• Title/Summary/Keyword: whole-cell activity

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Modulatory Activity of Bifidobacterium sp. BGN4 Cell Fractions on Immune Cells

  • Kim Nam-Ju;Ji Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.584-589
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    • 2006
  • Bifidobacteria has been suggested to exert health promoting effects on the host by maintaining microbial flora and modulating immune functions in the human intestine. We assessed modulatory effects of the different cell fractions of Bifidobacterium sp. BGN4 on macrophage cells and other immune cells from the spleen and Peyer's patches (PP) of mouse. Cell free extracts (CFE) of the BGN4 fractions induced well-developed morphological changes in the macrophages and increased the phagocytic activity more effectively than other fractions in the mouse peritoneal cells. Nitric oxide (NO) production was significantly reduced by both the cell walls (CW) and CFE in the cultured cells from the spleen and PP. The production of interleukin-6 (IL-6) and interleukin-10 (IL-10) was eminent in the spleen cells treated with experimental BGN4 cell fractions. However, in the PP cells, IL-6 was slightly decreased by the treatment with the whole cell (WC) and CW, whereas IL-10 was significantly increased by the treatment with the CW and CFE. These results suggest that different types of bifidobacterial cell fractions may have differential immunomodulatory activities depending on their location within the host immune system.

Studies on Whole Cell Immobilized Glucose Isomerase - I. Preparation and Properties of Whole Cell Immobilized Glucose Isomerase - (포도당 이성화 효소의 세포 고정화에 관한 연구 - I. 세포 고정화 효소의 제조와 성질 -)

  • Ahn, Byung-Yoon;Byun, Si-Myung
    • Korean Journal of Food Science and Technology
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    • v.11 no.3
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    • pp.192-199
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    • 1979
  • With cells of Streptomyces spp K-45 isolated from soil, the immobilization of glucose isomerase by a series of treatments ; heat, carefully manipulated drying, extrusion with a thickening agent, and glutaraldehyde-induced crosslinking, was presented. This was aimed to obtain a mechanically stable form of whole cell containing glucose isomerase. The resulted pellet form had a good mechanical strength, compared with a commercial product, and showed 26 % of the activity recovery. The specific activity was 48.1 units per g of the dry material. The immobilized glucose isomerase generally showed properties similar to those of the soluble enzyme ; optimal pH at $7.5{\sim}9.0$, optimal temperature at $80{\sim}85^{\circ}C$, activation energy of 10.9 kcal/mole, and $K_m$ for glucose of 10.9M. The immobilized enzyme was very thermostable and pH stable.

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Biodegradation of marine microplastics by the whole-cell catalyst overexpressing recombinant PETase (PET분해효소(PETase) 과발현 전세포 촉매의 해양미세플라스틱 생분해 활성 연구)

  • Hyunji, Kim;Jong-Ha, Park;Ae-Ran, Park;Dae-Hee, Lee;Joonho, Jeon;Hyuk Taek, Kwon;Sung In, Lim
    • Journal of Marine Bioscience and Biotechnology
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    • v.14 no.2
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    • pp.133-142
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    • 2022
  • The increased production and consumption of polyethylene terephthalate (PET)-based products over the past several decades has resulted in the discharge of countless tons of PET waste into the marine environment. PET microparticles resulting from the physical erosion of general PET wastes end up in the ocean and pose a threat to the marine biosphere and human health, necessitating the development of new technologies for recycling and upcycling. Notably, enzyme-mediated PET degradation is an appealing option due to its eco-friendly and energy-saving characteristics. PETase, a PET-hydrolyzing enzyme originating from Ideonella sakaiensis, is one of the most thoroughly researched biological catalysts. However, the industrial application of PETase-mediated PET recycling is limited due to the low stability and poor reusability of the enzyme. Here we developed the whole-cell catalyst (WCC) in which functional PETase is attached to the outer membrane of Escherichia coli. Immunoassays are used to identify the surface-expressed PETase, and we demonstrated that the WCC degraded PET microparticles most efficiently at 30℃ and pH 9 without agitation. Furthermore, the WCC increased the PET-degrading activity in a concentration-dependent manner, surpassing the limited activity of soluble PETase above 100 nM. Finally, we demonstrated that the WCC could be recycled up to three times.

Studies on the Cell Immobilization of Alkalophilic Streptomyces sp. B-2 for the Glucose Isomerization (포도당 이성화를 위한 Alkalophilic Streptomyces sp. B-2의 균체 고정화에 관한 연구)

  • 이은숙
    • The Korean Journal of Food And Nutrition
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    • v.11 no.3
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    • pp.319-322
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    • 1998
  • The whole cell of alkalophilic Streptomyces sp. B-2 which produce glucose isomerase was immobilized by entrapment method for the effective production of high fructose syrup. The highest immobilized activity was achieved when the enzyme was bound to 2% $textsc{k}$-carrageenan. Immobilized glucose isomerase the pH optimum was about pH 7.5~8.5. Immobilization of alkalophilic Streptomyces sp. B-2 on 2% $textsc{k}$-carrageenan at 7$0^{\circ}C$ showed an increase in glucose isomerase activity. GI activity of immobilized cells was maximum Co2+ concentration 10-3M, Mg2+ concentration 10-3M.

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RELATIONSHIP BETWEEN VIRULENCE, METABOLIC ACID AND GENETIC HETEROGENEITY OF PORPHYROMONAS GINGIVALIS (Porphyromonas gingivalis의 독성, 대사산물 및 유전자이종성과의 관련성)

  • Kim, Kang-Ju;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.1-15
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    • 1993
  • P. gingivalis has been implicated as a strong pathogen in periodontal disease and known to have three serotypes of P. gingivalis. The purpose of this study is to investigate on the relationship between virulence, metabolic acids and genetic heterogeneity of P. gingivalis. P. gingivalis W50 standard strain and five strains of P. gingivalis serotype b Korean isolates were used in this study. For in vitro virulence test, lyophilized whole cell P. gingivalis were suspended, and sonicated with ultrasonic dismembranometer. Sonicated samples were applied to cultured cells derived from periodontal ligament, and cell activity was assayed with growth and survival assay. The metabolic acids were also extracted, and determined by High Performance Liquid Chromatography. Pst I-digested bacterial genomic DNA was electrophoresed, and densitometric analysis was performed to study the genetic heterogeneity. All of the P. gingivalis serotype b produced butyric acid. In cell activity study, butyric acid inhibited the cell activity irrespective of its concentration. Densitometric analysis showed restriction fragment length polymorphism. These results suggested that there existed heterogeneity of the metabolic acids and the virulence of P. gingivalis and such heterogeneity might be related to genetic heterogeneity.

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Optimization of Microbial Cell-Based Spectrometric Assay for the Analysis of Epoxide Hydrolase Activity (미생물 세포 기반의 에폭사이드 가수분해효소 활성 측정을 위한 분광학적 분석법 최적화)

  • Kim Hee Sook;Lee Eun Yeol
    • Journal of Life Science
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    • v.15 no.1 s.68
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    • pp.136-140
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    • 2005
  • Microbial cell-based UV spectrometric assay for the quantitative measurement of epoxide hydrolase activity was evaluated and optimized for the efficient screening of whole cell activity of novel epoxide hydrolase. Epoxide hydrolase activity was determined by measuring the increase of the oxidized product, benzaldehyde. The effects of the concentrations of phenyl-1,2-ethanediol, sodium metaperiodate and cells were optimized for epoxide hydrolase-catalyzed hydrolysis of styrene oxide. The relevant kinetic parameters of Km and $V_{max}$ for the hydrolysis of (R)-styrene oxide by Rhodotorula glutinis were determined from Lineweaver-Burk plot as 41.2 nmol/min$\cdot$mg dcw and 7.5 mM respectively, and coincided well with those from GC analysis.

Effects of the Antitumor Component, F-D-P, Isolated from Elfvingia applanata on the Immune Response

  • Kim, Young-So;Ryu, Ku-Hyun;Mo, Young-Keun;Lee, Chong-Kil;Han, Seong-Sun
    • Korean Journal of Pharmacognosy
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    • v.25 no.4 s.99
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    • pp.348-355
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    • 1994
  • An antitumor component, F-D-P, was purified from the hot water extract of the carpophores of Elfvingia applanata by precipitation with ethanol, dialysis, and passage through a column of DEAE-cellulose ion exchange. F-D-P inhibited the growth of Sarcoma 180 in mice showing the tumor inhibition ratio of 88.3% in doses of 20 mg/kg for ten days. Chemical analysis of F-D-P showed that it was composed of polysaccharide(65.3%) and protein(6.5%0, and that the monosaccharides consisting of the polysaccharide was glucose(89.1%) and mannose(10.9%). The immunomodulatory activities of F-D-P were explored by determining its effect on the proliferation of the whole and subpopulations of lymphocytes, and on the generation of natural killer(NK) cell activity in vitro. F-D-P was mitogenic to total lymphocytes and B cells, but not to purified T cells, even in the presence of accessory cells. F-D-P did not increase NK cell activity when added to cultures of resting lymphocytes. From these results, it is clear that F-D-P modulates primarily the humoral immune responeses.

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Spontaneous Electrical Activity in Cerebellar Purkinje Neurons of Postnatal Rats

  • Nam, Sang-Chae
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.355-366
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    • 1997
  • Although cerebellar Purkinje cells display spontaneous electrical activity in vivo and in slice experiments, the mechanism of the spontaneous activity generation has not been clearly understood. The aim of this study was to investigate whether cerebellar Purkinje cells of postnatal rats generate spontaneous electrical activity without synaptic inputs. Dissociated cerebellar Purkinje cells were used for reducing synaptic inputs in the present study. Cerebellar Purkinje cells with dendrites were dissociated from postnatal rats using enzymatic treatment followed by mechanical trituration. Spontaneous electrical activities were recorded from dissociated cells without any stimulus using whole-cell patch clamp configuration. Two types, spontaneously firing or quiescent, of dissociated Purkinje cells were observed in postnatal rats. Both types of cells were identified as Purkinje cells using immunocytochemical staining technique with anti-calbindin after recording. Spontaneously active cells displayed two patterns of firing, repetitive and burst firings. Two thirds of dissociated Purkinje cells displayed repetitive firing and the rest of them did burst firing under same recording condition. Repetitive firing activities were maintained even after further isolation using either physical or pharmacological techniques. Neither high magnessium solution nor excitatory synaptic blockers, AP-5 and DNQX, block the spontaneous activity. These results demonstrate that spontaneous electrical activity of isolated cerebellar Purkinje cells in postnatal rats is generated by intrinsic membrane properties rather than synaptic inputs.

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Antioxidant and Anti-inflammatory Effects of Taraxacum hallaisanense Nakai Extracts (좀민들레(Taraxacum hallaisanense Nakai) 추출물의 항산화 및 항염증 활성 효과)

  • Nan, Li;Choo, Byung-Kil
    • Korean Journal of Organic Agriculture
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    • v.26 no.3
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    • pp.501-514
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    • 2018
  • Taraxacum hallaisanense (pr), a species of the family Compositae is a perennial herb plants that inhabit to Jeju Island. In the study, we performed to determine the anti-inflammatory effects in LPS-induced RAW 264.7 cells and antioxidant activity of ethanol extracts from T. hallaisanense whole plants. The antioxidant activity of extracts was measured by contents of polyphenol and flavonoid, DPPH radical scavenging, and reducing power activity. The anti-inflammatory effect of T. hallaisanense extracts was measured by NO and $IL-1{\beta}$ production inhibitory activity and the expression of pro-inflammatory in lipopolysaccharide (LPS)-induced Raw 264.7 cells. Also, the expression of pro-inflammatory genes such as iNOS, COX-2 and NF-kB protein were reduced. In the cytotoxicity measurement by cytotoxicity kit, the extract was exhibited Raw 264.7 cell viabilities as nontoxic result in concentration of $25{\sim}400{\mu}g/ml$. These results indicated that ethanol extracts of T. hallaisanense whole plants expected development possibility as nutrial additives through high anti-inflammatory effects and antioxidant activity.

Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi (김치로부터 분리된 Lactobacillus paraplantarum KNUC25가 만드는 항균 물질의 특성)

  • Kim, Ma-Rie;Lee, Su-Jin;Seul, Keyung-Jo;Park, Yu-Mi;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.24-32
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    • 2009
  • The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at $30^{\circ}C$.