Nashrieh Shimi va Mohandesi Shimi Iran

Nashrieh Shimi va Mohandesi Shimi Iran

Statistical Analysis and Optimization of Operation Conditions of Lipase Production by Rhizopus Arrhizus

Document Type : Research Article

Authors
Shabnam Ghanbarzade
Mahsa Esmaelzade
Zeynab Salehi
Pejman Ghelich
School of Chemical Engineering, College of Engineering, University of Tehran, Tehran, I.R. IRAN
Abstract
In this study, in order to optimize the production of lipase from Rhizopus oryzae in the liquid culture medium, the design of the experiment was carried out by using the response surface methodology to the central operating state by the design expert 7.0.0 software. Fermentation was performed by selecting the parameters including the mass ratio of carbon source to nitrogen source, the agitation speed, and the initial pH in the range of variation (0.2-1), (100-250), (5-8) respectively. The experiments were carried out by using the olive oil stimulus as a carbon source at 32 ° C. The activity amount of the lipase enzyme was measured by titration of the free fatty acids (NaOH 0.1 molar as a titrant) and the mass of biomass was calculated by the gravity method. (Responses: Enzyme activity and the bulk of mass both produced after 48 and 96 hours). The statistical analysis of the experimental data was obtained from measuring the activity of the lipase enzyme, indicates that the models presented with 95% confidence for enzyme activity after 48 hours of cultivation have the desired effect of increasing the agitation and the alkalinity of the culture. It also shows that It is desirable to increase the agitation of the stirrer and reduce the mass ratio of the carbon source to the nitrogen source for the activity of the lipase enzyme after 96 hours of cultivation. Optimum fermentation conditions of lipase enzyme, respectively, the ratio of carbon source to nitrogen source 0.4, agitation 100 (rpm) and initial pH 8, with the production of the highest activity of the extracellular lipase enzyme (2.48, 4.89)(U/ml) was obtained after 48 and 96 h cultivation which shows a more efficient process than previous similar researches.
Keywords
Optimization
Rhizopus oryzae
Statistical
Operation Conditions
Response surface methodology

Subjects

[1] Vaquero, M.E., et al., Heterologous Expression of a Fungal Sterol Esterase/Lipase in Different Hosts: Effect on Solubility, Glycosylation and Production. Journal of Bioscience and Bioengineering, 120(6): 637-643. (2015).
[2] Bussamara, R., et al., Isolation of a Lipase-Secreting Yeast for Enzyme Production in a Pilot-Plant Scale Batch Fermentation. Bioresource Technology, 101(1): 268-275. (2010).
[3] Marzuki, N.H.C., et al., Sustainable Production of the Emulsifier Methyl Oleate by Candida Rugosa Lipase Nanoconjugates. Food and Bioproducts Processing, 96: 211-220. (2015).
[4] Ashjari, M., Mohammadi, M. R. Badri, Selective Concentration of Eicosapentaenoic Acid and Docosahexaenoic Acid from Fish Oil with Immobilized/Stabilized Preparations of Rhizopus Oryzae Lipase. Journal of Molecular Catalysis B: Enzymatic, 122: 147-155. (2015).
[5] Sathishkumar, R., et al., A Statistical Approach for Optimization of Alkaline Lipase Production by Ascidian Associated—Halobacillus Trueperi RSK CAS9. Biotechnology Reports, 8: 64-71. (2015).
[6] Singh, A.K., Mukhopadhyay, M. Overview of Fungal Lipase: A Review. Applied Biochemistry and Biotechnology, 166(2): 486-520. (2012).
[7] Colla, L.M., et al., Surface Response Methodology for the Optimization of Lipase Production Under Submerged Fermentation by Filamentous Fungi. Brazilian Journal of Microbiology, 47(2): 461-467. (2016).
[8] Sivaramakrishnan, R. Incharoensakdi, A. Purification and Characterization of Solvent Tolerant Lipase from Bacillus Sp. for Methyl Ester Production from Algal Oil. Journal of Bioscience and Bioengineering, 121(5): 517-522. (2016).
[9] Ungcharoenwiwat, P. Aran, H. Purification and Characterization of Lipase from Burkholderia Sp. EQ3 Isolated from Wastewater from a Canned Fish Factory and Its Application for the Synthesis of Wax Esters. Journal of Molecular Catalysis B: Enzymatic, 115: 96-104. (2015).
[10] زارع باقی آباد، وجیهه؛ طباطبایی یزدی، فریده؛ مرتضوی، علی؛ وریدی، مهدی؛ بهینه سازی تولید آنزیم لیپاز مخمرCryptococcus albidus با استفاده از رویه سطح پاسخ، نشریه جهاد دانشگاهی، 13 (1395).
[11] Salihu, A., et al., Lipase Production: an Insight in the Utilization of Renewable AgriculturalResidues. Resources, Conservation and Recycling, 58: 36-44. (2012).
[12] Ülker, S. Karaoğlu, Ş.A. Purification and Characterization of an Extracellular Lipase from Mucor Hiemalis F. Corticola Isolated from Soil. Journal of Bioscience and Bioengineering, 114(4): 385-390. (2012).
[13] Potumarthi, R., et al., Surface Immobilization of Rhizopus Oryzae (ATCC 96382) for Enhanced Production of Lipase Enzyme by Multiple Responses Optimization. Asia‐Pacific Journal of Chemical Engineering, 7(S3): (2012).
[14] Liu, C.-H., et al., Optimizing Lipase Production from Isolated Burkholderia sp. Journal of the Taiwan Institute of Chemical Engineers, 43(4): 511-516. (2012).
[15] Teng, Y., Y. Xu, Wang, D. Production and Regulation of Different Lipase Activities from Rhizopus Chinensis in Submerged Fermentation by Lipids. Journal of Molecular Catalysis B: Enzymatic, 57(1): 292-298. (2009).
[16] Maldonado, R.R., et al., Evaluation of Lipase Production by Geotrichum Candidum in Shaken Flasks and Bench-Scale Stirred Bioreactor Using Different Impellers. Biocatalysis and Agricultural Biotechnology, 1(2): 147-151. (2012).
[17] JAVED, M.M., optimization or Cultural Conditions for the Production or Lipase by Submerged Culture or Rhizopus Oligosporous rUV-31. Pak. J. I ’0t, 35(4): 519-525. (2003).
[18] Rajendran, A. Thangavelu, V. Statistical Experimental Design for Evaluation of Medium Components for Lipase Production by Rhizopus Arrhizus MTCC 2233. LWT-Food Science and Technology, 42(5): 985-992. (2009).
[19] Yang, X., et al., Production of Lipase by Repeated Batch Fermentation with Immobilized Rhizopus Arrhizus. Process Biochemistry, 40(6): 2095-2103. (2005).
[20] Йорданова, М., et al., Влияние на Компонентите на Хранителната Среда Върху Биосинтезата на Липаза от Микромицетен Щам Rhizopus Arrhizus КБ-2.
[21] Elibol, M. Ozer, D. Response Surface Analysis of Lipase Production by Freely Suspended Rhizopus Arrhizus. Process Biochemistry, 38(3): 367-372. (2002).
[22] Tan, T., et al., Screening of High Lipase Producing Candida Sp. and Production of Lipase by Fermentation. Process Biochemistry, 39(4): 459-465. (2003).
[23] Salihu, A., et al., Optimization of Lipase Production by Candida Cylindracea in Palm Oil Mill Effluent Based Medium using Statistical Experimental Design. Journal of Molecular Catalysis B: Enzymatic, 69(1): 66-73. (2011).
[24] Teng, Y. Xu, Y. Culture Condition Improvement for whole-Cell Lipase Production in Submerged Fermentation by Rhizopus Chinensis using Statistical Method. Bioresource technology, 99(9): 3900-3907. (2008).
[25] Kantak, J.B., et al., Isolation, Identification and Optimization of a New Extracellular Lipase Producing Strain of Rhizopus Sp. Applied Biochemistry and Biotechnology, 164(7): 969-978. (2011).
[26] Papagora, C., Roukas, T. Kotzekidou, P. Optimization of Extracellular Lipase Production by Debaryomyces Hansenii Isolates from Dry-Salted Olives using Response Surface Methodology. Food and Bioproducts Processing, 91(4): 413-420. (2013).
[27] Zandi-Atashbar, N., Ensafi, A.A. Ahoor, A.H. Nano-CeO2/SiO2 as an Efficient Catalytic Conversion of Waste Engine Oil Into Liquid Fuel. Journal of Cleaner Production, 166: 1010-1019. (2017).
[28] Massart, D.L., et al., Handbook of Chemometrics and Qualimetrics: Part A. Elsevier Science Inc. 639-647. (1997).
[29] عالم زاده، ایران؛ وثوقی، منوچهر؛ وریدی، مهدی؛ صفری، هانیه؛ بهینه سازی محیط کشت حاوی شیره خرما جهت تولید لیپازتوسط آسپرژیلوس نایجر به روش پاسخ عکس العمل سطح، نشریه جهاد دانشگاهی، 1396. 14.
[30] Rajendran, A. Thangavelu, V. Application of Central Composite Design and Artificial Neural Network for the Optimization of Fermentation Conditions for Lipase Production by Rhizopus Arrhizus MTCC 2233. J. Bioproces. Biotechniq, 2(3): 1. (2012).
[31] Zhu, S., et al., Optimization of Lipase-Catalyzed Synthesis of Acetylated EGCG by Response Surface Methodology. Journal of Molecular Catalysis B: Enzymatic, 97: 87-94. (2013).
[32] Barrigón, J.M., Montesinos, J.L. Valero, F. Searching the Best Operational Strategies for Rhizopus Oryzae Lipase Production in Pichia Pastoris Mut+ Phenotype: Methanol Limited or Methanol Non-Limited Fed-Batch Cultures? Biochemical Engineering Journal, 75: 47-54. (2013).
[33] Brígida, A.I., et al., Lipase from Yarrowia Lipolytica: Production, Characterization and Application as an Industrial Biocatalyst. Journal of Molecular Catalysis B: Enzymatic, 101: 148-158. (2014).
[34] Colla, L.M., et al., Production and Characterization of Lipases by Two New Isolates of Aspergillus Through Solid-State and Submerged Fermentation. BioMed Research International, 2015. (2015).
[35] Hiol, A., et al., Purification and Characterization of an Extracellular Lipase from a Thermophilic Rhizopus Oryzae Strain Isolated from Palm Fruit. Enzyme and Microbial Technology, 26(5-6): 421-430. (2000).
[36] Li, E. Mira de Orduña, R. A Rapid Method for the Determination of Microbial Biomass by Dry Weight Using a Moisture Analyser with an Infrared Heating Source and an Analytical Balance. Letters In Applied Microbiology, 50(3):  283-288. (2010).
[37] Lima, V.M., et al., Effect of Nitrogen and Carbon Sources on Lipase Production by Penicillium Aurantiogriseum. Food Technology and Biotechnology, 41(2): 105-110. (2003).
[38]  سلماسی, د.ش.ف.م., "کاربرد آمار و طراحی آزمایش ها در رشته های علوم و مهندسی". چاپ اول، انتشارات دانشگاه تهران. 1396. 166-167.
[39] Montesinos, J., E. Dalmau, and C. Casas, Lipase Production in Continuous Culture of CandidaRugosa. Journal of Chemical Technology and Biotechnology, 78(7): 753-761. (2003).
[40] Ghelich, P., Salehi, Z., Mohajerzedeh, S., & Jafarkhani, M. Experimental and Numerical Study on a Novel Microfluidic Method to Fabricate Curcumin Loaded Calcium Alginate Microfibres. The Canadian Journal of Chemical Engineering, (2018).