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A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments

By: Hassan Mustafa Arif, Zulfiqar Ali Malik, Muhammad Irfan, Fakhra Hanif, Fariha Hasan, Samiullah Khan, Malik Badshah, Aamer Ali Shah

Key Words: L-asparaginase, Bacillus sp., Taguchi Design of Experiments, Acute lymphoblastic leukemia, Acrylamide.

Int. J. Biosci. 18(3), 276-289, March 2021.

DOI: http://dx.doi.org/10.12692/ijb/18.3.276-289

Certification: ijb 2021 0196 [Generate Certificate]

Abstract

The current study suggests the screening of soil samples for L-asparaginase producing microorganisms and optimization of enzyme production by Taguchi Design of Experiments as a suitable model for finding alternate L-asparaginase enzyme to the ones used in current therapeutic methods. A bacterium designated as strain MH01 was isolated that could effectively breakdown L-asparagine. It was found to be specie of genus Bacillus based on morphological, biochemical analysis and 16S rRNA gene sequencing, resulting in 96% similarity with Bacillus marcorestinctum strain LQQ (GQ900516). Enzyme production was statistically optimized using Taguchi design of experiment. The maximum specific activity 0.8 IU/µg was obtained at 30°C temperature, 2:1 inoculum to substrate ratio and a pH 5.0. L-asparaginase produced under optimized conditions was partially purified by size exclusion column chromatography using sephadex G-100. The molecular weight was estimated to be 41.88 kDa. The enzyme was purified 1.79 folds with 31.98% yield and 1.586 IU/µg specific activity with an optimum temperature of 30°C and pH of 8.0-8.5. The enzyme had Km value of 0.389 mM, Vmax value of 30395 µML-1 min-1 and Kcat value of 429.36 s-1 which indicates its efficiency and potential for further study and proving the success of our model in finding an alternate L-asparaginase enzyme.

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A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments

Narta UK, Kanwar SS, Azmi W. 2007. Pharmacological and clinical evaluation of L-asparaginase in the treatment of leukemia, Crit. Rev. Oncol. Hematol ogy 61(3),  208-21.

Arif HM, Hussain Z. 2014. Important sources and medicinal applications of L-asparaginase, International Journal of Pharmaceutics Science Reviews 3, 35-45.

Nagarethinam S, Nagappa AN, Udupa N, Rao V. 2012. Microbial L-Asparaginase and its future prospects, Asian Journal of  Medical Research 1(4), 159-168.

Vrooman LMSupko JGNeuberg DSAsselin BLAthale UHClavell L, Kelly KM, Laverdière CMichon BSchorin MCohen HJSallan SESilverman LB. 2010. Erwinia asparaginase after allergy to E. coli asparaginase in children with acute lymphoblastic leukemia, Pediatric. blood. & cancer 54(2), 199-205.

Chan WK, Lorenzi PL, Anishkin A, Purwaha P, Rogers DM, Sukharev S, Rempe SB, Weinstein JN. 2014. The glutaminase activity of L-asparaginase is not required for anticancer activity against ASNS-negative cells, Blood  123(23), 3596-3606.

Kwon YMChung HSMoon CYockman JPark YJGitlin SDDavid AEYang VCL. 2009, Asparaginase encapsulated intact erythrocytes for treatment of acute lymphoblastic leukemia (ALL), Journal of Controlled Release 139(3), 182-189.

Domenech CThomas XChabaud SBaruchel AGueyffier FMazingue F. 2011. L‐asparaginase loaded red blood cells in refractory or relapsing acute lymphoblastic leukaemia in children and adults: results of the GRASPALL 2005‐01 randomized trial, British Journal of Haematology 153(1), 58-65.

Gökmen V, Acrylamide Formation in Foods. 2016. Role of Composition and Processing, in Emerging and Traditional Technologies for Safe, Healthy and Quality Food, Springer, p 67-80.

Stadler RHBlank IVarga NRobert FHau J, Guy PA. 2002. Food chemistry: acrylamide from Maillard reaction products, Nature 419(6906), 449-

450.

Yaylayan VA, Stadler RH. 2005. Acrylamide formation in food: a mechanistic perspective Journal

of AOAC International 88(1), 262-267.

Anese M, Quarta B, Peloux L, Calligaris S. 2011. Effect of formulation on the capacity of l-asparaginase to minimize acrylamide formation in short dough biscuits, Food Research International 44(9), 2837-2842.

Mahajan RVSaran SSaxena RKSrivastava AK. 2013. A rapid, efficient and sensitive plate assay for detection and screening of l-asparaginase-producing microorganisms, FEMS. Microbiol letters, 341(2), 122-126.

Imada A, Igarasi S, Nakahama K, Isono M. 1973. Asparaginase and glutaminase activities of micro-organisms Microbiology 76(1), 85-99.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193(1),  265-75.

Holt JG. 1977. The shorter Bergey’s manual of determinative bacteriology. The shorter Bergey’s manual of determinative bacteriology, 8th edition.

Kumar S, Stecher G, Tamura K, MEGA7. 2016. Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular. biol. evolution 33(7), 1870-1874.

Tamura K, Nei M. 1993. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees, Molecular Biology and Evolution 10(3), 512-526.

Dehnad K., Quality control, robust design, and the Taguchi method. 2012. Springer Science & Business Media.

Nandal P, Ravella SR, Kuhad RC. 2013. Laccase production by Coriolopsis caperata RCK2011: optimization under solid state fermentation by Taguchi DOE methodology. Scientific reports, 3.

Roy RK. Design of experiments using the Taguchi approach: 16 steps to product and process improvement. 2001: John Wiley & Sons.

Andrew P. 2006. Estimation of molecular size and molecular weights of biological compounds by gel filtration. Methods of Biochemical Analysis, Volume 18, p 1-53.

Andrews P. 1964. Estimation of the molecular weights of proteins by Sephadex gel-filtration, Biochemical Journal 91(2), 222.

Batool TMakky EAJalal MYusoff MM. 2016, A comprehensive review on L-asparaginase and its applications, Applied biochemistry and biotechnology 178(5), 900-923.

Zuo S, Xue D,  Zhang T,  Jiang B,  Mu W. 2014.  Biochemical characterization of an extremely thermostable l-asparaginase from Thermococcus gammatolerans EJ3, J. Molecular Catalysis B: Enzymatic 122-129.

Derst C, Henseling J, Röhm KH. 2000. Engineering the substrate specificity of Escherichia coli asparaginase II. Selective reduction of glutaminase activity by amino acid replacements at position 248, Protein Science 9(10), 2009-2017.

Sanches M, Sanches MBarbosa JAde Oliveira RTAbrahão Neto JPolikarpov I. 2003. Structural comparison of Escherichia coli L-asparaginase in two monoclinic space groups, Acta Crystallographica Section D: Biological Crystallography 59(3), 416-422.

Wriston Jr JC, Yellin TL. 1973. L-asparaginase: a review. Adv Enzymol Relat Areas Journal of Molecular Biology 39, 185-248.

Belur PRKPD. 2010. Partial purification and characterization of. Pediatr, Blood Cancer 54, 199-205.

Phillips RM, Saleem MU, Williams A, Morgan J. 2013. Kinetic analysis of the glutaminase activity of l-asparaginases derived from Erwinia chrysanthemi (Erwinase) and Eshericia coli (Kidrolase), AACR Annual Meeting., (Abstract C154.).

Hassan Mustafa Arif, Zulfiqar Ali Malik, Muhammad Irfan, Fakhra Hanif, Fariha Hasan, Samiullah Khan, Malik Badshah, Aamer Ali Shah.
A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments.
Int. J. Biosci. 18(3), 276-289, March 2021.
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