• 한국미생물·생명공학회지
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• 제30권1호
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• pp.39-45
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• 2002

실크 피브로인의 가수분해를 통해 생산된 펩타이드 성분에 의해 murine macrophage RAW264.7 세포에 의한 nitric oxide 생성이 촉진됨을 발견하였다. 산 및 효소적 가수분해물의 가수분해도를 비교한 결과 실크 피브로인 단백질은 산 가수분해에 의해 가장 효과적으로 분해되었으며 효소적 가수분해의 경우에는 pepsin, trypsin, Alcalase의 순으로 가수분해도가 높았다. 산 가수분해물을 단독으로 macrophage에 처리하였을 때 처리농도에 따라 NO 생성촉진활성이 높아졌으나 이 활성은 가수분해물 내의 펩타이드 성분들과 오염되어 혼재하는 LPS 성분의 상호작용에 의한 것임이 확인되었다. 함유된 LPS 성분들을 한외여과에 의해 제거한 효소적 가수분해물들의 NO 생성촉진활성은 peptic hydrolysate가 가장 높았고 tryptic-, Alcalase hydrolysate 순이었다. 이러한 활성의 차이는 가수분해물 내의 고분자량 펩타이드 분포가 많을수록 활성이 높다는 관계에 기인하였으나 산 가수분해물의 경우에는 예외적으로 나타났다. 각 가수분해물의 아미노산 조성을 분석한 결과 arginine, lysine의 함량이 높을수록 활성이 높으며 alanine의 glycine에 대한 비율이 커질수록 활성이 높아졌다. 산 가수분해물의 경우에는 낮은 분자량의 펩타이드들이 많이 분포하지만 arginine 및 alanine의 함량이 높아 비교적 높은 NO 생성 촉진활성을 나타내는 것으로 확인되었다.

• 한국식품저장유통학회지
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• 제24권4호
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• pp.550-558
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• 2017

The purpose of current study is to investigate the beneficial effect of enzyme (Alcalase) hydrolysates of silk protein in rat. Alcalase-treated silk protein hydrolysate (ATSH) itself did not show any cytotoxicity on the hepatic tissues and blood biochemistry, similar to the normal condition. ATSH played a protective role in tert-butyl hydroperoxide (t-BHP)-induced hepatotoxicity and liver damage. The values of AST (aspartate aminotransferase) and ALT (alanine aminotransferase), which are the indicators of the liver function, were effectively alleviated with the ATSH treatment in a dose dependent manner. The level of Lactate dehydrogenase (LDH) and Malondialdehyde (MDA), which were increased with t-BHP treatment, were significantly reduced by ATSH. High dose of ATSH (2 g/kg) reduced the t-BHP-induced LDH release by 48%. Antioxidant and antioxidant enzymes in liver cells were significantly increased by ATSH treatment in their level and activities. ATSH (2 g/kg) increased glutathione (GSH), an intracelluar antioxidant, by 2.5-fold compared with the t-BHP treated group. The activities of glutathione-s-transferase (GST), superoxide dismutase (SOD), and catalase were also elevated by 38%, 60%, and 45%, respectively, with ATSH (2 g/kg) treatment. The antioxidative effect of ATSH was recapitulated to the protection from t-BHP induced liver damages in hematoxylin and eosin (H&E) staining. Thus, ATSH might be used as a hepatoprotective agent.

• 한국축산식품학회지
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• 제34권3호
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• pp.362-371
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• 2014

This study utilized commercially available proteolytic enzymes to prepare egg-white protein hydrolysates (EPHs) with different degrees of hydrolysis. The antioxidant effect and functionalities of the resultant products were then investigated. Treatment with Neutrase yielded the most ${\alpha}$-amino groups (6.52 mg/mL). Alcalase, Flavourzyme, Protamex, and Ficin showed similar degrees of ${\alpha}$-amino group liberation (3.19-3.62 mg/mL). Neutrase treatment also resulted in the highest degree of hydrolysis (23.4%). Alcalase and Ficin treatment resulted in similar degrees of hydrolysis. All hydrolysates, except for the Flavourzyme hydrolysate, had greater radical scavenging activity than the control. The Neutrase hydrolysate showed the highest 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity ($IC_{50}=3.6mg/mL$). Therefore, Neutrase was identified as the optimal enzyme for hydrolyzing egg-white protein to yield antioxidant peptides. During Neutrase hydrolysis, the reaction rate was rapid over the first 4 h, and then subsequently declined. The $IC_{50}$ value was lowest after the first hour (2.99 mg/mL). The emulsifying activity index (EAI) of EPH treated with Neutrase decreased, as the pH decreased. The EPH foaming capacity was maximal at pH 3.6, and decreased at an alkaline pH. Digestion resulted in significantly higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ABTS radical scavenging activity. The active peptides released from egg-white protein showed antioxidative activities on ABTS and DHHP radical. Thus, this approach may be useful for the preparation of potent antioxidant products.

• Fisheries and Aquatic Sciences
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• 제25권6호
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• pp.335-349
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• 2022

To optimize the hydrolysis conditions in the production of antioxidant hydrolysates from tuna cooking juice concentrate (TC) to maximize the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, TC containing 48.91% protein was hydrolyzed with Alcalase 2.4 L, and response surface methodology (RSM) was applied. The optimum hydrolysis conditions included a 2.2% (w/v) Alcalase concentration and 281 min hydrolysis time, resulting in the highest DPPH radical scavenging activity of 66.49% (0.98 µmol Trolox/mg protein). The analysis of variance for RSM showed that hydrolysis time was an important factor that significantly affected the process (p < 0.05). The effects of different drying methods (freeze drying, hot air drying, and vacuum drying) on the DPPH radical scavenging activity and amino acid (AA) profiles of TC hydrolysate (TCH) were evaluated. Vacuum-dried TCH (VD) exhibited an increase in DPPH radical scavenging activity of 81.28% (1.20 µmol Trolox/mg protein). The VD samples were further fractionated by ultrafiltration. The AA profiles and antioxidant activities in terms of the DPPH radical scavenging activity, 2,2'-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acid (ABTS) radical scavenging activity, ferric reducing antioxidant power, and ferrous ion chelating activity were investigated. Glutamic acid, glycine, arginine, and cysteine were the major AAs found in the TCH fractions. The highest DPPH radical scavenging activity was found in the VD-1 fraction (< 5 kDa). The VD-3 fraction (> 10 kDa) exhibited the highest ABTS radical scavenging activity and ferric reducing antioxidant power. The ferrous ion chelating activity was the highest in VD-1 and VD-2 (5 to 10 kDa). In conclusion, this study provided the optimal conditions to obtain high antioxidant activities through TCH production, and these conditions could provide a basis for the future application of TCH as a functional food ingredient.

• Journal of Applied Biological Chemistry
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• 제59권3호
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• pp.265-271
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• 2016

해마는 아시아 등지에서 이미 약재로 사용이 되어지고 있지만 그와 관련된 연구는 부족한 실정이다. 이에 본 연구는 해양생물인 해마의 간 보호 효능을 확인하고자 하였다. 해마를 단백질 가수분해효소인 alcalase를 이용하여 가수분해 후 얻은 가수분해물(ALC)을 얻은 후 실험을 수행하였다. Chang 세포에 ALC를 1시간 전처리 후 에탄올 800 mM을 가하여 24시간 후 세포생존율을 확인했을 때, 세포가 알코올 독성으로부터 보호되는 것을 확인할 수 있었다. 그리고 Chang 세포에 ALC를 2시간 전처리 후 TGF-${\beta}$ 10 ng/mL을 처리하였을 때도, TGF-${\beta}$에 의해 증가된 vimentin, ${\alpha}$-SMA, slug의 발현이 억제되는 것을 확인하였다. 또한 에탄올을 이용한 급성과 만성 In vivo 조건에서도 ALC에 의한 간 보호 효과를 확인할 수 있었다. 알코올 투여에 의한 간 무게 감소와 혈청 GOT 및 GPT 활성 증가가 해마 가수분해물 처리에 의해 억제되었으며, 만성 알코올 독성 실험의 경우에는 실험동물의 무게와 식이섭취량도 해마 가수분해물 처리에 의해 정상군과 유사한 수준으로 회복되는 것을 확인할 수 있었다. 따라서 추가적인 연구를 통해 해마가 간 보호 효능의 기능성 식품소재로 활용 가능할 수 있을 것이라 사료된다.

• 현장농수산연구지
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• 제14권1호
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• pp.93-105
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• 2012

서해안 멸치를 이용한 저염 천연조미료를 생산하고자 시판 효소 3종을 사용하여 효소가수분해물을 제조하고 이의 품질특성을 파악하였다. 멸치 효소가수분해물의 주요 유리아미노산은 글루타민산과 라이신이었으며, 전체 유리아미노산의 함량은 글루타민산(12.6%), 리신(8.56%), 발린(7.06%), 아스파탐(5.73%)순으로 나타났다. 휘발성 정미성분은 전체 30개의 성분이 검출되었으며, aldehyde, ketone, cyclic compound 등으로 나타났다. 가수분해물 제조과정 중 히스타민 함량의 변화는 17.1-18.1 mg/100g으로 변패에 대해 안전한 수준이었다.

• Preventive Nutrition and Food Science
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• 제15권4호
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• pp.316-321
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• 2010

Krill byproduct was hydrolyzed with Alcalase 2.4L to produce functional ingredients for high antioxidative activities against 1,1-dimethyl-2-picryl-hydrazyl (DPPH) radical and Fe. The objective of this study was to investigate the optimum condition for degree of hydrolysis and antioxidative activity of enzymatic hydrolysate produced with the commercial Alcalase using response surface methodology (RSM) with a central composite rotatable design (CCRD). The ranges of independent variables were pH 7.6~10.4 for initial pH and $50.9{\sim}79.1^{\circ}C$ for hydrolysis temperature and their dependent variables were degree of hydrolysis, Brix, amount of phenolic compounds, DPPH-scavenging activity and Fe-chelating activity. RSM with CCRD was well designed to investigate the optimum condition for functional ingredients with high antioxidative activities using Alcalase 2.4L because of their high $R^2$ values of the range of 0.93~0.99 except the $R^2$ value of 0.50 for the amount of total phenolic compounds. The optimum hydrolysis conditions were pH 9.5 and $62^{\circ}C$ for degree of hydrolysis (DH) and pH 9.1 and $64^{\circ}C$ for DPPH-scavenging activity by response surface methodology. The yield of DH and DPPH-scavenging activity were $14.1{\pm}0.5%$ and $10.5{\pm}0.2%$, respectively. It is advantageous to determine the optimum hydrolysis conditions of krill and its by-products for the creation of different kinds of food products, as well as to increase the usage of marine protein sources.

• KSBB Journal
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• 제14권5호
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• pp.600-605
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• 1999

본 연구는 도축 폐기물인 가축혈액을 이용하여 항고혈압 기능성 식품소재로서의 angiotensin I converting enzyme 저해 펩타이드 분획을 생산하기 위한 조건과 가능성을 조사하기 위하여 수행되었다. 산업적으로 이용 가능한 단백분해효소 중 Alcalase가 혈장 원액 및 그로부터 분리된 albumin에 대하여 가장 높은 활성의 가수분해물을 생성하였다. 특히 albumin의 Alcalase 가수분해물과 이를 gel chromatography를 통새 분획하여 얻은 고활성 분획의 $IC_50$값은 각각 0.5 및 0.02 mg/mL로서 지금까지 보고된 식품단백질 유래 펩타이드 혼합물들과 비교할 때 활성이 매우 높은 것에 속함을 알 수 있었다. 또한 이 고활성 펩타이드 분획은 혈장 원액으로부터 단순한 한외여과만을 거쳐도 얻을 수 있음을 확인함으로써 산업적 실용화 가능성이 높은 공정임을 알게 되었다.

• ALGAE
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• 제19권1호
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• pp.59-68
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• 2004

Hizikia fusiformis hydroysates by five carbohydrases (Viscozyme, Celluclast, Termamyl and Ultraflo) and five proteases (Protamex, Kojizyme, Neutrase, Flavourzyme and Alcalase) were investigated for their extraction efficacy (yield and total total polyphenolic content) and antioxidative activity (DPPH radical and hydrogen peroxide scavenging activity). Termamyl and Ultraflo of the carbohydrases and Flavourzyme and Alcalase of proteases were selected by their high eficacy of extraction and antioxidative activity. Selected enzymes were used to investigate the optimum enzymatic reaction time and dosage (enzyme/substrate ratio) suitable for hydorolysis. Optimum reaction time for the enzymatic hydrolysis was 3 days and optimum dosage of hydrolysis was observed as 5%. Simultaneously, Ultraflo of the two carbohydrases and Alcalse of the two proteases were selected as the most effective enzymes. Combination of Ultraflo and Alcalase under optimum hydrolysis conditions could intensify the extraction efficacy of antioxidative materials form H. fusiformis. The hydrolysate obtained by combining the enzymes was separated into four different molecular weight fractions (<1kD, 1-10 kD, 10-30 kD and >30 kD) and recorded the polyphenolic content distribution and respective antioxidative ability. The fraction <1kD was identified as less effective and those fractions > 1kD indicated comparatively higher antioxidative activities related to their polyphenolic content.

• Asian-Australasian Journal of Animal Sciences
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• 제21권5호
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• pp.732-737
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• 2008

This study developed a natural ingredient as a functional food possessing properties of attenuation of hypertension and cardiovascular hypertrophy. In a previous study hydrolysates obtained from chicken leg bone protein using Alcalase strongly inhibited angiotensin I converting enzyme (ACE) in vitro. In particular, hydrolysate (A4H) from four hours of incubation exhibited the highest ACE inhibitory activity (IC50 = 0.545 mg/ml). A4H was selected as a potent ACE inhibitor and orally administrated to spontaneously hypertensive rats (SHR) for eight weeks to investigate attenuating effects on age-related development of hypertension and cardiovascular hypertrophy. Results showed that treatment with A4H of SHRs attenuated the development of hypertension as effectively as the clinical antihypertensive drug captopril. Moreover, a significantly lower heart to body weight ratio and thinness of coronary arterial wall was observed in SHRs that had been treated with A4H or captopril. The results suggest that A4H can be utilized in developing an ACE inhibitor as a potential ingredient of functional foods to alleviate hypertension and cardiovascular hypertrophy.