Characterization of Ammonium Sulphate Fraction Tripsin Isolated from Intestine of Little Tuna Trypsin from little tuna
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Accepted
27 August 2020
Published
4 September 2020
Keywords:
-
ammonium sulphate fraction, BAPNA, viscera, little tuna, trypsin enzyme
Abstract
Intestine is part of the digestive tract of fish having neutral pH. Thus, fish intestine is a potential source of typsin and can be an alternative source to the commercial trypsin enzyme derived from pork and cattle. This study was aimed to analyze the characteristics of the ammonium sulphates fraction of the trypsin enzyme from tuna intestine. The research began with the extraction of the trypsin enzyme from the little tuna intestine and followed up by fractionation using ammonium sulphates (0-80%). Optimum temperature and pH as well as effects of metal ions, stability to NaCl, optimum substrate and reaction kinetics were determined. Enzyme activity was measured using N-α-benzoyl-DL-arginine-p-nitroanilide (BAPNA). The results showed that the trypsin crude extract had an activity of 0.205 U / mL. The highest enzyme activity was found in the ammonium sulphates fraction of 40-50% . The enzyme worked optimally at 60°C and pH 9. Metal ions including ZnCl2 and CaCl2 inhibited trypsin activity, while MnCl2, CuCl2, and NaCl increased trypsin activity. Trypsin activity was stable at 5-30% NaCl. The enzyme had a Vmax value of 0.42 mmol/s and a Km value of 1.12 mM.
References
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Silva JF, Esposito TS, Marchuschi M, Ribeiro K, Cavalli RO, Oliveira V, Bezerra RS. 2011. Purification and partial characterization of a trypsin from the processing waste of the silver mojarra (Diapterus rhombeus). Food Chemistry. 129(3): 777-782.
Sinaga M, Nugroho TT, Dahliaty A. 2014. Pemekatan enzim selulase Penicillium sp. LBKURCC20 dengan pengendapan amonium sulfat 80%. Jurnal Online Mahasiswa Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Riau.1(2): 283-288.
Suhartono MT. 1989. Enzim dan Bioteknologi. Bogor (ID): Departemen Pendidikan dan Kebudayaan, Direktorat Jenderal Pendidikan Tinggi Antar Universitas Bioteknologi, Institut Pertanian Bogor.
Sulistyowati E, Salirawati D, Amanatie. 2016. Karakterisasi beberapa ion logam terhadap aktivitas enzim tripsin. Jurnal Penelitian Saintek. 21(2): 107-119.
Sumardjo D. 2006. Pengantar Kimia: Buku Panduan Kuliah Mahasiswa Kedokteran dan Program Satrata I Fakultas Bioeksakta. Jakarta (ID): Penerbit Buku Kedokteran EGC.
Teich N, Nemoda Z, Kohler H, Heinritz W, Mossner J, Keim V, Sahin-Toth M. 2005. Gene conversion between functional tripsinogen genes PRSS1 and PRSS2 associated with chronic pancreatitis in a six-year-old girl. Human Mutatation. 25(4): 343-347.
Wahyuni S. 2017. Biokimia Enzim dan Karbohidrat. Lhokseumawe (ID): Unimal Press
Whitaker JR. 1994. Principles of Enzymology for the Food Science 2nd Edition. New York (USA): Marcel Dekker Inc.
Wiseman A. 1985. Handbook of Enzyme Biotechnology Second Edition. Chicester (US): Ellis Hordwood Limited.
Yazid E. 2006. Penuntun Praktikum Biokimia Untuk Mahasiswa Analisis. Jakarta (ID): Graha Ilmu.
Barkia A, Bougatef A, Nasri R, Fetoui E, Balti R, Nasri M. 2010. Trypsin from the viscera of Bogue (Boop boops): isolation and characterization. Fish Physiology Biochemistry.36: 893-902.
Benjakul S, Morrissey MT. 1997. Protein hydrolysates from pacific whiting solid wastes. Journal of Agricultural Food Chemistry. 45 (9): 3423-3430.
Bougatef A, Souissi N, Fakhfakh N, Ellouz-triki Y, Nasri M. 2007. Purification and characterization of trypsin from the viscera of sardine (Sardina pilchardus). Food Chemistry. 102: 343-350.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry.72: 248-254.
Chodrijah U, Hidayat T, Noegroho T. 2013. Estimasi parameter populasi ikan tongkol komo (Euthynnus affinis) di perairan laut jawa. BAWAL. 5(3): 167-174.
Falch EA. 1991. Industrial enzymes developments in production and application. Biotechnology advances. 9(4): 643-658.
Green AA, Hughes WL. 1955. Protein solubility on the basic of solubility in aqueous solutions of salts and organic solvent. Methods Enzymology.1: 67-90.
Green NM, Neurath H. 1953. The effect of divalent cations on trypsin. Journal of Biological Chemistry. 204(1): 379-390.
Grizlle JM, Roger WA. 2003. Anatomy and histology of the freshwater. (USA): Departement of Fisheries And Applied Aquaculture, Auburn University
Jatmiko PD. 2018. Karakteristik ekstrak kasar pepsin ikan tuna sirip kuning (Thunnus albacore) [skripsi]. Bogor (ID): Institut Pertanian Bogor.
[Kemerisetdikti] Kementrian Riset, Teknologi, dan Pendidikan Tinggi Republik Indonesia (ID). 2017. Kemandirian Produk Enzim Indonesia. https://www.ristekdikti.go.id/kabar/kemandirian-produk-enzim-indonesia/
Khaled HB, Jellouli K, Souissi N, Ghorbel S, Barkia A, Nasri M. 2011. Purification and characterization of three trypsin isoforms from viscera of sardinelle (Sardinella aurita). Fish Physiology and Biochemistry. 37(1): 123-133.
Kramer DL, Bryant MJ. 1995. Intestine leght in the fishes of a tropical stream: 1 Ontogenic allometry, 2 relation to diet the long and short of a convoluted issue. Environtmental Biology Fish. 42(2): 115-141.
Khandagale AS, Mundodi L, Sarojini BK. 2017. Isolation and characterization of trypsin from fish viscera of oil sardine (Sardinella longiceps).International Journal of Fisheries and Aquatic Studies. 5(2): 33-37.
Kim M, Jeong Y.2013. Purification and characterization of trypsin-like protease from faltfish (paralichthy olivaceus) intestine. Journal of Food Biochemistry. 37: 732-741.
Kita Y, Arakawa T, Lin TY, Timasheff S. 1994. Contribution of the surface free energy perturbations to protein solvent interactions. Biochemistry. 33: 15178-15189.
Kosim M. 2009. Pengaruh suhu pada protease dari Bacillus subtilis [skripsi]. Surabaya (ID): Institut Teknologi Sepuluh Nopember.
Lehninger AL. 1997. Dasar-Dasar Biokimia. Thenawidjaja M, penerjemah; Suwarman, editor. Jakarta (ID): Penerbit Erlangga. Terjemahan dari: Principle of Biochemistry.
Li ZY, Youravong w, H-Kittikun A. 2006. Separation of protease from yellowfin tuna spleen by ultrafiltration. Bioresource Technology. 97(18): 2364-2370.
Miniharapon T, Pattipeilohy F. 2016. Pemanfaatan daging merah dari limbah tuna loin dalam pengolahan kecap ikan. Majalah Bahan Alam, Industri, Aneka Pangan, Minyak Atsiri. 12(1): 27-31.
Muallifa AY. 2017. Mengurai hadis tahnik dan gerakan anti vaksin. Jurnal Living Hadis. 2(2): 253-269.
Nurkhotimah. 2017. Pengaruh suhu dan pH terhadap aktivitas enzim fosfatase bakteri termofilik sungai gendol pasca erupsi merapi. Jurnal Prodi Biologi. 6(8): 465-471.
Oktavianto D, Susilo U, Priyanto S. 2014. Respon aktivitas amylase dan protease ikan gurami Osphronemus gouramy Lac. terhadap perbedaan temperature air. Scripta Biologica. 1(4): 14-18.
Parfiit K. 1999. Martindale and Complete Drug Reference. Ed ke-13. (USA): Pharmaceutical Press.
Ranasasmta R. 2015. Kehalalan produk obat-obatan, terutama obat herbal. Di dalam: Ranasasmita R, Roswiem AP, editor. Prosiding Simposium Penelitian Bahan Obat Alami XIV [Internet]. [Waktu dan tempat pertemuan tidak diketahui].Bogor(ID): Institut Pertanian Bogor. Hlm 552-559; [diunduh 2018 Nov 4] Tersedia pada: http://xdocs.net/previewmakalah-halal-5bf85c45
Santos CWVd, Marques MEdS, Tenoroi HdA, Miranda ECd, Pereria HJV. 2016. Purification and characterization of trypsin Luphiosilurus alexandri. Biochemistry and Biophysics Report .8: 29-33.
Silva JF, Esposito TS, Marchuschi M, Ribeiro K, Cavalli RO, Oliveira V, Bezerra RS. 2011. Purification and partial characterization of a trypsin from the processing waste of the silver mojarra (Diapterus rhombeus). Food Chemistry. 129(3): 777-782.
Sinaga M, Nugroho TT, Dahliaty A. 2014. Pemekatan enzim selulase Penicillium sp. LBKURCC20 dengan pengendapan amonium sulfat 80%. Jurnal Online Mahasiswa Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Riau.1(2): 283-288.
Suhartono MT. 1989. Enzim dan Bioteknologi. Bogor (ID): Departemen Pendidikan dan Kebudayaan, Direktorat Jenderal Pendidikan Tinggi Antar Universitas Bioteknologi, Institut Pertanian Bogor.
Sulistyowati E, Salirawati D, Amanatie. 2016. Karakterisasi beberapa ion logam terhadap aktivitas enzim tripsin. Jurnal Penelitian Saintek. 21(2): 107-119.
Sumardjo D. 2006. Pengantar Kimia: Buku Panduan Kuliah Mahasiswa Kedokteran dan Program Satrata I Fakultas Bioeksakta. Jakarta (ID): Penerbit Buku Kedokteran EGC.
Teich N, Nemoda Z, Kohler H, Heinritz W, Mossner J, Keim V, Sahin-Toth M. 2005. Gene conversion between functional tripsinogen genes PRSS1 and PRSS2 associated with chronic pancreatitis in a six-year-old girl. Human Mutatation. 25(4): 343-347.
Wahyuni S. 2017. Biokimia Enzim dan Karbohidrat. Lhokseumawe (ID): Unimal Press
Whitaker JR. 1994. Principles of Enzymology for the Food Science 2nd Edition. New York (USA): Marcel Dekker Inc.
Wiseman A. 1985. Handbook of Enzyme Biotechnology Second Edition. Chicester (US): Ellis Hordwood Limited.
Yazid E. 2006. Penuntun Praktikum Biokimia Untuk Mahasiswa Analisis. Jakarta (ID): Graha Ilmu.
Authors
NurhayatiT., NugrahaR., & Ningdya LihuanaD. (2020). Characterization of Ammonium Sulphate Fraction Tripsin Isolated from Intestine of Little Tuna: Trypsin from little tuna. Jurnal Pengolahan Hasil Perikanan Indonesia, 23(2), 372-382. https://doi.org/10.17844/jphpi.v23i2.32221
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