• Title/Summary/Keyword: Extracellular

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Production and Characteristics of an Extracellular Pigment through the Submerged Cultivation of Phellinus Sp. (Phellinus SP.의 액체배양에 의한 세포외 색소물질의 생산 및 특성)

  • Lee, Dong-Ki;Lee, Chul-Won;Lee, Shin-Young
    • KSBB Journal
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    • v.23 no.1
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    • pp.23-30
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    • 2008
  • An extracellular pigment production of three Phellinus sp. (Phellinus 421, P. linteus and P. hartigil) through submerged cultivation was investigated. The maximum brown pigment from culture broth was obtained from the precipitate by addition of 10% 1M HCI solution. This precipitate showed absorption characteristics with ${\lambda}_{max}$ of 360nm. The maximum production of extracellular pigment obtained at optimum medium and culture condition was 3.54 ($A_{360}$). The precipitate was fractionated by Sephadex G-75 gel chromatography, and the isolated brown pigment contained a large amount of polyphenol and the small amounts of sugar and protein. The brown pigment fraction was stable in temperature range of $30{\sim}60^{\circ}C$, pH range of $4{\sim}6$, sugar addition ranges of $1{\sim}5%$ and salt addition concentration of 3 molarity. Antioxidative activity of the brown pigment by TBA method was better than that of vitamin E (${\alpha}$-tocopherol).

Distribution of Extracellular Proteases from Various Vibrio Species (비브리오속 균주들에서외 세포의 효소의 분포)

  • 차재호;김윤희;정초록;김수광;양지영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.2
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    • pp.222-227
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    • 2001
  • The members of the genus Vibrio include harmless aquatic strain as well as strains capable of causing infections in human and fish. Pathogenic mechanisms are only understood for Vibrio cholerae O1 and O139 and not for the majority of Vibrio species. Twelve clinical and nonclinical strains were examined by in vitro and in vivo experiments for the importance of extracellular enzymes as a virulence determinant of Vibrio species. In vivo cytotoxicity assay was performed by injecting approximately $10^{8}$ cells/mL into mice (BALB/c). V. harvyi and V. vulnificus showed 100% lethality within 3hr after bacterial injection. V. fluvialis and four strains of V. parahaemolyticus showed 50% lethality within 4hr. V. mimicus, V. alginolyticus and V. furnissii revealed 30% lethality within 9hr. Nonclinical strains, V. campbellii and V. ordalii, did not show any lethality. In vitro protease and hemolytic activities were also good indicators for clinical and nonclinical strains of Vibrio species. The clinical strains showed much higher activities than nonclinical strains. The activity of some clinical strains of re-isolates was evidently increased. Most clinical strains had $\beta$ hemolytic activity. The results demonstrate that the prevalent distribution of extracellular proteases in pathogenic Vibrio sp. implies their importance as a virulence determinant.

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Apolar growth of Neurospora crassa leads to increased secretion of extracellular proteins

  • Lee, In-Hyung;Rodney G. Walline;Michael Plamann
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.78-89
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    • 2000
  • Protein secretion in filamentous fungi has been shown to be restricted to actively growing hyphal tips. To determine whether an increase in the amount of growing surface area of a fungus can lead to an increase in the amount of protein secretion, we examined secretion in a temperature-sensitive Neurospora crassa mcb mutant that shows a loss of growth polarity when incubated at restrictive-temperature. Incubation of the mcb mutant at restrictive-temperature results in a three- to five-fold increase in the level of extracellular protein and a 20- fold increase in carboxymethyl cellulase activity relative to a wild-type strain. A mutation in the cr-l gene has been shown previously to suppress the apolar growth phenotype of the mcb mutant, and we find that the level of extracellular protein produced by a mcb; cr-l double mutant was reduced to that of the wild-type control. Immunolocalization of a secreted endoglucanase revealed that proteins are secreted mainly at hyphal tips in hyphae exhibiting polar growth and over the entire surface area of bulbous regions of hyphae that are produced following a shift of the mcb mutant to restrictive-temperature. These results support the hypothesis that secretion of extracellular protein by a filamentous fungus can be significantly increased by mutations that alter growth polarity.

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Biomimetics of the extracellular matrix: an integrated three-dimensional fiber-hydrogel composite for cartilage tissue engineering

  • Coburn, Jeannine;Gibson, Matt;Bandalini, Pierre Alain;Laird, Christopher;Mao, Hai-Quan;Moroni, Lorenzo;Seliktar, Dror;Elisseeff, Jennifer
    • Smart Structures and Systems
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    • v.7 no.3
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    • pp.213-222
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    • 2011
  • The native extracellular matrix (ECM) consists of an integrated fibrous protein network and proteoglycan-based ground (hydrogel) substance. We designed a novel electrospinning technique to engineer a three dimensional fiber-hydrogel composite that mimics the native ECM structure, is injectable, and has practical macroscale dimensions for clinically relevant tissue defects. In a model system of articular cartilage tissue engineering, the fiber-hydrogel composites enhanced the biological response of adult stem cells, with dynamic mechanical stimulation resulting in near native levels of extracellular matrix. This technology platform was expanded through structural and biochemical modification of the fibers including hydrophilic fibers containing chondroitin sulfate, a significant component of endogenous tissues, and hydrophobic fibers containing ECM microparticles.

Protein A of Staphylococcus hyicus subsp hyicus isolated from pigs (돼지에서 분리한 Staphylococcus hyicus subsp hyicus의 protein A)

  • Kim, Do-kyoung;Yeo, Sang-geon
    • Korean Journal of Veterinary Research
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    • v.30 no.2
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    • pp.187-192
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    • 1990
  • The Presence and quantity of protein A in Staphylococcus hyicus subsp hyicus isolates from pigs were determined by indirect hemagglutination test and enzyme-linked immunosorbent assay(ELISA). Cell-bound and extracellular protein A was demonstrated in 87.7% and 36.0% of 489 isolates, respectively, by indirect hemagglutination test. When contents of the protein A were estimated by ELISA method, all of the isolates that were positive to protein A in indirect hemagglutination test produced extracellular protein A of less than 1ng/ml. Most of the isolates produced cell-bound protein A of less than 1ng/ml, whereas 28 isolates produced 1 to 35ng/ml and 11 isolates produced 25 to 108ng/ml.

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The Protective Effect of Uncariae Ramulus Et Uncus Extract on Middle Cerebral Artery Occlusion(MACO) (조구등이 뇌허혈을 유발시킨 백서에서의 뇌신경보호효과)

  • 강상렬;강승준;금현수;전연이;이은주;박치상;박창국;허진화;양재하
    • The Journal of Korean Medicine
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    • v.23 no.1
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    • pp.1-10
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    • 2002
  • Objectives: Uncariae Ramulus Et Uncus extract is clinically used in Korea to treat ischemic cerebral damage. The present study was undertaken to study the neuroprotective effect of Uncariae Ramulus Et Uncus extract in middle cerebral artery occlusion (MCAO) rats. Methods: Changes of extracellular levels of dopamine, DOPAC, HVA. and HIAA in striatum were collected at 20 minutes interval by in vivo microdialysis and then analyzed by HPLC (high performance liquid chromatography) in rats subjected to permanent focal cerebral ischemia induced by 2 hours of MCAO. Uncariae Ramulus Et Uncus extract was orally administrated before MCAO. Different animals were used for measurement of cerebral infarction volume induced by 24 hours of MCAO with TIC staining and image analysis. Results: Extracellular levels of dopamine decreased after treatment with Uncariae Ramulus Et Uncus extract, while extracellular levels of DOPAC and HVA significantly increased. Cerebral infarction volume also significantly decreased after treatment with Uncariae Ramulus Et Uncus extract. Conclusions: These results provided evidence that Uncariae Ramulus Et Uncus extract can produce a neuroprotective effect on cerebral ischemia by regulating extracellular excitatory neurotransmitters.

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Production of Extracellular Protein from Bacillus sp. (Bacillus sp.에 의한 균체외 단백질의 생산)

  • Lee, Jae-Sook;Kim, Chan-Jo;Lee, Jong-Soo
    • Applied Biological Chemistry
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    • v.31 no.2
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    • pp.187-192
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    • 1988
  • Extracellular protein was produced from bacterium, which was classified as a strain belong to the genus Bacillus. This bacterium produced 1.4mg/ml of extracellular protein when grown on chemically defined medium containing 3.0% of glucose, 1.3% of urea and 2.5% of calcium carbonate at $30^{\circ}C$ for 48 hours. The addition of $250{\mu}g/ml$ of cephalexin was effective for it and was produced 2.0mg/ml of extracellular protein for 96 hours.

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Perspectives on Bovine Milk-Derived Extracellular Vesicles for Therapeutic Applications in Gut Health

  • Mun, Daye;Oh, Sangnam;Kim, Younghoon
    • Food Science of Animal Resources
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    • v.42 no.2
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    • pp.197-209
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    • 2022
  • Extracellular vesicles (EVs) are nanosized vesicles secreted from cells into the extracellular environment and are composed of a lipid bilayer that contains cargos with biological activity, such as lipids, proteins, mRNAs, and noncoding microRNAs (miRNAs). Due to their biological activity and their role in cell-to-cell communication, interest in EVs is rapidly increasing. Bovine milk is a food consumed by people of all ages around the world that contains not only a significant amount of nutrients but also EVs. Milk-derived EVs also exhibit biological activity similar to other source-derived EVs, and studies on bovine milk EVs have been conducted in various research fields regarding sufficient milk production. In particular, not only are the effects of milk EVs themselves being studied, but the possibility of using them as drug carriers or biomarkers is also being studied. In this review, the characteristics and cargo of milk EVs are summarized, as well as their uptake and stability, efficacy and biological effects as carriers, and future research directions are presented.

Evaluation of Extracellular Enzyme Activity of Fungi from Freshwater Environment in South Korea (담수환경에서 분리한 곰팡이의 세포외분해효소 활성 탐색)

  • Hye Yeon Mun;Yoosun Oh;Jaeduk Goh
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.265-276
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    • 2023
  • This study aimed to isolate and characterize fungi from freshwater environments in South Korea and evaluate their extracellular enzyme activities. Fungal strains were collected from various freshwater sources and identified using phylogenetic analysis. The isolated fungi included known aquatic hyphomycetes and previously unreported species. Extracellular enzyme, including those of protease, amylase, lipase, cellulase, laccase, and chitinase, activities were evaluated. Among the isolated strains, several showed high enzyme activity, suggesting their potential role in organic matter decomposition in freshwater ecosystems. This research expands our knowledge of the diversity and enzyme activities of the fungi in freshwater environments, contributing to our understanding of their ecological roles.

Increased Yield of High-Purity and Active Tetrameric Recombinant Human EC-SOD by Solid Phase Refolding

  • Ryu, Kang;Kim, Young-Hoon;Kim, Young-Hwa;Lee, Joon-Seok;Jeon, Byeong-Wook;Kim, Tae-Yoon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.10
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    • pp.1648-1654
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    • 2008
  • Superoxide dismutase (SOD) removes damaging reactive oxygen species from the cellular environment by catalyzing the dismutation of two superoxide radicals to hydrogen peroxide and oxygen. Extracellular superoxide dismutase (EC-SOD) is a tetramer and is present in the extracellular space and to a lesser extent in the extracellular fluids. Increasing therapeutic applications for recombinant human extracellular superoxide dismutase (rEC-SOD) has broadened interest in optimizing methods for its purification, with a native conformation of tetramer. We describe a solid phase refolding procedure that combines immobilized metal affinity chromatography (IMAC) and gel filtration chromatography in the purification of rEC-SOD from Escherichia coli. The purified rEC-SOD tetramer from the $Ni^{2+}$-column chromatography is refolded in Tris buffer. This method yields greater than 90% of the tetramer form. Greater than 99% purity is achieved with further purification over a Superose 12PC 3.2/30 column to obtain the tetramer and specific activities as determined via DCFHDA assay. The improved yield of rEC-SOD in a simple chromatographic purification procedure promises to enhance the development and therapeutic application of this biologically potent molecule.