• Title/Summary/Keyword: peptide production

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Antimicrobial and Antioxidant Peptide from Bacillus Strain CBS73 Isolated from Korean Food

  • Kim, Miri;Khan, Md Maruf;Yoo, Jin Cheol
    • Journal of Integrative Natural Science
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    • v.10 no.3
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    • pp.154-161
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    • 2017
  • An antimicrobial peptides-producing Bacillus strain CBS73 was isolated from fermented food (kimchi) that produces low-molecular-weight proteins with broad-spectrum antimicrobial activity. Our goal was to explore the therapeutic potential of antimicrobial substances produced by Bacillus species. Peptide CBS73 was purified from Bacillus subtilis subsp. subtilis with identity of 99.79%. It was found to be stable at pH 4.0-10.0 and temp $20-60^{\circ}C$. A protein band around 5.2 kDa was detected in tricine-SDS-PAGE and band was confirmed by MALDI-TOF test. Peptide CBS73 showed antimicrobial activity against MDR bacteria. The minimal inhibitory concentration (MIC) of peptide CBS73 for vancomycin-resistant S. aureus (VRSA), vancomycin resistant Enterococci (VRE) and Salmonella typhimurium ranged from $10-40{\mu}g/mL$. The antioxidant activity of peptide CBS73 was measured by DPPH scavenging, reducing power activity and total phenolic content. Cell viability and NO production result showed less cytotoxic effect upto $12{\mu}g/mL$. Peptide CBS73 could be a promising antimicrobial agent for clinical application.

Metallo-collagenase production by Arthrobacter creatinolyticus KP015744

  • Savita A. Kate;Madhuri Sahasrabudhe;Archana Pethe
    • Advances in environmental research
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    • v.11 no.1
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    • pp.1-16
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    • 2022
  • Amongst 27 isolates from deteriorated leather samples, Arthrobacter creatinolyticus KP015744 zzx28 was found to be an efficient collagenase producer. Collagenase production of 13.33 µmoles/min was shown at an optimum temperature at 37℃ after 72h and at pH 7.5 by using 2 ml/dL inoculum in 10 mg/ml collagen peptide type I as a substrate. In presence of Hg2+, EDTA and 𝛽-mercaptoethanol the collagenase production by the isolate was strongly inhibited however Fe2+, Ca2+and DMSO enhanced production of the enzyme. Specific activity was found to be 19.46×103 U/mg and molecular weight 66 kD by SDS PAGE. Isolate also has potential to hydrolyze keratin which is another important protein found in leather. Experimental results propose that collagenase can be effectively used as a tool for collagen and keratin rich solid waste treatment.

Rapid Identification of Bioactive Compounds Reducing the Production of Amyloid β-Peptide (Aβ) from South African Plants Using an Automated HPLC/SPE/HPLC Coupling System

  • Kwon, Hak-Cheol;Cha, Jin-Wook;Park, Jin-Soo;Chun, Yoon-Sun;Moodley, Nivan;Maharaj, Vinesh J.;Youn, Sung-Hee;Chung, Sung-Kwon;Yang, Hyun-Ok
    • Biomolecules & Therapeutics
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    • v.19 no.1
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    • pp.90-96
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    • 2011
  • Automated HPLC/SPE/HPLC coupling experiments using the Sepbox system allowed the rapid identification of four bioactive principles reducing the production of amyloid $\beta$-peptide ($A{\beta}$) from two South African plants, Euclea crispa subsp. crispa and Crinum macowanii. The structures of biologically active compounds isolated from the methanol extract of Euclea crispa subsp. crispa were assigned as 3-oxo-oleanolic acid (1) and natalenone (2) based on their NMR and MS data, while lycorine (3) and hamayne (4) were isolated from the dichloromethane-methanol (1:1) extract of Crinum macowanii. These compounds were shown to inhibit the production of $A{\beta}$ from HeLa cells stably expressing Swedish mutant form of amyloid precursor protein (APPsw).

Anti-inflammatory and Wrinkle Improvement Effects of Peptides from Ginseng Berry Amino Acidic Complex (진생베리 아미노산 복합체로부터 분리한 펩타이드의 항염, 주름개선 효과)

  • Kang, Sang Moon;Park, Chung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.3
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    • pp.299-306
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    • 2019
  • Ginseng berry (GB) contains Ginsenoside Re and has anti-inflammatory and anti-wrinkle properties. In this study, TLC fractions 1, 2, and 4 of the ginseng berry amino acid complex were identified and analyzed by HPLC. And identified a peptide (AP-1) by LC/MASS analysis of fraction 1. The anti-inflammatory activity was confirmed by investigating the inhibitory effect of AP-1 on NO production. In addition, collagen synthesis using procollagen type I C-peptide (PIP) ELISA kit was 50% higher effective than that of the control group. From these results, the peptide isolated from ginseng berry amino acid complex is considered to have anti-inflammatory and anti-wrinkle effect, and may be useful as an anti-inflammatory and anti-aging cosmetic raw material.

Effects of Avocado Yoghurt, Three Beans Cookie, and Tiger Nut Galsu Drink on Fibroblasts (Hs68) and Keratinocytes (HaCaT)

  • Park, Seung-Yong;Jung, Eun-Kyung;Joo, Nami;Shin, Hyun-Soo;Shin, Ji-Sun;Myoung, Da-Bin;Lee, Kyung-Tae
    • Journal of Dairy Science and Biotechnology
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    • v.37 no.2
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    • pp.136-153
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    • 2019
  • The present study aimed to investigate the effects of skin care foods on the synthesis of pro-collagen type I C peptide and suppression of matrix metalloproteinase (MMP)-1 secretion through an in vitro study using fibroblasts (Hs68 cells) and keratinocytes (HaCaT cells). Among the three skin care foods developed in this study, three beans cookie and avocado yoghurt influenced the production of pro-collagen type I C peptide and suppressed MMP-1 secretion; however, tiger nut Galsu drink did not exhibit these effects. All skin care foods, including three beans cookie and plain yoghurt ($50{\mu}g/mL$, p<0.001) influenced the suppression of MMP-1 in addition to other commercially available breast milk production support foods examined, such as Heath Heather ($50{\mu}g/mL$, p<0.001), Happy Mama ($50{\mu}g/mL$, p<0.01), BioLys ($50{\mu}g/mL$, p<0.001), Enfamama ($25{\mu}g/mL$, p<0.0001), and Pregnagen ($25{\mu}g/mL$, p<0.001). Avocado fruit yoghurt ($25{\mu}g/mL$, p<0.05), avocado fruit jam yoghurt ($50{\mu}g/mL$, p<0.01), Enfamama ($100{\mu}g/mL$, p<0.05), and Pregnagen ($100{\mu}g/mL$, p<0.05) influenced the production of pro-collagen type I C peptide and suppressed MMP-1 secretion. This result indicates that only avocado jam yoghurt significantly influenced both the prevention of skin keratinization and acceleration of recovery of skin fibrous structure. Therefore, avocado is a favorable ingredient for nutrition-balanced dietary foods or an essential ingredient in products for revitalization of human skin.

Production of Peptides Enhancing Calcium Solubility in the Presence of Phosphate Ions In Vitro (In Vitro 상에서인 이온 존재 하에서의 칼슘 용해도를 증대시키는 펩타이드의 생산)

  • 이윤동;이현수
    • The Korean Journal of Food And Nutrition
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    • v.10 no.4
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    • pp.485-490
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    • 1997
  • Gluten peptide was produced from corn gluten by enzymatic hydrolysis. This peptide had an ability to increased the solubility of calcium owing to protect calcium ions from forming precipitates of calcium phosphate in the presence of phosphate ions. The solubility of calcium was increased 5.2 times in the presence of 8.3 mg peptide produced by the treatment of papain. These peptides contained high acidic amino acids and fractionated by Delta pack column into fractions No. 1, No. 2 and No. 3. Among them the fraction No. 3 had the highest calcium binding capacity.

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Identification of a Novel Single Nucleotide Polymorphism in Porcine Beta-Defensin-1 Gene

  • Pruthviraj, D.R.;Usha, A.P.;Venkatachalapathy, R.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.3
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    • pp.315-320
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    • 2016
  • Porcine beta-defensin-1 (PBD-1) gene plays an important role in the innate immunity of pigs. The peptide encoded by this gene is an antimicrobial peptide that has direct activity against a wide range of microbes. This peptide is involved in the co-creation of an antimicrobial barrier in the oral cavity of pigs. The objective of the present study was to detect polymorphisms, if any, in exon-1 and exon-2 regions of PBD-1 gene in Large White Yorkshire (LWY) and native Ankamali pigs of Kerala, India. Blood samples were collected from 100 pigs and genomic DNA was isolated using phenol chloroform method. The quantity of DNA was assessed in a spectrophotometer and quality by gel electrophoresis. Exon-1 and exon-2 regions of PBD-1 gene were amplified by polymerase chain reaction (PCR) and the products were subjected to single strand conformation polymorphism (SSCP) analysis. Subsequent silver staining of the polyacrylamide gels revealed three unique SSCP banding patterns in each of the two exons. The presence of single nucleotide polymorphisms (SNPs) was confirmed by nucleotide sequencing of the PCR products. A novel SNP was found in the 5'-UTR region of exon-1 and a SNP was detected in the mature peptide coding region of exon-2. In exon-1, the pooled population frequencies of GG, GT, and TT genotypes were 0.67, 0.30, and 0.03, respectively. GG genotype was predominant in both the breeds whereas TT genotype was not detected in LWY breed. Similarly, in exon-2, the pooled population frequencies of AA, AG, and GG genotypes were 0.50, 0.27, and 0.23, respectively. AA genotype was predominant in LWY pigs whereas GG genotype was predominant in native pigs. These results suggest that there exists a considerable genetic variation at PBD-1 locus and further association studies may help in development of a PCR based genotyping test to select pigs with better immunity.

Anti-Inflammatory Activity of Antimicrobial Peptide Allomyrinasin Derived from the Dynastid Beetle, Allomyrina dichotoma

  • Lee, Joon Ha;Seo, Minchul;Lee, Hwa Jeong;Baek, Minhee;Kim, In-Woo;Kim, Sun Young;Kim, Mi-Ae;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.687-695
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    • 2019
  • In a previous work, we performed de novo RNA sequencing of Allomyrina dichotoma using next generation sequencing and identified several antimicrobial peptide candidates based on transcriptome analysis. Among them, a cationic antimicrobial peptide, allomyrinasin, was selected bioinformatically based on its physicochemical properties. Here, we assessed the antimicrobial and anti-inflammatory activities of allomyrinasin against microorganisms and mouse macrophage Raw264.7 cells. Allomyrinasin showed antimicrobial activities against various microbes and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Furthermore, quantitative RT-PCR and ELISA revealed that allomyrinasin reduced cytokine expression levels in the Raw264.7 cells. We also identified inducible nitric oxide synthase, cyclooxygenase-2 expression, and $PGE_2$ production through western blot analysis and ELISA. We confirmed that allomyrinasin bound to bacterial cell membranes via a specific interaction with lipopolysaccharides. Taken together, these data indicate that allomyrinasin has antimicrobial and anti-inflammatory activities as exemplified in lipopolysaccharide-induced Raw264.7 cells. We have provided a potentially useful antimicrobial peptide candidate that has both antimicrobial and anti-inflammatory activities.

Effect of peptide on the mixed fermentation of Lactobacillus helveticus YM-1 and Streptococcus lactis $ML_3$ in skim milk (Lactobacillus helveticus YM-1과 Streptococcus lactis $ML_3$의 혼합발효에 미치는 peptide의 영향)

  • 박정길;류인덕;유주현
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.487-493
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    • 1986
  • Lactobacillus helveticus YM-1and Streptococcus lactis Ml$_3$ were inoculated together in reconstituted non-fat skim milk medium, and then their proteolytic activity and stimulatory compound for acid production were investigated. Significant difference between Lactobacillus helveticus YM-1 and Streptococcus lactis Ml$_3$was observed in the proteolytic activities. The proteolytic activity of Lactobacillus helveticus YM-1 and Streptococcus lactis Ml$_3$ was 105 $\mu\textrm{g}$/$m{\ell}$ and 30 $\mu\textrm{g}$/$m{\ell}$ when converted the amounts of hydrolysates of milk protein determined by Folin Ciocaleau phenol method into their tyrosine equivalent Stimulatory compounds in cell-free filtrate of Lactobacillus helveticus YM-1were identified as peptide with a molecular weight of approximately 4, 300 for the acid production by Streptococcus lactis Ml$_3$. Some kinds of amino acids, such as histidine, lysine, arginine and glutamic acid, were rich in acid hydrolysates of peptide. Among amino acids, histidine, glutamic acid and phenylalanine stimulated acid production, on the contrary isoleucine inhibited.

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Recombinant Production and Antimicrobial Activity of an Antimicrobial Model Peptide (Uu-ilys-CF) Derived from Spoon Worm Lysozyme, Uu-ilys (개불 라이소자임 유래 항균성 모델 펩타이드(Uu-ilys-CF)의 재조합 단백질 생산 및 항균 활성)

  • Oh, Hye Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • v.31 no.1
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    • pp.83-89
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    • 2021
  • Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.