The addition of dipeptides and 1,2-bis (aminoacyl) hydrazine derivatives a level of 250 ppm to corn oil resulted in the retardation of the oxidaitive deterioration of the oil when it was stored in the oven at 70℃ during 5 days. Their antioxidant activities were investigated by UV absorbance of the corn oil at the wavelength of 234nm. 1,2-bis (aminoacyl) hydrazine derivatives showed higher antioxidant activity than normal dipeptides. Dipeptides containing phenyl ring with which is conjugable a-carbon radical showed antioxidant activity.

아세틸 헥사펩타이드 8 (AH8)은 보톡스 메커니즘을 응용한 주름 개선 펩타이드 소재로, 보톡스의 타겟인 synatosomal-associated protein 25 (SNAP25) N말단 서열을 모방하여 개발되었다. 주름 개선 효과가 보고되고 있지만 큰 분자량과 친수성 성질에 의하여 피부 흡수는 잘 되지 않는다는 문제가 있다. 따라서 피부 보습 성분 중에서 AH8의 피부 흡수를 증가시켜 줄 수 있는 물질을 탐색하였는데, 히알루론산(HA)이 AH8의 피부 흡수를 증가시켰다. 형광물질로 표지한 AH8만 MicropigⓇ skin 에 발라주면 대부분 각질을 투과하지 못하고 각질층에 존재하였다. 반면, HA를 함께 도포한 경우에는 각질층을 투과하여 표피, 진피로 흡수된 AH8가 증가하는 것을 형광 이미지 분석을 통해 확인했다. 특히 5 kDa 저분자량 HA가 500 kDa, 2000 kDa HA보다 피부 흡수를 더 많이 증가시켰다. HA가 피부 각질층에 미치는 영향을 푸리에변환 적외 분광법(Fourier-transform infrared spectroscopy, FTIR)으로 분석해보니, 고분자량 HA는 각질 수분 함량을 증가시키고, 저분자량 HA는 지질층의 유동성을 증가시키는 경향성이 있었다. 따라서 HA는 AH8의 피부 흡수를 증가시켜 주름 개선 효과를 향상시켜 줄 수 있을 것으로 기대된다.

Coronary artery complications are the most severe complications in Kawasaki disease (KD). Its cause is unclear but superantigens are considered to influence KD. This study aimed to determine whether coronary artery complications and level of Nterminal pro-brain natriuretic peptide (NT-proBNP) are associated with Mycoplasma pneumoniae (MP) in patients with KD. We studied 142 pediatric patients diagnosed with KD. MP immunoglobulin M (IgM) antibody examination was conducted. All patients underwent echocardiography, and coronary artery dilatation was defined as a coronary Z-score >2.0. We also evaluated their NT-proBNP findings. The independent t-test and Pearson chi-squared test were used to analyze betweengroup differences; a p-value <0.05 was considered statistically significant. Forty children were MP IgM positive. MP IgMpositive patients were older than MP IgM-negative patients. There was no significant difference in the clinical manifestations between the groups. Comparison of the mean Z-score of the coronary artery revealed that only the Z-score of the left anterior descending artery was significantly different between the groups. However, the number of patients with coronary artery dilatation was not significantly different between the groups. Our findings demonstrated no relationship between MP infection and coronary artery dilatation or NT-proBNP levels in patients with KD.

콜라겐 펩타이드(collagen peptide)는 단백질의 가수 분해물로서 주름 완화, 보습력 증대, 탄력 개선 등의 특정 피부 효능을 나타낼 수 있어 화장품 혹은 피부 개선 기능식품으로서 활용되고 있다. γ -aminobutyric acid (GABA)는 척추 동물의 뇌, 척수에 존재하는 신경전달 물질로서 수면의 질과 양을 개선해 준다고 알려져 있다. 본 연구에서는 콜라겐 펩타이드와 GABA의 복합물이 수면 장애가 있는 여성에게 8주 경구 섭취를 통해서 수면 및 피부 상태를 개선할 수 있는가에 관하여 확인하였다. 복합물(J85091900)은 8주간 연속 적으로 섭취 시, 수면장애지수(PSQI)가 유의적으로 감소하였으며, 수면 시간을 7% 증가시켰다. 또한, 피부 거 칠기, 눈가 주름 및 피부 수분량(capacitance)을 유의적으로 개선하였다. 이상의 결과에서 콜라겐과 GABA의 복합물은 복합 수면 장애에 따른 피부의 노화 현상으로부터 피부를 보호할 수 있음을 확인하여 먹는 화장품의 핵심 소재로 활용 가능함을 확인하였다.

In this study, a peptide exhibiting antioxidant activity was isolated from sandfish (Arctoscopus japonicus) roe hydrolysate (SRH) in order to evaluate their practical uses as materials for manufacturing functional foods. The A. japonicus roe protein was hydrolyzed using Collupulin MG, and isolation of antioxidant peptide was performed using ultrafiltration (UF), prep-HPLC, and RP-HPLC. The SRH with a molecular weight below 3 kDa constituted about 38% of the whole hydrolysate, and the fraction with a molecular weight below 3 kDa showed significantly greater antioxidant activity compared to the original SRH and other fractions. The isolation fold of the antioxidant peptide isolated from SRH throughout the four-step procedure was 7.11-fold, and protein yield was 14.8%. The DPPH radical scavenging activity of isolated antioxidant peptide was above 90% at a concentration of 1.0 mg/mL, which was similar to that of the Trolox at a concentration of 0.1 mg/mL. These results suggested that the antioxidant peptide derived from A. japonicus roe could be a useful additive for producing functional foods and protein supplements. However, it is necessary to perform further study the structural characteristics of this antioxidant peptide isolated from A. japonicus roe.

In this study, we produced the recombinant lunasin peptide using E. coli and P. pastoris, and evaluated biological activity of the recombinant lunasin peptide. Lunasin peptide was produced from E. coli transfected with pPGEX-lunasin expression vector and P. pastoris GS115 transfected with pPIC-lunasin expression vector. These recombinant lunasin peptides were similar to the synthetic lunasin peptide in the identification by LC-ESI-MS. In addition, the recombinant lunasin peptide from E. coli and P. pastoris was bound in the chromatin, and inhibited histone acetylation and the activity of histone acetyltransferase. These findings suggest that the production of the lunasin peptide using E. coli and P. pastoris will be useful for industrial utilization of lunasin peptide.

In this study, we produced the recombinant lunasin peptide using E. coli and P. pastoris, and evaluated biological activity of the recombinant lunasin peptide. Lunasin peptide was produced from E. coli transfected with pPGEX-lunasin expression vector and P. pastoris GS115 transfected with pPIC-lunasin expression vector. These recombinant lunasin peptides were similar to the synthetic lunasin peptide in the identification by LC-ESI-MS. In addition, the recombinant lunasin peptide from E. coli and P. pastoris was bound in the chromatin, and inhibited histone acetylation and the activity of histone acetyltransferase. These findings suggest that the production of the lunasin peptide using E. coli and P. pastoris will be useful for industrial utilization of lunasin peptide.

본 연구에서는 파이토케미컬 펩타이드 유도체를 포함하는 니오좀 및 리포좀 제형의 안정성을 조사하였 다. 여러 가지 조건에서 레시틴 혹은 계면활성제를 이용하여 제조된 리포좀 중에서 sodium palmitoyl sarcosinate와 마카다미아넛 오일을 이용한 니오좀의 안정성이 제일 우수하였다. 파이토케미컬 펩타이드 함유 니오좀 (N9)의 형태는 TEM으로 확인하였으며, 입자크기는 95.7 nm로 관찰되었다. N9의 안정성은 Turbiscan 결과 (0°와 45°)와 제타포텐셜(－78.19 mV)로 확인하였다. N9의 펩타이드 봉입률은 BCA assay 결과 65.2%로 관찰되었다.

The aim of this study was to investigate and development collagen peptide materials from skate skin. Protein and fat content of collagen peptide showed about 95% and 0.1%, respectively. Average molecular weight of collagen peptide was measured as 1,015. In the analysis of amino acid, glycine and hydroxy proline content in collagen peptide was 19.32% and 16.25%, respectively, showing a typical characteristics of the collagen peptide. In obese db/db mice ingested 500 mg/day of collagen peptide for 18 days, the amounts of food and water intake were decreased considerably, contents of triglyceride, total cholesterol were decreased significantly in white adipose tissue of db/db mice. The final yield of collagen peptide was 17.23% in the optimized process for mass production. These results indicate that collagen peptide from skate skin may serve as candidates of fat reduction in adipose tissue and could be used as functional food and cosmetic ingredients.

Neurokinin B (NKB) and neurokinin B related peptide (NKBRP) belong to tachykinin peptide family. They act as a neurotransmitter and/or neuromodulator. Mutation of NKB and/or its cognate receptor, NK3R resulted in hypogonadotropic hypogonadism in mammals, implying a strong involvement of NKB/NK3R system in controlling mammalian reproduction. Teleosts possess NKBRP as well as NKB, but their roles in fish reproduction need to be clarified. In this study, NKB and NKBRP coding gene (tac3) was cloned from Nile tilapia and sequenced. Based on the sequence, Nile tilapia NKB and NKBRP peptide were synthesized and their biological potencies were tested in vitro pituitary culture. The synthetic NKBRP showed direct inhibitory effect on the expression of GTH subunits at the pituitary level. This inhibitory effect was confirmed in vivo by means of intraperitoneal (ip) injection of synthetic NKB and NKBRP to mature female tilapia (20 pmol/g body weight [BW]). Both NKB and NKBRP had no effect on the plasma level of sex steroids, E2 and 11-KT. However, NKBRP caused declines of expression level of GnRH I, Kiss2 and tac3 mRNAs in the brain while NKB seemed to have no distinct effect. These results indicate some inhibitory roles of NKBRP in reproduction of mature female Nile tilapia, although their exact functions are not clear at the moment.

피부 노화는 피부 표면에 나타나는 현상으로 피부 표면의 미세구조와 연관이 있다. 피부 미세구조의 변화는 주름형성, 피부 톤 저하 등 다양한 피부 노화 현상을 유발하는 요인이 된다. 본 연구는 주름 기능성 성분인 세드롤(Cedrol)과 콜라겐 유래 펩타이드 성분을 이용하여 시험관 수준(in vitro)에서 타입Ⅲ 콜라겐 합성 시너지 효과를 평가하였다. 여성 피시험자를 대상으로 4주 동안 상기 원료가 함유된 크림제품을 사용한 후 피부 미세구조, 스타 형상(star configurations), 보습, 탄력, 피부 밝기, 윤기, 피부 톤, 투명도를 평가하였다. 시험관 수준의 세드롤과 펩타이드를 동시에 처리하면 우수한 수준의 타입Ⅲ 콜라겐 합성 시너지효과를 나타내었다. 그리고 크림제형을 이용한 인체적용 시험에서는 사용 4주 후부터 피부 미세구조, 스타형상, 윤기, 피부 톤, 보습, 탄력을 개선시켰으나, 피부 밝기 개선 효능은 나타나지 않았다. 본 연구를 통하여 기존 주름 기능성 성분인 세드롤과 콜라겐 유래 펩타이드는 세포 수준에서 타입Ⅲ 콜라겐 합성 시너지 효과가 있음을 확인하였고 이를 화장품에 적용하여 시험한 결과 피부 노화 개선에 도움을 주는 성분임을 확인하였다.

Sorghum seed is traditionally used as secondary food sources in addition to rice in Korea. While the hypoglycemia regulating phytochemicals have been found in sorghum seed, peptides related with hypoglycemia never been studied before. To obtain the peptide characteristics and the specifically high-expressed peptides in hypoglycemic sorghum seed, peptide profiles of seven hypoglycemic and five nonhypoglycemic sorghum lines bred in RDA were determined using surface-enhanced laser desorption/ionization time-offlight mass spectrometry (SELDI-TOF MS). The twelve sorghum lines exhibited 104 peptides on CM10 protein chip array (weak cation exchange) and 95 peptides on Q10 (weak cation exchange) in the molecular mass range from 2,000 to 20,000 Da. Heat map via supervised hierarchical clustering of the significantly different peptides (p < 0.01) in peak intensity among the 12 lines effectively revealed the specifically upregulated peptides in each line and distinguished between 7 hypoglycemic and 5 non-hypoglycemic lines. Through the comparison with hypoglycemic and non-hypoglycemic lines, 10 peptides including 2231.6, 2845.4, 2907.9, 3063.5, 3132.6, 3520.8, 4078.8, 5066.2, 5296.5, 5375.5 Da were specifically high-expressed in hypoglycemic lines at p < 0.00001. This study characterized seed peptides of 12 sorghums and found ten peptides highly expressed for hypoglycemic sorghum lines, which could be used as peptide biomarkers for identification of hypoglycemic sorghum.

SELDI-TOF MS를 활용한 저분자 펩타이드 분석을 위해 서는 분석시료에 대한 최적화 분석조건을 확립하는 것이 필수적으로 선행되어야 한다. 본 연구는 수수 종자 내 존재하는 10 kDa 이하의 저분자 펩타이드를 프로파일링하기 위하여 활용된 SELDI-TOF MS의 최적화 분석조건을 확립하는데 있다. 분석조건은 (1) 프로테인 칩: CM10(weak cationexchanger), Q10(strong anion exchanger), (2) 바인딩 버퍼의 희석배수: 1/2, 1/5, 1/10, 1/20, 1/50, 1/100, 1/200, (3)Q10의 바인딩 버퍼 강도: 10 mM, 100 mM, (4) 단백질 추출버퍼: sodium borate, sodium borate + acetone, phenol,TCA 버퍼로 하였다.1. 바인딩 버퍼의 희석배수는 CM10과 Q10 모두 1/20과1/50이 최적화로 나타났다.2. Q10의 바인딩 버퍼 강도는 농도가 약한 10 mM에서 더많은 피크가 검출되었다.3. SELDI-TOF MS 분석에 적합한 수수 종자단백질 추출버퍼로는 2~10 kDa 범위에서는 sodium borate 버퍼와10~20 kDa 범위에서는 phenol 버퍼로 분석되었다.

최근에는 콜라겐 펩타이드가 항상화효과를 통해 피부의 재생을 촉진시키는 효과를 한다는 연구가 보고되고있다. 본 구는 저분자 경구 콜라겐 펩타이드가 1550 nm fractional photothermolysis 레이저 치료 후 피부재생에 미치는 영향을 알아보고자 한다. 본 실험에 참여한 환자는 총 10명으로 실험군과 대조군 각 5명씩 무작위배정되었다. 대조군과 실험군 모두 동일하게 fractional photothermolysis 레이저를 시술받았다. 실험군에서는 경구 콜라겐 펩타이드를 하루에 1,000 mg씩 총 8주간 복용하였다. 치료 전후 모든 환자에서 피부의 탄성도, 경표피수분손실도, 홍반지수를 측정하였다. 피부탄성도는 실험군에서 대조군에 비해 통계적으로 유의하게 피부의 탄력성이 증가하였음을 확인하였다(p < 0.05). 경표피수분손실도는 두군간에 유의한 차이가 없었고, 홍반지수는 실험군에서 더 빨리 감소하는 결과를 보였다. 환자의 주관적 만족도평가에서 시술 4주 후 환자의 만족도는 실험군에서 평균 2.0점, 대조군에서 1.2점으로 나타났다. 저분자 콜라겐 펩타이드는 1550 nm fractional photothermolysis 레이저 치료 후 피부회복 촉진에 도움이 된다.

This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.

Targeted gene silencing is an essential component of plant biotechnology. RNA interference is often employed for targeted gene silencing in plants. However, it suffers from off-target effects and unstable gene suppression in many cases. In recent years, engineered nuclease-based tools, such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs), have been developed to induce site-specific genome modifications. However, these approaches require much time and labor for extensive screening of mutants. We have recently reported that the activities of dimeric transcription factors are competently suppressed by genome-encoded small interfering peptides (siPEPs) that competitively form nonfunctional heterodimers in plants. In addition, some splice variants of transcription factors also act in a similar manner to negatively regulate the activities of specific transcription factors. We designated the siPEP-mediated suppression of transcription factors peptide interference (PEPi). Based on our previous observations, we also developed an artificial siPEP (a-siPEP) approach and evaluated its application for the targeted inactivation of transcription factors in the dicot model, Arabidopsis, and monocot model, Brachypodium. We designed a series of potential a-siPEPs of two representative transcription factors SUPPRESSOR OF OVEREXPRESSOR OF CONSTANS 1 (SOC1) and AGAMOUS (AG) that function in flowering induction and floral organogenesis, respectively. Transgenic plants overproducing a-siPEPs displayed phenotypes comparable to those of gene-deficient mutants. Collectively, our data demonstrate that the siPEP tool is an efficient protein knockout system for inactivating specific transcription factors, and other multimeric enzymes and membrane transporters as well, in plants. We will discuss about the global application of the siPEP toll to other plant species and potential advantages over other gene manipulation tools.

Cereal seeds, sorghum, foxtail millet, hog millet, adlay, and corn are traditionally used as health assistant as well as energy supplying food in Korea. While beneficial phytochemicals to human have revealed in cereals, the information on peptides from cereals is far less accumulated than major reserve protein. Here, we analyzed peptide profiles using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS) in cereal seeds for construction of peptide information and attempted to develop peptide biomarkers for cereal identification. To optimize the analysis condition of SELDI-TOF MS, the effect of dilution factor on binding affinity to protein chips was tested using CM10 and Q10 arrays. Peptide clusters were significantly different at the level of 0.01 p-value. Peak spectra were the most stable in 1:50 of dilution factor in both chip arrays. Numbers of detected peak of 5 cereal seeds were 131 in CM10 and 74 in Q10 array. Each cereal was grouped as a cluster and well discriminated into different cluster in the level of 0.01 p-value. Numbers of potentially identified peptide biomarkers are 11, 13, 9, 5 and 12 in sorghum, foxtail millet, hog millet, adlay and corn, respectively. This study demonstrates that each cereal seed have own distinguishable specific peptides although their function are not identified yet in this study. In addition, the proteomic profiling using SELDI-TOF MS techniques could be a useful and powerful tool to discover peptide biomarker for discrimination and assess crop species, especially under 20 kDa.

Discovery, identification, and informatics of low molecular weight peptide are extensively rising in the field of proteomics research. In this study, we analyzed protein profiles to discover peptide based biomarker for twelve different soybean seeds with three different agronomic types using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). For optimization of SELDI-TOF MS in soybean seed proteome analysis, four different extraction buffers were tested with urea solubilization buffer, thiourea/urea solubilization buffer, phenol extraction buffer, and modified trichloroacetic acid (TCA)/acetone precipitation/urea solubilization extraction buffer. Two different type of ProteinChip arrays, cation exchange (CM10) and anion exchange (Q10), applied to profile peptides. Among the four different extraction buffers, phenol extraction was selected to protein extraction methodology. Numbers of detected peak cluster in twelve soybean seeds were 125 at CM10 and 90 at Q10 array in the mass range from 2 to 40 kDa. Among them, 82 peak clusters at CM10 and 33 peak clusters at Q10 array showed significantly different peak clusters at p<0.00004 (CM10) and p<0.00005 (Q10) among twelve different soybean cultivars. Moreover, 29 peak clusters at CM10 and 17 peak clusters at Q10 array were detected in all cultivars as an ‘universally existed peptide’. In comparison with three different agronomic types, total of 55 peak clusters (CM10) and 23 peak clusters (Q10) were significantly different peak clusters at p<0.00004 and p<0.0001, respectively. In these probability levels, soybean seeds were well discriminated into different cultivar and different type with each other. Also we could find several specific peptide biomarkers for agronomic type.

The objectives of present study were to characterize the peptides which were isolated from Korean fermented soybean paste, chungkukjang, and to determine their antioxidant activities. Four fractions were collected from the methanol extract of chungkukjang by using a recycling preparative HPLC. Among fractions, Fr-2 was identified to be highly potent free radical scavenging activity in the assay of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and nitroblue tetrazolium(NBT)-reduction inhibition. Base on antioxidant effects, fraction Fr-2 was employed for the refraction with a prep-column and separated into five fractions of which two fractions were identified to have higher antioxidant activity. To confirm the amino acid constituents of antioxidant fractions Fr-2-2 and Fr-2-3 were analyzed, and eight kinds of amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, lysine, histidine, and arginine were identified as the constituent amino acids. Antioxidant activities of the separated peptides were further assessed cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT), and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H2O2). Chungkukjang peptides have shown their ability to protect H4IIE rat hepatoma cells against H2O2- induced oxidative stress by concentration and time-dependent manner. Therefore, These results indicated that fermented soybean paste chungkukjang will be promoted the antioxidant and radical scavenging activities, and beneficial for health. The antioxidant peptide fractions Fr-2-2 and Fr-2-3 were denominated as P-NICS-1 and P-NICS-2, respectively. However, further studies were required to clarify their amino acid sequences and molecular properties, and physiological significances.