Fractionation and Enzymatic Characterization of Endoprotease and Exopeptidase from Crude Extracts of Cuttlefish Sepia officinalis Hepatopancreas
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참고문헌
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초록
This study examines the optimal fractionation method and conditions for the isolation of endoprotease- and exopeptidase-active fractions from crude extracts of cuttlefish hepatopancreas (HP) using four fractionation methods: ammonium sulfate fractionation (ASF), polyethylene glycol fractionation (PGF), ion exchange chromatography (IEC), and gel filtration chromatography (GFC). Total endoprotease activity highest in the fraction II (concentrate of fractions 34-42; 842.60 U) of GFC, followed by fraction III (40-60% ammonium sulfate fraction; 670.25 U) of ASF, fraction I (concentrate of fractions 8-12; 436.89 U) of IEC, and fraction II (10-20% polyethylene glycol; 307.31 U) of PGF. Total exopeptidase activity of these fractions was highest in fraction II (2,704.70U) of GFC, fraction III (2,110.50 U) of ASF, fraction III (1,605.60 U) of PGF, and fraction II (concentrate of fractions 38-44;1,196.22 U) of IEC. These results showed that fraction II of GFC had the highest activity toward both exopeptidase and endoprotease,with exopeptidase activity being 3.21 times higher than of endoprotease. These results suggest cuttlefish HP could be used as a potential source for the extraction of exopeptidase, an enzyme capable of catalyzing the cleavage of N- and C-terminal amino acids in polypeptides, Like endoprotease, the most efficient method for separating exopeptide-active fractions was GFC.
Abstract
This study examines the optimal fractionation method and conditions for the isolation of endoprotease- and exopeptidase-active fractions from crude extracts of cuttlefish hepatopancreas (HP) using four fractionation methods: ammonium sulfate fractionation (ASF), polyethylene glycol fractionation (PGF), ion exchange chromatography (IEC), and gel filtration chromatography (GFC). Total endoprotease activity highest in the fraction II (concentrate of fractions 34-42; 842.60 U) of GFC, followed by fraction III (40-60% ammonium sulfate fraction; 670.25 U) of ASF, fraction I (concentrate of fractions 8-12; 436.89 U) of IEC, and fraction II (10-20% polyethylene glycol; 307.31 U) of PGF. Total exopeptidase activity of these fractions was highest in fraction II (2,704.70U) of GFC, fraction III (2,110.50 U) of ASF, fraction III (1,605.60 U) of PGF, and fraction II (concentrate of fractions 38-44;1,196.22 U) of IEC. These results showed that fraction II of GFC had the highest activity toward both exopeptidase and endoprotease,with exopeptidase activity being 3.21 times higher than of endoprotease. These results suggest cuttlefish HP could be used as a potential source for the extraction of exopeptidase, an enzyme capable of catalyzing the cleavage of N- and C-terminal amino acids in polypeptides, Like endoprotease, the most efficient method for separating exopeptide-active fractions was GFC.
참고문헌 (32)
- cAewsiri T, "Functional properties of gelatin from cuttlefish (Sepia pharaonis) skin as affected by bleaching using hydrogen peroxide", Food Chem, 115, pp.243~249 (2009)
- Agriculture and Fishery Statistics Department, "Statistical Report of Fish Catches" (2010), Statistics Korea
- Bezerra RS, "Alkaline proteinase from intenstine of Nile tilapia (Oreochromis niloticus)", Process Biochem, 40, pp.1829~1834 (2005)
- Bougatef A, "Purification and characterization of trypsin from the viscera of sardine (Sardina pilchardus)", Food Chem, 102, pp.343~350 (2007)
- Bumberger E, "Bitter taste of enzymatic hydrolysates of casein", Z Lebensm Unters Forsch, 197, pp.14~19 (1993)
- Chiou TK, "Purification and properties of an aminopeptidase from mullet, Mugil cephalus, roe", Agric Biol Chem, 52, pp.235~242 (1988)
- Clegg KM, "The structure of a bitter peptide derived from casein by digestion with papain", J Dairy Res, 41, pp.283~287 (1974)
- Dawson RMC, "Data for Biochemical Research. 3rd ed", pp.417~441 (1986), Oxford University Press
- Deejing S, "Purification and characterization of hyperthermotolerant leucine aminopeptidase from Geobacillus thermoleovorans 47b", J Ind Microbiol Biotechnol, 32, pp.269~276 (2005)
- Fu XY, "Study of a highly alkaline protease extracted from digestive tract of sea cucumber (Stichopus japonicus)", Food Res Int, 38, pp.323~329 (2005)
- Garcíia-Carreñno FL, "Characterization of proteinase classes in langostilla (Pleuroncodes planipes) and crayfish (Pacifastacus astacus) extracts", J Food Biochem, 17, pp.97~113 (1993)
- Gupta R, "Bacterial alkaline proteases: molecular approaches and industrial applications", Appl Microbiol Biotechnol, 59, pp.15~32 (2002)
- Haard NF, "Enzymes from food myosystems", J Muscle Foods, 1, pp.293~338 (1990)
- Hernáandez-Cortées, "Purification and characterization of chymotrypsin from Penaeus vannamei (Crustacea: Decapoda)", J Food Biochem, 21, pp.497~514 (1997)
- 허민수, "Extraction, fractionation and activity characteristics of proteases from shrimp processing discards", Journal of Food Biochemistry, 27(3), pp.221~236, Food and Nutrition Press, Inc.
- Hoque MS, "Effect of heat treatment of film-forming solution on the properties of film from cuttlefish (Sepia pharaonis) skin gelatin", J Food Eng, 96, pp.66~73 (2010)
- Ivankovic H, "Preparation of highly porous hydroxyapatite from cuttlefish bone", J Mater Sci Mater Med, 20, pp.1039~1046 (2009)
- Izawa N, "Debittering of protein hydrolysates using Aeromonas caviae aminopeptidase", J Agric Food Chem, 45, pp.543~545 (1997)
- Kechaou ES, "Enzymatic hydrolysis of cuttlefish (Sepia officinalis) and sardine (Sardina pilchardus) viscera using commercial proteases: effects on lipid distribution and amino composition", J Biosci Bioeng, 107, pp.158~164 (2009)
- 김혜숙, "원양산 오징어(Illex argentinus) 내장으로부터 Endoprotease", 한국수산과학회지, 41(3), pp.176~181 (2008), 한국수산과학회
- Kishimura H, "Characteristics of two trypsin isozymes from the viscera of Japanese anchovy (Engraulis japonica)", J Food Biochem, 29, pp.459~469 (2005)
- Lowry OH, "Protein measurement with the folin phenol reagent", J Biol Chem, 193, pp.265~275 (1951)
- Matoba T, "Isolation of bitter peptides from tryptic hydrolysate of casein and their chemical structure", Agric Biol Chem, 34, pp.1235~1243 (1970)
- Minagawa E, "Debittering mechanism in bitter peptides of enzymatic hydrolysates from milk casein by aminopeptidase T", J Food Sci, 54, pp.1225~1229 (1989)
- Nagai T, "Isolation and characterisation of collagen from the outer skin waste material of cuttlefish (Sepia lycidas)", Food Chem, 72, pp.425~429 (2001)
- 박정현, "Development of Refined Cuttlefish (Todarodes pacificus) Oil and Its Use as Functional Ingredients", Food Science and Biotechnology, 20(2), pp.389~394 (2011), 한국식품과학회
- Raksakulthai R, "Purification and characterization of aminopeptidase fractions from squid (Illex illecebrosus) hepatopancreas", J Food Biochem, 23, pp.123~144 (1999)
- Rao MB, "Molecular and biotechnological aspects of microbial proteases", Microbiol Mol Biol Rev, 62, pp.597~635 (1998)
- Souissi N, "Preparation and use of media for protease-producing bacterial strains based on by-products from cuttlefish (Sepia officinalis) and wastewaters from marine-products processing factories", Microbiol Res, 163, pp.473~480 (2008)
- Starky PM, "Elastase and cathepsin G: the serine proteinases of human neutrophil leucocytes and spleen. In: Proteinases in Mammalian Cells and Tissues", pp.57~89 (1977), North-Holland Publishing Co
- Umetsu H, "Purification and properties of an aminopeptidase from the mid-gut gland of scallop (Patinopecten yessoensis)", Comp Biochem Physiol B Biochem Mol Biol, 136, pp.935~942 (2003)
- Vo VT, "Purification and some properties of two aminopeptidases from sardines", Agric Biol Chem, 47, pp.245~2459 (1983)