for maximizing L-Asparaginase production from endophytic fungi Fusarium Sp. LCJ273. Biosciences Biotechnology Research Asia 15(4):887‒898. http://dx.doi.org/10.13005/bbra/2699
Jiao L, Chi H, Lu Z, Zhang C, Chia SR, Show PL, Lu F (2020) Characterization of a novel type I L-Asparaginase from Acinetobacter soli and its ability to inhibit acrylamide formation in potato chips. Journal of bioscience and bioengineering 129(6):672‒678. https://doi.org/10.1016/j.jbiosc.2020.01.007
Kalyanasundaram I, Nagamuthu J, Srinivasan B, Pachayappan A, Muthukumarasamy S (2015) Production, purification and characterisation of extracellular L-Asparaginase from salt marsh fungal endophytes. World Journal of Pharmacy and Pharmaceutical Science 4(3):663‒677.
Krishnapura PR, Belur PD (2020) L-Asparaginase production using solid-state fermentation by an endophytic Talaromyces pinophilus isolated from rhizomes of Curcuma amada. Journal of Pure and Applied Microbiology 14:307‒318. https://doi.org/10.22207/JPAM.14.1.32
Li PJ, Xia JL, Shan Y, Nie ZY, Su DL, Gao QR, Ma YL (2015) Optimizing production of pectinase from orange peel by Penicillium oxalicum PJ02 using response surface methodology. Waste and Biomass Valorization 6(1):13‒22.
Meghavarnam AK, Sahu R, Janakiraman S (2022) Fungal L-asparaginases-A mini review. Kavaka 58(2):37-52. https://doi.org/10.36460/Kavaka/58/2/2022/37-52
Mishra A (2006) Production of L-Asparaginase, an anticancer agent, from Aspergillus niger using agricultural waste in solid state fermentation. Applied Biochemistry and Biotechnology 135(1):33‒42.
Mohamed SA, Elshal MF, Kumosani TA, Aldahlawi AM (2015) Purification and characterization of asparaginase from Phaseolus vulgaris seeds. Evidence-Based Complementary and Alternative Medicine 2015:1‒6. https://doi.org/10.1155/2015/309214
Nunes JC, Cristóvão RO, Freire MG, Santos-Ebinuma VC, Faria JL, Silva CG, Tavares AP (2020) Recent strategies and applications for L-asparaginase confinement. Molecules 25(24):5827. https://doi.org/10.3390/molecules25245827
Oza VP, Parmar PP, Patel DH, Subramanian RB (2011) Cloning, expression and characterization of L-Asparaginase from Withania somnifera L. for large scale production. 3 Biotech 1(1):21-26. https://doi.org/10.1007/s13205-011-0003-y
Priya SR, Subhashini A (2022) Characterization and optimization of fungal L-Asparaginase isolated from soil and medicinal plants. Journal of Pure and Applied Microbiology 16(1):453‒460. https://doi.org/10.22207/JPAM.16.1.43
Pundir RK, Yadav D, Jain P (2020) Production, optimization and partial purification of L-Asparaginase from endophytic fungus Aspergillus sp., Isolated from Cassia fistula. Applied Biological research 22:26–33. http://dx.doi.org/10.5958/0974-4517.2020.00008.7
Ramya LN, Doble M, Rekha VPB, Pulicherla KK (2012) L-Asparaginase as potent anti-leukemic agent and its significance of having reduced glutaminase side activity for better treatment of acute lymphoblastic leukaemia. Applied biochemistry and biotechnology 167:2144‒2159. https://doi.org/10.1007/s12010-012-9755-z
Shah IJ, Gami PN, Shukla RM, Acharya DK (2014) Optimization for α-amylase production by Aspergillus oryzae using submerged fermentation technology. Basic Research Journal of Microbiology 1(4):1‒10.
Udayan E, Gnanadoss JJ (2023) Isolation and characterization of L-Asparaginase producing endophytic fungi from medicinal plants of Rutaceae family. Bioscience Biotechnology Research Asia 20(1).
Uzma F, Narasimha Murthy K, Srinivas C (2016) Optimization of physiological conditions for L-Asparaginase production by endophytic fungi (Fusarium solani) isolated from Tinospora cordifolia (Willd.) Hook. F & Thomson. European Journal of Experimental Biology 6(3):37‒45.
Vimal A, Kumar A (2022) Optimized production of medically significant enzyme L-Asparaginase under submerged and solid-state fermentation from agricultural wastes. Current Microbiology 79(12):394. https://doi.org/10.1007/s00284-022-03095-x
Yap LS, Lee WL, Ting ASY (2021) Optimization of L-Asparaginase production from endophytic Fusarium proliferatum using OFAT and RSM and its cytotoxic evaluation. Journal of Microbiological Methods 191:106358. https://doi.org/10.1016/j.mimet.2021.106358
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