• 한국수산과학회지
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• 제42권6호
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• pp.537-544
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• 2009

This study was conducted to improve yield and functional properties of autoclave-treated salmon frame extracts (SFETA) using commercial enzymes (Alcalase 2.4 L FG, Flavourzyme 500 MG, Neutrase 0.8 L and Protamex 1.5 MG). Yield and angiotensin I converting enzyme (ACE) inhibitory activity of all enzymatic hydrolysates improved compared to those of control (undigested extracts), which were the highest in hydrolysates incubated with Protamex 1.5 MG for 4 hrs (P4-treated hydrolysates) and 2 hrs (P2-treated hydrolysates), respectively. However, antioxidant activities of all enzymatic hydrolysates showed less than 29%. According to the trichloroacetic acid soluble-N, volatile component intensity and sensory evaluation, when compared to control, taste of P4-treated hydrolysates improved, while its fish odor strongly smelt. Therefore, for efficient use of P4-hydrolysates, the fish odor should be improved by Maillard reaction of extracts or pre-treatment of salmon frame.

• 대한화장품학회지
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• 제40권4호
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• pp.341-347
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• 2014

본 연구에서는 노무라입깃해파리(Nemopilema nomurai) 가수분해 추출물의 미백활성에 대하여 검색하였다. 가수분해 시료(NHE, N. nomurai Hydrolized Extract)는 노무라입깃해파리를 상업용 단백질분해효소(Protamex, Alcalase, Flavourzyme, Neutrase)를 이용하여 $55^{\circ}C$에서 pH 5-8 조건으로 처리하는 과정을 거쳐 얻었다. 미백활성은 B16-F1 멜라노마 세포에서 멜라닌 색소 합성 저해능 평가를 통하여 확인하였다. 실험결과, Neutrase로 처리된 시료(N-NHE)에서 $100{\mu}g/mL$의 농도에서 멜라닌 생성이 가장 효율적으로 89.9% 감소되는 것을 관찰하였다. 이 시료는 또한 tyrosinase와 TRP-1 (tyrosinase related protein-1) 단백질의 발현을 억제하였다. 이상의 연구 결과는 노무라입깃해파리 가수분해 추출물의 화장품 미백 소재로의 개발 가능성을 보여주고 있다.

• 생명과학회지
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• 제22권1호
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• pp.117-125
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• 2012

본 연구는 굴 가수분해물의 간 보호 효과를 확인하는 것을 목표로 하였다. 굴은 많은 기능적 요소를 가지고 있다고 알려져 있으며 특히, 효소에 의해 생산된 가수분해물은 우수한 기능적 물질을 포함한다. 타크린으로 유발한 간세포 독성에 대한 효소별 굴 가수분해물의 보호 효과를 확인하기 위하여 HepG2세포를 이용하여 in vitro상에서 확인하였다. 사용한 샘플은 Neutrase, Flavourzyme, Protamex을 이용하여 효소 가수분해한 것이다. 타크린으로 손상을 유발한 간세포에서는 GOT와 LDH가 증가하게 된다. 굴 효소 가수분해물을 처리한 실험군에서는 아무런 처리를 하지 않은 실험군에 비하여 높은 세포 생존율을 확인할 수 있었다. 또한 GOT와 LDH 역시 감소하는 것을 알 수 있었다. 본 연구를 통하여 타크린 독성에 대한 신약, 식품의 기초 자료를 제공할 수 있을 것으로 기대된다.

• 한국수산과학회지
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• 제39권3호
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• pp.269-277
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• 2006

This study prepared functional oyster hydrolysates using two-step enzymatic hydrolysis and investigated their functional properties. To prepare two-step enzymatic hydrolysates (TSEH), oysters were hydrolyzed using 1% Protamex (PR) at $40^{\circ}C$ and pH 6.0 for 1 hr before sequential treatment with one of the following enzymes for 1 hr: Alcalase (AL), Flavourzyme (FL), Neutrase (NE), pepsin (PE), and trypsin (TR). The PRAL, PRNE and PRTR hydrolysates had significantly greater angiotensin I converting enzyme (ACE) inhibitory activity than did PR and the other TSEHs. Only the antioxidant activity of the PRNE hydrolysate was significantly different (p<0.05), while none of the TSEHs had antimicrobial activity. The oyster hydrolysate prepared by sequential treatment with Protamex and Neutrase (PRNE) had the best ACE inhibitory activity and antioxidant activity, with $IC_{50}$ values of 0.40 and 0.94 mg/mL, respectively. The PRNE hydrolysate was processed through an ultrafiltration (UF) series with molecular weight cut-off (MWCO) membranes of 3, 5, 10, and 30 kDa, and the ACE inhibitory, antioxidant, and antimicrobial activities of the permeates were determined. The permeate through the 3-kDa MWCO membrane had greater ACE inhibitory activity and antioxidant activity than did the other PRNE permeates, with $IC_{50}$ values of 0.11 and 0.40 mg/mL, respectively.

• Food Science and Biotechnology
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• 제16권4호
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• pp.565-571
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• 2007

This study was carried out to investigate an optimum condition for the high angiotensin-l converting enzyme (ACE) inhibitory activity and the yield on enzyme concentration, casein concentration, and hydrolysis time. The optimum condition was performed by response surface methodology for acquirement of casein hydrolysate of milk which shows high ACE inhibitory activity, Among 8 tested enzymes, Protamex showed the highest activation degree with 77.03 unit/g from casein. Their hydrolysis degrees of flovourzyme 500MG, protamex, mixture from 1% casein were 85.5, 88.5, and 93.5%, respectively. The ranges of enzyme concentration (0.25-1.25%), casein concentration (2.5-12.5%), and hydrolysis time (20-100 min) as 3 independent variables through preliminary experiments of the yield of casein hydrolysate and ACE inhibitory activity, and it shows optimum response surface at a saddle point. It shows enzyme concentration (0.64%), casein concentration (8.38%), and hydrolysis time (55.81 min) in the yield aspect and showed the highest activity at enzyme concentration (0.86%), casein concentration (5.97%), and hydrolysis time (63.86 min) in ACE inhibitory aspect. The $R^2$ value of a fitted optimum formula on the hydrolysis yield was 0.9751 as the significant level of 1%. The $R^2$ value of a fitted optimum formula on ACE inhibitory activity is 0.8398, and the significance is recognized in the range of 5%.

• 한국식품영양과학회지
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• 제44권11호
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• pp.1645-1652
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• 2015

해양생물 유래 피부건강 기능성의 소재를 발굴할 목적으로 굴 단백질을 Protamex와 Neutrase로 가수분해 한 후 생성된 굴 가수분해물 펩타이드의 주름 개선 효과를 조사하였다. 굴 가수분해물은 약 7.9%의 유리아미노산을 포함하며 urea, taurine, alanine, glycine 등의 아미노산의 함량이 높았다. 굴 가수분해물은 collagenase 저해 활성을 가지고 있었고, 피부세포인 CCD986sk에 대한 독성은 없었다. 분획물의 수율은 1,000 Da 미만이 40%였으며, 5,000 Da 미만이 60.4%를 차지하였다. 1,000~3,000 Da인 분획물이 항산화 활성, procollagen의 생성능 및 MMP-1 효소 저해 활성이 가장 높았다. 굴 가수분해물과 5,000 Da 미만의 분획물은 주름 개선을 위한 피부건강 기능성 소재로서 화장품 등의 제품에 응용 가능함을 확인하였다.

• Preventive Nutrition and Food Science
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• 제5권3호
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• pp.148-152
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• 2000

Mussel adhesive protein (MAP) was extracted from Korean Mytilus edulis and then partially purified using Sephacryl S-300 gel permeation chromatography and reversed-phase high performance liquid chromatography. As an indicator of adhesiveness, is 3,4-dihydroxyphenylalanine (DOPA) content was determined. Its DOPA/protein ratio of 0.19 was higher than those of other reports, indicating a good adhesive. The partially purified MAP was confirmed by acid-urea polyacrylamide gel electrophoresis using cetylpiridinium bromide as a cationic detergent. Sea mussel hydrolysates were prepared using three commercial proteases to provide value-added functional materials and their angiotensin converting enzyme (ACE) inhibitory activities were determined. Among hydrolysates of sea mussel, Protamex was the best and further purification would improved ACE inhibitory activity.

• Nutrition Research and Practice
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• 제4권3호
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• pp.183-190
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• 2010

Blueberry was enzymatically hydrolyzed using selected commercial food grade carbohydrases (AMG, Celluclast, Termamyl, Ultraflo and Viscozyme) and proteases (Alcalase, Flavourzyme, Kojizyme, Neutrase and Protamex) to obtain water soluble compounds, and their protective effect was investigated against $H_2O_2$-induced damage in Chinese hamster lung fibroblast cell line (V79-4) via various published methods. Both AMG and Alcalase hydrolysates showed higher total phenolic content as well as higher cell viability and ROS scavenging activities, and hence, selected for further antioxidant assays. Both AMG and Alcalase hydrolysates also showed higher protective effects against lipid peroxidation, DNA damage and apoptotic body formation in a dose-dependent fashion. Thus, the results indicated that water soluble compounds obtained by enzymatic hydrolysis of blueberry possess good antioxidant activity against $H_2O_2$-induced cell damage in vitro.

• ALGAE
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• 제18권4호
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• pp.341-347
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• 2003

The potential antioxidative activity of water-soluble enzymatic hydrolyzates from a kelp, Ecklonia cava was evaluated by free radical scavenging and lipid peroxidation assays. To prepare water-soluble hydrolyzates from E. cava the seaweed was enzymatically hydrolyzed by five carbohydrases (Viscozyme, Celluclast, AMG, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase). Among all the hydrolyzates, Celluclast hydrolyzate effectively scavenged free radicals released from DPPH (1,1-diphenyl-2- pricrylhydrazyl) and recorded around 73% scavenging activity at the concentration of 4 mg ${\cdot}ml^{-1}$. This hydrolyzate was thermally stable and DPPH radical scavenging activity remained 80% or higher at heating temperatures of 40 and 60$^{\circ}C$ up to 12 h and around 80% at 100$^{\circ}C$ up to 8 h. AMG and Ultraflo hydrolyzate inhibited the lipid peroxidation of fish oil as that of $\alpha$-tocopherol. These results suggested that an enzymatic extraction will be an effective way for the production of a potential antioxidant from seaweeds.

• 한국축산식품학회지
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• 제32권5호
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• pp.652-658
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• 2012

본 연구는 유단백질 유래 카제인염을 상업용 단백질가수분해 효소로 처리하여 효소의 종류와 가수분해 시간에 따른 ACE 저해효과를 살펴보고 주요 성인병인 고혈압의 예방을 위한 혈압강하 효과가 높은 가수분해물을 제조하고자 수행하였다. 카제인염을 6종의 단백질 분해효소로 기질대 효소비 1000:1로 첨가하여 8시간 가수분해했을 때 가수분해도는 2.54-4.25%로 나타났고, 다시 가수분해물을 기질대 효소비 500:1로 처리하여 4시간 동안 2차 가수분해하였을 때 4.30-5.22%의 가수분해가 일어났다. 효소별 가수분해물의 ACE 저해효과는 가수분해가 진행됨에 따라 $IC_{50}$의 수치는 감소하였고 저해율을 증가하였다. 1차 가수 분해시 8시간 가수분해한 가수분해물의 $IC_{50}$ 수치는 Protamex 처리군이 $516{\mu}g/mL$로 가장 낮았고 Flavourzyme이 $866{\mu}g/mL$로 가장 높았다. 1차 가수분해물을 Flavourzyme로 4시간 2차 가수분해를 하였을 때 1차 가수분해물 보다 $IC_{50}$ 수치가 감소되어 ACE저해 효과가 증가하였으며 Neutrase로 처리하였을 때 $282{\mu}g/mL$로 가장 낮았고 Protease M이 $570{\mu}g/mL$로 가장 효과가 적었다. 가장 효과가 좋은 Neutrase 2차 가수분해물을 소화효소인 pepsin, trypsin, ${\alpha}$-chymotrypsin으로 가수분해 하였을 경우 $IC_{50}$ 수치는 $597{\mu}g/mL$로 1차 가수분해물보다 저해효과가 유의적으로 감소되었다(p<0.05). Neutrase 2차 가수분해물을 MW 10,000로 한외여과하였을 때 MW 10,000 미만 permeate의 $IC_{50}$ 수치는 $273{\mu}g/mL$로 저해효과는 유의차가 없었으나, 10,000 이상의 retentate는 $IC_{50}$ 수치가 $635{\mu}g/mL$로 유의적 수준에서 저해효과가 감소하였다(p<0.05). 이에 효소의 종류와 가수분해 시간의 조합에 의한 ACE 저해활성을 측정하고 분리공정을 최적화하기 위한 추가 연구를 수행한다면 주요 유단백질인 카제인 유래 기능성 식품의 산업적 대량생산이 가능할 것으로 사료된다.