بهینه‌سازی فعالیت آنزیم لیپاز و متغیرهای شرایط تخمیری حاوی سوبسترای شیره انگور در محیط کشت آسپرژیلوس نایجر
محورهای موضوعی :
نانوبیوتکنولوژی
نیما زرگران
1
,
رضا حبیبی پور
2
,
نرگس قبادی
3
1 - کارشناسی ارشد، گروه میکروبیولوژی، دانشکده علوم پایه، واحد همدان، دانشگاه آزاد اسلامی، همدان، ایران.
2 - دانشیار، گروه میکروبیولوژی، دانشکده علوم پایه، واحد همدان، دانشگاه آزاد اسلامی، همدان، ایران
3 - استادیار، گروه میکروبیولوژی، دانشکده علوم پایه، واحد همدان، دانشگاه آزاد اسلامی، همدان، ایران
تاریخ دریافت : 1400/11/03
تاریخ پذیرش : 1400/09/30
تاریخ انتشار : 1401/01/01
کلید واژه:
آسپرژیلوس نایجر,
آنزیم لیپاز,
شیره انگور,
محیط کشت,
چکیده مقاله :
هدف: آنزیم لیپاز از آنزیم‌‌های با اهمیت در صنایع غذایی، مواد شوینده و نساجی می‌‌باشد. همچنین امروزه صنایع وابسته به این آنزیم نظیر فرآوری مواد آلی، سنتز بیوسورفکتنت‌‌ها، نوشیدنی‌‌ها و صنایع آرایشی، دارویی و غذایی گسترش چشمگیری داشته‌‌اند. در این راستا، هدف پژوهش حاضر بهینه‌سازی فعالیت آنزیم لیپاز و متغیرهای شرایط تخمیری حاوی سوبسترای شیره انگور در محیط کشت آسپرژیلوس نایجر است.
مواد و روش‌ها: در این پژوهش برای اولین بار جهت دستیابی به بیشترین فعالیت آنزیم در محیط کشت حاوی منبع کربنی شیره انگور، متغیرهای موثر در فرآیند تخمیر قارچ آسپرژیلوس نایجر یعنی دما، اسیدیته، غلظت شیره انگور و منبع نیتروژنی (عصاره مخمر-پپتون) با کمک نرم‌افزار Design Expert بهینه‌سازی گردید.
یافته‌ها: اسیدیته معادل 5/7، مقدار غلظت شیره انگور و ترکیب عصاره مخمر و پپتون (با نسبت دو به یک) به ترتیب با میزان 5/1 و 75/0 درصد بدست آمد. فعالیت لیپاز پس از بهینه‌سازی شرایط محیط کشت برابر با U/ml 694/17 بود.
نتیجه‌گیری: آسپرژیلوس نایجر در محیط کشت با فاکتورهای دما، غلظت شیره انگور و عصاره مخمر و پپتون در میزان بهینه خود و اسیدیته معادل 5/7، فعالیت لیپاز قابل توجهی ثبت می‌‌نماید. بنابراین منابع کربنی و نیتروژنی بکار رفته در این پژوهش، منابع مناسبی برای استفاده در محیط کشت این میکروارگانیسم جهت تولید آنزیم لیپاز می‌‌باشند.
چکیده انگلیسی:
Objective: Lipase enzyme is one of the important enzymes in food, detergent and textile industries. Also, today, industries related to this enzyme such as processing of organic materials, synthesis of biosurfactants, beverages and cosmetic, pharmaceutical and food industries have expanded significantly. In this regard, the aim of the current research is to optimize the activity of lipase enzyme and the variables of fermentation conditions containing grape juice substrate in Aspergillus niger culture medium.
Materials and methods: In this research, for the first time, in order to achieve the maximum activity of the enzyme in the culture medium containing the carbon source of grape juice, the effective variables in the fermentation process of Aspergillus niger, i.e. temperature, acidity, concentration of grape juice and nitrogen source (yeast extract-peptone) with the help of Design Expert software was optimized.
Findings: The acidity equal to 7.5, the amount of concentration of grape juice and the combination of yeast extract and peptone (with a ratio of two to one) were obtained with 1.5 and 0.75%, respectively. Lipase activity after optimizing the conditions of the culture medium was equal to 17.694 U/ml.
Conclusion: Aspergillus niger records significant lipase activity in the culture medium with the factors of temperature, concentration of grape juice and yeast extract and peptone in its optimal amount and acidity equivalent to 7.5. Therefore, the carbon and nitrogen sources used in this research are suitable sources for use in the culture medium of this microorganism to produce lipase enzyme.
منابع و مأخذ:
Bussamara R, Fuentefria AM, Oliveira ESD, Broetto L, Simcikova M, Valente P, Schrank A & Vainstein MH. Isolation of a lipase-secreting yeast for enzyme production in a pilot-plant scale batch fermentation. Bioresource Technology. 2010; 101: 268–2
Nwuche CO & Ogbonna, JCh. Isolation of lipase producing fungi from palm oil mill effluent (POME) dump sites at Nsukka. Brazilian Archives of Biology and Technology. 2011; 54(1): 113-116.
Whitaker JR, Voragen AG & Wong DW. Handbook of food enzymology. 1st, 2003.
Ghamari M, Tabataba I, Yazdi F, Alam Zadeh I, Vosoghi M, Varidi M & Safari H. Optimization of Physical Parameters effecting lipase production by Aspergillus niger PTCC 5010 in culture medium containing date waste and Studying the properties of the produced enzyme. Food Science and Technology. 2016; 13(57): 159-167. (In Persian)
Rekha KS & et al. Production and optimization of lipase from Candida rugosa using groundnut oil cake under solid state fermentation. International Journal Research in Engineering and Technology. 2012; 1(4): 571-577.
Thakur S. Lipases, its sources, properties and applications: A Review. International Journal of Scientific & Engineering Research. 2012; 3(7): 147-154
Ahmadpour F & et al. Cloning and expression of Bacillus pumilus lipase gene in Pichia pastoris. Genetics in the Millennium. 2012; 1: 2997-3.
Haliru M & Bukola Ch. Screening of microorganisms isolated from different environmental samples for extracellular lipase production. Assumption University Journal of Technology. 2012; 15(3): 179-186.
Tiwari P, Upadhyay MK, Silawat N & Verma HN. Optimization and characterization of a thermo tolerant lipase from Cryptococcus albidus. Der pharma Chemica. 2011; 3(4): 501-508.
Thabet HM, Pasha C, Ahmad MM & Linga VR. Isolation of Novel Lipases Producing Sporobolomyces salmonicolor OVS8 from oil mill spillage and enhancement of lipase production. Jordan Journal of Biological Sciences. 2012; 5(4): 301-306
Coelho MAZ, Amaral PFF & Belo I. Yarrowia lipolytica: an industrial workhorse. Technology and education topics in applied microbiology and microbial biotethnology. 2010; 14: 930-944.
Ghamari M, Tabatabaie F, Alemzade I & Safari H. Investigation of lipase properties of Aspergillus niger. Food Science and Technology Research Institue; 2013.
Faostat F., Food and Agriculture Organization of the United Nations (2017).
Ghamari M, Tabatabaie F, Alemzade I, Vosooghi M & Varidi M. Optimization of culture medium containing date waste for lipase production by Aspergillus niger using RSM method. Food Science and Technology. 2015; 14(65): 85-96
Fernández-Lorente G & et al. Purification of different lipases from Aspergillus niger by using a highly selective adsorption on hydrophobic supports. Bioeng. 2005; 92: 773-779.
Kazem NA & Salehi M. Introduction of Taguchi experimental design and a practical example. Neda. 2007; 1(2): 11-20. (In Persian)
Ghobadi N, Ogino C & Ohmura Characterizations of the Submerged Fermentation of Aspergillus oryzae Using a Fullzone Impeller in a Stirred Tank Bioreactor. Journal of Bioscience and Bioengineering. 2016; 123(1): 1-8. DOI: 10.1016/j.jbiosc.2016.07.001
Elibol M & Ozer D. Influence of Oxygen Transfer on Lipase Production by Rhizopus arrhizus. Process Biochem. 2000; 36: 325-329.
Burkert JFM, Maugeri F & Rodrigues MI. Optimization of extracellular lipase production by Geotrichum sp. Using factorial design. Bioresour. Technol. 2004; 91: 77-84
Amoozegar MA & Saleh Ghamari E. Optimization of lipase production in Salinivibrio SA2 by Taguchi design. Nova Biol. Rep. 2017; 3: 288-294.
Falony G, Armas JC, Dustet Mendoza JC & Martínez Hernández JL. Production of Extracellular Lipase from Aspergillus niger by Solid-State Fermentation. Food Technol. Biotechnol. 2006; 44(2): 235-240.
Plascencia-Jatomea M, Viniegra G, Olayo R, Castillo-Ortega MM, & Shirai K. Effect of Chitosan and Temperature on Spore Germination of Aspergillus niger. Macromolecular Bioscience. 2003; 3(10): 582-586.
Parvaneh V. Quality control and the chemical analysis of food. University of Tehran press; 2006.
Adham NZ & Ahmed EM. Extracellular lipase of Aspergillus niger NRRL3 production, partial purification and properties. J Microbiol. 2009; 49: 77-83.
Acikel U, Ersana M & Acikel YS. The effects of the composition of growth medium and fermentation conditions on the production of lipase by delemar . Turk J Biol. 2011; 35: 35-44.
Macris JB, Kourentzi E & Hatzinikolaou DG. Studies on the localization and regulation of lipase production by Aspergillus niger. Process Biochem. 1996; 31: 807-812.
Sangal A & Behbahani M. Isolation and optimization of pectinase production of a useful industrial enzyme from Aspergillus niger, Rhizopus ores and Penicillium chrysogenum. Biology of microorganisms. 2016; 5(17): 121-140
Pimentel M & et al. Lipase from a Brazilian strain of Penicillium citrinum. Applied biochemistry and biotechnology.1994; 49(1): 59-74.
Onishi Y, Yoshida J & Sekiguchi J. Lipase production of Aspergillus oryzae. Bioeng. 1994; 77: 490-495.
Abraham S, Kamini NR & Gowthaman MK. Process strategies for alkaline lipase production using Aspergillus niger MTCC 2594. Journal of Applied pharmacy. 2022; 1(3): 126-133.
Bornscheuer UT (ed.). Enzymes in lipid modification Weinheim. Wiley-VCH; 2000.
Banu A, Kalpana M, Gnanaprabhal RG, Pradeep VB & Palaniswamy M. Production and characterization of pectinase enzyme from penicillium chrysogenum. Indian Journal of Science and Technology. 2012; 3(4): 377-81.
Huwang F, Kim KS, Zimmerman W & Fiechter A. Pectinolytic enzymes from actinomycetes for the degumming of ramie bast fibers. Apply Environment. 1994; 60(7): 2107-12.
Naghavi NS & et al. Optimization of exopectinase activity of Monilia isolated from tangerine in immersed fermentation. Biology of microorganisms. 2012; 1(1): 23-30
Reddy LP & Sreeramulu A. Isolation, identification and screening of pectinolytic fungi from different soil samples of chitoor district. Int J Life Sc Bt & pharm Res. 2012; 1: 186-93.
Ardestani F & Businessman R. Simulation of protozoan protein production process from Aspergillus niger based on non-structural kinetic models. Quarterly Journal of Food Science and Technology. 2015; 4: 308-319.
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Bussamara R, Fuentefria AM, Oliveira ESD, Broetto L, Simcikova M, Valente P, Schrank A & Vainstein MH. Isolation of a lipase-secreting yeast for enzyme production in a pilot-plant scale batch fermentation. Bioresource Technology. 2010; 101: 268–2
Nwuche CO & Ogbonna, JCh. Isolation of lipase producing fungi from palm oil mill effluent (POME) dump sites at Nsukka. Brazilian Archives of Biology and Technology. 2011; 54(1): 113-116.
Whitaker JR, Voragen AG & Wong DW. Handbook of food enzymology. 1st, 2003.
Ghamari M, Tabataba I, Yazdi F, Alam Zadeh I, Vosoghi M, Varidi M & Safari H. Optimization of Physical Parameters effecting lipase production by Aspergillus niger PTCC 5010 in culture medium containing date waste and Studying the properties of the produced enzyme. Food Science and Technology. 2016; 13(57): 159-167. (In Persian)
Rekha KS & et al. Production and optimization of lipase from Candida rugosa using groundnut oil cake under solid state fermentation. International Journal Research in Engineering and Technology. 2012; 1(4): 571-577.
Thakur S. Lipases, its sources, properties and applications: A Review. International Journal of Scientific & Engineering Research. 2012; 3(7): 147-154
Ahmadpour F & et al. Cloning and expression of Bacillus pumilus lipase gene in Pichia pastoris. Genetics in the Millennium. 2012; 1: 2997-3.
Haliru M & Bukola Ch. Screening of microorganisms isolated from different environmental samples for extracellular lipase production. Assumption University Journal of Technology. 2012; 15(3): 179-186.
Tiwari P, Upadhyay MK, Silawat N & Verma HN. Optimization and characterization of a thermo tolerant lipase from Cryptococcus albidus. Der pharma Chemica. 2011; 3(4): 501-508.
Thabet HM, Pasha C, Ahmad MM & Linga VR. Isolation of Novel Lipases Producing Sporobolomyces salmonicolor OVS8 from oil mill spillage and enhancement of lipase production. Jordan Journal of Biological Sciences. 2012; 5(4): 301-306
Coelho MAZ, Amaral PFF & Belo I. Yarrowia lipolytica: an industrial workhorse. Technology and education topics in applied microbiology and microbial biotethnology. 2010; 14: 930-944.
Ghamari M, Tabatabaie F, Alemzade I & Safari H. Investigation of lipase properties of Aspergillus niger. Food Science and Technology Research Institue; 2013.
Faostat F., Food and Agriculture Organization of the United Nations (2017).
Ghamari M, Tabatabaie F, Alemzade I, Vosooghi M & Varidi M. Optimization of culture medium containing date waste for lipase production by Aspergillus niger using RSM method. Food Science and Technology. 2015; 14(65): 85-96
Fernández-Lorente G & et al. Purification of different lipases from Aspergillus niger by using a highly selective adsorption on hydrophobic supports. Bioeng. 2005; 92: 773-779.
Kazem NA & Salehi M. Introduction of Taguchi experimental design and a practical example. Neda. 2007; 1(2): 11-20. (In Persian)
Ghobadi N, Ogino C & Ohmura Characterizations of the Submerged Fermentation of Aspergillus oryzae Using a Fullzone Impeller in a Stirred Tank Bioreactor. Journal of Bioscience and Bioengineering. 2016; 123(1): 1-8. DOI: 10.1016/j.jbiosc.2016.07.001
Elibol M & Ozer D. Influence of Oxygen Transfer on Lipase Production by Rhizopus arrhizus. Process Biochem. 2000; 36: 325-329.
Burkert JFM, Maugeri F & Rodrigues MI. Optimization of extracellular lipase production by Geotrichum sp. Using factorial design. Bioresour. Technol. 2004; 91: 77-84
Amoozegar MA & Saleh Ghamari E. Optimization of lipase production in Salinivibrio SA2 by Taguchi design. Nova Biol. Rep. 2017; 3: 288-294.
Falony G, Armas JC, Dustet Mendoza JC & Martínez Hernández JL. Production of Extracellular Lipase from Aspergillus niger by Solid-State Fermentation. Food Technol. Biotechnol. 2006; 44(2): 235-240.
Plascencia-Jatomea M, Viniegra G, Olayo R, Castillo-Ortega MM, & Shirai K. Effect of Chitosan and Temperature on Spore Germination of Aspergillus niger. Macromolecular Bioscience. 2003; 3(10): 582-586.
Parvaneh V. Quality control and the chemical analysis of food. University of Tehran press; 2006.
Adham NZ & Ahmed EM. Extracellular lipase of Aspergillus niger NRRL3 production, partial purification and properties. J Microbiol. 2009; 49: 77-83.
Acikel U, Ersana M & Acikel YS. The effects of the composition of growth medium and fermentation conditions on the production of lipase by delemar . Turk J Biol. 2011; 35: 35-44.
Macris JB, Kourentzi E & Hatzinikolaou DG. Studies on the localization and regulation of lipase production by Aspergillus niger. Process Biochem. 1996; 31: 807-812.
Sangal A & Behbahani M. Isolation and optimization of pectinase production of a useful industrial enzyme from Aspergillus niger, Rhizopus ores and Penicillium chrysogenum. Biology of microorganisms. 2016; 5(17): 121-140
Pimentel M & et al. Lipase from a Brazilian strain of Penicillium citrinum. Applied biochemistry and biotechnology.1994; 49(1): 59-74.
Onishi Y, Yoshida J & Sekiguchi J. Lipase production of Aspergillus oryzae. Bioeng. 1994; 77: 490-495.
Abraham S, Kamini NR & Gowthaman MK. Process strategies for alkaline lipase production using Aspergillus niger MTCC 2594. Journal of Applied pharmacy. 2022; 1(3): 126-133.
Bornscheuer UT (ed.). Enzymes in lipid modification Weinheim. Wiley-VCH; 2000.
Banu A, Kalpana M, Gnanaprabhal RG, Pradeep VB & Palaniswamy M. Production and characterization of pectinase enzyme from penicillium chrysogenum. Indian Journal of Science and Technology. 2012; 3(4): 377-81.
Huwang F, Kim KS, Zimmerman W & Fiechter A. Pectinolytic enzymes from actinomycetes for the degumming of ramie bast fibers. Apply Environment. 1994; 60(7): 2107-12.
Naghavi NS & et al. Optimization of exopectinase activity of Monilia isolated from tangerine in immersed fermentation. Biology of microorganisms. 2012; 1(1): 23-30
Reddy LP & Sreeramulu A. Isolation, identification and screening of pectinolytic fungi from different soil samples of chitoor district. Int J Life Sc Bt & pharm Res. 2012; 1: 186-93.
Ardestani F & Businessman R. Simulation of protozoan protein production process from Aspergillus niger based on non-structural kinetic models. Quarterly Journal of Food Science and Technology. 2015; 4: 308-319.
