EXTRACTION, PURIFICATION AND CHARACTERIZATION OF LIPASE FROM THE DIGESTIVE DUCT OF COMMON CARP CYPRINUS CARPIO L.
DOI:
https://doi.org/10.36103/ijas.v53i5.1615Keywords:
lipase activity, Ion exchange, gel filtration, molecular weight.Abstract
This study was aimed to use digestive duct of common carp Cyprinus Carpio L. as economic source for fish lipase extraction . The enzyme was extracted using potassium phosphate buffer (pH 7 , 0.2 M ). The crude extract then precipitated at the saturation range between 40 – 80 % and dialyzed . Further purified carried out through ion exchange chromatography using DEAE-Cellulose column (2.5 × 30 cm) and gel filtration chromatography by Sephadex G-100 column (1.5 x 60 cm). The purified lipase had a molecular weight of 37.150KDa ,and optimum pH and temperature for enzyme activity about 45◦C and 6.5 using olive oil as substrate .The specific activity, number of purification folds and yield of purified Lipase were 146.48U/mg, 9.76 and 40.50% respectively.
References
Abadi, F. A. and J. M. Naser. 2019. Effect of wet gluten addition on staling characteristics of barley bread. The Iraqi Journal of Agricultural Sciences.50(1):390-397.
Al-Aithy, Musa. 2012. Extraction of β-Glucan from barley bran and assessment of some functional properties and molecular weight. The Iraqi Journal of Agricultural Sciences.43(2):100-108.
Al- Mousawi, A. J .2007. Purification and Characterization of Polymorph Nuclear Lipases from Mastitis Milk and Effect of Somatic Cell on Lipolysis in Cheddar Cheese. Ph. D. Dissertation. University of Baghdad.
Bradford, M. M. 1976. A rapid and sensitive method for the quantities of micro gram quantities of protein utilizing the principle of protein –dye binding. Analytical Biochemistry.72 :248-254.
Bier, M.1955. Lipase. In methods in Enzymology .Vol. 1(ed. By Sidney , P. Colowich and Nathano, O. Kaplan) Academic press, New York, San Francisco, London.pp:245-276.
Borkar, P. S.; R. G. Bodade; S. R. Rao and C. N. Khobragade .2009. Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT 9. Brazilian Journal of Microbiology, 40(2), 358-366.
Chen, X.; C. Wang ; J. Xu; F. Wang ; Y. Jiang ; Y. Chen and X. Zhao. 2020. Purification and characterization of a novel lipase from antarctic krill. Journal of Ocean University of China. 19(1), 209-215.
Colakoglu, A.S. and H. Özkaya. 2012. Potential use of exogenous lipases for DATEM replacement to modify the reological and thermal properties of wheat fl our dough. Journal of Cereal Science, 55: 397–404.
Emmanuel, M. B.; E. C. Evans; A. Abubakar; L. M. Labaran; A. V. Ali; and M. Zabe. 2020 . Production, Partial Purification and Characterization of Lipase Enzyme Expressed by Klebsiella Pnemoniae of Vegetable Oil Contaminated Soil.pp:122-134.
Gupta, R.; A. Kumari; P. Syal and Y. Singh. 2015. Molecular and functional diversity of yeast and fungal lipases: Their role in biotechnology and cellular physiology. Progress in Lipid Research, 57: 40–54.
Georlette, D; V. Blaise; T. Collins ; S. D’Amico; E. Gratia; A. Hoyoux; J. C. Marx; G. Sonan; G. Feller and C. Gerday. 2004. Some like it cold: biocatalysis at low temperatures. FEMS Microbiol Rev 28:25–42
Görgün, S. and M. A. Akpınar. 2012. Purification and characterization of lipase from the liver of carp, Cyprinuscarpio L.(1758), living in Lake Tödürge (Sivas, Türkiye). Turkish Journal of Fisheries and Aquatic Sciences, 12(2), 207-215.
Iqbal, S. A. and A. Rehman. 2015. Characterization of Lipase from Bacillus subtilis I-4 and Its Potential Use in Oil Contaminated Wastewater. Brazilian Archives of Biology and Technology, 58(5), 789-797.
Iijima , N. ; S. Tanaka and Y. Ota. 1998. Purification and characterization of bile salt– activated lipase from the hepatopancreas of red sea bream, Pagrus major. Fish Physiology and Biochemistry .18:59-69.
Kameshwar Sharma, Y. V.; N. Boora and P. Tyagi.2014. Isolation, purification and characterization of secondary structure and kinetic study of lipase from Indian major Carp, Catla catla (Catla). Enzyme Engineering .pp: 120-131.
Kurtovic I; S. N. Marshall ; X. Zhao and B.K. Simpson.2009. Lipases from mammalsand fishes. Reviews in Fisheries Science 17: 18-40.
Kurtovic, I.; S. N. Marshall; X. Zhao; and B. K. Simpson. 2010. Purification and properties of digestive lipases from Chinook salmon (Oncorhynchustshawytscha) and New Zealand hoki (Macruronusnovaezelandiae). Fish Physiology and Biochemistry, 36(4), 1041-1060.
Macedo, G.A.; M. M. S. Lozano and G. M. Pastore. 2003. Enzymatic synthesis of short chain citronellyl esters by a new lipase from Rhizopus sp. Electronic Journal of Biotechnology, 6(1): 72–75.
Naser, J.M. 2010. Improving of Baking Quality and Shelf Life of Loaf by Partially Purified ɑ-Amylase from Hepatopancreas of Common Carp (CyprinusCarpiol L.). Ph.D. Disseration Collage of Agriculture. University of Baghdad.pp:44-60.
Price, N. C. and L. Stevens. 1988. Fundamentals of Enzymology. Oxford University Press.pp:133-157.
21. Reed, G. 1975. Carbohydrases. In: Enzymes in food processing , 2nded., Academic press.Newyork.pp:33-57.
Sae-Leaw, T., and S. Benjakul. 2018. Lipase from liver of seabass (Latescalcarifer): Characteristics and the use for defatting of fish skin. Food Chemistry, 240, 9-15.
Segel, I. H. 1976. Biochemical Calculations. 2nded.,John Wiley and Ssons.Inc. New York .
Sethi, B. K.; P. K. Nanda and S. Sahoo. 2016. Characterization of biotechnologically relevant extracellular lipase produced by Aspergillus terreus NCFT 4269.10. Brazilian Journal of Microbiology, 47(1), 143-149.
Tako, M.; A. Kotogán; T. Papp; S. Kadaikunnan; N. S. Alharbi and C. Vágvölgyi. 2017. Purification and properties of extracellular lipases with trans esterification activity and 1, 3-regioselectivity from Rhizomucor miehei and Rhizopus oryzae. Journal of Microbiology and Biotechnology, 27(2), 277-288.
Verma, N.; S. Thakur and A. K. Bhatt. 2012. Microbial Lipases: Industrial Applications and Properties (a review). International Research Journal of Biological Sciences, 1(8): 88–92.
Wang, Q.; Y. Hou; Y. Ding and P. Yan. 2012. Purification and biochemical characterization of a cold-active lipase from Antarctic sea ice bacteria Pseudoalteromonas sp. NJ 70. Molecular biology reports, 39(9), 9233-9238.
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
How to Cite
2.jpg)
https://orcid.org/0000-0002-5774-5906 2.jpg)
