In vitro investigation of some plant-associated fungi as biological synthesizers of L-asparaginase

Articles in Press

Document Type : Original Article

Authors

Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agricultural Science and Natural Resources, Gorgan, Golestan, 4918943464, Iran.

Abstract

Fungi are promising sources of novel bioactive metabolites due to their abundant extracellular enzyme production, easy extraction, and purification. This study aimed to investigate the capability of some plant-associated fungi to produce L-asparaginase enzymes. Twenty plant-associated fungi were inoculated into a modified Czapex Dox broth medium containing 10.0 g/L L-asparagine and 0.3 mL of 2.5% phenol red (pH 7), and maintained at 25 ± 2 °C, for 7 days. The L-asparaginase enzyme activity was evaluated by measuring the amount of free ammonia released through the hydrolysis of L-asparagine through UV-visible spectrophotometry analysis at 450 nm. Seventy percent of the studied fungi could produce the L-asparaginase enzyme. Trichoderma atroviride, Aspergillus flavus, and T. harzianum demonstrated the highest production of L-asparaginase enzyme by 0.47, 0.35, and 0.24 U/mL, respectively. Furthermore, Cladosporium cladosporioides, C. ramotenellum and Verticillium dahliae with 0.022, 0.009 and 0.015 U/mL, respectively, showed the lowest production of the L-asparaginase enzyme among the others. This research also reports the first successful production of the L-asparaginase enzyme from Bipolaris oryzae, Curvularia trifolii, T. atroviride, T. harzianum, and T. virens. Plant-associated fungi represent a promising resource for the pharmaceutical industry due to their ability to produce bioactive compounds such as L-asparaginase. While their use may contribute to more sustainable and potentially cost-effective production, successful large-scale application depends on multiple factors beyond enzyme production, including cultivation feasibility, purification processes, and regulatory considerations.
 

Keywords

Main Subjects

References

Abdel-Hamid NS, Abdel-Khalek HH, Ramadan E, Mattar ZA, Abou-Taleb KA. 2022. Optimization of L-asparaginase production from Fusarium oxysporum f-s3 using irradiated pomegranate peel under solid-state fermentation. Egyptian Journal of Chemistry. 65(6): 381-397. https://doi.org/10.21608/ejchem.2021.104129.4817
Akbari Oghaz N, Hatamzadeh S, Rahnama K, Moghaddam MK, Vaziee S ... et al. 2022. Adjustment and quantification of UV–visible spectrophotometry analysis: an accurate and rapid method for estimating Cladosporium spp. spore concentration in a water suspension. World Journal of Microbiology and Biotechnology. 38(10): 183. https://doi.org/10.1007/s11274-022-03356-8
Akbari Oghaz N, Rahnama K, Habibi R. 2021. First Report of Isolation and Identification of Fusarium wilt and Root Rot of Garlic Fields from Sari Area. Journal of Iranian Plant Protection Research. 35(2): 197-202. https://doi.org/10.22067/jpp.2021.32779.0
Akbari Oghaz N, Rahnama K, Habibi R, Razavi S, Farias A. 2024. Endophytic and rhizospheric Trichoderma spp. associated with cucumber plants as potential biocontrol agents of Fusarium oxysporum f. sp. cucumerinum. Asian Journal of Mycology. 7(1): 31-46.
Ali SS, Rai V, Soni K, Kulshrestha P, Lal SK. 1993. Antineoplastic activity in L-asparaginase of Cladosporium cladosporiodies. Indian Journal of Clinical Biochemistry. 8(1): 68-70. https://doi.org/10.1007/BF02867729
Baltussen TJ, Zoll J, Verweij PE, Melchers WJ. 2020. Molecular mechanisms of conidial germination in Aspergillus spp. Microbiology and Molecular Biology Reviews. 84(1): e00049-19. https://doi.org/10.1128/mmbr.00049
Castro D, Marques ASC, Almeida MR, de Paiva GB, Bento HBS ... et al. 2021. L-asparaginase production review: bioprocess design and biochemical characteristics. Applied microbiology and biotechnology. 105 4515-4534. https://doi.org/10.1007/s00253-021-11359-y
Chand S, Mahajan RV, Prasad JP, Sahoo DK, Mihooliya KN ... et al. 2020. A comprehensive review on microbial l‐asparaginase: Bioprocessing, characterization, and industrial applications. Biotechnology and applied biochemistry. 67(4): 619-647. https://doi.org/10.1002/bab.1888
da Cunha MC, dos Santos Aguilar JG, de Melo RR, Nagamatsu ST, Ali F ... et al. 2019. Fungal L-asparaginase: Strategies for production and food applications. Food Research International. 126 108658. https://doi.org/10.1016/j.foodres.2019.108658
Darvishi F, Jahanafrooz Z, Mokhtarzadeh A. 2022. Microbial L-asparaginase as a promising enzyme for treatment of various cancers. Applied microbiology and biotechnology. 106(17): 5335-5347. https://doi.org/10.1007/s00253-022-12086-8
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.
Elsaba YM, Salama WH, Soliman ERS. 2022. In silico and biochemical analysis on a newly isolated Trichoderma asperellum l-asparaginase. Biocatalysis and Agricultural Biotechnology. 40 102309. https://doi.org/10.1016/j.bcab.2022.102309
Elshafei A, El-Ghonemy DH. 2021. Extracellular glutaminase-free L-asparaginase from Trichoderma viride f2: Purification, biochemical characterization and evaluation of its potential in mitigating acrylamide formation in starchy fried food. Journal of microbiology, biotechnology and food sciences. 11(2): e4336-e4336. https://doi.org/10.15414/jmbfs.4336
Gulati R, Saxena RK, Gupta R. 1997. A rapid plate assay for screening L‐asparaginase producing micro‐organisms. Letters in applied microbiology. 24(1): 23-26. https://doi.org/10.1046/j.1472-765X.1997.00331.x
Habibi R, Rahnama K, Taghinasab M. 2015. Evaluating the effectiveness of native Trichoderma species in production of extracellular enzymes during interaction with plant pathogenic fungus Fusarium oxysporum. Journal of Applied Research in Plant Protection. 4(2): 73-85.
Habibi R, Rahnama K, Taghinasab M. 2018. Investigation on the effect of different levels of temperature and pH of some Trichoderma isolates for biological control of tomato wilt disease under laboratory and greenhouse conditions. Journal of Iranian Plant Protection Research. 32(2): 199.
Hatamzadeh S, Akbari Oghaz N, Rahnama K, Noori F. 2024. Comparison of the antifungal activity of chlorine dioxide, peracetic acid and some chemical fungicides in post-harvest management of Penicillium digitatum and Botrytis cinerea infecting sweet orange and strawberry fruits. Agricultural Research. 13(1): 72-84. https://doi.org/10.1007/s40003-023-00677-4
Hatamzadeh S, Rahnama K, Nasrollahnejad S, Fotouhifar KB, Hemmati K ... et al. 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
Hatamzadeh S, Rahnama K, White JF, Akbari Oghaz N, Nasrollahnejad S ... et al. 2023. Investigation of some endophytic fungi from five medicinal plants with growth promoting ability on maize (Zea mays L.). Journal of Applied Microbiology. 134(1): lxac015. https://doi.org/10.1093/jambio/lxac015
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
Jahantigh S, Akbari Oghaz N, Rahnama K, Hatamzadeh S. 2023. Application of Lactobacillus spp. for the biological management of green mold (Penicillium digitatum) on sweet orange fruit under in vitro and storehouse conditions. Biocontrol Science and Technology. 33(6): 567-581. https://doi.org/10.1080/09583157.2023.2208781
Klosterman SJ, Atallah ZK, Vallad GE, Subbarao KV. 2009. Diversity, pathogenicity, and management of Verticillium species. Annual review of phytopathology. 47 39-62. https://doi.org/10.1146/annurev-phyto-080508-081748
Lincoln L, Niyonzima FN, More SS. 2015. Purification and properties of a fungal L-asparaginase from Trichoderma viride pers: SF GREY. The Journal of Microbiology, Biotechnology and Food Sciences. 4(4): 310-316. 10.15414/jmbfs.2014.4.4.310-316
Maghsoudlou R, Taheri AAH, Rahnama K. 2007. Identification and pathogenicity of Fusarium spp. Isolated from root and crown of wheat in Gorgan area. Journal of Agricultural Sciences and natural resources. 14(2): 175-189.
Möller M, Stukenbrock EH. 2017. Evolution and genome architecture in fungal plant pathogens. Nature Reviews Microbiology. 15(12): 756-771. https://doi.org/10.1038/nrmicro.2017.76
Nakahama K, Imada A, Igarasi S, Tubaki K. 1973. Formation of L-asparaginase by Fusarium species. Microbiology. 75(2): 269-273. https://doi.org/10.1099/00221287-75-2-269
Naser S, Saber W, El-Metwally M, Moustafa M, El-Kott A. 2020. Fungal assembly of L-asparaginase using solid-state fermentation: A review. Biocell. 44(2): 147-155. http://dx.doi.org/10.32604/biocell.2020.09522
Noori F, Rahnama K, Mashayekhi K, Akbari Oghaz N, Naeini SAM ... et al. 2023. Investigation of the symbiotic relationship between Serendipita indica and ZP684 hybrid cultivar of maize plant under a combined condition of root rot disease (Fusarium proliferatum) and drought stress. Biological Control.  105282. https://doi.org/10.1016/j.biocontrol.2023.105282
Sanei S, Razavi S. 2018. In vitro antifungal activities of saffron (Crocus sativus L.) stigmas against Aspergillus species and toxin production. Iranian Journal of Medicinal and Aromatic Plants Research. 34(1): 77-86.
Sanei SJ, Razavi SE. 2012. Survey of olive fungal disease in north of Iran. Annual Review and Research in Biology. 2(1): 27-36.
Sanei SJ, Razavi SE, Okhovvat SM. 2012. Rice brown spot disease and indentification of the causal agent fungi in Golestan province. Cereal Research. 1(1): 65-74.
Shuker WH. 2019. Study the effect of some growth factor on the production of L-Asparaginase enzyme by fungal isolate Aspergillus flavus. Journal of Global Pharma Technology. 11(5): 196-201.
Singh R, Kumar M, Mittal A, Mehta PK. 2016. Microbial enzymes: industrial progress in 21st century. 3 Biotech. 6 174. https://doi.org/10.1007/s13205-016-0485-8
Vakilizarj Z, Rahnama K, Rahnema M. 2013. The Comparative growth and determination of the isolates reaction of Sclerotinia sclerotiorum on oil seed rape cultivars. International Journal of Agronomy and Plant Production. 4(5): 928-935.
Van Trimpont M, Peeters E, De Visser Y, Schalk AM, Mondelaers V ... et al. 2022. Novel insights on the use of L-asparaginase as an efficient and safe anti-cancer therapy. Cancers. 14(4): 902. https://doi.org/10.3390/cancers14040902
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.org/10.3897/mycokeys.108.128983.
Mycologia Iranica

Articles in Press, Accepted Manuscript
Available Online from 06 May 2025

Files

History

  • Receive Date: 26 February 2025
  • Revise Date: 30 April 2025
  • Accept Date: 03 May 2025
  • Publish Date: 06 May 2025

Share

How to cite

Statistics