• Title/Summary/Keyword: Functional peptide

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FUNCTIONAL EXPRESSION OF A PEPTIDE TRANSPORTER IN XENOPUS OOCYTES

  • Oh, Doo-Man
    • Journal of Pharmaceutical Investigation
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    • v.23 no.3
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    • pp.31-40
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    • 1993
  • It is essential to clone the peptide transporter in order to obtain better understanding of its molecular structure, regulation, and substrate specificity. Characteristics of an endogenous peptide transporter in oocytes were studied along with expression of an exogenous proton/peptide cotransporter from rabbit intestine. And further efforts toward cloning the transporter were performed. The presence of an endogenous peptide transporter was detected in Xenopus laevis oocytes by measuring the uptake of $0.25\;{\mu}M\;(10\;{\mu}Ci/ml)\;[^3H]-glycylsarcosine$ (Gly-Sar) at pH 5.5 with or without inhibitors. Uptake of Gly-Sar in oocytes was significantly inhibited by 25 mM Ala-Ala, Gly-Gly, and Gly-Sar (p<0.05), but not by 2.5 mM of Glu-Glu, Ala-Ala, Gly-Gly, Gly-Sar and 25 mM glycine and sarcosine. This result suggests that a selective transporter is involved in the endogenous uptake of dipeptides. Collagenase treatment of oocytes used to strip oocytes from ovarian follicles did not affect the Gly-Sar uptake. Changing pH from 5.5 to 7.5 did not affect the Gly-Sar uptake significantly, suggesting no dependence of the endogenous transporter on a transmembrane proton gradient. An exogenous $H^+/peptide$ cotransporter was expressed after microinjection of polyadenylated messenger ribonucleic acid $[poly\;(A)^+-mRNA]$ obtained from rabbit small intestine. The Gly-Sar uptake in mRNA-injected oocytes was 9 times higher than that in water-injected oocytes. Thus, frog oocytes can be utilized for expression cloning of the genes encoding intestinal $H^+/peptide$ cotransporters. Using the technique size fractionation of mRNA was sucessfully obtained.

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Purification and characterization of β-secretase inhibitory peptide from sea hare (Aplysia kurodai) by enzymatic hydrolysis

  • Lee, Jung Kwon;Kim, Sung Rae;Byun, Hee-Guk
    • Fisheries and Aquatic Sciences
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    • v.21 no.5
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    • pp.13.1-13.8
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    • 2018
  • Amyloid plaque, also called senile plaque, the product of aggregation of ${\beta}$-amyloid peptides ($A{\beta}$), is observed in brains of the patients with Alzheimer's disease (AD) and is one of the key factors in etiology of the disease. In this study, hydrolysates obtained from the sea hare (Aplysia kurodai) were investigated for ${\beta}$-secretase inhibitory peptide. The sea hare's muscle protein was hydrolyzed using six enzymes in a batch reactor. Trypsin hydrolysate had highest ${\beta}$-secretase inhibitory activity compared to the other hydrolysates. ${\beta}$-secretase inhibitory peptide was separated using Sephadex G-25 column chromatography and high-performance liquid chromatography on a C18 column. ${\beta}$-secretase inhibitory peptide was identified as eight amino acid residues of Val-Ala-Ala-Leu-Met-Leu-Phe-Asn by N-terminal amino acid sequence analysis. $IC_{50}$ value of purified ${\beta}$-secretase inhibitory peptide was $74.25{\mu}M$, and Lineweaver-Burk plots suggested that the peptide purified from sea hare muscle protein acts as a competitive inhibitor against ${\beta}$-secretase. Results of this study suggest that peptides derived from sea hare muscle may be beneficial as anti-dementia compounds in functional foods or as pharmaceuticals.

Purification of Angiotensin I-Converting Enzyme Inhibitory Peptide from Squid Todarodes pacificus Skin (오징어(Todarodes pacificus) 껍질로부터 Angiotensin I 전환효소 저해 펩티드의 분리 정제)

  • Lee, Jung-Kwon;Jeon, Joong-Kyun;Byun, Hee-Guk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.2
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    • pp.118-125
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    • 2011
  • In this study, an angiotensin I-converting enzyme (ACE) inhibitor from squid skin was purified and characterized. Squid (Todarodes pacificus) skin protein isolates were hydrolyzed using six commercial proteases: alcalase, ${\alpha}$-chymotrypsin, neutrase, papain, pepsin, and trypsin. The peptic hydrolysate had the highest ACE inhibitory activity. The ACE inhibitory peptide was purified using Sephadex G-25 column chromatography and reverse phase high-performance liquid chromatography (HPLC) with a $C_{18}$ column. The purified ACE inhibitory peptide was identified and sequenced, and found to consist of seven amino acid residues: Ser-Ala-Gly-Ser-Leu-Val-Pro (657Da). The $IC_{50}$ value of the purified ACE inhibitory peptide was 766.2 ${\mu}M$, and Lineweaver-Burk plots suggested that the purified peptide acts as a noncompetitive ACE inhibitor. These results suggest that the ACE inhibitory peptide purified from the peptic hydrolysate of squid skin may be of benefit in developing antihypertensive drugs and functional foods.

Anti-proliferative Effect of a Novel Anti-oxidative Peptide in Hanwoo Beef on Human Colorectal Carcinoma Cells

  • Kim, Hye-Jin;Yang, Se-Ran;Jang, Aera
    • Food Science of Animal Resources
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    • v.38 no.6
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    • pp.1168-1178
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    • 2018
  • The present study aimed to characterise anti-oxidant peptides from water-soluble protein extracts of Hanwoo beef and evaluate their anti-proliferative effect on human colorectal carcinoma cells (HCT116). Antioxidant peptides were purified from the low-molecular-weight fraction (<3 kDa) of Hanwoo beef extract. Antioxidant activity of peptide fractions was determined using the oxygen radical absorbance capacity (ORAC) assay. Purified peptide (P3) displayed higher ORAC activity than the low-molecular-weight fraction ($202.66{\mu}M\;TE/g$ vs $167.38{\mu}M\;TE/g$ of dry matter, respectively) (p<0.05). The peptide sequence of P3 was Cys-Cys-Cys-Cys-Ser-Val-Gln-Lys (888.30 Da). The novel peptide P3, at $250{\mu}g/mL$, also significantly inhibited HCT116 cell proliferation up to 25.24% through phosphorylation of ERK, JNK, and p38 kinase (p<0.05). Hence, antioxidant peptide P3 from Hanwoo beef extract can be used as an antioxidative and anticancer agent in the functional food industry.

Development of High Functional Collagen Peptide Materials using Skate Skins (홍어껍질을 이용한 고기능성 콜라겐 펩타이드 소재 개발)

  • Baek, Jang-Mi;Kang, Keon-Hee;Kim, Sang-Ho;Noh, Jeong-Sook;Jeong, Kap-Seop
    • Journal of Environmental Science International
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    • v.25 no.4
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    • pp.579-588
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    • 2016
  • 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.

A novel nicotinoyl peptide, nicotinoyl-LVH, for collagen synthesis enhancement in skin cells

  • Kim, Hyoung Shik;Ryu, Seung Hwan;Kim, Hye In;Cho, Seung Hee;Lee, Jeong Hun;Han, Byung Suk;Lee, Kyung Rok;Moh, Sang Hyun
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.239-242
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    • 2016
  • A novel Nicotinoyl fused peptide, Nicotinoyl-LVH, was synthesized by solid phase peptide synthesis method, purified, and tested in cultured skin cells. Nicotinoyl-LVH enhanced the expression level of collagen mRNA and its fragments in fibroblasts. These data show that this novel Nicotinoyl peptide is a promising biomaterial in the anti-aging functional cosmetic market.

The Novel Angiotensin I Converting Enzyme Inhibitory Peptide from Rainbow Trout Muscle Hydrolysate

  • Kim, Sung-Rae;Byun, Hee-Guk
    • Fisheries and Aquatic Sciences
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    • v.15 no.3
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    • pp.183-190
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    • 2012
  • The purpose of this study was the purification and characterization of an angiotensin I converting enzyme (ACE) inhibitory peptide purified from enzymatic hydrolysates of rainbow trout Oncorhynchus mykiss muscle. After removal of lipid, the approximate composition analysis of the rainbow trout revealed 24.4%, 1.7%, and 68.3% for protein, lipid, and moisture, respectively. Among six hydrolysates, the peptic hydrolysate exhibited the highest ACE inhibitory activity. We attempted to purify ACE inhibitory peptides from peptic hydrolysate using high performance liquid chromatography on an ODS column. The $IC_{50}$ value of purified ACE inhibitory peptide was $63.9{\mu}M$. The amino acid sequence of the peptide was identified as Lys-Val-Asn-Gly-Pro-Ala-Met-Ser-Pro-Asn-Ala-Asn, with a molecular weight of 1,220 Da, and the Lineweaver-Burk plots suggested that they act as a competitive inhibitor against ACE. Our study suggested that novel ACE inhibitory peptides purified from rainbow trout muscle protein may be beneficial as anti-hypertension compounds in functional foods.

A Clinical Study for the Efficacy and Safety of Functional Cosmetics Containing peptide Scolopendrasin-I isolated from the Scolopendra subspinipes mutilans in Patients with Dry Skin due to Mild Atopic Dermatitis (아토피성 피부로 건조함을 가진 대상자에 대한 왕지네로부터 분리한 펩타이드 Scolopendrasin-I 함유 기능성 화장품의 기능성 및 안전성을 평가하기 위한 임상적 연구)

  • Park, Hye-Su;Kim, Tae-Jun;Park, Shin-Yong;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.32 no.3
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    • pp.23-36
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    • 2019
  • Objectives : The purpose of this study is to confirm the efficacy and safety of "Functional cosmetics containing peptide Scolopendrasin-I isolated from the Scolopendra subspinipes mutilans" on dry skin due to mild atopic dermatitis. Methods : A total of 24 patients who visited Semyung Oriental Medical Center from November 1st, 2018 to December 20th, 2018 were included in the study. In this study, the patients were treated with Functional cosmetics containing peptide Scolopendrasin-I isolated from the Scolopendra subspinipes mutilans. For 4 weeks of gross examination, instrumental assessment were made before and after the study to evaluate how well the products for treatment group in recovering the dry skin barriers by mild atopic dermatitis. Results : 1. In the primary endpoint, Skin Hydration showed a statistically significant increase in treatment group in baseline, 2 weeks and 4 weeks. 2. In the primary endpoint, Transepidermal Water Loss(TEWL) showed a statistically significant decrease in treatment group between Baseline and 4 weeks. 3. In the secondary endpoint, Skin pH showed a statistically significant decrease in treatment group between baseline and 4 weeks. 4. To evaluate the safety of the products for the human body, Adverse events, SCORAD Index Assessment were conducted; There were no severe adverse events during this study. And SCORAD Index showed a statistically significant decrease in treatment group in baseline, 2 weeks and 4 weeks. Therefore, it is suggested that products, if used for certain period, should be safe for the human body. Conclusions : According to the above experiments, it is suggested that "Functional cosmetics containing peptide Scolopendrasin-I isolated from the Scolopendra subspinipes mutilans" should be effective for dry skin due to mild atopic dermatitis.

Structural and Functional Study of Antimicrobial Peptide Using NMR Spectroscopy

  • Suh, Jeong-Yong;Lee, Young-Tae;Park, Byong-Seok
    • Proceedings of the Korean Biophysical Society Conference
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    • 1997.07a
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    • pp.14-14
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    • 1997
  • Structure of potent derivatives of gaegurin, an antimicrobial peptide from Korean frog, is studied by CD and NMR spectroscopy. Gaegurin did not show any secondary structure in aqueous environment, but adopted ${\alpha}$-helix in aqueous TFE solution, SDS and liposome buffer. NMR study showed distinct difference in stability near proline residue in helix.(omitted)

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Interactions of Membrane and PMAP-23 Studied by $^{31}P$ solid-state NMR Spectroscopy

  • Kim, Si-Won;Kim, Suhk-Mann
    • Journal of the Korean Magnetic Resonance Society
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    • v.11 no.2
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    • pp.110-114
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    • 2007
  • [ $^{31}P$ ] powder pattern spectra were measured to investigate the aspects of the interaction between the MLV (Multilamellar vesicle) and PMAP-23, a membrane of cathelicidin family and then CSAs(chemical shift anisotropy) were calculated to indentify the extent of perturbation of phospholipid mobility by the peptides. We found that acidic phospholipid interacts strongly with PMAP-23, and the analogues which modified to increase the amphipathic property showed that larger change of CSA. The analogue which introduced positive charge showed the same effects with amphipathic property.

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