• Title/Summary/Keyword: Peptide extracts

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Production and Characterization of an Anti-Angiogenic Agent front Saccharomyces cerevisiae K-7

  • Jeong, Seung-Chan;Lee, Dae-Hyoung;Lee, Jong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1904-1911
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    • 2006
  • The cell-free extracts of 250 yeasts were screened for their in vitro anti-angiogenic activity, to develop a new cancer metastasis inhibitor. Saccharomyces cerevisiae K-7 was selected as the producer of the anti-angiogenic agent, because it had the highest anti-angiogenic activity. The anti-angiogenic agent was produced maximally from hydrolysates of Saccharomyces cerevisiae K-7, when the yeast was cultured in yeast extract-peptone-dextrose medium at 30$^{\circ}C$ for 24 h, and cell-free extracts were than digested with pepsin for 4 h at 37$^{\circ}C$. The anti-angiogenic agent was further purified by ultrafiltration, Sephadex G-25 gel permeation chromatography and reverse-phase HPLC, and the anti-angiogenic activity of the final purified preparation was 72.7% at 10 $\mu$M/egg. The purified anti-angiogenic agent was found to originate from the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) molecule of Saccharomyces cerevisiae K-7, and its peptide sequence was Val-Ser-Trp-Tyr-Asp-Asn-Glu-Tyr-Gly-Tyr-Ser-Thr-Arg-Val-Val-Asp. In the MTT assay, the shape of the HT-l 080 cell was clearly changed to a circular type at 0.2 mM purified anti-angiogenic agent. This result indicated that the growth of the HT-I080 cell was significantly inhibited at 0.2 mM of the purified anti-angiogenic agent. The MMP activity of the treated HT-l080 cells was not affected, evidenced by the gelatin zymography, indicating that the anti-angiogenic mechanism of the purified anti-angiogenic agent is not mediated through MMP activity.

Suppression of β-Secretase (BACE1) Activity and β-Amyloid Protein-Induced Neurotoxicity by Solvent Fractions from Petasites japonicus Leaves

  • Hong, Seung-Young;Park, In-Shik;Jun, Mi-Ra
    • Preventive Nutrition and Food Science
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    • v.16 no.1
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    • pp.18-23
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    • 2011
  • Alzheimer's disease (AD) is a neurodegenerative disorder characterized by neuronal loss and extracellular senile plaques containing $\beta$-amyloid peptide (A$\beta$). The deposition of the A$\beta$ peptide following proteolytic processing of amyloid precursor protein (APP) by $\beta$-secretase (BACE1) and $\gamma$-secretase is a critical feature in the progression of AD. Among the plant extracts tested, the ethanol extract of Petasites japonicus leaves showed novel protective effect on B103 neuroblastoma cells against neurotoxicity induced by A$\beta$, as well as a strong suppressive effect on BACE1 activity. Ethanol extracts of P. japonicus leaves were sequentially extracted with methylene chloride, ethyl acetate and butanol and evaluated for potential to inhibit BACE1, as well as to suppress A$\beta$-induced neurotoxicity. Exposure to A$\beta$ significantly reduced cell viability and increased apoptotic cell death. However, pretreatment with ethyl acetate fraction of P. japonicus leaves prior to A$\beta$ (50 ${\mu}M$) significantly increased cell viability (p<0.01). In parallel, cell apoptosis triggered by A$\beta$ was also dramatically inhibited by ethyl acetate fraction of P. japonicus leaves. Moreover, the ethyl acetate fraction suppressed caspase-3 activity to the basal level at 30 ppm. Taken together, these results demonstrated that P. japonicus leaves appear to be a useful source for the inhibition and/or prevention of AD by suppression of BACE1 activity and attenuation of A$\beta$ induced neurocytotoxicity.

Characterization of an Indigenous Antimicrobial Substance-producing Paenibacillus sp. BCNU 5011 (항균물질을 생산하는 토착 미생물 Paenibacillus sp. BCNU 5011의 특성화)

  • Choi, Hye-Jung;Kim, Ya-Ell;Bang, Ji-Hun;Kim, Dong-Wan;Ahn, Cheol-Soo;Jeong, Young-Kee;Joo, Woo-Hong
    • KSBB Journal
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    • v.26 no.2
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    • pp.100-106
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    • 2011
  • Strain BCNU 5011 was isolated from forest soil samples collected in the Taebaek mountain in the Gangwon province, Korea. The biochemical, physiological and 16S rRNA sequence analysis strongly indicated that this isolate was most closely related to Paenibacillus polymyxa. A maximum production level of antimicrobial substances of Paenibacillus sp. BCNU 5011 was achieved under aerobic incubation at $30^{\circ}C$ for 3 days in SST broth.Paenibacillus sp. BCNU 5011 showed a broad spectrum of activity against Gram positive and Gram negative bacteria, including methicllinresistant Staphylococcus aureus (MRSA). Paenibacillus sp. BCNU 5011 was also shown to inhibit the growth of different potential human pathogenic bacteria and fungi in vitro. Peptide extract showed better antimicrobial activity than solvent extracts. But active antimicrobial compounds might be included in both peptide extract and solvent extracts. Further separation, purification and identification of active principles leads project to develop antimicrobial agents and anti-MRSA agents.

Korean Curcuma longa L. induces lipolysis and regulates leptin in adipocyte cells and rats

  • Song, Won-Yeong;Choi, Jeong-Hwa
    • Nutrition Research and Practice
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    • v.10 no.5
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    • pp.487-493
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    • 2016
  • BACKGROUND/OBJECTIVES: Turmeric (Curcuma longa L.) has been reported to have many biological functions including anti-obesity. Leptin, peptide hormone produced by adipocytes and its concentration is increased in proportion to the amount of the adipocytes. In the present study, we examined the effects of Korean turmeric on the regulation of adiposity and leptin levels in 3T3-L1 adipocytes and rats fed a high-fat and high-cholesterol diet. MATERIALS/METHODS: Leptin secretion, free fatty acid and glycerol contents in 3T3-L1 adipocytes were measured after incubation of cells with turmeric for 24 hours. Rats were divided into four experimental groups: a normal diet group (N), a high-fat and high-cholesterol diet group (HF), a high-fat and high-cholesterol diet group supplemented with 2.5% turmeric extracts (TPA group) and a high-fat and high-cholesterol diet group supplemented with 5% turmeric extracts (TPB group). Serum samples were used for the measurement of leptin concentration. RESULTS: Contents of free fatty acid and glycerol showed concentration dependent increase in response to turmeric extracts. Effects of turmeric extracts on reduction of lipid accumulation in 3T3-L1 cells were examined by Oil Red O staining. Treatment with turmeric extracts resulted in increased expression levels of adipose triglyceride lipase and hormone-sensitive lipase mRNA. The concentration of leptin from 3T3-L1 adipocytes was significantly decreased by turmeric. Proportional abdominal and epididymal fats weights of the turmeric 5% supplemented group, TPB has significantly decreased compared to the HF group. The serum levels of leptin in the TPA and TPB groups were significantly lower than those of the HF group. CONCLUSIONS: Based on these results, we suggested that Korean turmeric may contribute to the decreasing of body fat and regulating leptin secretion.

Screening of Positive Inotropic Effect from Herbal Extracts in Beating Rabbit Atria (박동관류 심방모델에서 강심효과를 나타내는 단미 한약재 검색)

  • Lee, Yun Jung;Kwon, Oh Jeong;Kim, Hye Yoom;Namgung, Seung;Lee, Jae Yun;Yu, Yun Cho;Kang, Dae Gill
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.30 no.1
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    • pp.40-46
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    • 2016
  • Many medicinal plants have been used for the treatment of edema, jaundice, and gonorrhea in traditional Oriental medicine. This screening study was designed to search the positive inotropic effects of herbal extracts in beating rabbit atria. Aquarius extracts of twenty six herbs were examined in atrial mechanical dynamics such as pulse pressure and stroke volume and atrial natriuretic peptide (ANP), one of the main hormones involved in the regulation of the body fluid and blood pressure homeostasis in perfused beating rabbit atria. Sophora flavescens Ait., Rheum officinale Baill., Acorus gramineus Sol., Chelidonium majus L., Pulsatilla koreana Nakai., Reynoutria japonica Houtt., Euphorbia lathyris L., Pyrrosia lingua (Thunb.) Farwell, Poncirus trifoliata Rafin., Anemarrhena asphodeloides Bunge, Kochia scoparia Schrad. significantly increased stroke volume and pulse pressure. However, those herbal extracts were not induced ANP secretion. We clarified the eleven herbal extracts for the positive inotropic effect independent of ANP secretion in beating rabbit atria. Thus these results provide a beneficial data for the treatment of the impairment of body fluid and blood pressure in traditional Oriental medicine.

Screening of Inhibitors of Extracellular Serine Protease of Acanthamoeba castellanii from Mushroom Extracts (버섯 추출물로부터 Acanthamoeba castellanii의 세포외 Serine 단백질분해효소 저해제 탐색)

  • Lee, Seung-Eun;Sancheti, Sandesh;Sancheti, Shruti;Choi, Mie-Young;Seo, Sung-Yum
    • The Korean Journal of Mycology
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    • v.36 no.2
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    • pp.178-182
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    • 2008
  • Although the number of patients with Acanthamoeba keratitis has increased dramatically since the widespread use of contact lens, it is still very hard to cure the disease. The proteases from the Acanthamoeba were reported to play important role in the pathogenesis of keratitis. In this study, the inhibitors for extracellular serine proteases of A. castellanii were screened from the extracts of 230 mushroom samples collected from various regions of Korea. The mushrooms were extracted with methanol and water ($65^{\circ}C$). Filtered and concentrated extracts (0.3 mg/ml) were preincubated with proteases before addition of peptide substrate N-succinyl-ala-ala-pro-phe p-anilide. The selected extracts showing strong inhibitory effects were characterized. Although inhibition with single extract was not so high enough, the complete inhibition was achieved with combination of two extracts. The selected extract showed little effect on other serine proteases such as thrombin (human and bovine) and on general protease such as protease K.

Extracellular Products from Cyanobacteria (시아노박테리아의 세포외산물에 대한 연구)

  • Kwon, Jong-Hee;Kim, Gi-Eun
    • KSBB Journal
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    • v.23 no.5
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    • pp.398-402
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    • 2008
  • Cyanobacteria havebeen identified as one of the most promising group producing novel biochemically active natural products. Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Though many useful natural products have been identified in cyanobacterial biomass, cyanobacteria produce also extracellular proteins related with NRPS/PKS. Detection of unknown secondary metabolites in medium was carried in the present study by a screening of 98 cyanobacterial strains. A degenerated PCR technique as molecular approaches was used for general screening of NRPS/PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%) and by A3/A7 primer in 26 strains (26.5%). HPLC analysis for a detection of natural products was performed in extracts from medium in which cyanobacteria containing putative PKS or NRPS were cultivated. CBT57, CBT62, CBT590 and CBT632 strains were screened for a production of extracellular natural products. 5 pure substances were detected from medium of these cyanobacteria.

The Effects of Chicken Leg Bone Extract on Antioxidative Properties under Different Heating Condition

  • Cheng, Fu-Yuan;Wan, Tien-Chun;Huang, Chao-Wei;Tominaga, Kana;Lin, Liang-Chuan;Sakata, Ryoichi
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1815-1820
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    • 2008
  • The aim of this study was to extract chicken leg bone, which is a by-product of industrial poultry processing, using different heating temperatures (80, 90 and $100^{\circ}C$) and durations (5, 10 and 15 min). The pH value, soluble protein content, peptide content and antioxidative properties, including superoxide anion scavenging ability, 1, 1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging ability, reducing capacity and inhibitory activity of linoleic acid peroxidation, were measured. The results showed no significant differences (p>0.05) in pH value among all treatments. Decreased soluble protein content and peptide content were observed in chicken leg bone extract obtained under higher heating temperatures (90 or $100^{\circ}C$) and longer heating durations (10 or 15 min). In antioxidative properties, the extracts which were heated at 90 or $100^{\circ}C$ for 15 min exhibited significantly higher superoxide anion scavenging ability, DPPH free radical scavenging ability, reducing capacity and inhibitory activity of linoleic acid peroxidation (p<0.05).

Increase in antifungal activity by the combination of tolaasin and its analogue peptides (톨라신류 펩티드 혼합처리에 의한 항진균 활성의 증가)

  • Yun, Yeong-Bae;Lee, Hyoung-Jin;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.69-73
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    • 2018
  • Oak mushroom (Lentinus edodes) is cultivated by using oak logs and sawdust medium. Green mold (Trichoderma) infection on these media severely suppresses the growth of oak mushroom. Usages of antibiotics and chemicals are not generally allowed to control of green mold since the mushroom is a fresh food. Tolaasin and its analogues, peptide toxins secreted by Pseudomonas tolaasii, have the antifungal activity and they have been successful to control the green mold disease. When the green mold, Trichoderma harzianum H1, was cultured in the presence of these toxins, the growth of fungus was effectively suppressed. In sawdust media, when the bacterial culture supernatants were sprayed on the aerial hyphae of green molds, the fungal growth was completely suppressed. Particularly, the antifungal activity was greatly increased by the combined culture extracts of P. tolaasii 6264 and HK11 strains. Therefore, these bacterial strains and their peptide toxins were able to suppress the growth of green molds and these can be good candidates to prevent from Trichoderma disease in oak mushroom cultivation.

Isolation of Angiotensin Converting Enzyme Inhibitor from Doenjang (전통된장으로부터 Angiotensin Converting Enzyme 저해물질의 분리)

  • Kim, Seung-Ho;Lee, Yun-Jin;Kwon, Dae-Young
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.848-854
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    • 1999
  • Inhibitory compounds of angiotensin converting enzyme (ACE) were separated from Doenjang (traditional Korean fermented soybean paste). Water extracts from Doenjang which showed ACE inhibitory activity were separated with gel permeation chromatography (GPC), in which two fractions with high ACE inhibitory activities were obtained. The first fraction from GPC was further isolated by semi-preparative reverse phase preparative-HPLC (high performance liquid chromatography) and 2-dimensional electrophoresis/thin layer chromatography (TLC). The purified spot had molecular weight of 759 daltons and ninhydrin-positive non-peptide. The second fraction from GPC was also further isolated by semi-preparative reverse phase HPLC and $NH_2-column$ HPLC. One fraction with high ACE inhibitory activity was purified and characterized. Molecular weight of this fraction by LC-MS was 272.34 daltons. The active fraction was identified as Arg-Pro with ACE $IC_{50}$ of $92\;{\mu}M$.

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