• Title/Summary/Keyword: enzyme resource

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Xanthone attenuates mast cell-mediated allergic inflammation

  • AYE, AYE;Jeon, Yong-Deok;Song, Young-Jae;Jin, Jong-Sik
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.123-123
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    • 2019
  • Xanthone is a kind of polyphenolic compounds that contain a distinctive chemical structure with a tricyclic aromatic ring found in a few higher plant families e.g. gentian root. This compound had a variety of biological activity, for instance antioxidant, antibacterial, anti-inflammatory, and anticancer effects. However, the effect of xanthone on mast cell-mediated allergic inflammation and its associated mechanism have not been elucidated. Therefore, the aim of this study was to elucidate the anti-allergic inflammatory effects and the underlying molecular mechanism of xanthone in PMACI-stimulated human mast cells-1 (HMC-1). In this result, xanthone treatment decreased the production of histamine, pro-inflammatory cytokines such as tumor necrosis factor-a (TNF-${\alpha}$), IL-6, and IL-8 and expressions of TSLP in PMACI-stimulated HMC-cells. In addition, xanthone significantly suppressed the phosphorylation of MAPKs and the activation of NF-${\kappa}B$ signal pathway in activated mast cells. Furthermore, xanthone inhibited the activation of caspase-1, an IL-$1{\beta}$ converting enzyme, in PMACI-stimulated HMC-1 cells. These findings provide evidence that xanthone could be a potential therapeutic agent for allergy-related inflammatory disorders.

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Development of Bile Salt-Resistant Leuconostoc citreum by Expression of Bile Salt Hydrolase Gene

  • Cho, Seung Kee;Lee, Soo Jin;Shin, So-Yeon;Moon, Jin Seok;Li, Ling;Joo, Wooha;Kang, Dae-Kyung;Han, Nam Soo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.12
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    • pp.2100-2105
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    • 2015
  • Probiotic bacteria must have not only tolerance against bile salt but also no genes for antibiotic resistance. Leuconostoc citreum is a dominant lactic acid bacterium in various fermented foods, but it is not regarded as a probiotic because it lacks bile salt resistance. Therefore, we aimed to construct a bile salt-resistant L. citreum strain by transforming it with a bile salt hydrolase gene (bsh). We obtained the 1,001 bp bsh gene from the chromosomal DNA of Lactobacillus plantarum and subcloned it into the pCB4170 vector under a constitutive P710 promoter. The resulting vector, pCB4170BSH was transformed into L. citreum CB2567 by electroporation, and bile salt-resistant transformants were selected. Upon incubation with glycodeoxycholic acid sodium salt (GDCA), the L. citreum transformants grew and formed colonies, successfully transcribed the bsh gene, and expressed the BSH enzyme. The recombinant strain grew in up to 0.3% (w/v) GDCA, conditions unsuitable for the host strain. In in vitro digestion conditions of 10 mM bile salt, the transformant was over 67.6% viable, whereas only 0.8% of the host strain survived.

Production of Minor Gisenosides from Gypenoside V (Gypenoside V로부터 minor ginsenosides의 생산)

  • Son, Na-Ri;Min, Jin-Woo;Jang, Mi;Kim, Hyo-Yeon;Jeon, Ji-Na;Yang, Deok-Chun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.10a
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    • pp.20-20
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    • 2010
  • Panax ginseng C.A Meyer is frequently taken orally as a traditional herbal medicine in Asian countries. The major components of ginseng are ginsenoside, which are pharmaceutical activity. The six major ginsenosides, including Rb1, Rb2, Rc, Rd, Re and Rg1 account for 90% of total ginsenosides. Even though the minor ginsenosides, including Rg3, Rh2 and compound K has high pharmacetical activities, the price of minor ginsenosides is too high. Therefore we isolated the gypenoside V and made it converted to minor ginsenosides. In the plant Gynostemma pentaphyllum Makino, gypenosdie V was presented as dominant saponin (content about 2.4%), and was similar to protopanaxadol type ginsenosides such as ginsenoside Rb1. In this study, we confirmed that the coversion of gypenoside V to minor ginsenosides after using the various treatment such as heating, acid treatment, commercial edible enzyme, and lactobacillus. Consequently, we optimizied the transformation of gypenoside V to minor ginsenoside using Thin Layer Chromatography (TLC), High Performance Liquid Chromatography (HPLC), Time-of-flight Mass Spectrometry (LC/TOF/MS).

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A Culture-Based Study of the Bacterial Communities within the Guts of Nine Longicorn Beetle Species and their Exo-enzyme Producing Properties for Degrading Xylan and Pectin

  • Park, Doo-Sang;Oh, Hyun-Woo;Jeong, Won-Jin;Kim, Hyang-Mi;Park, Ho-Yong;Bae, Kyung-Sook
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.394-401
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    • 2007
  • In this study, bacterial communities within the guts of several longicorn beetles were investigated by a culture-dependent method. A total of 142 bacterial strains were isolated from nine species of longicorn beetle, including adults and larvae. A comparison of their partial 16S rRNA gene sequences showed that most of the bacteria constituting the gut communities can typically be found in soil, plants and the intestines of animals, and approximately 10% were proposed as unreported. Phylogenetic analysis demonstrated that the bacterial species comprised 7 phyla, and approximately half were Gammaproteobacteria. Actinobacteria were the second most populous group (19%), followed by Firmicutes (13%) and Alphaproteobacteria (11%). Betaproteobacteria, Flavobacteria, and Acidobacteria were minor constituents. The taxonomic compositions of the isolates were variable according to the species of longicorn beetle. Particularly, an abundance of Actinobacteria existed in Moechotypa diphysis and Mesosa hirsute, which eat broadleaf trees; however, no Actinobacteria were isolated from Corymbia rubra and Monochamus alternatus, which are needle-leaf eaters. Considerable proportions of xylanase and pectinase producing bacteria in the guts of the longicorn beetles implied that the bacteria may play an important role in the digestion of woody diets. Actinobacteria and Gammaproteobacteria were the dominant xylanase producers in the guts of the beetles.

Characterization of an Extracellular Lipase in Burkholderia sp. HY-10 Isolated from a Longicorn Beetle

  • Park, Doo-Sang;Oh, Hyun-Woo;Heo, Sun-Yeon;Jeong, Won-Jin;Shin, Dong-Ha;Bae, Kyung-Sook;Park, Ho-Yong
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.409-417
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    • 2007
  • Burkholderia sp. HY-10 isolated from the digestive tracts of the longicorn beetle, Prionus insularis, produced an extracellular lipase with a molecular weight of 33.5 kDa estimated by SDS-PAGE. The lipase was purified from the culture supernatant to near electrophoretic homogenity by a one-step adsorption-desorption procedure using a polypropylene matrix followed by a concentration step. The purified lipase exhibited highest activities at pH 8.5 and $60^{\circ}C$. A broad range of lipase substrates, from $C_4\;to\;C_{18}$ p-nitrophenyl esters, were hydrolyzed efficiently by the lipase. The most efficient substrate was p-nitrophenyl caproate ($C_6$). A 2485 bp DNA fragment was isolated by PCR amplification and chromosomal walking which encoded two polypeptides of 364 and 346 amino acids, identified as a lipase and a lipase foldase, respectively. The N-terminal amino acid sequence of the purified lipase and nucleotide sequence analysis predicted that the precursor lipase was proteolytically modified through the secretion step and produced a catalytically active 33.5 kDa protein. The deduced amino acid sequence for the lipase shared extensive similarity with those of the lipase family 1.2 of lipases from other bacteria. The deduced amino acid sequence contained two Cystein residues forming a disulfide bond in the molecule and three, well-conserved amino acid residues, $Ser^{131},\;His^{330},\;and\;Asp^{308}$, which composed the catalytic triad of the enzyme.

Functional Properties of Peptides in Mixed Whey and Soybean Extracts after Fermentation by Lactic Acid Bacteria

  • Dong-Gyu Yoo;Yu-Bin Jeon;Se-Hui Moon;Ha-Neul Kim;Ji-Won Lee;Cheol-Hyun Kim
    • Journal of Dairy Science and Biotechnology
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    • v.41 no.3
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    • pp.113-125
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    • 2023
  • In this study, we explored the synergistic effects of whey protein concentrate (WPC) and soybean protein components after fermentation with lactic acid bacteria isolated from kimchi, and identified several peptides with desirable physiological functions, proteolysis, and immune effects. Antioxidant activity was determined using 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid, 1,1-diphenyl-2-picrylhydrazyl, ferric-reducing antioxidant power, and hydroxyl radical scavenging assays, followed by cross-validation of the four antioxidant activities. These assays revealed that samples with a 8:2 and 9:1 whey to soy ratio possessed higher antioxidant activity than the control samples. Antibacterial potency testing revealed high antibacterial activity in the 9:1 and 8:2 samples. Cytotoxicity testing of samples using 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide revealed that only the 10:0, 1:9, and 0:10 samples had <80% viable cells, indicating no significant cytotoxicity. Nitric oxide (NO) assays revealed that NO expression was reduced in 8:2, 5:5, and 0:10 protein ratio fermentations, indicating low inflammatory reaction stimulatory potential. Cytokine expression was confirmed using an enzyme-linked immunosorbent assay kit. The 8:2 sample had the lowest inflammatory cytokine (interleukin [IL]-1α, IL-6, and tumor necrosis factor-α) levels compared with the lipopolysaccharide-treated group. Amino acid profiling of the 8:2 sample identified 17 amino acids. These results suggest that inoculating and fermenting Lactobacillus plantarum DK203 and Lactobacillus paracasei DK209 with an 8:2 mixture of WPC and soybean protein releases bioactive peptides with excellent anti-inflammatory and antioxidant properties, making them suitable for functional food development.

Enzymatic Hydrolysis of Cellulose (섬유소(纖維素)의 당화(糖化))

  • Lee, Kye-Joon
    • Korean Journal of Pharmacognosy
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    • v.7 no.2
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    • pp.85-93
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    • 1976
  • Since cellulose is the only organic material that is annually replenishable in very large quantities, we must explore ways to utilize it as a source of energy, food and chemicals. For the utilization of this resource, it is first enzymatic hydrolyzed to glucose, then the glucose can be used as a food, converted single cell protein by microorganism, fermented to clean burning fuel and other chemicals. Cellulolytic enzyme, cellulase, consists of two or three major components, $C_1-cellulase$, $C_x-cellulase$ and ${\beta}-glucosidase$. $C_x-cellulase$ are fairly common but $C_1-cellulase$ are quite rare. Trichoderma viride is the best source of active cellulose, especially $C_1-enzyme$. Saccharification rate of cellulose in greatly influenced by the degree of crystallinity and extent of lignification. But by the pretreatment the substrate with cellulose swelling agent, delignifying reagent and physical treatment, the degree of saccharification is enhanced. Thus, glucose syrups of 2 to 10% concentration are realized from milled newspaper. The enzymatic hydrolysis of such energy rich material, such as cellulose, to glucose is technically feasible and practically achievable on a very large scale.

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Lysosomal Storage Disorders in India: A Mini Review

  • Gupta, Neerja;Aggarwal, Bhawana;Kabra, Madhulika
    • Journal of mucopolysaccharidosis and rare diseases
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    • v.4 no.1
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    • pp.1-6
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    • 2018
  • Lysosomal storage disorders are a group of rare inherited metabolic disorders with protean manifestations and variable severity ranging from attenuated forms to severe ones. It is necessary to diagnose and manage these disorders timely before irreversible damage occurs. Prior to the era of enzyme replacement therapy and newer therapeutics, only treatment option available was palliative care. Over the past two decades, extensive research in the lysosomal storage disorders has led to substantial expansion of our understanding about them. This mini review focusses on the spectrum, challenges faced in the diagnosis and therapy and remedial actions taken so far in lysosomal storage disorders in resource constrained country like India.

Enzymatic Hydrolysis of Marine Algae Hizikia fusiforme (해조류 톳 (Hizikia fusiforme)의 효소 가수분해)

  • Song, Bu-Bok;Kim, Sung-Koo;Jeong, Gwi-Taek
    • KSBB Journal
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    • v.26 no.4
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    • pp.347-351
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    • 2011
  • In this study, we investigated the effect of reaction factors on enzymatic hydrolysis of Hizikia fusiforme, which is brown algae in marine biomass resource, using commercial enzymes. The composition of H. fusiforme is 38.9% of reducing sugar, 4.8% of moisture, 17.8% of ash, and 38.5% of others. In the condition of 1-5% substrate, the increase of substrate concentration enhanced the increase of reducing sugar formation; however, the hydrolysis yield did not increase after 24 h. After reaction of 75 h, conversion yield of reducing sugar were obtained to 16.45%, 17.99%, and 14.55% at 1, 2.5, and 5% substrate, respectively. As a result of effect of enzyme amount, the formation of reducing sugar did not show considerable change at 1% substrate. However, in the condition of 2.5% substrate, the great change of reducing sugar formation was observed by the increase of enzyme amount. The conversion yields of reducing sugar were obtained to 18.77% and 22.83% at 1% and 2.5% substrate with 30% enzyme, respectively. As a result of heat treatment of biomass, the high yield was obtained in 2.5% substrate and the yields were increased to 0.06-7.2% by the heat treatment. This result will provide the basic information for production process of biofuels and chemicals from marine biomass H. fusiforme.

Expression, Purification, and Crystallization of D-Psicose 3-Epimerase from Agrobacterium tumefaciens

  • Kim Kwang-Soo;Kim Hye-Jung;Oh Deok-Kun;Cheong Jong-Joo;Rhee Sang-Kee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.647-650
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    • 2006
  • D-Psicose 3-epimerase (DPE) catalyzes the interconversion of D-fructose to D-psicose by epimerizing the carbon-3 position. The DPE from Agrobacterium tumefaciens was cloned and expressed in Escherichia coli. The expressed enzyme was purified by affinity chromatography on an IMAC, gel filtration chromatography on a Sephacryl S-300 HR, and anion-exchange chromatography on a RESOURCE Q. The molecular mass of the purified enzyme was estimated to be about 135 kDa by Superdex 200 gel filtration chromatography, corresponding to a homotetramer. The enzyme produced crystals suitable for X-ray diffraction to a $2.0{\AA}$ resolution at 100 K. The crystals were found to belong to the orthorhombic space group $P2_12_12_1$, with unit-cell parameters a=102.4, b=113.0, and $c=131.8{\AA}$. In addition, the calculated packing parameter $(V_m)$ was $2.79{\AA}^3/Da$, the solvent content was 55.92%, and an asymmetric unit consisted of four monomers.