BACKGROUND: The present study focused on the production of L-asparaginase using Solid State Fermentation (SSF) by Myroides gitamensis. METHODS: Initially, five significant parameters (Carbon source; Nitrogen source, temperature, pH and incubation period) were identified that affect the production process of L-asparaginase using Classical One Factor at a Time (OFAT) optimization. An optimized L-asparaginase specific activity obtained by OFAT was recorded as 85.7 IU. Central Composite Design (CCD) was also employed successively to optimize the multiple parameters at a time and their results were compared. RESULTS: Maximum L-asparaginase enzyme specific activity obtained by CCD method was 295.6 IU under the hold values of carbon source (wheat bran) 12 g/L, nitrogen source (yeast extract) 7 g/L, temperature 37°C, pH=7.5 and incubation period 47 hr. Upon validation, the obtained results proved that there was a good relation existing between the experimental and the predicted model (p<0.05). L-asparaginase activity was enhanced in statistical method up to 3.4 folds compared to that of classical method. CONCLUSION: Utilization of wheat bran as a low cost carbon source in SSF for the production of L-asparaginase enzyme makes the process economical and in turn reduces the environmental pollution by biotransformation to commercially useful bio product.

Department of Biotechnology College of Natural and Computational Science University of Gondar Gondar Ethiopia

Department of Biotechnology GITAM Institute of Science GITAM University Visakhapatnam 530 045 Andhra Pradesh India

Department of Biotechnology University of Chemistry and Technology Prague Technicka 5 CZ 16628 Prague 6 Czech Republic

Zobrazit více v PubMed

Mishra A. Production of L-asparaginase, an anticancer agent from Aspergillus niger using agricultural waste in solid state fermentation. Appl Biochem Biotechnol 2006; 135(1):33–42. PubMed

Verma N, Kumar K, Kaur G, Anand S. L-asparaginase: a promising chemotherapeutic agent. Crit Rev Biotechnol 2007;27(1):45–62. PubMed

Teodor E, Litescu SC, Lazar V, Somoghi R. Hydrogelmagnetic nanoparticles with immobilized L-asparaginase for biomedical applications. J Mater Sci Mater Med 2009;20(6):1307–1314. PubMed

Narta UK, Kanwar SS, Azmi W. Pharmacological and clinical evaluation of l-asparaginase in the treatment of leukemia. Crit Rev Oncol Hematol 2007;61(3):208–221. PubMed

Pedreschi F, Kaack K, Granby K. The effect of asparaginase on acrylamide formation in French fries. Food Chem 2008;109(2):386–392. PubMed

Offman MN, Krol M, Patel N, Krishnan S, Liu J, Saha V, et al. Rational engineering of L-asparaginase reveals importance of dual activity for cancer cell toxicity. Blood 2011;117(5):1614–1621. PubMed

Savitri N, Asthana N, Azmi W. Microbial L-asparaginase: a potent antitumour enzyme. Indian J Biotechnol 2003;2:184–194.

El-Shishtawy RM, Mohamed SA, Asiri AM, Gomaa AB, Ibrahim IH, Al-Talhi HA. Solid fermentation of wheat bran for hydrolytic enzymes production and saccharification content by a local isolate Bacillus megatherium. BMC Biotechnol 2014;14:29. PubMed PMC

Stevenson L, Phillips F, O’sullivan K, Walton J. Wheat bran: its composition and benefits to health, a European perspective. Int J Food Sci Nutr 2012;63(8):1001–1013. PubMed PMC

Irfan M, Nadeem M, Syed Q. One-factor-at-a-time (OFAT) optimization of xylanase production from Trichoderma viride-IR05 in solid-state fermentation. J Radiation Res Applied Sci 2014;7(3):317–326.

Talluri VP, Bhavana M, Siva Kumar K, Anil Kumar P, Rajagopal SV. Myroides gitamensis sp.nov., L-asparaginase producing bacteria isolated from slaughter house soil sample in Visakhapatnam, India. J Microb Biochem Technol 2014;6(3):144–147.

Gulati R, Saxena RK, Gupta R. A rapid plate assay for screening L-asparaginase producing micro-organisms. Lett Appl Microbiol 1997;24(1):23–26. PubMed

Mashburn LT, Wriston JC., Jr Tumor inhibitory effect of Lasparaginase from E.coli. Arch Biochem Biophys 1964;105:450–452. PubMed

Lowry OH, Rosebrough NL, Farr AL, Randall RJ. Protein measurement with the folin reagent. J Biol Chem 1951; 193(1):265–275. PubMed

Zhang H, Wang Z, Xu SY. Optimization of processing parameters for cloudy ginkgo (Ginkgo biloba Linn.) juice. J Food Eng 2007;80(4):1226–1232.

Charles P, Devanathan V, Anbu P, Ponnuswamy MN, Kalaichelvan PT, Hur BK. Purification, characterization and crystallization of an extracellular alkaline protease from Aspergillus nidulans HA-10. J Basic Microbiol 2008;48:347–352. PubMed

Adinarayana K, Ellaiah P, Prasad DS. Purification and partial characterization of thermostable serine alkaline protease from a newly isolated Bacillus subtilis PE-11. AAPS PharmSciTech 2003;4(4):E56. PubMed PMC

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259):680–685. PubMed

Shahnoor HM, Abul Kalam A, Abu Sayem SM, Mostafa G, Md. Mozammel Hoq. Production and partial characterization of feather-degrading keratinolytic serine protease from Bacillus licheniformis MZK-3. J Biol Sci 2007; 7(4):599–606.

Rajesh MJ, Leelavathy R, Vanapalli VS, Thirumurugan G, Rajaram MS. Effect of inducers and physical parameters on the production of L-asparaginase using Aspergillusterrus. J Bioprocess Biotech 2011;1:1–6.

Manna S, Sinha A, Sadhukhan R, Chakrabarty SL. Purification, characterization and antitumor activity of L-asparaginase isolated from Pseudomonas stutzeri MB-405. Curr Microbiol 1995;30(5):291–298. PubMed

Moorthy V, Ramalingam A, Sumantha A, Shankaranaya RT. Production, purification and characterisation of extracellular L-Asparaginase from a soil isolate of Bacillus sp. Afr J Microbiol Res 2010;4(18):1862–1867.

Hosamani R, Kaliwal BB. Isolation, molecular identification and optimization of fermentation parameters for the production of L-asparaginase, an anticancer agent by Fusarium equiseti. Int J Microbiol Res 2011;3(2):108–119.

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): 191–198. PubMed

Rani GB, Chiranjeevi T, Chandel AK, Satish T, Radhika K, Narasu ML, et al. Optimization of selective production media for enhanced production of xylanases in submerged fermentation by Thielaviopsis basicola MTCC 1467 using L16 orthogonal array. J Food Sci Technol 2014;51(10):2508–2516. PubMed PMC

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. PubMed PMC

Potumarthi R, Jacques L, Harry W, Michael D. Surface immobilization of Rhizopus oryzae (ATCC 96382) for enhanced production of lipase enzyme by multiple responses optimization. Asia Pac J Chem Eng 2012;7(S3): S285–S295.

Madruga MS, Camara FS. The chemical composition of Multimistura as a food supplement. Food Chem 2000;68 (1):41–44.

Beg QK, Bhushan B, Kapoor M, Hoondal GS. Enhanced production of thermostable xylanase from Streptomyces sp. QG11-3 and its application in biobleaching of eucalyptus kraft pulp. Enzyme Microb Technol 2000;27(7): 459–466. PubMed

Ellaiah P, Srinivasulu B, Adinarayana K. Optimization studies on neomycin production by a mutant strain of Streptomyces marinensis in solid state fermentation. Process Biochem 2004;39(5):529–534.

Krishna C. Solid-state fermentation systems-an overview. Crit Rev Biotechnol 2005;25(1–2):1–30. PubMed