• Title/Summary/Keyword: polylysine

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Separation of ε-poly-L-lysine from the fermentation broth of Streptomyces albulus (Streptomyces albulus 배양액으로부터 ε-poly-L-lysine의 분리)

  • Sun, Heung-Suk;Park, Chan-Young
    • Journal of the Korean Society of Industry Convergence
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    • v.2 no.1
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    • pp.77-83
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    • 1999
  • Grown in the secondary broth of production media, the strain Streptomyces albulus has increased more the production of its metabolite ${\varepsilon}$-poly-L-lysine, one of poly(amino acid)s used as disinfecting food additives, than the strain in the primary culture of growth nutrients. Having the strain removed, the large concentrate obtained by ultrafiltrating the secondary culture broth. The concentrated production broth exchanged into followed by detecting in UV flowcell at 220nm the peptide bond of the components eluting the adsorbed proteins and polylysine with NaCl salt of gradient concentration, and has separated into five components. Among them the component in the fourth peak fraction has proved to be the pure ${\varepsilon}$-poly-L-lysine after the portion being hydrolyzed the fraction with HCl into amino acid followed by being the composing amino acid analysis.

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Enhanced delivery of protein fused to cell penetrating peptides to mammalian cells

  • Moon, Jung-Il;Han, Min-Joon;Yu, Shin-Hye;Lee, Eun-Hye;Kim, Sang-Mi;Han, Kyuboem;Park, Chang-Hwan;Kim, Chun-Hyung
    • BMB Reports
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    • v.52 no.5
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    • pp.324-329
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    • 2019
  • Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols.

Evaluation of Antifungal Activity of Natural Antimicrobials in Fuctional Beverage (기능성 음료의 곰팡이 제어를 위한 천연항균제 활성 평가)

  • Lee, Hee-Seok;Yeon, Ji-Hye;Ha, Sang-Do;Park, Chul-Soo;Woo, Moon-Joe;Lee, Sang-Hun;Kim, Jin-Soo;Lee, Chan
    • Journal of Food Hygiene and Safety
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    • v.24 no.3
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    • pp.262-266
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    • 2009
  • Antifungal activity of seven natural antimicrobials, such as $\varepsilon$-polylysine, extract of Yucca shidigera, TLS (vitamin B1 derivative), BMB-FS, chitooligosaccharide, KDSP 001, and KDSP 002 were investigated for their applications in functional beverage. Five fungi including Aspergillus niger, Penicillium citrinum, Rhizopus oryzae, Fusarium moniliforme, and Mucor rouxii were applied as test fungi and mininum inhibitory concentrations (MICs) of antimicrobials were examined. TLS exhibited the strongst antifungal activity among tested antimicrobials and the growth of all fungi was inhibited at 100 ppm. The antifungal activity of BMB-FS appeared different. The growth of Fusarium moniliforme was inhibited by BMB-FS at 100 ppm, but it exhibit antifungal activity on P. citrinum, and M. rouxii at 1000ppm. MICs of TLS appeared to be 60 ppm for four test fungi except A. niger (100 ppm). These results indicate the possible usage of TLS and BMB-FS as natural antimicrobials in functional beverage.

Evaluation of the Natural Antimicrobials against Yeasts in Functional Beverages to Control Quality loss (효모에 의한 기능성 음료 변질 제어를 위한 천연항균물질 항균력 평가)

  • Yeon, Ji-Hye;Lee, Ji-Young;Lee, Hee-Seok;Ha, Sang-Do;Park, Chul-Soo;Woo, Moon-Jea;Lee, Sang-Hun;Kim, Jin-Soo;Lee, Chon
    • Journal of Food Hygiene and Safety
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    • v.24 no.3
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    • pp.273-276
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    • 2009
  • We investigated eight active natural antimicrobials for preservation of functional beverages that are usually degraded by yeasts rather than by bacteria due to a high sugar content and a low pH. Five strains of yeasts (S. cerevisiae, Z. bailii, P. membranaefaciens, C. albicans, and P Anomala) were tested with eight natural antimicrobial agents ($\varepsilon$-polylysine, yucca extract, vitamin $B_1$ derivative, scutellaria baicalensis extract, chitooligosaccharid, allyl isothiocyanate, sucrose-fatty acid ester, and oligosaccharide). The lowest minimal inhibitory concentrations (MIC) were 10 ppm for oligosaccharide and sucrose-fatty acid ester against S. cerevisiae and Z. bailii, 10 ppm for allyl isothiocynate against P. membranaefaciens and C. albican, and 10 ppm for allyl isothiocynate and oligosaccharide against P. anomala. No growth were observed for five kinds of yeasts in functional beverages containing sodium benzonate at concentration of 0.015% or higher. The resistance of S. cerevisiae, Z. bailii, and P. Anomala against natural antimicrobial agents was lower than those of P. membranaefaciens and C. albican. Allyl isothiocyanate, oligosaccharide, and sucrose-fatty acid ester showed the highest antimicrobial activities among the eight tested antimicrobials. These results can be applied to develop new natural antimicrobial agents to improve microbial quality of functional beverages.

Status, Antimicrobial Mechanism, and Regulation of Natural Preservatives in Livestock Food Systems

  • Lee, Na-Kyoung;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.36 no.4
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    • pp.547-557
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    • 2016
  • This review discusses the status, antimicrobial mechanisms, application, and regulation of natural preservatives in livestock food systems. Conventional preservatives are synthetic chemical substances including nitrates/nitrites, sulfites, sodium benzoate, propyl gallate, and potassium sorbate. The use of artificial preservatives is being reconsidered because of concerns relating to headache, allergies, and cancer. As the demand for biopreservation in food systems has increased, new natural antimicrobial compounds of various origins are being developed, including plant-derived products (polyphenolics, essential oils, plant antimicrobial peptides (pAMPs)), animal-derived products (lysozymes, lactoperoxidase, lactoferrin, ovotransferrin, antimicrobial peptide (AMP), chitosan and others), and microbial metabolites (nisin, natamycin, pullulan, ε-polylysine, organic acid, and others). These natural preservatives act by inhibiting microbial cell walls/membranes, DNA/RNA replication and transcription, protein synthesis, and metabolism. Natural preservatives have been recognized for their safety; however, these substances can influence color, smell, and toxicity in large amounts while being effective as a food preservative. Therefore, to evaluate the safety and toxicity of natural preservatives, various trials including combinations of other substances or different food preservation systems, and capsulation have been performed. Natamycin and nisin are currently the only natural preservatives being regulated, and other natural preservatives will have to be legally regulated before their widespread use.

Low Molecular Weight PEI Conjugated Pluronic Copolymer: Useful Additive for Enhancing Gene Transfection Efficiency

  • Cho Kyung-Chul;Choi Seung-Ho;Park Tae-Gwan
    • Macromolecular Research
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    • v.14 no.3
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    • pp.348-353
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    • 2006
  • For enhancing the gene delivery efficiency of polyplexes, a new formulation was developed using PEI conjugated Pluronic F127 copolymer as an effective additive. Low molecular weight, branched polyethylenimine Mw 600 (LMW BPEI 600) was conjugated to the terminal end of Pluronic F127. The PEI-modified Pluronic copolymers formed a micellar structure in aqueous solution, similar to that of unmodified Pluronic copolymer. PEI modification of Pluronic copolymer increased the size of micelles while concomitantly raising the critical micelle concentration (CMC). The PEI-modified Pluronic copolymer was used as a micellar additive to enhance the gene transfection efficiency of pre-formulated polyelectrolyte complex nanoparticles composed of luciferase plasmid DNA and branched PEI Mw 25k (BPEI 25k) or polylysine Mw 39k (PLL 39k). The luciferase gene expression levels were significantly enhanced by the addition of the BPEI-modified Pluronic copolymer for the two formulations of BPEl and PLL polyplexes. The results indicated that the BPEI-modified Pluronic copolymer micelles ionically interacted on the surface of DNA/BPEI (PLL) polyplexes which might facilitate cellular uptake process.

Evaluation of Antimicrobial Activities of Sequential Spray Applications of Decontamination Treatments on Chicken Carcasses

  • Benli, Hakan;Sanchez-Plata, Marcos X.;Ilhak, Osman Irfan;De Gonzalez, Maryuri T. Nunez;Keeton, Jimmy T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.3
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    • pp.405-410
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    • 2015
  • The objective of this study was to evaluate the effects of sequential applications of ${\varepsilon}$-polylysine (EPL) or lauramide arginine ethyl ester (LAE) sprays followed by an acidic calcium sulfate (ACS) spray on inoculated chicken carcasses to reduce Salmonella (Salmonella enterica serovars including Salmonella typhimurium and Salmonella enteritidis) contamination during 6 days of storage ($4.4^{\circ}C$). Secondly, reductions of the resident microflora were studied on uninoculated chicken carcasses following the sequential application of the treatments, chilling and 10 days of storage at $4.4^{\circ}C$. The treatment of Salmonella inoculated carcasses with 300 mg/L EPL followed by 30% ACS (EPL300-ACS30) sprays reduced Salmonella counts initially by 1.5 log cfu/mL and then by 1.2 log cfu/mL (p<0.05) following 6 days of storage at $4.4^{\circ}C$. Likewise, 200 mg/L LAE followed by 30% ACS (LAE200-ACS30) treatment reduced initial Salmonella counts on poultry carcasses by 1.8, 1.4 and 1.8 log cfu/mL (p<0.05), respectively, after 0, 3, and 6 days storage. Immediately after the treatments, EPL300-ACS30 and LAE200-ACS30 both reduced Escherichia coli counts significantly by 2.6 and 2.9 log cfu/mL, respectively. EPL300-ACS30 and LAE200-ASC30 were effective in lowering psychrotroph counts by 1 log cfu/mL on day 10 when compared to the control and distilled water treatments. This study demonstrated that EPL300-ACS30 and LAE200-ACS30 were effective in reducing Salmonella on inoculated chicken carcasses both after treatment and during the storage at $4.4^{\circ}C$ for up to 6 days. In addition, reductions in psychrotroph counts indicated that these treatments might have the potential to increase the shelf-life of poultry carcasses.

Rat Duodenal Mucosa Inositol Monophosphatase; Novel Enzyme of Which Properties are Distinct from Brain Enzyme

  • Kwon, Hyeok-Yil;Lim, Bong-Hee;Park, Hyung-Seo;Lee, Yun-Lyul;Lee, Eun-Hee;Choi, Soo-Young;Park, Hyoung-Jin
    • BMB Reports
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    • v.31 no.3
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    • pp.274-280
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    • 1998
  • An inositol monophosphatase (IMPase) was purified to homogeneity from rat duodenal mucosa for the first time and its enzymatic properties were investigated. Rat duodenal mucosa peculiarly exhibited the highest IMPase activity among various rat tissues examined. By means of ammonium sulfate precipitation, followed by Q-Sepharose, polylysine agarose, reactive-red agarose column chromatography, Uno-Q FPLC, and Bio-Silect FPLC, duodenal IMPase was purified 223-fold to a specific activity of 13.6 U/mg protein. The molecular mass of the native enzyme was estimated to be 48,000 Da on gel filtration. The subunit molecular mass was determined by SDS-PAGE to be 24,000 Da. These results indicate that duodenal IMPase is a dime ric protein made up of identical subunits. Rat duodenal IMPase has distinct properties from brain IMPase. It has a broad spectrum of substrate specificity and is insensitive to $Li^+$. Duodenal IMPase does not absolutely require $Mg^{2+}$ for its catalytic activity. Furthermore, duodenal IMPase is less stable to heat than brain enzyme. It is suggested that the rat duodenal mucosa needs a large amount of IMPase whose properties are quite different from that of the brain enzyme.

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Purification and Characterization of Biopolymer by Bacillus coagulans CE-74 (Bacillus coagulans CE-74가 생산하는 Biopolymer의 분리 및 정제)

  • Lee, Seon-Ho;Seung, Tae-Su;Choi, Cheong
    • Applied Biological Chemistry
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    • v.43 no.2
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    • pp.81-85
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    • 2000
  • Screening was performed to isolate biopolymer-producing microorganisms from natural sources. The bacteriological characteristics of this strain and physicochemical properties of the biopolymer produced were investigated. The bacterial strain was identified as a Bacillus coagulans. Crude biopolymer treated with ethanol and actone was purified to fraction I and II by ion exchange chromatography and gel chromatography (Shephadex G-100 and Shepharose CL-2B). Analysis of chemical composition and various color reaction revealed that the polymer is composed of amino acids. It was confirmed that fraction II is a homopolymer of glutamic acid and fraction I is a homopolymer of lysine by analysis data of amino acid analyzer, GC and IR.

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Preparation and Characterization of Paclitaxel-loaded PLGA Nanoparticles Coated with Cationic SM5-1 Single-chain Antibody

  • Kou, Geng;Gao, Jie;Wang, Hao;Chen, Huaiwen;Li, Bohua;Zhang, Dapeng;Wang, Shuhui;Hou, Sheng;Qian, Weizhu;Dai, Jianxin;Zhong, Yanqiang;Guo, Yajun
    • BMB Reports
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    • v.40 no.5
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    • pp.731-739
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    • 2007
  • The purpose of this study was to develop paclitaxel-loaded poly(lactide-co-glycolide) (PLGA) nanoparticles coated with cationic SM5-1 single-chain antibody (scFv) containing a polylysine (SMFv-polylys). SM5-1 scFv (SMFv) is derived from SM5-1 monoclonal antibody, which binds to a 230 kDa membrane protein specifically expressed on melanoma, hepatocellular carcinoma and breast cancer cells. SMFv-polylys was expressed in Escherichia coli and purified by cation-exchange chromatography. Purified SMFv-polylys was fixed to paclitaxel-loaded PLGA nanoparticles to form paclitaxel-loaded PLGA nanoparticles coated with SMFv-polylys (Ptx-NP-S). Ptx-NP-S was shown to retain the specific antigen-binding affinity of SMFv-polylys to SM5-1 binding protein-positive Ch-hep-3 cells. Finally, the cytotoxicity of Ptx-NP-S was evaluated by a non-radioactive cell proliferation assay. It was demonstrated that Ptx-NP-S had significantly enhanced in vitro cytotoxicity against Ch-hep-3 cells as compared with non-targeted paclitaxel-loaded PLGA nanoparticles. In conclusion, our results suggest that cationic SMFv-polylys has been successfully generated and may be used as targeted ligand for preparing cancer-targeted nanoparticles.