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http://dx.doi.org/10.5657/FAS.2012.0283

Fractionation and Enzymatic Characterization of Endoprotease and Exopeptidase from Crude Extracts of Cuttlefish Sepia officinalis Hepatopancreas  

Kim, Min Ji (Department of Seafood Science and Technology, Gyeongsang National University)
Kim, Hyeon Jeong (Department of Seafood Science and Technology, Gyeongsang National University)
Kim, Ki Hyun (Department of Seafood Science and Technology, Gyeongsang National University)
Heu, Min Soo (Department of Food Science Nutrition, Gyeongsang National University)
Lee, Jung Suck (Industry-Academic Cooperation Foundation, Jeju National University)
Kim, Jin-Soo (Department of Seafood Science and Technology, Gyeongsang National University)
Publication Information
Fisheries and Aquatic Sciences / v.15, no.4, 2012 , pp. 283-291 More about this Journal
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 (2,704.70 U) 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.
Keywords
Cuttlefish; Sepia officinalis; Hepatopancreas; Exopeptidase; Endoprotease;
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1 Aewsiri T, Benjakul S and Visessanguan W. 2009. Functional properties of gelatin from cuttlefish (Sepia pharaonis) skin as affected by bleaching using hydrogen peroxide. Food Chem 115, 243-249.   DOI   ScienceOn
2 Agriculture and Fishery Statistics Department. 2010. Statistical Report of Fish Catches. Statistics Korea, Seoul, KR.
3 Bezerra RS, Lins EJF, Alencar RB, Paiva PMG, Chaves MEC, Coelho LCBB and Carvalho LB Jr. 2005. Alkaline proteinase from intenstine of Nile tilapia (Oreochromis niloticus). Process Biochem 40, 1829-1834.   DOI   ScienceOn
4 Bougatef A, Souissi N, Fakhfakh N, Ellouz-Triki Y and Nasri M. 2007. Purification and characterization of trypsin from the viscera of sardine (Sardina pilchardus). Food Chem 102, 343-350.   DOI   ScienceOn
5 Bumberger E and Belitz HD. 1993. Bitter taste of enzymatic hydrolysates of casein. Z Lebensm Unters Forsch 197, 14-19.   DOI
6 Chiou TK, Matsui T and Konosu S. 1988. Purification and properties of an aminopeptidase from mullet, Mugil cephalus, roe. Agric Biol Chem 52, 235-242.   DOI
7 Clegg KM, Lim CL and Manson W. 1974. The structure of a bitter peptide derived from casein by digestion with papain. J Dairy Res 41, 283-287.   DOI
8 Dawson RMC, Elliott DC, Elliott WH and Jones KM. 1986. Data for Biochemical Research. 3rd ed. Oxford University Press, Oxford, GB, pp. 417-441.
9 Deejing S, Yoshimune K, Lumyong S and Moriguchi M. 2005. Purification and characterization of hyperthermotolerant leucine aminopeptidase from Geobacillus thermoleovorans 47b. J Ind Microbiol Biotechnol 32, 269-276.   DOI
10 Fu XY, Xue CH, Miao BC, Li ZJ, Yang WG and Wang DF. 2005. Study of a highly alkaline protease extracted from digestive tract of sea cucumber (Stichopus japonicus). Food Res Int 38, 323-329.   DOI   ScienceOn
11 Garcia-Carreno FL and Haard NF. 1993. Characterization of proteinase classes in langostilla (Pleuroncodes planipes) and crayfish (Pacifastacus astacus) extracts. J Food Biochem 17, 97-113.   DOI   ScienceOn
12 Gupta R, Beg QK and Lorenz P. 2002. Bacterial alkaline proteases: molecular approaches and industrial applications. Appl Microbiol Biotechnol 59, 15-32.   DOI   ScienceOn
13 Haard NF. 1990. Enzymes from food myosystems. J Muscle Foods 1, 293-338.   DOI
14 Hernandez-Cortes P, Whitaker JR and Garcia-Carreno FL. 1997. Purification and characterization of chymotrypsin from Penaeus vannamei (Crustacea: Decapoda). J Food Biochem 21, 497-514.   DOI   ScienceOn
15 Izawa N, Tokuyasu K and Hayashi K. 1997. Debittering of protein hydrolysates using Aeromonas caviae aminopeptidase. J Agric Food Chem 45, 543-545.   DOI   ScienceOn
16 Heu MS, Kim JS, Shahidi F, Jeong Y and Jeon YJ. 2003. Extraction, fractionation and activity characteristics of proteases from shrimp processing discards. J Food Biochem 27, 221-236.   DOI   ScienceOn
17 Hoque MS, Benjakul S and Prodpran T. 2010. Effect of heat treatment of film-forming solution on the properties of film from cuttlefish (Sepia pharaonis) skin gelatin. J Food Eng 96, 66-73.   DOI   ScienceOn
18 Ivankovic H, Gallego Ferrer G, Tkalcec E, Orlic S and Ivankovic M. 2009. Preparation of highly porous hydroxyapatite from cuttlefish bone. J Mater Sci Mater Med 20, 1039-1046.
19 Kechaou ES, Dumay J, Donnay-Moreno C, Jaouen P, Gouygou JP, Berge JP and Amar RB. 2009. 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, 158-164.   DOI   ScienceOn
20 Kim HS, Kim JS and Heu MS. 2008. Fractionation of endoprotease from viscera of the argentina shortfin squid Illex argentinus. J Korean Fish Soc 41, 176-181.   과학기술학회마을   DOI   ScienceOn
21 Kishimura H, Hayashi K, Miyashita Y and Nonami Y. 2005. Characteristics of two trypsin isozymes from the viscera of Japanese anchovy (Engraulis japonica). J Food Biochem 29, 459-469.   DOI   ScienceOn
22 Lowry OH, Rosebrough NJ, Farr AL and Randall RJ. 1951. Protein measurement with the folin phenol reagent. J Biol Chem 193, 265-275.
23 Matoba T, Hayashi R and Hata T. 1970. Isolation of bitter peptides from tryptic hydrolysate of casein and their chemical structure. Agric Biol Chem 34, 1235-1243.   DOI
24 Raksakulthai R and Haard NF. 1999. Purification and characterization of aminopeptidase fractions from squid (Illex illecebrosus) hepatopancreas. J Food Biochem 23, 123-144.   DOI
25 Minagawa E, Kaminogawa S, Tsukasaki F and Yamauchi K. 1989. Debittering mechanism in bitter peptides of enzymatic hydrolysates from milk casein by aminopeptidase T. J Food Sci 54, 1225- 1229.   DOI
26 Nagai T, Yamashita E, Taniguchi K, Kanamori N and Suzuki N. 2001. Isolation and characterisation of collagen from the outer skin waste material of cuttlefish (Sepia lycidas). Food Chem 72, 425-429.   DOI   ScienceOn
27 Park JH, Lee H, Lee GY, Kim HR, Kim YW and Lee YB. 2011. Development of refined cuttlefish (Todarodes pacificus) oil and its use as functional ingredients. Food Sci Biotechnol 20, 389-394.   과학기술학회마을   DOI
28 Rao MB, Tanksale AM, Ghatge MS and Deshpande VV. 1998. Molecular and biotechnological aspects of microbial proteases. Microbiol Mol Biol Rev 62, 597-635.
29 Souissi N, Ellouz-Triki Y, Bougatef A, Blibech M and Nasri M. 2008. 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, 473-480.   DOI   ScienceOn
30 Starky PM. 1977. Elastase and cathepsin G: the serine proteinases of human neutrophil leucocytes and spleen. In: Proteinases in Mammalian Cells and Tissues. Barrett AJ, ed. North-Holland Publishing Co., Amsterdam, NL, pp. 57-89.
31 Umetsu H, Arai M, Ota T, Kudo R, Sugiura H, Ishiyama H and Sasaki K. 2003. Purification and properties of an aminopeptidase from the mid-gut gland of scallop (Patinopecten yessoensis). Comp Biochem Physiol B Biochem Mol Biol 136, 935-942.   DOI   ScienceOn
32 Vo VT, Kusakabe I and Murakami K. 1983. Purification and some properties of two aminopeptidases from sardines. Agric Biol Chem 47, 2453-2459.   DOI