The effects of the composition of growth medium and fermentation conditions on the production of lipase by R. delemar
Lipases (triacylglycerol hydrolases) are hydrolytic enzymes that can catalyze the hydrolysis of the ester bond of long-chain acylglycerols at the oil-water interface. The present study investigated the effects of inducers, surface-active materials, activators, and inhibitors in the fermentation medium on lipase activity in Rhizopus delemar. In the presence of certain commercial oils and tributyrin as an inducer, lipase activity decreased in the order of sunflower oil > soybean oil > hazelnut oil > corn oil > tributyrin > olive oil. In addition, the effects of the stirring and aeration rates on lipase activity were investigated. To investigate the effect of surface-active materials on lipase activity 2 different surface-active materials, Tween-80 and rhamnolipid (biosurfactant), were used as comparatives with tributyrin. Maximum lipase activity was observed in the fermentation medium containing Tween-80 as both an inducer and surface-active material. The combined effects of molasses and sucrose on lipase production by R. delemar were also investigated. Lipase activity in the presence of activators decreased in the order of NaCl > KCl > CaCl2 > MgCl2 > gum arabic > EDTA, and maximum lipase activity of 964.55 U/L was obtained. The presence of heavy metal ions in the fermentation media severely inhibited lipase activity. Lipase activity in the presence of heavy metal ions decreased in the order of Fe2+ > Mn2+> Co2+> Ni2+.
The effects of the composition of growth medium and fermentation conditions on the production of lipase by R. delemar
Lipases (triacylglycerol hydrolases) are hydrolytic enzymes that can catalyze the hydrolysis of the ester bond of long-chain acylglycerols at the oil-water interface. The present study investigated the effects of inducers, surface-active materials, activators, and inhibitors in the fermentation medium on lipase activity in Rhizopus delemar. In the presence of certain commercial oils and tributyrin as an inducer, lipase activity decreased in the order of sunflower oil > soybean oil > hazelnut oil > corn oil > tributyrin > olive oil. In addition, the effects of the stirring and aeration rates on lipase activity were investigated. To investigate the effect of surface-active materials on lipase activity 2 different surface-active materials, Tween-80 and rhamnolipid (biosurfactant), were used as comparatives with tributyrin. Maximum lipase activity was observed in the fermentation medium containing Tween-80 as both an inducer and surface-active material. The combined effects of molasses and sucrose on lipase production by R. delemar were also investigated. Lipase activity in the presence of activators decreased in the order of NaCl > KCl > CaCl2 > MgCl2 > gum arabic > EDTA, and maximum lipase activity of 964.55 U/L was obtained. The presence of heavy metal ions in the fermentation media severely inhibited lipase activity. Lipase activity in the presence of heavy metal ions decreased in the order of Fe2+ > Mn2+> Co2+> Ni2+.
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- 1. Ellaiah P, Prabhakar T, Ramakrishna B et al. Production of lipase by immobilized cells of Aspergillus niger. Process Biochem 39: 525-528, 2004.
- 2. Wisdom RA, Dunnill P, Lilly MD. Enzymic interesterification of fats: laboratory and pilot-scale studies with immobilized lipase from Rhizopus arrhizus. Biotechnol Bioeng 29: 1081-1085, 1987.
- 3. Tweddell RJ, Kermasha S, Combes D et al. Esterification and interesterification activities of lipases from Rhizopus niveus and Mucor miehei in three different types of organic media: a comparative study. Enzyme Microb Tech 22: 439-445, 1998.
- 4. Chowdary GV, Ramesh MN, Prapulla SG. Enzymic synthesis of isoamyl isovalerate using immobilized lipase from Rhizomucor miehei: a multivariate analysis. Process Biochem 36: 331-339, 2000.
- 5. Sharma R, Chisti Y, Banerjee UC. Production, purification, characterization, and applications of lipases. Biotechnol Adv 19: 627-662, 2001.
- 6. Hasan F, Shah AA, Hameed A. Industrial applications of microbial lipases. Enzyme Microb Tech 39: 235-251, 2006.
- 7. Stransky K, Zarevucka M, Kejik Z et al. Substrate specificity, regioselectivity and hydrolytic activity of lipases activated from Geotrichum sp. Biochem Eng J 34: 209-216, 2007.
- 8. Joseph B, Ramteke PW, Thomas G. Cold active microbial lipases: Some hot issues and recent developments. Biotechnol Adv 26: 457-470, 2008.
- 9. Li N, Zeng QM, Zong MH. Substrate specificity of lipase from Burkholderia cepacia in the synthesis of 3’-arylaliphatic acid esters of floxuridine. J Biotechnol 142: 267-270, 2009.
- 10. Couto SR, Sanroman MA. Application of solid-state fermentation to food industry - A review. J Food Eng 76: 291- 302, 2006.
- 11. Liu R, Jiang X, Mou H et al. A novel low-temperature resistant alkaline lipase from a soda lake fungus strain Fusarium solani N4-2 for detergent formulation. Biochem Eng J 46: 265-270, 2009.
- 12. Gotor-Fernandez V, Brieva R, Gotor V. Lipases: Useful biocatalysts for the preparation of pharmaceuticals. J Mol Catal B-Enzym 40: 111-120, 2006.
- 13. Jaeger KE, Reetz TM. Microbial lipases from versatile tools for biotechnology. Trends Biotechnol 16: 396-403, 1998.
- 14. Perez JPH, Lopez-Cabarcos E, Lopez-Ruiz B. The application of methacrylate-based polymers to enzyme biosensors. Biomol Eng 23: 233-245, 2006.
- 15. Umare SS, Chandure AS. Synthesis, characterization and biodegradation studies of poly(ester urethane)s. Chem Eng J 142: 65-77, 2008.
- 16. Dizge N, Aydiner C, Imer DY et al. Biodiesel production from sunflower, soybean, and waste cooking oils by transesterification using lipase immobilized onto a novel microporous polymer. Bioresource Technol 100: 1983-1991.
- 17. Salameh M, Wiegel J. Lipases from extremophiles and potential for industrial applications. Adv Appl Microbiol 61: 253-283.
- 18. Dharmsthiti S, Kuhasuntisuk B. Lipase from Pseudomonas aeruginosa LP602: biochemical properties and application for wastewater treatment. J Ind Microbiol Biot 21: 75-80, 1998.
- 19. Asses N, Ayed L, Bouallagui H et al. Use of Geotrichum candidum for olive mill wastewater treatment in submerged and static culture. Bioresource Technol 100: 2182-2188, 2009.
- 20. Haas MJ, Cichowicz DJ, Bailey DG. Purification and characterization of an extracellular lipase from the fungus Rhizopus delemar. Lipids 27: 571-576, 1992.
- 21. Joerger RD, Haas MJ. Overexpression of a Rhizopus delemar lipase gene in Escherichia coli. Lipids 28: 81-88, 1993.
- 22. Joerger RD, Haas MJ. Alteration of chain length selectivity of a Rhizopus delemar lipase through site-directed mutagenesis. Lipids 29: 377-384, 1994.
- 23. Giuseppin MLF. Effects of dissolved oxygen concentration on lipase production by Rhizopus delemar. Appl Microbiol Biot 20: 161-165, 1984.
- 24. Nadeem M, Qazi JI, Syed Q et al. Optimization of Process Parameters for Alkaline Protease Production by Bacillus licheniformis N-2 and Kinetics Studies in Batch Fermentation. Turk J Biol 32: 243-251, 2008.
- 25. Wonderwülbecke T, Kieslich K, Erdman H. Comparison of lipases by different assays. Enzyme Microb Tech 14: 631-639, 1992.
- 26. Li D, Wang B, Tan T. Production enhancement of Rhizopus arrhizus lipase by feeding oleic acid. J Mol Catal B-Enzym 43: 40-43, 2006.
- 27. Elibol M, Özer D. Lipase production by immobilised Rhizopus arrhizus. Process Biochem 36: 219-223, 2000.
- 28. Rodriguez JA, Mateos JC, Nungaray J et al. Improving lipase production by nutrient source modification using Rhizopus homothallicus cultured in solid state fermentation. Process Biochem 41: 2264-2269, 2006.
- 29. Espinosa E, Sanchez S, Farres A. Nutritional factors affecting lipase production by Rhizopus delemar CDBB H313. Biotechnol Lett 12: 209-214, 1990.
- 30. Li CY, Cheng CY, Chen TL. Fed-batch production of lipase by Acinetobacter radioresistens using Tween-80 as the carbon source. Biochem Eng J 19: 25-31, 2004.
- 31. Labuschagne RB, Tonder AV, Litthauer D. Flavobacterium odoratum lipase: Isolation and characterization. Enzyme Microb Tech 21: 52-58, 1997.
- 32. Iızumı T, Nakamura K, Fukase T. Purification and characterization of a thermostable lipase from newly isolated Pseudomonas sp. KWI-56. Agric Biol Chem 54: 1253-1258, 1990.
- 33. Sharon C, Nakazato M, Ogawa HI et al. Lipase-induced hydrolysis of castor oil: effect of various metals. J Ind Microbiol Biotechnol 21: 292-295, 1998.
- 34. Hiol A, Jonzo MD, Rugani N et al. Purification and characterization of an extracellular lipase from a thermophilic Rhizopus oryzaestrain isolated from palm fruit. Enzyme Microb Tech 26: 421-430, 2000.
- 35. Chandan RC, Shahani KM. Role of sulfhydryl groups in the activity of milk lipase. J Dairy Sci 48: 1413-1418, 1965.
- 36. Liebeton K, Zacharias A, Jaeger KE. Disulfide bond in Pseudomonas aeruginosa lipase stabilizes the structure but is not required for interaction with its foldase. J Bacteriol 183: 597- 603, 2001.