• 한국임상수의학회지
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• 제37권5호
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• pp.249-254
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• 2020

Neutrophil extracellular trap (NET) formation is an immune response for the invasion of microbes. The purpose of this study is to examine the effect of zinc on NET formation of porcine peripheral blood polymorphonuclear cells (PMNs). The NET formation of PMNs was measured by fluorescence microplate reader. The production of tumor necrosis factor (TNF)-α in the culture supernatants from zinc-treated peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay (ELISA). Zinc itself did not have no effect on NET formation. However, the NET formation of PMNs was increased by culture supernatants from PBMCs treated with zinc. Also, the NET formation of PMNs was increased by recombinant porcine (rp) TNF-α. The production of TNF-α in PBMCs culture supernatants was shown to increase upon zinc treatments. These NET formations of PMNs increased by either culture supernatant from PBMCs treated with zinc or rpTNF-α were inhibited by treatment of anti-rpTNF-α polyclonal antibody (pAb). These results suggested that zinc has an immunostimulating effect on the NET formation of PMNs, which is mediated by TNF-α released from zinc-treated PBMCs. Therefore, zinc may play an important role for NET formation in the defense of porcine inflammatory diseases.

• IMMUNE NETWORK
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• 제5권1호
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• pp.30-35
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• 2005

Background: p38 and extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase (MAPK) signaling are thought to have critical role in lipopolysaccharide (LPS)-induced immune response but the molecular mechanism underlying the induction of these signaling are not clear. Methods: Specific inhibitors for p38, SB203580, and for ERK, PD98059 were used. Cells were stimulated by LPS with or without specific MAPK inhibitors. Results: LPS activated inducible nitric oxide synthase (iNOS), subsequent NO productions, and pro-inflammatory cytokine gene expressions (TNF-${\alpha}$, IL-$1{\beta}$, IL-6, and IL-12). Treatment of both SB203580 and PD98059 decreased LPS-induced NO productions. Concomitant decreases in the expression of iNOS mRNA and protein were detected. SB203580 and PD98059 decreased LPS-induced gene expression of IL-$1{\beta}$ and IL-6. SB203580 increased LPS-induced expression of TNF-${\alpha}$ and IL-12, and reactive oxygen species production, but PD98059 had no effect. Conclusion: These results indicate that both p38 and ERK pathways are involved in LPS-stimulated NO synthesis, and expression of IL-$1{\beta}$ and IL-6. p38 signaling pathways are involved in LPS-induced TNF-${\alpha}$ and IL-12, and reactive oxygen species plays an important role in these signaling in macrophage.

• Journal of Microbiology
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• 제40권1호
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• pp.20-25
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• 2002

An extracellular PHB depolymerase was purified from P. simplicissimum LAR13 cultural medium by Sepharose CL-6B chromatography. When the fungus was grown in a basal salt medium with poly(3-hydroxybutyrate) (PHB) as the sole carbon source, PHB depolymerase production reached maximum at its stationary phase. The mycelial growth rate was higher at 37$^{\circ}C$ than at 30$^{\circ}C$ and even higher than at 25$^{\circ}C$, However, the enzyme production was lower at 37$^{\circ}C$ than 30$^{\circ}C$ or 25$^{\circ}C$. The isolated enzyme is composed of a single polypeptide chain with a molecular mass of about 36 kDa as determined by SDS-PAGE. The optimum conditions for the enzyme activity are pH 5.0 and 45$^{\circ}C$. The enzyme was stable for 30 min at a temperature lower than 50$^{\circ}C$, and stable at pH higher than 2.0 but it was unstable at pH 1.0.1 mM Fe$\^$2+/ reduced the enzyme activity by 56% and the enzyme was inhibited almost completely by 4 mM Fe$\^$2+/ . The enzyme was partially inhibited by phenylmethylsulfonyl fluoride and was very sensitive to diazo-DL-norleucine methyl esters dithiothreitol and mercuric ion. However, N-p - tosyl - L - Iysinechloromethyl ketone, p -hydroxymercuricbenzoate and N- acetylimidazole had no influence upon its activity.

• 한국미생물·생명공학회지
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• 제24권6호
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• pp.756-762
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• 1996

In order to develop the method for mass production of natural food colorant from Monascus anka, optimum cultivation conditions for producing red and yellow pigments by cultiva- ting the mold in a jar fermenter and their color characteristics were investigated. The mold produced red and yellow pigments both intracellularly and extracellularly. These pigments showed unique light absorption characteristics with maximum absorption of 494, 380, 506, and 388 nm for extracellular red pigment (ERP), extracellular yellow pigment (EYP), intracellular red pigment (IRP), and intracellular yellow pigment (IYP), respectively. Optimum conditions for producing red pigments were found to be temperature 30$\circ$C, initial pH 6.0, rice powder 3-5%, peptone 0.05%, magnesium sulfate 0.25%, aeration rate 0.1 vvm. Optimum temperature for producing yellow pigments was around 35$\circ$C which is higher than that of producing red pigments. The initial pH and rice powder concentration for producing yellow pigments were the same as those of producing red pigments. The higher concentration of nitrogen source and inorganic salt, aeration rate, the more the yellow pigments were produced. The optimum agitation speed was 100 - 300 rpm for pigment production.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.590-600
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• 2003

The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

• 미생물학회지
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• 제30권6호
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• pp.501-506
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• 1992

An alkalophilic Vibrio sp. RH530 showing high proteolytic activity was isolated form soil samples by enrichment culture. The activity staining using gelatin SDS- polyacrylamide gel electrophoresis (PAGE ) revealed that the strain produced an alkaline major protease (Apr B) with a size of 27 kDa, and at least six minor proteases. The apparent sizes of four of the minor proteases were approximately 45, 28, 22 and 19 kDa. Apr B and five of the minor proteases were inhibited by serine protease inhibitors including PMSF and DFP, suggesting that they are serine proteases. One of the minor proteases was inhibited by metalloprotease inhibitors, not by serine protease inhibitors, indicating it to be a metalloprotease. Furthermore, the activities of Apr B and Prt 3 were not inhibited by SDS in the reaction mixture. The production of Apr B and some of the minor proteases was specifically affected by culture temperature (30 to 37.deg.C) and pH (7 to 10). The production of Apr B. Prt 2, Prt 5 and Prt 6 was mainly affected by culture temperature, while Prt 4 by culture pH. Prt 1 and Prt 3 were not affected by neither of these factors.

• 한국미생물·생명공학회지
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• 제34권3호
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• pp.216-220
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• 2006

본 균의 생육 및 효소생산에 유용한 탄소원으로서 자연계의 식물에 풍부한 펙틴을 탄소원으로 할 경우, 그 생육도는 전분보다 뛰어났으며, alcohol oxidase와 catalase의 생산량도 높아지는 것으로 나타났다. 특히 alcohol oxidase의 경우는 전분의 15배 이상의 생산량을 보여 본 균과 펙틴 이용성과의 관계를 시사하였고, 세포외 pectin esterase, pectinase등의 높은 활성이 검출되어 이를 증명하였다. 또한 alcohol oxidase 반응에서 생성되는 물질인 formaldehyde를 산화하는 formaldehyde dehydrogenase와, formate를 산화하여 $CO_2$를 생성하는 formate dehydrogenase의 반응을 발견하여, 본 균의 pectin 이용성과 관련한 일련의 에너지 대사계의 존재를 추정할 수 있었다.

• 미생물학회지
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• 제12권3호
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• pp.115-130
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• 1974

Searches for the nutrition requirements of three strains of Brevibacterium ammoniagenes reported in the previous paper were carried out with an aim of achieving the striking accumulation of L-glutamic acid and the large multipication of cells. It was recognized that all three strains required both biotin and thiamine, together with amino acids such as histidine or cysteine, for their good growth and extracellular L-glutamic acid accumulation. The quantity of biotin required for remarkable growth of these microorganisms was quite different from that for the maximum production of L-glutamic acid. This result, however, did not apply in the case of thiamine. It was also confirmed that, of 18 amino acids, histidine and cysteine were the msot effective organic nitrogen sources, while the most available inorganic ammonium salt resulting in a large amount of L-glutamic acid-production and considerable cell gorwth was found to be only urea. Maximum accumulation of extracellular L-glutamic acid, more than 50%(w/w) of the initial sugar content, could be obtained from fermentation in the medium containing wheat-bran extract(Brev. ammoniagenes T-1 and Brev.ammoniagenes Y-2) or rice-bran extract(Brev. ammoniagenes YR-2), which confirmed us a possibility that these bacteria might be employed for industrial fermentation of L-glutamic acid.

• 미생물학회지
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• 제31권1호
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• pp.79-84
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• 1993

Infection of Chinook Salmon Embryo (CHSE) cells with IPNV resulted in a significant decrease in intracellular free calcium concentration ([$Ca^{2+}$]i) compared to mock-infected cells. The degree of the decrease in [Ca$^{2+}$]i was dependent on the amount of input virus, and treatment of IPNV-infected CHSE cells with metabolic inhibitors such as cyloheximide cordycepin partially reversed the decrease in [$Ca^{2+}$]i in IPNV-infected cells. Inactiation of PINV with UV also abolished IPNV-induced decrease in [$Ca^{2+}$]i. These data suggest an active role of IPNV in the decrease of [Ca$^{2+}$]i in the infected CHSE cells. The importance of the decrease in [$Ca^{2}$i] could be supported by the finding that the production of IPNV plaques increased in the cells treated with verapamil, a calcium influex blocker, and by lowering the concentration of extracellular calcium. Decreased production of IPNV plaques was observed by elevating the extracellular calcium. Thus, it is suggested that IPNV induced a decreased in [$Ca^{2+}$]i and the decrease in [$Ca^{2+}$]i may plan an importat role in efficient replication of IPNV.ation of IPNV.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1316-1323
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• 2017

Geosmin and 2-methylisoborneol (2-MIB), responsible for earthy or musty smell, are a major concern for safe drinking water supplies. This study investigated the effects of environmental factors on odorous compound production and cell growth in cyanobacterial strains. Anabaena sp. FACHB-1384, a 2-MIB producer, was sensitive to low temperature (<$20^{\circ}C$). However, geosmin producers, Anabaena sp. Chusori and Anabaena sp. NIER, were sensitive to high light intensity (>$100{\mu}mol/m^2/sec$), but not to low temperature. Geosmin concentrations increased under higher nitrate concentrations, being linearly proportional to cell density. A P-limited chemostat showed that P-stress decreased the geosmin productivity and extracellular geosmin amount per cell in Anabaena sp. NIER. However, only 2-MIB productivity was reduced in Planktothrix sp. FACHB-1374 under P-limitation. The extracellular 2-MIB amount per cell remained constant at all dilution rates. In conclusion, high light intensity and P-stress can contribute to the lower incidence of geosmin, whereas 2-MIB reduction could be attainable at a lower temperature.