• Title/Summary/Keyword: peptide production

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Identification of duck liver-expressed antimicrobial peptide 2 and characterization of its bactericidal activity

  • Hong, Yeojin;Truong, Anh Duc;Lee, Janggeun;Lee, Kyungbaek;Kim, Geun-Bae;Heo, Kang-Nyeong;Lillehoj, Hyun S.;Hong, Yeong Ho
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.1052-1061
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    • 2019
  • Objective: This study was conducted to identify duck liver-expressed antimicrobial peptide 2 (LEAP-2) and demonstrate its antimicrobial activity against various pathogens. Methods: Tissue samples were collected from 6 to 8-week-old Pekin ducks (Anas platyrhynchos domesticus), total RNA was extracted, and cDNA was synthesized. To confirm the duck LEAP-2 transcript expression levels, quantitative real-time polymerase chain reaction was conducted. Two kinds of peptides (a linear peptide and a disulfide-type peptide) were synthesized to compare the antimicrobial activity. Then, antimicrobial activity assay and fluorescence microscopic analysis were conducted to demonstrate duck LEAP-2 bactericidal activity. Results: The duck LEAP-2 peptide sequence showed high identity with those of other avian species (>85%), as well as more than 55% of identity with mammalian sequences. LEAP-2 mRNA was highly expressed in the liver with duodenum next, and then followed by lung, spleen, bursa and jejunum and was the lowest in the muscle. Both of LEAP-2 peptides efficiently killed bacteria, although the disulfide-type LEAP-2 showed more powerful bactericidal activity. Also, gram-positive bacteria was more susceptible to duck LEAP-2 than gram-negative bacteria. Using microscopy, we confirmed that LEAP-2 peptides could kill bacteria by disrupting the bacterial cell envelope. Conclusion: Duck LEAP-2 showed its antimicrobial activity against both gram-positive and gram-negative bacteria. Disulfide bonds were important for the powerful killing effect by disrupting the bacterial cell envelope. Therefore, duck LEAP-2 can be used for effective antibiotics alternatives.

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.

Effect of Exogenous Porcine GH Administration on GH Responses to GH-releasing Peptide-2 and GH-releasing Hormone in Swine

  • Nou, V.;Tomoshi, K.;Inoue, H.;Matsunaga, N.;Kuwayama, H.;Hidari, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1188-1192
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    • 2003
  • Negative feedback on GH responses to GH-releasing hormone (GHRH) and GH-releasing peptides (GHRPs) has been reported and this action has been suggested to act through an increase in somatostatin. To determine whether the acute administration of porcine GH (pGH) inhibits GH responsiveness to GHRP-2 and GHRH in swine, swine were intravenously administered with pGH (5${\mu}g$/kg BW) or placebo followed 180 min later by a second intravenous administration of saline, GHRP-2 (30 ${\mu}g$/kg BW), GHRH (1${\mu}g$/kg BW) and a combination of GHRP-2 and GHRH. Plasma GH concentration was measured by radioimmunoassay. Administration of pGH caused a significant increase in GH area under curve and GH peak concentrations (p<0.001) over placebo-treated group. Plasma GH concentrations peaked at 15 min and returned to baseline level within 90 min. Pretreatment of pGH abolished (p<0.01) GH response to GHRH and attenuated (p<0.05) GH response to GHRP-2 and GHRH combined, without affecting GH response to GHRP-2. These results demonstrate that negative feedback action on GH releasing effect of GHRH occurs in swine, and that GHRP-2 has ability to interact in this action.

Effect of Medium Components on the Production of Cyclosporin A by Immobilized Fungal Cell, Tolypocladium inflatum (배지성분이 고정화 곰팡이 세포를 이용한 Cyclosporin A 생산에 미치는 영향)

  • 이태호;장용근전계택
    • KSBB Journal
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    • v.11 no.5
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    • pp.613-621
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    • 1996
  • The effects of important medium components such as carbon, nitrogen sources and amino acids on the production of cyclosporin A(CyA) were investigated in an immobilized fungal cell fermentation using Tolypocladium inflatum. As carbon sources in the synthetic medium, fructose and maltose stimulated CyA production remarkably compared to glucose when ammonium sulfate was supplemented as a nitrogen source. In the absence of ammonium sulfate in the medium, however, CyA biosynthesis was reduced considerably without regard to C-sources tested. Ammonium sulfate was found to be the best N-source, and also ammonium phosphate and ammonium citrate showed some positive effects on CyA production. Optimum concentration of ammonium sulfate was 10g/L, and supplementation of ammonium sulfate at the start of fermentation was found to be the most efficacious for maximal production of CyA. Among the constituent amino acids of cyclic peptide, CyA, L-valine had the most significant effect on the biosynthesis of CyA, and maximum CyA production was observed when 10 g/L of L-valine was initially added.

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Separation of Heme-iron by Dialysis (투석법에 의한 Heme-iron의 분리)

  • Kang, In-Kyu;In, Man-Jin;Oh, Nam-Soon
    • Applied Biological Chemistry
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    • v.46 no.2
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    • pp.130-133
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    • 2003
  • A method for separating heme-iron from hemoglobin (Hgb) hydrolysate by dialysis was developed. Recovery of heme-iron increased with increasing Hgb concentration, whereas rejection of peptide and separation effciency expressed by HP ratio (heme-iron/peptide) did not show significant differences. HP ratio increased with increases in the degree of hydrolysis of Hgb and $KH_2PO_4$ concentrations of dialysis solution. Recovery of heme-iron decreased with increase in the pH of dialysis solution due to wash-out of heme-iron across the dialysis membrane caused by increase in solubility of heme-iron. Rejections of peptide were 74.5 and 87.5% (2 and 5 kDa of cut off size, respectively), whereas recovery of heme-iron decreased from 86.5 (2 kDa) to 63.1% (25 kDa). Amounts of heme-iron and peptide of dried heme-iron product were 21.7 and 77.0%, and HP ratio and production yield were 28.2 and 6.5%, respectively.

Production and Separation of Angiotension Converting Enzyme Inhibitor during Natto Fermentation (납두 발효과정 중 Angiotensin Converting Enzyme 저해물질의 생성 및 분리)

  • Cho, Young-Je;Cha, Woen-Suep;Bok, Su-Kyung;Kim, Myung-Uk;Chun, Sung-Sook;Choi, Ung-Kyu;Kim, Soon-Hee;Park, Kyung-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.29 no.4
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    • pp.737-742
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    • 2000
  • As functionality investigation of a soybean fermentation food, a angiotensin converting enzyme inhibitory peptide was separated during natto fermentation by Bacillus natto and inhibitory effect was investigated. After incubation at each 2$0^{\circ}C$, 3$0^{\circ}C$, 4$0^{\circ}C$, 5$0^{\circ}C$, 6$0^{\circ}C$ for the 0~72 hr, protein content, protease activity and angiotensin converting enzyme inhibition were determined. The protein content and protease activity were increased and reached maximum at 60 hr fermentation with 4$0^{\circ}C$ and decreased after the 60 hr fermentation during natto fermentation. The optimum condition for angiotensin converting enzyme inhibitors was appeared at fermentation for 60 hr at 4$0^{\circ}C$. Crude extract of natto was partially purified by Amicon membrane YM-3 and Sephadex G-10, G-25 gel filtration, stepwise. The inhibitory rate was increased in a concentration dependent manner, espcially the most potent activity about 74.74% at 1.0 mg peptide content. The most prominent amino acid of the peptide from natto was alanine, followed by phenylalnine, histidine.

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Regulation of Class II Bacteriocin Production by Cell-Cell Signaling

  • Quadri, Luis E.N.
    • Journal of Microbiology
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    • v.41 no.3
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    • pp.175-182
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    • 2003
  • Production of ribosomally synthesized antimicrobial peptides usually referred to as bacteriocins is an inducible trait in several gram positive bacteria, particularly in those belonging to the group of lactic acid bacteria. In many of these organisms, production of bacteriocins is inducible and induction requires secretion and extracellular accumulation of peptides that act as chemical messengers and trigger bacteriocin production. These inducer peptides are often referred to as autoinducers and are believed to permit a quorum sensing-based regulation of bacteriocin production. Notably, the peptides acting as autoinducers are dedicated peptides with or without antimicrobial activity or the bacteriocins themselves. The autoinducer-dependent induction of bacteriocin production requires histidine protein kinases and response regulator proteins of two-component signal transduction systems. The current working model for the regulation of class II bacteriocin production in lactic acid bacteria and the most relevant direct and indirect pieces of evidence supporting the model are discussed in this minireview.

The Review on the Study related to Anti-inflammatory Mechanism of Bee Venom Therapy (봉독요법(蜂毒療法)의 항염증(抗炎症) 기전(機轉) 연구(硏究)에 관(關)한 고찰(考察))

  • Choi, Jung-Sik;Park, Jang-Woo;Oh, Min-Seok
    • Journal of Haehwa Medicine
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    • v.15 no.1
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    • pp.141-160
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    • 2006
  • The obtained results are summarized as follows 1. New findings are reporting year by year as for the study related to Anti-inflammatory mechanism of Bee Venom therapy. 2. The Anti-inflammatory effect of Bee Venom therapy is achieved through counterirritation, stimulations to adrenal cortex, immuno-regulation, antioxidation, removal of free radicals, modulation of AGP gene induction. 3. The chief components of Bee Venom related to Anti-inflammatory effect are Melittin, MCD peptide, Apamin, Adolapin etc. 4. Melittin binds to secretory phospholipase A2 and inhibits its enzymatic activity. 5. Melittin blocks neutophil O2-production. 6. MCD peptide(Peptide 401) stimulates the mast cell secrets histamine, Anti-inflammatory effect caused by this is 'conterirritation'. 7. Melittin & Apamin have an anti-inflammatory effect by inducing cortisone secretion. 8. MCD peptide & Apamin increase immunologic fuction by stimulating hypophysis & adrenal cortex and have an anti-inflammatory effect by inhibiting synthesis of prostaglandin from arachidonic acid. 9. Adolapin have an anti-inflammatory effect by inhibiting COX. 10. Bee Venom have an anti-inflammatory effect by suppressing AGP($\alpha$-acid glycoprotein). 11. Bee Venom have an anti-inflammatory effect by inhibiting NO, iNOS, PLA2, COX-2, TNF-$\alpha$, IL-1, NF-${\kappa}B$, MAP kinase.

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Production and characterization of cross-reactive anti-Porphyromonas gingivalis heat shock protein 60 monoclonal antibody (항-Porphyromonas gingivalis heat shock protein 60 단클론항체의 생성과 특성 규명)

  • Lee, Ji-Young;Lee, Ju-Youn;Kim, Seong-Jo;Choi, Jeom-Il
    • Journal of Periodontal and Implant Science
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    • v.38 no.4
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    • pp.565-578
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    • 2008
  • Purpose: Porphyromonas gingivalis(P. gingivalis) heat shock protein (HSP)60 may play a role in the immunopathogenesis of periodontitis as well as atherosclerosis by modulating autoimmune reaction due to its high level of sequence homology between bacteria and human counterpart. The purpose of this study was to identify immunodomiant epitope of P. gingivalis HSP60 that is reactive exclusively to the homologous bacteria without reacting with human HSP. Materials and methods: The present study was performed to identify the peptide specifically recognized by anti-P. gingivalis HSP60 monoclonal antibodies mono-reactive to P. gingivalis HSP60. Results: Four different hybridomas were cloned producing monoclonal IgG antibodies exclusively to P. gingivalis HSP60. Thirty seven synthetic peptides (20-mer with 5-amino acid overlapping) were synthesized. All of these peptide were subject to SDS-PAGE for immunblot analysis. One peptide (TVPGGGTTYIRAIAALEGLK) and the other peptide (TLVVNRLRGSLKICAVKAPG) were recognized by all and one of the four monoclonal antibodies, respectively, that reacted solely with P. gingivalis HSP60. Immunohistochemistry to identify the localization of the HSP60 in the diseased gingival tissues revealed that all of the four monoclonal antibodies were highly reacted with the diseased gingival tissue than normal gingival tissue. Conclusion: The P. gingivalis HSP60 peptides (TVPGGGTTYIRAIAALEGLK and TLVVNRLRGSLKICAVKAPG, respectively) are positively involved in the immunopathologic process of periodontal disease. The peptide may potentially be developed as vaccine candidates. Further investigations are under way to identify more clones producing monoclonal antibodies reactive to P. gingivalis HSP and to other periodontopathogenic bacteria as well, while maintaining specificities to human counterpart.

Glyceryl Esterification of Fibroin Peptide by Papain (파파인을 이용한 피브로인 펩타이드의 글리세릴 에스터화 반응)

  • Jeong, Jae-Ho;Lee, Shin-Young;Hur, Won
    • KSBB Journal
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    • v.25 no.4
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    • pp.395-400
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    • 2010
  • Papain hydrolysate of fibroin was found to be mainly composed of several even-numbered peptides that can be produced at a large scale and can be used as a precursor for biological fine-chemicals such as peptide detergents. Thus, the hydrolysate was further modified to synthesize a peptide mixture of glyceryl esters using the identical enzyme for the production of such chemicals. Formation of glyceryl ester of each peptide was confirmed by identifying peaks of the nominal mass shift of +74 Da in mass spectrometry. Analysis of the mass spectra indicated that glyceryl esters of di- and tetra-peptides were the major constituents of the mixture and that alanylglycine was most preferentially esterified. It also suggests that papain prefers dipeptide to tetrapeptide and alanine to serine or tyrosine at $P_2$ position as substrate for glyceryl esterification. The glyceryl esters were recovered using ion exchange resin and the yield of glyceryl esterification recorded was 17.8% by weight.