• Journal of Life Science
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• 제12권2호
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• pp.53-56
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• 2002

The baculovirus expression vector system (BEVS) is one of the powerful heterologous protein expression systems using insect cells. As a result this has become a hot issue in the fleld of biotechnology. The advantage of the BEVS is that the large-scale production of heterologous proteins, which undergo posttranslational modification in the endoplasmic reticulum (ER), can be accomplished. Altrough posttranslational modification of heterologous proteins in insect cells is more similar to mammalian cells than yeast, it is not always identical. Therefore, aggregation and degradation can sometimes occur in the ER. To produce a high level of bioactive heterologous proteins using BEVS in insect cells, the prerequisite is to completely understand the posttranslational conditions that determine how newly synthesized polypeptides are folded and assembling with ER chaperones in the ER lumen. Here, we provide information on current BEVS problems and the possibility of successful heterologous protein production from mammalian cells.

• 한국축산식품학회지
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• 제44권4호
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• pp.885-898
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• 2024

Ovomucin (OM), which has insoluble fractions is a viscous glycoprotein, found in egg albumin. Enzymatic hydrolysates of OM have water solubility and bioactive properties. This study investigated that the immunostimulatory effects of OM hydrolysates (OMHs) obtained by using various proteolytic enzymes (Alcalase^®, bromelain, α-chymotrypsin, Neutrase^®, pancreatin, papain, Protamax^®, and trypsin) in RAW 264.7 cells. The results showed that OMH prepared with pancreatin (OMPA) produced the highest levels of nitrite oxide in RAW 264.7 cells, through upregulation of inducible nitric oxide synthase mRNA expression. The production of pro-inflammatory cytokines such as tumor necrosis factor-α and interleukin-6 were increased with the cytokines mRNA expression. The effect of OMPA on mitogen-activated protein kinase signaling pathway was increased the phosphorylation of p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase in a concentration-dependent manner. Therefore, OMPA could be used as a potential immune-stimulating agent in the functional food industry.

• Journal of Ginseng Research
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• 제40권4호
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• pp.400-408
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• 2016

Background: Ginsenoside Rh2 (GRh2) is the main bioactive component in American ginseng, a commonly used herb, and its antitumor activity had been studied in previous studies. PDZ-binding kinase/T-LAK cell-originated protein kinase (PBK/TOPK), a serine/threonine protein kinase, is highly expressed in HCT116 colorectal cancer cells. Methods: We examined the effect of GRh2 on HCT116 cells ex vivo. Next, we performed in vitro binding assay and in vitro kinase assay to search for the target of GRh2. Furthermore, we elucidated the underlying molecular mechanisms for the antitumor effect of GRh2 ex vivo and in vivo. Results: The results of our in vitro studies indicated that GRh2 can directly bind with PBK/TOPK and GRh2 also can directly inhibit PBK/TOPK activity. Ex vivo studies showed that GRh2 significantly induced cell death in HCT116 colorectal cancer cells. Further mechanistic study demonstrated that these compounds inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 (ERK1/2) and (H3) in HCT116 colorectal cancer cells. In vivo studies showed GRh2 inhibited the growth of xenograft tumors of HCT116 cells and inhibited the phosphorylation levels of the extracellular regulated protein kinases 1/2 and histone H3. Conclusion: The results indicate that GRh2 exerts promising antitumor effect that is specific to human HCT116 colorectal cancer cells through inhibiting the activity of PBK/TOPK.

• Asian-Australasian Journal of Animal Sciences
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• 제32권2호
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• pp.265-273
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• 2019

Objective: Terminalia arjuna plant, specially its leaves, bark, and roots, are widely used in traditional herbal medicine due to presence of bioactive components and being a rich source of natural antioxidants. But its fruit has not been used for any such purposes despite its potential to retard oxidation. Hence, the antioxidant potential of Arjuna fruit extract (AFE) in retarding lipid and protein oxidation of raw ground pork was evaluated during refrigerated storage for 9 days. Methods: The AFEs were prepared using different solvents viz. ethanol (EH), water, ethanol: water (60:40) and methanol:hot water (60:40). The AFEs were analysed for total phenolic content (TPC), 2, 2-diphenyl-1-picrylhydrazyl radical scavenging activity and reducing power. Water extract (WE) and ethanol-water extract (EH-WE) were selected and incorporated at 1.0% into freshly minced pork meat and compared with a synthetic antioxidant, in retarding lipid and protein oxidation during storage. Results: The TPC in AFEs using different solvents ranged from 11.04 to 16.53 mg gallic acid equivalents/g and extracts exhibited appreciable scavenging activity ranging from 50.02% to 58.62%. Arjuna extracts significantly (p<0.05) improved the colour score of meat samples by reducing the formation of metmyoglobin during storage. Both the AFEs (WE and EH-WE) significantly (p<0.05) lowered the thiobarbituric acid reactive substances value, peroxide formation and formation of protein carbonyls in raw pork than control sample during storage. Upon sensory evaluation of all samples, it was found that AFE treatment could prolong the storage period of meat samples, without influencing the colour and odour score, up to 6 days. Conclusion: AFEs used at 1% improved the oxidative stability, colour and odour score and prolonged the refrigerated shelf life of ground pork up 6 days. Therefore, AFE could be explored as an alternative natural antioxidant in retarding lipid and protein oxidation in meat products.

• 대한화학회지
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• 제51권1호
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• pp.43-50
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• 2007

리포솜은 수십~수백 나노미터 크기의 소포체(small vesicle)로서 약물전달에 유용한 구조체이나 체내 혈류 순환계(blood circulatory system)에서 구조적 불안정성에 의해 체내 순환시간이 짧고 세망내피계(reticuloendothelial system)에 의해 소실되어 이를 개선하기 위한 다양한 연구가 시도되고 있다. 본 연구에서는 이당류(disaccharide)로서 락토오스와 슈크로스가 1,2-디스테아로일-sn-글리세로-3-포스포에탄올아민(1,2-Distearoylsn-glycero-3-phosphoethanolamine, DSPE)과 공유결합된 새로운 당-DSPE 유도체를 합성하고 이를 리포솜의 구성성분으로 하는 리포솜을 제조함으로써 리포솜의 표면이 이당으로 수식된 리포솜을 제조하였다. 제조된 리포솜의 입자크기는 대략 100 nm였으며, 표면전하 값은 당이 수식되지 않은 대조군 리포솜이 -10 mV를 나타내었으나 당-DSPE 유도체를 함유한 리포솜의 경우 리포솜 표면에 존재하는 당의 수산화기에 의하여 표면전하의 값은 -25 mV를 나타내었다. 리포솜 내부에 모델약물인 독소루비신(doxorubicin)의 로딩효율은 약 90%를 나타내었다. 당-DSPE 유도체 함유 리포솜의 in vitro 안정성은 혈청 내에서 단백흡착량의 변화 및 리포솜의 입자크기를 관찰하여 평가하였다. 당-인지질 유도체를 함유한 리포솜은 당-인지질 유도체를 함유하지 않은 리포솜 또는 폴리에틸렌글리콜(PEG) 수식 리포솜에 비해 단백흡착의 양과 혈청 내 입자크기의 변화가 적은 것으로 관찰되었다. 이당이 결합된 DSPE와 이를 함유한 리포솜은 혈액내 안정성이 향상되어 체순환계 내에서 장시간 순환이 가능한 약물전달체로서 유용할 것이라 사료된다.

• ALGAE
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• 제33권3호
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• pp.279-290
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• 2018

Microalgae have been utilized in diverse industries including aquaculture. Among the microalgae, dinoflagellates are known to have various bioactive compounds, and thus the interest in their application to industry has increased. In order to test their potential as food materials for aquaculture animals, the crude protein contents and compositions of amino acids of six dinoflagellates Heterocapsa rotundata (family Heterocapsaceae), Ansanella granifera (Suessiaceae), Alexandrium andersonii (Ostreopsidaceae), Takayama tasmanica (Brachidiniaceae), Takayama helix, and Gymnodinium smaydae (Gymnodiniaceae) belonging to diverse families were analyzed. The percentage of the amount of the crude protein relative to dry weight of T. tasmanica was the highest (65%) and that of A. andersonii was the lowest (26%). However, the highest percentage of total detected amino acids in crude protein was found in A. andersonii (98.2%). In all six dinoflagellates, glutamic acid was the most dominant amino acid in crude protein. However, the second main amino acid was aspartic acid for H. rotundata, A. granifera, T. helix, and G. smaydae, but were arginine and leucine for A. andersonii and T. tasmanica, respectively. Furthermore, T. tasmanica and T. helix did not have taurine and gamma-aminobutyric acid, whereas the other dinoflagellates possessed them. The percentages of essential amino acid contents of the dinoflagellates met the requirement levels for juvenile shrimps. In addition, the dinoflagellates were not toxic to the brine shrimp Artemia salina. Compared with the other microalgae reported so far, H. rotundata and A. andersonii can be used for arginine-rich diets, T. tasmanica for valine and leucine-rich diets, A. granifera for histidine-rich diets, T. helix for threonine-rich diets, and G. smaydae for lysine-rich diets. Therefore, based on their biochemical composition and toxicity to Artemia, the dinoflagellates could be used as essential amino acid sources for cultivating animals in the aquaculture industry.

• Journal of Dairy Science and Biotechnology
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• 제41권3호
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• pp.113-125
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• 2023

In this study, we explored the synergistic effects of whey protein concentrate (WPC) and soybean protein components after fermentation with lactic acid bacteria isolated from kimchi, and identified several peptides with desirable physiological functions, proteolysis, and immune effects. Antioxidant activity was determined using 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid, 1,1-diphenyl-2-picrylhydrazyl, ferric-reducing antioxidant power, and hydroxyl radical scavenging assays, followed by cross-validation of the four antioxidant activities. These assays revealed that samples with a 8:2 and 9:1 whey to soy ratio possessed higher antioxidant activity than the control samples. Antibacterial potency testing revealed high antibacterial activity in the 9:1 and 8:2 samples. Cytotoxicity testing of samples using 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide revealed that only the 10:0, 1:9, and 0:10 samples had <80% viable cells, indicating no significant cytotoxicity. Nitric oxide (NO) assays revealed that NO expression was reduced in 8:2, 5:5, and 0:10 protein ratio fermentations, indicating low inflammatory reaction stimulatory potential. Cytokine expression was confirmed using an enzyme-linked immunosorbent assay kit. The 8:2 sample had the lowest inflammatory cytokine (interleukin [IL]-1α, IL-6, and tumor necrosis factor-α) levels compared with the lipopolysaccharide-treated group. Amino acid profiling of the 8:2 sample identified 17 amino acids. These results suggest that inoculating and fermenting Lactobacillus plantarum DK203 and Lactobacillus paracasei DK209 with an 8:2 mixture of WPC and soybean protein releases bioactive peptides with excellent anti-inflammatory and antioxidant properties, making them suitable for functional food development.

• 생명과학회지
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• 제20권11호
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• pp.1718-1724
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• 2010

누에분말을 과일(키위, 파파야, 파인애플 및 배) 단백질 분해 효소로 반응시켜 생리활성작용 및 이화학적 특성을 조사하기 위하여 pH, 산도, 단백질 함량, 미네랄 함량, 지방산 조성, 단백질 패턴, 항산화 및 혈전용해 활성을 측정하였다. 누에분말은 각 과일 단백질 분해 효소로 $60^{\circ}C$에서 24시간 반응시켰다. 단백질 농도는 과즙액 처리 누에분말에서 약간 높은 것으로 나타났다. 누에분말의 주요 미네랄은 칼륨, 마그네슘, 칼슘 및 아연이였으며, 주요 지방산 조성은 linolenic acid, oleic acid 및 palmitic acid 였다. SDS-PAGE상의 단백질 패턴 분석에서 66-97 kDa 정도 크기의 누에 단백질 밴드가 파인애플, 파파야 및 배 반응에 의해서는 대부분 분해가 일어났지만, 키위 반응에 의해서는 거의 분해가 일어나지 않은 것으로 관찰되었다. 혈전용해 활성은 파파야 및 배 반응에 의한 누에분말에서만 나타났다. 항산화 활성은 0.1% 처리 농도에서 과일 단백질 분해 효소 반응에 의한 누에분말에서 미반응 누에분말 보다 증가하였으나, 시판 항산화제 BHT 처리보다는 활성이 많이 낮았다. 따라서 과즙액 처리 누에분말의 경우 반응 전 누에분말 보다 생리활성작용 및 이화학적 특성이 강화됨으로써 건강기능식품 소재로서의 가치가 증가된 것으로 사료되어 진다.

• 생약학회지
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• 제50권1호
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• pp.37-45
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• 2019

Portulaca oleracea L. (PL) has been used in traditional medicine herb for treatment of various diseases, such as diarrhea, dysentery, and skin inflammation. Previous studies have shown that the PL regulates the inflammation by inhibition of pro-inflammatory cytokines. Although PL might have improvement effects of intestinal function and bioactive effects, there are not enough studies to demonstrate. This study investigated the effects of KDC16-2 on the improvement of intestinal function and anti-inflammatory effects in vivo and in vitro. The improvement effect of intestinal function was measured fecal amount, water content and intestinal transit rate in KDC16-2 treated ICR mice. As results, compared with the control group, the KDC16-2 group showed a significant increase in wet fecal weight, dry fecal weight and fecal water content. The intestinal transit rate of KDC16-2 group was significantly increased. Based on the results, KDC16-2 is considered to have effects on improving intestinal function. The effect of anti-inflammatory demonstrated by using dextran sulfate sodium (DSS)-induced colitis mice. The mice were administered 3% DSS along with KDC16-2 (100, 300 mg/kg) for 14 days. DSS-induced colitis mice were significantly ameliorated in KDC16-2 treated group, including body weight loss, colon length shortening, tight junction protein of colon and histological colon injury. The levels of inflammatory mediators (IgG2a, IgA, C-reactive protein and Myeloperoxidase) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-${\alpha}$, Interleukin (IL)-6) which are involved in inflammatory responses were increased in the DSS-treated group as compared to those in the control group, and the levels were significantly decreased in the KDC16-2 groups. In addition, we investigated the impact of KDC16-2 on lipopolysaccharide (LPS)-induced inflammatory responses in J774A.1 cells. KDC16-2 inhibited production of prostaglandin E2 (PGE2) and reactive oxygen species (ROS). These results suggested that the KDC16-2 could effectively alleviate the dysfunction of intestinal and inflammatory mediators. Thus, these KDC16-2 can be potentially used as health functional food of intestinal.

• 대한화장품학회지
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• 제47권2호
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• pp.123-131
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• 2021

해양 홍조류 참도박(Grateloupia elliptica, G. elliptica)을 흑효모(Aureobasidium pullulans, A. pullulans)로 발효시킨 추출물들의 인간 피부각질세포주(HaCaT)에 대한 항염증 효과를 조사하였다. 흑효모로 발효한 참도박 추출물은 참도박 추출물에 비하여 총 폴리페놀(total polyphenol content)은 최대 2.7 배로 증가되었으며 총 플라보노이드(total flavonoid conten)는 최대 2.4 배 증가되었다. 인간 피부각질세포를 lipopolysaccharide (LPS) 또는 H₂O₂로 염증을 유도 한 후 참도박 추출물과 참도박 발효 추출물을 100 ㎍/mL 농도로 처리 한 결과 LPS 또는 H₂O₂ 만 처리한 세포군에 비하여 약 3 ~ 10% 정도 세포생존율이 증가되었다. 또한 염증 유발과 관련된 단백질 발현 변화를 조사한 결과 참도박 발효추출물이cyclooxygenase-2 및 70 kDa heat shock protein 의 발현을 현저히 감소시켰다. 따라서 흑효모로 발효한 참도박은 항염효과를 가진 피부수렴용 화장품 소재로 유용하다.