• Korean Journal of Fisheries and Aquatic Sciences
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• v.41 no.3
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• pp.176-181
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• 2008

To evaluate the effective use of endoprotease from squid viscera as a food processing aid, various methods of fractionating endoprotease from viscera of the Argentina shortfin squid (Illex argentinus) were evaluated. The endoprotease-positive fractions of each fractionation were fraction II (30-40%, w/w) with cold acetone, fraction IV (50-60% saturation) with ammonium sulfate, fraction UF with anion exchange chromatography, and fraction II (15-24 kDa) with gel filtration. The specific activities (approximately 25 U/mg) of the fractions using ammonium sulfate and gel filtration were higher than the others. Total azocaseinolytic activity and recovery of the positive fraction using gel filtration were 806.95U and 37.82%, respectively, and were the highest among the positive fractions. Based on the results, gel filtration was the most efficient method for fractionating endoprotease from the viscera of Illex argentinus.

• Fisheries and Aquatic Sciences
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• v.15 no.4
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• pp.283-291
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• 2012

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.

• Applied Biological Chemistry
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• v.50 no.4
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• pp.308-315
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• 2007

For the effective use of squid processing by-products as food resources, the distribution and the extraction condition of endoprotease and exopeptidase from viscera of Illex argentinus were investigated. Crude protein and lipid contents of viscera of Illex argentinus were 17.2% and 16.9%, respectively. Regardless of kinds of extraction solution (water, 1% NaCl, 1% KCl and 1% NaCl- KCl) and extraction times (1-20 h), endoprotease activities from viscera of Illex argentinus on Hb, casein and azocasein (pH 6.0) were higher than those on casein and azocasein of the other pHs, thus indicating that the distribution of protein hydrolysing protease is distinctive in the weak acid pH range. Exopeptidase activities against LeuPNA and ArgPNA at pH 7.5 were relatively higher than endoprotease activity of the same pH. The results suggested that exopeptidase among proteases from viscera of Illex argentinus was reasonable for application in food industry compared to endoprotease. The activity in enzymes from viscera of Illex argentinus was the highest in the exopeptidase extracted with deionized distilled water at room temperature for 6-8 h. The optimal reaction conditions of crude enzyme from viscera of Illex argentinus were 7.5 for pH and $50-55^{\circ}C$ for temperature.

• Journal of Plant Biology
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• v.37 no.3
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• pp.357-363
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• 1994

An expreiment with non-nodulating alfalfa (Medicago sativa L.) plants was designed to investigate the changes in protein contents and the activities of proteolytic enzymes during a regrowth period of 24 d. Shoot removal caused a depression of root growth and significantly reduced protein contents in roots. An initial decline of root proteins for the first 10 d was followed by a rapid recovery from d 11 to 24. The major increase of regrowing shoot weight occurred also from d 11. The activities of aminopeptidase and endoprotease slightly decreased in regrowing leaves, while protein contents remains stable after shoot removal. Roots exhibited source behaviour with a rapid increase of endoprotease activities for the first 10 d of regrowth; about a 370% increase over the initial level was observed. Increase in endoprotease activity in roots coincided with the time of protein remobilization after shoot removal, indicating the important role of endoproteases in protein degradation.

• Korean Journal of Microbiology
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• v.24 no.2
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• pp.119-126
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• 1986

A periplasmic endoprotease, named protease Pi, was purified to homogeneity from Escherkchia coli by conventional procedure with insulin as substrate. This enzyme degrades insulin and glucagon to trichloroacetic acid-soluble meterials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. Its molecular weight was 110, 000 when determined by gel filtration on Sephacryl S-300 and was 105, 000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, it appears to be single polypeptide. This snzyme is metalloprotease, since it is completely inhibited by o-phenanthroline and can be activated by addition of divalent metal cations, such as $Mg^{2+}\;and\;Co^{2+}$. It is destinct from protease Ci, a cytoplasmic insulin degrading enzyme, since protease Pi is localized to the periplasm. Since protease Pi selectively degrades GTP cyclohydrolase I, it appears to play a role in the regulation of pteridine biosynthesis.

• Korean Journal of Food Science and Technology
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• v.29 no.6
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• pp.1125-1130
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• 1997

The effects of enzyme combination, pH, acid washing and enzyme treatment sequence were investigated in the hydrolysis of soy protein. Comparing Alcalase vs. Neutrase/Alcalase, it appeared that Neutrase/Alcalase was more efficient than Alcalase alone, as the highest degree of hydrolysis (DH) was seen in Neutrase/Alcalase. A surprisingly high DH (more than 60%) was observed with Flavourzyme in the second hydrolysis. The separation of insolubles from the first hydrolysis had little effect on the second hydrolysis. When the washing water from the first hydrolysis was reused in the next hydrolysis, the DH and protein recovery were increased. The addition of calcium ion showed not so much positive effects by the stabilization of Neutrase on the Protein hydrolysis. The use of carbohydrase and repeated acid washing gave positive effects on DH. The simultaneous treatment using endoprotease and exoprotease with pH adjustment improved DH significantly.

• Proceedings of the Zoological Society Korea Conference
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• 1999.10b
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• pp.240.2-240
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• 1999

No Abstract, See Full Text

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.8
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• pp.1265-1273
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• 2005

The purpose of this study was to prepare accelerated salt-fermented anchovy sauce using a shrimp processing byproducts (head, shell and tail) as a fermenting accelerator, and to investigate its physicochemical and enzymatic properties. Four types of sauces were prepared with 0, 10, 20, and 30$\%$ (w/w) addition of shrimp byproduct and fermented at 24$\pm$2$^{\circ}C$ for 360 days. During fermentation, all four type sauces decreased moisture content (67.5$\%$68.0$\%$ to 64.0$\∼$64.8$\%$) and pH (5.52$\∼$7.10 to 5.03$\∼$6.58), but showed increase in their crude protein (7.0$\∼$8.2 to 10.8$\%$) and volatile basic nitrogen contents (40$\∼$75 to 180$\∼$200 mg/100 g of sauce). The ratio of amino nitrogen to total nitrogen contents of control (0$\%$) and sauce with 10$\%$ shrimp byproducts (10$\%$ sauce) were maximized at 270 days, whereas 20$ \% $ and 30$\%$ added sauces were at 180 days. Endoprotease and exoprotease activities of anchovy sauces added with 20$\%$ and 30$\%$ of shrimp byproducts tend to be higher than those of control (0$\%$) and 10$\%$ addition. Proteolytic activities of sauces at pH 9 were about 2 times higher than those at pH 6. Amidolytic activities for LeuPNA decreased remarkably during fermentation, and control (0$\%$) almost lost their activity at 180 days, while additional sauces were relatively stable. These suggest that alkaline pretense of anchovy and shrimp byproducts as a endoprotease mainly contributed to the fermentation of salt-fermented sauces. The protein molecular weight distribution of sauces indicated 2 groups of peaks (peak 1,>70,000 da and peak 2, 3,000$\∼$29,000 da). As the fermentation proceeded, peak 1 tended to decrease in all of sauces, but peak 2 increased rapidly from 30 to 270 days. Optimum fermentation periods of control and 10$\%$ sauces were 270 days and those of 20$\%$ and 30$\%$ sauce were 180 days. The results suggest that shrimp byproduct can be used as accelerator of salt-fermented sauce.

• Fisheries and Aquatic Sciences
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• v.15 no.3
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• pp.197-202
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• 2012

This study examined and compared the exopeptidase and endoprotease activities of the hepatopancreas (HP) of cuttlefish, squid, and octopus species. The protein concentration in crude extract (CE) from octopus HP was 3,940 mg/100 g, lower than those in CEs from squid HP (4,157 mg/100 g) and cuttlefish HP (5,940 mg/100 g). With azocasein of pH 6 as a substrate, the total activity in HP CE of octopus was 31,000 U, lower than the values for cuttlefish (57,000 U) and squid (69,000 U). Regardless of sample type, the total activities of the CEs with azocasein as the substrate were higher at pH 6 (31,000-69,000 U) than at pH 9 (19,000-34,000 U). With L-leucine-p-nitroanilide (LeuPNA) of pH 6 as the substrate, the total activity of the HP CE from octopus was 138,000 U, higher than values from both cuttlefish HP (72,000 U) and squid HP (63,000 U). Regardless of sample type, the total activities of the CEs with LeuPNA as the substrate were higher at pH 6 (63,000-138,000 U) than at pH 9 (41,000-122,000 U). With LeuPNA as the substrate, the total activities of the CEs from octopus HP and cuttlefish HP were higher at pH 6 than at pH 9. However, there was no difference in total activity between pH 6 and 9 for squid HP CE with LeuPNA as the substrate. These results suggest that the octopus HP is superior to the cuttlefish HP and squid HP as a potential resource for extracting exopeptidases. Exopeptidases from octopus HP have potential industrial applications and their use might aid in reducing pollution related to the octopus industry.

• Parasites, Hosts and Diseases
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• v.54 no.6
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• pp.751-758
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• 2016

This study aimed at constructing a draft genome of the adult female worm Toxocara canis using next-generation sequencing (NGS) and de novo assembly, as well as to find new genes after annotation using functional genomics tools. Using an NGS machine, we produced DNA read data of T. canis. The de novo assembly of the read data was performed using SOAPdenovo. RNA read data were assembled using Trinity. Structural annotation, homology search, functional annotation, classification of protein domains, and KEGG pathway analysis were carried out. Besides them, recently developed tools such as MAKER, PASA, Evidence Modeler, and Blast2GO were used. The scaffold DNA was obtained, the N50 was 108,950 bp, and the overall length was 341,776,187 bp. The N50 of the transcriptome was 940 bp, and its length was 53,046,952 bp. The GC content of the entire genome was 39.3%. The total number of genes was 20,178, and the total number of protein sequences was 22,358. Of the 22,358 protein sequences, 4,992 were newly observed in T. canis. Following proteins previously unknown were found: E3 ubiquitin-protein ligase cbl-b and antigen T-cell receptor, zeta chain for T-cell and B-cell regulation; endoprotease bli-4 for cuticle metabolism; mucin 12Ea and polymorphic mucin variant C6/1/40r2.1 for mucin production; tropomodulin-family protein and ryanodine receptor calcium release channels for muscle movement. We were able to find new hypothetical polypeptides sequences unique to T. canis, and the findings of this study are capable of serving as a basis for extending our biological understanding of T. canis.