Research Article

Production of L-Asparaginase by Yemeni Filamentous Fungi

Maysoon A Al Zubairy*, Faten H Thamer, Habib Thabet and Khawlah Shrf Aldeen

Published: 23 October, 2023 | Volume 7 - Issue 1 | Pages: 018-023

Yemen with its diverse climatic regions represents a rich resource for bioactive compounds obtained from microorganisms. This study was designed to screen fungal isolates obtained from the Microbiology branch, Biological Sciences Department, Faculty of Science, Sana’a University for their ability to produce L-asparaginase enzyme. In preliminary screening for L-asparaginase, among 77 fungal isolates about 29 fungal isolates representing 37.66% were high producers of L-asparaginase. These fungal isolates belonged to the genera Aspergillus, Eupenicillium, Fusarium, Penicillium, and Stachyobotrys. These 29 fungal isolates were screened for their ability to produce L-asparaginase using the agar well diffusion method. 12 fungal isolates out of 29 showed the ability to produce extracellular L-asparaginase. These isolates belonged to 8 species which were: A. sulphurs, A. ustus, F. sacchari, P. chrysogenum, P. citrinum, P. corylophilum, P. melinii, and P. subturcoseum. Only 5 isolates were obtained for the determination of enzymatic activity, among them P. chrysogenum showed the highest activity (279.8696U ml-1) followed by A. ustus (170.9435U ml-1). This finding is the first report on the L-asparaginase production from filamentous fungi in Yemen.

Read Full Article HTML DOI: 10.29328/journal.abb.1001036 Cite this Article Read Full Article PDF


L-asparaginase; Filamentous fungi; Production; Yemen


  1. Ghasemi Y, Ebrahimminezhad A, Amini SR, Zarrini G, Ghoshoon MB, Raee MJ, Morowvat MH, Kafilzadeh F, Kazemi A. An optimized medium for screening of L-asparaginase production by Escherichia coli. Amer J Biochem Biotechnol. 2008; vol: 4(4): 422-24.
  2. Niharika Y, Supriya S. Production of L-Asparaginase by Fusarium Oxysporum Using Submerged Fermentation. International Journal of Pharmaceutical Science Invention. 2014; 3(6): 32-40.
  3. Zuo S, Zhang T, Jiang B, Mu W. Recent research progress on microbial L-asparaginases. Appl Microbiol Biotechnol. 2015 Feb;99(3):1069-79. doi: 10.1007/s00253-014-6271-9. Epub 2014 Dec 11. PMID: 25492420.
  4. Siddalingeshwara KG, Lingappa K. Production and characterization of L-Asparginase a tumor inhibitor.Int. J. Pharm Tech Res. 2011; 3: 314-319.
  5. Cachumba JJ, Antunes FA, Peres GF, Brumano LP, Santos JC, Da Silva SS. Current applications and different approaches for microbial l-asparaginase production. Braz J Microbiol. 2016 Dec;47 Suppl 1(Suppl 1):77-85. doi: 10.1016/j.bjm.2016.10.004. Epub 2016 Oct 27. PMID: 27866936; PMCID: PMC5156506.
  6. Sahu MK, Poorani E, Sivakumar K, Thangaradjou T, Kannan L. Partial purification and anti-leukemic activity of L-asparaginase enzyme of the actinomycete strain LA-29 isolated from the estuarine fish, Mugil cephalus (Linn.). J Environ Biol. 2007 Jul;28(3):645-50. PMID: 18380089.
  7. Dharmsthiti SC, Luechai S. Purification and characterization of asparaginase from solidstate culture of Aspergillus Niger International Journal of Biotechnology and Biochemistry. 2010; 6(7):1083-1092.
  8. Theantana T, Hyde KD. Lumyong S. Asparaginase production by endophytic fungi from Thai medicinal plants: cytotoxic properties. International Journal of Integrative Biology.2009; 7(1): 1-8.
  9. Kumar Jha S, Divya P, Rati KS, Hare RS, Vinod KN, Ambrish SV. Microbial l-asparaginase: ‘Review on current scenario and future prospects. International Journal of Pharmaceutical Science and Research. 2012; 3(9): 3076-3090.
  10. Trilokchandran B., Agrawal P, Krishna V. Screening studies of microbial species for L-asparaginase production. J. Environ. Res. Dev. 2016; 10(4): 712-716.
  11. Xu F, Oruna-Concha MJ, Elmore JS. The use of asparaginase to reduce acrylamide levels in cooked food. Food Chem. 2016 Nov 1; 210:163-71. doi: 10.1016/j.foodchem.2016.04.105. Epub 2016 Apr 22. PMID: 27211635.
  12. Pedreschi F, Kaack K, Granby K. The effect of asparaginase on acrylamide formation in French fries. Food Chem. 2008 Jul 15;109(2):386-92. doi: 10.1016/j.foodchem.2007.12.057. Epub 2007 Dec 28. PMID: 26003362.
  13. Warangkar SC, Khobragada CN, Dawane BS, Bhosale RB. Effect of dihydropyrimidine dervatives on kinetic parameters of carotovora L-asparginae. Int J Biotechnol. App. 2009; 1:5-13.
  14. Jha SK, Pasrija D, Sinha RK, Singh HR, Nigam VK, Vidyarthi AS. Microbial L-Asparaginase: A Review On Current Scenario and Future Prospects. International Journal of Pharmaceutical Sciences and Research. 2012; 3: 3076-3090.
  15. Arjun JK, Aneesh BP, Harikrishnan K. Sequencing and characterization of L-asparaginase (ansB) gene of Bacillus megaterium isolated from Western Ghats, Kerala, India. International Journal of Current Microbiology and Applied Sciences. 2015; 4: 753-760.
  16. El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM. Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly Isolated Streptomyces olivaceus NEAE-119 Using Response Surface Methodology. Biomed Res Int. 2015; 2015:627031. doi: 10.1155/2015/627031. Epub 2015 Jun 9. PMID: 26180806; PMCID: PMC4477217.
  17. Ghasemi A, Asad S, Kabiri M, Dabirmanesh B. Cloning and characterization of Halomonas elongata L-asparaginase, a promising chemotherapeutic agent. Appl Microbiol Biotechnol. 2017 Oct;101(19):7227-7238. doi: 10.1007/s00253-017-8456-5. Epub 2017 Aug 11. PMID: 28801829.
  18. Gurunathan B, Sahadaevan R. Optimization of media components and operating conditions for exogenous production of fungal L-asparaginase. Chiang Mai J Sci 2011; 38:270-279.
  19. Adamson RH, Fabro S. Antitumor activity and other biologic properties of L-asparaginase (NSC-109229)-a review. Cancer Chemother Rep. 1968 Oct;52(6):617-26. PMID: 4895425.
  20. Cooney DA, Handschumacher RE. L-asparaginase and L-asparagine metabolism. Annu Rev Pharmacol. 1970; 10:421-40. doi: 10.1146/annurev.pa.10.040170.002225. PMID: 4911021.
  21. Emmanuel E, Nzelibe HC, Onyike E. Isolation, Partial Purification and Characterization of L-Asparaginase from Hedgehog Serum. Journal of Microbial & Biochemical Technology. 2015; 7: 404-409.
  22. Savitri N, Asthana N, Azmi W. Microbial L-asparaginase a potent antitumor enzyme. Ind. J. Biotechnol. 2003; 2:184-194.
  23. Verma N, Kumar K, Kaur G, Anand S. L-asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol. 2007 Jan-Mar;27(1):45-62. doi: 10.1080/07388550601173926. PMID: 17364689.
  24. Mishra A. Production of L-asparaginase, an anticancer agent, from Aspergillus niger using agricultural waste in solid state fermentation. Appl Biochem Biotechnol. 2006 Oct;135(1):33-42. doi: 10.1385/abab:135:1:33. PMID: 17057254.
  25. Dange V, Peshwe S. Purification and biochemical characterization of L-asparaginase from Aspergillus niger and evaluation of its antineoplastic activity. International Journal of Science and Research. 2015; 564–569.
  26. Pedreschi F, Mariotti S, Granby K, Risum J. Acrylamide reduction in potato chips by using commercial asparaginase in combination with conventional blanching. LWT—Food Science and Technology. 2011; 44:1473–1476. DOI 10.1016/j.lwt.2011.02.004.
  27. Johnson LF, Curel EA. Methodes for research on ecology of soil-borne pathogens. Burgess Publishing Co., Minnepolis, M N, USA. 1973.
  28. Raper KB, Fennell DI. The genus Aspergillus. Krieger Publishing Company. Huntingon, New York, USA. 1977.
  29. Pitt JI. The genus Penicillium and its telemorphic states Eupenicillium and Academic Press. London. 1979.
  30. Moubasher AH. Soil fungi in Qatar and other Arab countries. The Centre of Scientific and Applied Research. University of Qatar. Doha, Qatar. 1993.
  31. Samson RA, Hoekstra ES, Frisvad JS. and Filtenborg, O. Introduction to food-borne fungi. Centraalbureau voor schimmelcultures Baarn Delft. 1995.
  32. Watanabe T. Pictorial atlas of soil and seed fungi. Morphological of cultured fungi and key to species. CRC Press LLC. USA. 2002.
  33. Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening L-asparaginase producing micro-organisms. Lett Appl Microbiol. 1997 Jan;24(1):23-6. doi: 10.1046/j.1472-765x.1997.00331x. PMID: 9024001.
  34. Imada A, Igarasi S, Nakahama K, Isono M. Asparaginase and glutaminase activities of micro-organisms. J Gen Microbiol. 1973 May;76(1):85-99. doi: 10.1099/00221287-76-1-85. PMID: 4723072.
  35. Heatley NG. A method for the assay of penicillin. Biochem J. 1944;38(1):61-5. doi: 10.1042/bj0380061. PMID: 16747749; PMCID: PMC1258024.
  36. Theantana TKD, Hyde S, Lumyong KMITL. Sci. Tech. J‖. 2007; 7: 13-18.
  37. LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265-75. PMID: 14907713.
  38. Bhosale H, As-Suhbani A. Screening of fungal endophytes isolated from medicinal plants for glutaminase free L-asparaginase activity. Journal of Experimental Biology and Agricultural Sciences. 2019; 7(4): 396-402.
  39. Vala A, Dudhagara D, Dave B. Enhanced L-asparaginase production by a marine-derived eurihaline Aspergillus niger strain AKV MKBU- a statical model. Indian Journal of Geo Marine Science. 2016; 47(06):1172-1179.
  40. Bacelar A, Maia A, Rueda J, Conte Vanzela A. Fungal production of the anti-leukemic enzyme L-asparaginase: from screening to medium development, Maringá. 2016; 38(4):387-394.
  41. Wade HE, Robinson HK, Phillips BW. Asparaginase and glutaminase activities of bacteria. J Gen Microbiol. 1971 Dec;69(3):299-312. doi: 10.1099/00221287-69-3-299. PMID: 5004160.
  42. Pinheiro IO, Araujo JM, Ximenes ECPA, Pinto JCS, Alves TLM. Production of L-asparaginase by Zymomonas mobilis strain CP4. Biomaterial and Diagnostic BD06. 2001; 243-244.
  43. Carta de-Angeli L, Pocchiari F, Russi S, Tonolo A, Zurita VE, Ciaranfi E, Perin A. Effect of L-asparaginase from Aspergillus terreus on ascites sarcoma in the rat. Nature. 1970 Feb 7;225(5232):549-50. doi: 10.1038/225549a0. PMID: 5411861.
  44. Arima K. Igarasi S, Nakahama K, Isonom M. Production of extracellular L-asparaginases from microorganisms. Agricul. Biol.Chem. 1972; 36:356-361.
  45. Al-Zubairy MA, Hussein K, Alkhyat SH, Al-Mahdi AY, Alghalibi SM, Al-Gheethi AA, Al-Shaibani MM, El Enshasy HA, Sidik NM. Antibacterial Activity of a Novel Oligosaccharide from Streptomyces californicsagainst Erwinia carotovora Carotovora. Molecules. 2022 Apr 7;27(8):2384. doi: 10.3390/molecules27082384. PMID: 35458585; PMCID: PMC9032947.
  46. Yusuf Q, Al-Maqtari M, Al-Mahdi A, Al-Awadhi O. New Records of Streptomyces and Non Streptomyces Actinomycetes Isolated from Soils Surrounding Sana'a High Mountain. International Journal of Research in Pharmacy and Biosciences. 2016; 3: (3): 19-3.


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