• 한국수산과학회지
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• 제41권3호
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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.

• 한국수산과학회지
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• 제47권2호
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• pp.135-143
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• 2014

Exopeptidase active fractions from the hepatopancreas of the Argentina shortfin squid Illex argentinus, were obtained with acetone (AC 30-40%), ammonium sulfate (AS 60-70% saturation), anion exchange chromatography (AE-II, 0.2 M NaCl) and gel filtration chromatography (GF-I, 30-50 kDa) fractionation methods. A bitter peptide solution that has a bitterness equivalent to that of 2% glycylphenylalanine and prepared by tryptic hydrolysis of milk casein, was treated with the exopeptidase active fractions. The GF-I fraction was the best based on aminopeptidase activity (35.3 U/mg), percentage of recovery (30.7%) and a sensory evaluation (1.7). The amount of released amino acids increased as incubation time increased, and the bitterness of the enzyme reaction mixtures decreased. Incubation with the GF-I fraction for 24 h resulted in the hydrolysis of several peptides as revealed by the reverse-phase high performance liguid chromatography profile, with three peaks (3, 5 and 6) decreasing in area (%) and three peaks (1, 2 and 4) increasing in area (%). Therefore, the GF-I fraction appeared to be ideally suited to reduce bitterness in protein hydrolysates by catalyzing the hydrolysis of bitter peptides.

• 한국식품영양과학회지
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• 제37권8호
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• pp.1009-1017
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• 2008

원양산 오징어 내장 조효소로부터 아세톤법, 황산암모늄법, 음이온교환 크로마토그래피법 및 겔 여과법을 이용하여 exopeptidase를 분획한 다음, 각 분획방법별 fraction들의 효소 활성(총활성 및 비활성), 정제도 및 회수율 등을 검토하여 최적 분획조건을 구명하고자 하였다. 아세톤 분획법의 경우 아세톤의 최종농도가 $30{\sim}40%$가 되도록 분획하는 것이, 황산암모늄 분획법의 경우 황산암모늄을 $60{\sim}70%$의 포화농도가 되도록 분획하는 것이, 음이온교환 크로마토그래피의 경우 0.2 M NaCl을 이용하여 분획하는 것이, 그리고, 겔 여과의 경우 분자량 $30{\sim}50\;kDa$ 범위의 것을 분획하는 것이 효율적이었다. 분획방법별 최적 활성 fraction간의 상호비교에서 LeuPNA 및 ArgPNA에 대한 총활성 및 회수율은 겔 여과가, 비활성 및 정제도는 황산암모늄에 의한 분획법이 가장 우수한 것으로 나타났다. 원양산 오징어 내장으로부터 탈 이온수로 조효소를 추출한 다음, 겔 여과로 분획한 exopeptidase의 식품가공소재로서 산업적 이용을 위해서는 겔 여과법의 특성을 기초로 하여 추출 조효소로부터 연속 및 대량처리를 위한 공정개발이 필요할 것으로 사료된다.