• 한국패류학회지
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• 제29권2호
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• pp.155-161
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• 2013

Cathepsins are lysosomal/cysteine proteases belong to papain family (C1 family) that is involved in intracellular protein degradation, antigen processing, hormone maturation, and immune responses. In this study, member of cathepsin family was identified from Manila clam (Mc-Cathepsin D) and investigated the immune response against brown ring disease (BRD) causing Vibrio tapetis challenge. The identified Mc-Cathepsin D gene encodes characteristic features typical for the cathepsin family including eukaryotic and viral aspartyl protease signature domain and two highly conserved active sites ($^{84}VVFDTGSSNLWV^{95}$ and $^{270}IADTGTSLLAG^{281}$). Moreover, MC-Cathepsin D shows higher identity values (-50-70%) and conserved amino acids with known cathepsin D members. Transcriptional results (by quantitative real-time RT-PCR) showed that Mc-Cathepsin D was expressed at higher levels in gills and hemocytes than mantle, adductor muscle, foot, and siphon. After the V. tapetis challenge under laboratory conditions, Mc-Cathepsin D mRNA was up-regulated in gills and hemocytes. Present study indicates that Mc-Cathepsin D is constitutively expressed in different tissues and potentially inducible when infecting BRD by V. tapetis. It is further suggesting that Mc-Cathepsin D may be involved in multiple role including immune response reactions against BRD.

• KSBB Journal
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• 제29권3호
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• pp.145-154
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• 2014

A fresh vegetable and fruit juice has become a new healthy food available for detoxification, dieting and health. This paper presents the useful information about the quality changes of fresh juice according to different juicer. Quality of fresh juice could be evaluated by several factors such as juice yield, enzyme activity, antioxidant activity, polyphenol contents, and anti-inflammatory activity. The juice yields of 12 different vegetables and fruits were compared using 6 different juicers and it was observed that the yield of slow juicer was better than that of conventional blender. Among 12 samples, the juice yield of grape is the best and the pH of the juice was in the acidic range of 3 and 4. Kiwi and grapefruit were the best in terms of protease enzyme activities by Hemoglobin units on the tyrosine basis and Spectrophotometric acid protease unit and papain units on the tyrosine basis of KFDA protocols. The total polyphenol contents were also high in kiwi and grapefruit. The antioxidant activity by diphenyl-1-picrylhydrazyl (DPPH), superoxide dismutase (SOD), and radical scavenging assay were high in the order of kiwi, grapefruit, grape, tomato, and orange. Anti-inflammatory activities were also assay for 12 samples with 6 juicers. It can be concluded that of fresh fruit and vegetable juice provides a source of antioxidant components and enzymes with high activity. And the enzyme activities could be used as one of the quality indicator of fresh juice. Concerning the juicers used in this study, slow juicer could be recommended to prepare the fresh juice in terms of the juice quality.

• 한방재활의학과학회지
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• 제23권4호
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• pp.23-37
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• 2013

Objectives This study was performed to investigate the effects of Saengkanggamchotang (SKT) on the monosodium iodoacetate (MIA) induced osteoarthritis in rats. Methods Osteoarthritis was induced by injection of MIA (50 ul, 60 mg/ml) into knee joints of rats. Rats are divided into a total of 4 groups (normal, control, positive comparison group, SKT treated group, each n=6). Normal group are not treated at all without inducing osteoarthritis whereas control group were induced for osteoarthritis by MIA and oral medicated with 20 ml of distilled water per day. Positive comparison group was injected with MIA and after 7 days, that was taken indomethacin (30 mg/kg/mouse). SKT treated group was injected with MIA and after 7 days that was taken SKT (30 mg/kg/mouse). Positive comparison group and SKT treated group were oral medicated for each substance a total of 4 weeks with one time per day. After experiments (from 1 week after injection of papain to 4 weeks elapsed), the functions of liver and kidney, Prostaglandin E2, inflammatory cytokine (IL-$1{\beta}$, IL-6, TNF-${\alpha}$), osteocalcin, TIMP-1, MMP-9 within serum. Knee joint structures were observed by H&E, safranin-O staining method, and amount of cartilage were measured by ${\mu}CT$-arthrography. Results 1) Hind paw weight bearing ability was significantly improved. 2) Functions of liver and kidney were not affected. 3) Prostaglandin E2, osteocalcin, TIMP-1, MMP-9 in serum were significantly decreased. 4) Inflammatory cytokine IL-$1{\beta}$ was significantly decreased, and IL-6, TNF-${\alpha}$ were decreased but had not significant. 5) In terms of histopathology, significantly reduced subsidence of cartilage and bone in H&E staining. And in Safranin O staining, proteoglycan content in synovial membrane was significantly increased compared with control group. 6) Destruction of cartilage on ${\mu}CT$-arthrography was significantly reduced. Conclusions Based on all results mentioned above, Saengkanggamchotang (SKT) is believed to be meaningful for suppressing the progress of osteoarthritis and its treatments.

• Fisheries and Aquatic Sciences
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• 제19권7호
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• pp.29.1-29.6
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• 2016

In the present study, we investigated to the antioxidant and angiotensin I-converting enzyme (ACE) inhibitory activities of the northern shrimp (Pandalus borealis) by-products (PBB) hydrolysates prepared by enzymatic hydrolysis. The antioxidant and ACE inhibitory activities of five enzymatic hydrolysates (alcalase, protamex, flavourzyme, papain, and trypsin) of PBB were evaluated by the 2, 2'-azino-bis [3-ethylbenzothiazoline-6-sulfonic acid] ($ABTS^+$) radical scavenging and superoxide dismutase (SOD)-like activities, reducing power and Li's method for ACE inhibitory activity. Of these PBB hydrolysates, the protamex hydrolysate exhibited the most potent ACE inhibitory activity with $IC_{50}$ value of $0.08{\pm}0.00mg/mL$. The PBB protamex hydrolysate was fractionated by two ultrafiltration membranes with 3 and 10 kDa (below 3 kDa, between 3 and 10 kDa, and above 10 kDa). These three fractions were evaluated for the total amino acids composition, antioxidant, and ACE inhibitory activities. Among these fractions, the < 3 kDa and 3-10 kDa fractions showed more potent $ABTS^+$ radical scavenging activity than that of > 10 kDa fraction, while the > 10 kDa fraction exhibited the significant reducing power than others. In addition, 3-10 kDa and > 10 kDa fractions showed the significant ACE inhibitory activity. These results suggested that the high molecular weight enzymatic hydrolysate derived from PBB could be used for control oxidative stress and prevent hypertension.

• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• 한국수산과학회지
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• 제53권5호
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• pp.707-716
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• 2020

Red sea cucumber Stichopus japonicus, was dried using three methods-far-infrared ray, vacuum, and freeze drying and then enzymatically hydrolyzed using nine proteases: Alcalase, Flavourzyme, Kojizyme, Neutrase, Protamex, trypsin, α-chymotrypsin, and papain. In addition, the potential ability of hydrolysates to inhibit lipid accumulation in 3T3-L1 adipocytes was evaluated. The yield of hydrolysates from red sea cucumbers dried using each method was higher than that of the distilled water extract, and protein contents were either similar or higher. The hydrolysates that exhibited inhibitory effects on lipid accumulation, as demonstrated via Oil red O staining, were those obtained by far-infrared ray drying coupled with Alcalase, Flavourzyme, Kojizyme, or Neutrase treatment. In addition to the advantages of far-infrared drying and the characteristics of Flavourzyme, the Flavourzyme hydrolysate of far-infrared-dried red sea cucumber showed the highest inhibitory effect on lipid accumulation. In addition, this hydrolysate significantly decreased the expression of the protein factor fatty acid-binding protein 4, which is related to the late differentiation of 3T3-L1 adipocytes. Taken together, these results suggest that Flavourzyme hydrolysates from farinfrared-dried red sea cucumber may be used as a functional food and/or a pharmaceutical ingredient for the inhibition of lipid accumulation.

• Fisheries and Aquatic Sciences
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• 제19권5호
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• pp.25.1-25.8
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• 2016

The fractionation of serine protease inhibitor (SPI) from fish roe extracts was carried out using polyethylene glycol-4000 (PEG4000) precipitation. The protease inhibitory activity of extracts and PEG fractions from Alaska pollock (AP), bastard halibut (BH), skipjack tuna (ST), and yellowfin tuna (YT) roes were determined against target proteases. All of the roe extracts showed inhibitory activity toward bromelain (BR), chymotrypsin (CH), trypsin (TR), papain-EDTA (PED), and alcalase (AL) as target proteases. PEG fractions, which have positive inhibitory activity and high recovery (%), were the PEG1 fraction (0-5 %, w/v) against cysteine proteases (BR and PA) and the PEG4 fraction (20-40 %, w/v) against serine proteases (CH and TR). The strongest specific inhibitory activity toward CH and TR of PEG4 fractions was AP (9278 and 1170 U/mg) followed by ST (6687 and 2064 U/mg), YT (3951 and 1536 U/mg), and BH (538 and 98 U/mg). The inhibitory activity of serine protease in extracts and PEG fractions from fish roe was stronger than that of cysteine protease toward common casein substrate. Therefore, SPI is mainly distributed in fish roe and PEG fractionation effectively isolated the SPI from fish roes.

• 한국식품과학회지
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• 제12권4호
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• pp.263-271
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• 1980

피마자박 단백질을 사료 또는 식품화 하기 위하여 탈지 피마자박으로부터 독성분이 완전하게 제거된 단백질을 만들었다. 이 피마자 단백질의 용해도는 ${\varepsilon}$-아미노기의 숙시닐화 및 아세틸화로 $pH\;7{\sim}8$에서 현저하게 증가하였다. 아미노산 분석결과, 황-함유 아미노산과 L-리신이 제한 아미노산이었고, 아실화 과정은 아미노산 함량에 약간의 손실을 주었다. 파파인을 이용한 1 단계법 plastein 반응으로 피마자 단백질 또는 아실화 피마자 단백질과 DL-메티오닌 에틸 에스테르로부터 L-메티오닌 강화 피마자 단백질을 합성하였고, 이 방법으로 L-메티오닌 도입율은 50%였다. 피마자 단백질 및 수식된 피마자 단백질의 펩신에 의한 소화율은 모두 92% 정도였으나, 트립신에 의한 소화율은 숙시닐화 및 아세틸화 단백질이 현저하게 떨어져서 각각 42% 및 26%였다. 피마자 단백질의 단백질 효율은 L-메티오닌 강화로 카제인의 단백질 효율의 90%까지 향상되었으나, 피마자 단백질을 숙시닐화 및 아세틸화 하면 단백질 효율은 감소되어, 각각 카제인의 55% 및 69%였다.

• 한국식품과학회지
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• 제29권4호
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• pp.752-757
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• 1997

Papaya 유액 추출물에서 protein을 정제 회수하기 위해 papaya 유액으로부터 추출한 추출액을 경제적이고도 처리가 비교적 간단한 ethanol 침전방법을 사용하였다. 실험계획법에 의하여 설정된 3수준의 부분요인 실험법에 의하여 실시된 각 처리구의 실험 결과를 다중 회귀분석 방법에 의해 다음과 같은 모델식 $Y=7.4554+1.2657X_{2}+1.0552X_{3}-1.7407X_{4}-0.01666X_{2}\;^{2}-0.1134X_{4}^{2}$을 산출하였으며 pH는 다중 회귀분석에 의해 제거되었다. 반응표면 분석법(RSM)을 이용한 결과 protein을 회수하기 위한 최적 조건으로는 protein 농도가 38.2 mg/ml, 40%의 ethanol 농도, 침전 온도는 $-8^{\circ}C$였다. 이러한 최적 조건에 의한 회수율의 실험치는 68.97%로 예측치인 77.28%에 근접한 결과였다. 또한 유의 수준을 검정하는 자유도와 F-value도 양호한 값(p<0.001)을 나타내어 종속변수와 독립변수와의 상관관계가 뛰어남을 알 수 있으며 따라서 다중 회귀분석에 얻어진 모델식의 각 변수들 사이에는 유의성이 높음을 알 수 있었다.

• 한국식품영양과학회지
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• 제27권4호
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• pp.603-608
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• 1998

Lipid oxidation in ascidian was studied when fresh, deshelled and sliced meats were fermented for 50 days at 5$\pm$2$^{\circ}C$ with 8%(w/w) salt and 0.1% papain. Antioxidative effects of butylated hydroxytoluene(BHT) and carotenoid extracts from ascidian tunic on lipid oxidation and oxidationrelated discoloration of ascidian meat during fermentation were investigated. Changes in peroxide value, carbonyl value, thiobarbituric acid value, fatty acids composition, the loss of total carotenoid and sensory evaluation were determined to assess the rancidity. Peroxide and carbonyl values in BHT and carotenoid extract treatments increased less than those of the control during fermentation. TBA value increased until 30 days, hereafter tended to decrease a little in the control during fermentation. TBA value increased until 30 days, hereafter tended to decrease a little in the control but it increased slowly until 40 days in cases of 0.02% BHT or 0.02% BHT with 0.05% carotenoid added. Fatty acids of fresh ascidian composed of polyenoic acid, saturated acid and monoenoic acid of 51.5%, 28.1% and 20.7%, respectively. Saturated fatty acids(C16:0, C14:0, C18:0) and monoenoic acids(C18:1, C16:1) increased while polyenoic acids(C20:5, C22:6) decreased during fermentation. Carotenoid was markedly degraded and discolored in the control during fermentation. But 0.02% BHT and 0.05% carotenoid treatments had bright color like fresh meat during 40 days. The results of sensory evaluation during the fermentation also convinced the retard of discoloration by the addition of BHT and carotenoid.