• Title/Summary/Keyword: Functional peptide

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Preparation for Calcium and Iron-binding Peptides from Rice Bran Protein Hydrolysates (미강 단백질 가수분해물로부터 Ca, Fe 결합된 peptide 제조)

  • Jeon, So-Jeong;Lee, Ji-Hye;Song, Kyung-Bin
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.174-178
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    • 2010
  • Calcium and iron binding peptides were prepared by enzymatic hydrolysis and ultrafiltration of rice bran protein (RBP), which was isolated from defatted rice bran by phytase and xylanase treatment and ultrasonication. The isolated RBP had a molecular weight in the range of 10-66 kDa. The extracted proteins were hydrolyzed using Flavourzyme for 6 hr. After ultrafiltration under 5 kDa as molecular weight, the peptides were fractionated into 4 peaks by Sephadex G-15 gel permeation chromatography, and each fraction was determined for calcium and iron binding activity. As the result, Fl and F2 fractions were the best candidate for calcium and iron chelation, respectively. These results suggest that the calcium and iron binding peptides can be used as functional food additives in food industry.

Expression of Antihypertensive Peptide, His-His-Leu, as Tandem Repeats in Escherichia coli

  • Jeong, Do-Won;Shin, Dong-Seok;Ahn, Chang-Won;Song, In-Sang;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.952-959
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    • 2007
  • His-His-Leu (HHL), a tripeptide derived from a Korean soybean paste, is an angiotensin-I-converting enzyme (ACE) inhibitor. We report here a method of producing this tripeptide efficiently by expressing tandem multimers of the codons encoding the peptide in E. coli and purifying the HHL after hydrolysis of the peptide multimers. The HHL gene, tandemly multimerized to a 40-mer, was ligated with ubiquitin as a fusion gene (UH40). UH40 was inserted into vector pET29b; the UH40 fusion protein was then produced in E. coli BL21. The recombinant UH40 protein was purified by cation-exchange chromatography with a yield of 17.3mg/l and analyzed by matrixassisted laser desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry and protein N-terminal sequencing. Leucine aminopeptidase was used to cleave a 405-Da HHL monomer from the UH40 fusion protein and the peptide was purified using reverse-phase high-performance liquid chromatography (HPLC) on a C18 HPLC column, with a final yield of 6.2mg/l. The resulting peptide was confirmed to be HHL with the aid of MALDI-TOF mass spectrometry, glutamine-TOF mass spectrometry, N-terminal sequencing, and measurement of ACE inhibiting activity. These results suggest that our production method is useful for obtaining a large quantity of recombinant HHL for functional antihypertensive peptide studies.

Identification of a Transferrin Receptor-binding Peptide from a Phage-displayed Peptide Library (파지-펩타이드 문고로부터 트랜스페린 수용체에 결합하는 펩타이드 탐색)

  • Kim, Sung-Il;Choi, Suk-Jung
    • Journal of Life Science
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    • v.18 no.3
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    • pp.298-303
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    • 2008
  • Using a phage peptide library approach, we have isolated a peptide ligand that binds to transferrin receptor on the surface of human melanoma cell, B16F10. The library was first screened twice by recovering internalized phages and was further screened three times by competitively eluting transferrin receptor-specific phages with human transferrin among the phages bound to the cell surface. The peptides displayed by the selected phages were fused to translocation and catalytic domain of Pseudomonas exotoxin to prepare recombinant toxins. After estimating cytotoxicity of each recombinant toxin toward B16F10 cell, seven clones were selected. Sequence analysis revealed that one of the clones displayed a peptide which had a significant sequence homology with human transferrin. The peptide was chemically synthesized and was shown to be functional in delivering cytotoxic agents into B16F10 cell via interaction with transferrin receptor.

Characterization of a New Anti-dementia β-secretase Inhibitory Peptide from Arctoscopus japonicus

  • Park, Seul Bit Na;Kim, Sung Rae;Byun, Hee-Guk
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.220-227
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    • 2018
  • Amyloid plaque is a product of aggregation of ${\beta}$-amyloid peptide ($A{\beta}$) and is an important factor in the pathogenesis of Alzheimer's Disease (AD). $A{\beta}$ is a major component of amyloid plaque and vascular deposits in the AD brain. The enzyme ${\beta}$-secretase is required for the production of $A{\beta}$; thus, prevention of the formation of $A{\beta}$ through the inhibition of ${\beta}$-secretase is a major focus in the study of the treatment of AD. In this study, we investigated ${\beta}$-secretase inhibitory activity of an Arctoscopus japonicus peptide. An Alcalase hydrolysate had the highest ${\beta}$-secretase inhibitory activity. A ${\beta}$-secretase inhibitory activity peptide was separated using ion exchange column chromatography (carboxy-methyl: CM, quaternary methyl ammonium: QMA) and reverse phase high performance liquid chromatography (RP-HPLC) on a C18 column. The $IC_{50}$ value of the purified peptide was $248.2{\pm}1.73{\mu}g/mL$. The ${\beta}$-secretase inhibitory peptide was identified as a six amino acid residue of Gly-Pro-Val-Gly-Ala-Pro (MW: 497.27 Da). In cell viability experiments, the final purified fraction, the carboxy-methyl ion exchange column fraction (CM-F1) showed no significant cytotoxic effect in SH-SY5Y cells at concentrations below $100{\mu}g/mL$ in 24 h. The results of this study suggest that peptides separated from Arctoscopus japonicus may be beneficial as ${\beta}$-secretase inhibitor compounds in functional foods.

Influence of Hermetia illucens Larvae-Derived Functional Feed Additives on Immune Function of Broilers (동애등에(Hermetia illucens)유충 유래 기능성 사료첨가제가 육계 면역기능에 미치는 영향)

  • Ji, Sang-Yun;Park, Kwan-Ho;Kim, Ki-Hyun;Lee, Heui-Sam;Choi, Gyu-Sung;Lim, Yeon-Ji;Yu, Ri;Choi, In-Hag;Chung, Tae-Ho
    • Journal of Environmental Science International
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    • v.27 no.12
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    • pp.1305-1307
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    • 2018
  • We investigate the effects of the immune function (HI titer) in broilers fed diets containing Hermetia illucens (H. illucens) peptide extract over a 40-day period. Twenty-four broiler chicks (Arbor Acres, 1 d old) were divided into four groups and fed different diets (control, 0.1%, 0.5%, and 1% H. illucens peptide extract). To evaluate HI titer, all broilers were vaccinated with H9H2 vaccine subcutaneously on the lateral thorax, according to the manufacturer's recommendations. Similar HI titer was observed with 1% H. illucens peptide extract treatment compared to the control after 40 days (p>0.01). Groups fed 0.5% H. illucens peptide extract demonstrated the most effective immune effects (p<0.01), followed by groups fed 0.1% H. illucens peptide extract. In conclusion, using 0.1% or 0.5% H. illucens peptide extract before or after vaccination improved HI titer immune function in broilers.

Ubiquitin Fusion System for Recombinant Peptide Expression and Purification: Application to the Cytoplasmic Domain of Syndecan-4

  • Chae, Young-Kee;Lee, Ha-Yan;Lee, Weon-Tae
    • Bulletin of the Korean Chemical Society
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    • v.28 no.9
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    • pp.1549-1552
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    • 2007
  • The cytoplasmic domain of syndecan-4, a type I transmembrane heparan sulfate proteoglycan, was overexpressed as a fused form with the ubiquitin molecule in Escherichia coli, and the fusion protein was purified using immobilized metal affinity chromatography (IMAC). The cytoplasmic domain was released from its fusion partner by using yeast ubiquitin hydrolase (YUH), and subsequently purified by reverse phase chromatography. The integrity of the resulting peptide fragment was checked by MALDI-TOF and NMR spectroscopy. The yield of the peptide was 3.0-1.5 mg per liter in LB or minimal medium, respectively. The recombinant expression and purification of this domain will enable us its structural and functional studies using multidimensional NMR spectroscopy.

Self-Assembled Peptide Structures for Efficient Water Oxidation

  • Lee, Jae Hun;Lee, Jung Ho;Park, Yong Sun;Nam, Ki Tae
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.280-280
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    • 2013
  • In green plants, energy generation is accomplished through light-harvesting photosystem, which utilize abundant visible light and multi-stepwise redox reaction to oxidize water and reduce NADP+, transferring electrons efficiently with active cofactors1. Inspired by natural photosynthesis, artificial solar water-splitting devices are being designed variously. However, the several approaches involving immobilization2, conjugation3, and surface modification4 still have limitations. We have made artificial photosynthesis templates by self-assembling tyrosine-based peptide to mimick photosystem II. Porphyrin sensitizer absorbing blue light strongly was conjugated with the templates and they were hybridized with cobalt oxide through the reduction of cobalt ions in an aqueous solution. The formation of hybrid templates was characterized using TEM, and their water oxidation performance was measured by fluorescence oxygen probe. Our results suggest that the bio-templated assembly of functional compounds has a great potential for artificial photosynthesis.

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In Vivo/In Vitro Properties of Novel Antioxidant Peptide from Pinctada fucata

  • Ma, Yongkai;Huang, Kehui;Wu, Yanyan
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.33-42
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    • 2021
  • Due to the potential of antioxidants to scavenge free radicals in human body, it is important to be able to prepare antioxidant peptides that meet the industrial requirements for cosmetics and food. Here, we determined in vivo/in vitro activities of antioxidant peptide from P. fucata (PFAOP) prepared by bio-fermentation method. The antioxidant property test results showed the DPPH, hydroxyl, superoxide radical-scavenging, and cellular antioxidant activity. EC50 values of PFAOPs were 0.018 ± 0.005, 0.126 ± 0.008, 0.168 ± 0.005, and 0.105 ± 0.005 mg/ml, respectively, exhibiting higher antioxidant activities than glutathione (p < 0.05). Moreover, anti-proliferation and cytotoxicity activity results illustrated PFAOP has a potent anti-proliferative activity against HepG2, Caco-2, and MCF-7 carcinoma cells with no cytotoxicity. Moreover, the protocols we developed in this work demonstrated several excellent advantages in PFAOP preparation compared to enzymatic hydrolysis or chemical synthesis methods and provide a theoretical foundation for higher-value application of marine-derived functional peptides.

The Effects of Combination of Functional Beverage(Garcinia Cambogia, L-Carnitine, and Soy Peptide) and Exercise on the Improvement of Body Fat

  • Kim, Kyung-Soo;Jung, Ju-Hae;Song, Chan-Hee;Sung, Bong-Ju
    • Biomolecules & Therapeutics
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    • v.11 no.2
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    • pp.99-108
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    • 2003
  • Background: There is abundance of studies on the decrease in body fat through limited calorie intake or exercise; however, studies focusing simultaneously on functional foods and exercise are rare. This study was aimed to identify the effects of combination of a functional beverage and exercise on body composition and biochemical metabolic profiles. Method: Eighty-one healthy volunteers (69 females aged 19 to 50 years and 12 males aged 19 to 55 years), who maintained their body weight stable with 23 or higher in BMI (kg/$m^2$) and 25% or higher body fat for the last three months, were recruited in the study through written advertisement. All the subjects gave their informed consent, and the study was conducted in accordance with the Declaration of Helsinki. The study design was a randomized double-blind placebo-controlled parallel group design. All participants were given 12-week programed-exercise, which was performed 3 times a week. One bottle (100 ml) of test (Garcinia cambogia 300 mg, L-carnitine 20 mg, Soy peptide 1,000 mg) or placebo solution was given daily 30 min before each session of programed-exercise. BMI (body mass index), %fat, local fat amount (visceral and mid-thigh), waist circumference, skin fold thickness and some biochemical metabolic parameters like glucose, insulin and free fatty acids, etc in the blood were measured and compared before and after 12-week intervention within groups as well as between groups according to the protocol Results: Twenty six volunteers were dropped out and fifty five volunteers completed the study. At the end of 4, 8, and 12 weeks, approximately 1.98%, 3.00% and 3.50% losses of initial body weight were observed, respectively, in the test group (P<0.01), and 0.29, % ,0.74%, 1.60%, respectively, in the placebo group (P>0.05). BMI changed by 2.40%, 3.41% and 4.46%, respectively, in the test group, and 0.38%, 0.95% and 1.75%, respectively, in the placebo group, at each period of time. The reductions of body weight and BMI were significantly higher in the test group than in the placebo group at each period of time (P<0.05). Conclusions: It is thought that the combination of functional beverage, which contains mostly garcinia cambogia, L-carnitine, and soy peptide, and exercise have synergy effects on reducing body fat.

A novel L-ascorbic acid and peptide conjugate with increased stability and collagen biosynthesis

  • Choi, Ho-Il;Park, Jong-Il;Kim, Heung-Jae;Kim, Dong-Won;Kim, Soung-Soo
    • BMB Reports
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    • v.42 no.11
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    • pp.743-746
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    • 2009
  • L-ascorbic acid (Vitamin C) and peptide are both useful compounds for collagen biosynthesis in cosmeceuticals (cosmetic and pharmaceutical fields). The instability of these compounds, however, limit their application in these industries. In this report, we describe the development of a novel compound, Stabilized Ascorbyl Pentapeptide (SAP), which physically is much more stable than L-ascorbic acid in water. The inhibitory effects of this SAP compound on tyrosinase and melanin synthesis is comparable to that of L-ascorbic acid. Importantly, the SAP compound displays no cytotoxicity at a high concentration (5 mM). The ability of SAP to promote collagen biosynthesis is greater than that of L-ascorbic acid or the KTTKS peptide alone. Considering the in vitro stability and functional effects, our data strongly suggest that the SAP compound is a good candidate not only as a cosmetic ingredient, but also as a wound healing agent.