• 제목/요약/키워드: resting cell

검색결과 208건 처리시간 0.025초

Optimal Conditions and Substrate Specificity for Trehalose Production by Resting Cells of Arthrobacter crystallopoietes N-08

  • Seo, Yi-Seul;Shin, Kwang-Soon
    • Preventive Nutrition and Food Science
    • /
    • 제16권4호
    • /
    • pp.357-363
    • /
    • 2011
  • Recently, we found that Arthrobacter crystallopoietes N-08 isolated from soil directly produces trehalose from maltose by a resting cell reaction. In this study, the optimal set of conditions and substrate specificity for the trehalose production using resting cells was investigated. Optimum temperature and pH of the resting cell reaction were $55^{\circ}C$ and pH 5.5, respectively, and the reaction was stable for two hours at $37{\sim}55^{\circ}C$ and for one hour at the wide pH ranges of 3~9. Various disaccharide substrates with different glycosidic linkages, such as maltose, isomaltose, cellobiose, nigerose, sophorose, and laminaribiose, were converted into trehalose-like spots in thin layer chromatography (TLC). These results indicated broad substrate specificity of this reaction and the possibility that cellobiose could be converted into other trehalose anomers such as ${\alpha},{\beta}$- and ${\beta},{\beta}$-trehalose. Therefore, the product after the resting cell reaction with cellobiose was purified by ${\beta}$-glucosidase treatment and Dowex-1 ($OH^-$) column chromatography and its structure was analyzed. Component sugar and methylation analyses indicated that this cellobiose-conversion product was composed of only non-reducing terminal glucopyranoside. MALDI-TOF and ESI-MS/MS analyses suggested that this oligosaccharide contained a non-reducing disaccharide unit with a 1,1-glucosidic linkage. When this disaccharide was analyzed by $^1H$-NMR and $^{13}C$-NMR, it gave the same signals with ${\alpha}$-D-glucopyranosyl-(1,1)-${\alpha}$-D-glucopyranoside. These results suggest that cellobiose can be converted to ${\alpha},{\alpha}$-trehalose by the resting cells of A. crystallopoietes N-08.

간헐적 정수압이 세포의 부착에 미치는 영향성 평가 (Effects of Intermittent Hydrostatic Pressure on the Cell Adhesive force on the substrate)

  • 김영직;이명곤;박수아;신호준;김인애;이용재;신지원;신정욱
    • 한국정밀공학회:학술대회논문집
    • /
    • 한국정밀공학회 2005년도 추계학술대회 논문집
    • /
    • pp.69-72
    • /
    • 2005
  • In this study, effects of IHPs with various resting times to cell adhesion were investigated through the measurements of cell adhesive force, number and area of focal contacts (stained vinculin spots), and projected cell area, perimeter and circularity. In addition correlation tests and curve estimations using the experimental results were performed fur the finding an essential factor for increment of cell adhesive force. Tn the results, immediately after mechanical stimuli (150 minutes after seeding) and one hour later (210 minutes after seeding), the average adhesive force of experimental group 5 (resting time: 15min) compared with that of control group at same culture time was increased significantly (p<0.05). Average projected area and perimeter of cells at Group 5 were increased significantly (p<0.05), while average circularity of cells at Group 5 incubated fur 210 minutes was decreased significantly (p<0.05). In the digital image analysis of focal contacts containing vinculins, area and numbers of focal contacts per cell at Group 5 were higher than those of the other groups. This study indicated that IHP with appropriate resting time could contribute in improving cell adhesive force, cell spreading, development of cytoskeleton and formation of focal contacts. And cell adhesive force was correlated to the morphological aspects of cell and development of focal contacts. Particularly, area of focal contacts was closely related to cell adhesive force.

  • PDF

세포 내 칼슐 농도의 변화에 따른 간헐적 정수압이 세포 부착력에 미치는 영향 (Contribution of intermittent hydrostatic pressure to the cell adhesive forces throught the changes in intracelluar $Ca^{2+}$ concentration)

  • 김동화;김영직;신지원;신정욱
    • 대한기계학회:학술대회논문집
    • /
    • 대한기계학회 2008년도 추계학술대회A
    • /
    • pp.1580-1581
    • /
    • 2008
  • We investigated the effects of intermittent hydrostatic pressure with various duration of resting period on changes in calcium ($Ca^{2+}$) concentration and adhesive forces of cells on substrates. The quantitive adhesive forces of cells were measured under various resting periods. When the pressure applied to the cells, the concentration of $Ca^{2+}$ increased. Under intermittent hydrostatic pressure, the concentration of $Ca^{2+}$ was maintained under a resting period of 15 min, while it was not decreased with other resting periods of less than 15 min. With a resting period of 15 min, the magnitudes of adhesive forces were significantly increase. In addition, the adhesive forces were measured with and without $Ca^{2+}$ chelating agents to evaluate the effect of $Ca^{2+}$ on cell adhesiveness. When $Ca^{2+}$ ions were chelated, the adhesive forces dramatically decreased, even under intermittent hydrostatic pressure. We conclude that $Ca^{2+}$ plays an crucial role in modulating the adhesive forces of cells, and that the concentration of $Ca^{2+}$ can be increased by intermittent hydrostatic stimuli.

  • PDF

Aspergillus niger의 휴지균체에 의한 Gluconic Acid생성에 관한 연구 (Studies on the Production of Gluconic Acid by Resting Cell System of Aspergillus niger)

  • 정지관;양호석;신규철;양한철
    • 한국미생물·생명공학회지
    • /
    • 제9권1호
    • /
    • pp.7-19
    • /
    • 1981
  • Aspergillus niger의 휴지균체법에 의한 glucose 로부터 gluconic acid 생산에 대해서 연구한 결과, gluconic acid 생산은 반응 pH, 온도, 기질농도, 통기조건, 금속 ions, 휴지균체의 보존상태 및 반응시간에 따라 크게 영향을 받았다. 휴지균체법에 의해서 glucose로부터 전환된 생성물질은 thin layer chromatography와 infrared spectrophotometer에 의해서 gluconic acid로 확인되었으며, glucose로부터 gluconic acid 생성반응에서는 $Mg^{++}$, S $n^{++}$의 첨가로 촉진효과를 보였으나, C $u^{++}$, H $g^{++}$, C$d^{++}$, A $g^{+}$ 및 Cyanide는 저해효과를 나타냈다. 휴지균체의 최적보존 조건으로는 0.05 M phosphate buffer(pH7.0)에서 -$25^{\circ}C$ 보존 시가제일 안정하였다. 휴지균체법에 의한 gluconic acid 생산은 발효법에 비해서 배양시간이 짧고 수율이 양호하고, 정제 시 회수가 간단할 뿐 아니라 균체의 반복사용이 가능하였다.

  • PDF

간헐적 정수압의 다양한 패턴에 따른 세포 부착력과 F-actin 및 vinculin 의 분포 변화 연구 (A Study on Changes of Cell Adhesive force and Distribution of F-actin and Vinculin under Various Intermittent Hydrostatic Pressure.)

  • 김영직;박수아;신호준;김인애;이용재;허수진;황영미;신정욱
    • 한국정밀공학회:학술대회논문집
    • /
    • 한국정밀공학회 2004년도 추계학술대회 논문집
    • /
    • pp.1215-1220
    • /
    • 2004
  • Cell adhesion to any material surface is considered to be fundamental and important phenomenon in the fields of tissue engineering. Cell adhesion molecules, mechanism, and attachment force have been studied and described a lot. However, the effects of mechanical stimuli on the adhesive forces still have been left much to be investigated. In this study, to investigate the changes in cell adhesive force due to resting time period during the intermittent hydrostatic pressurizing (IHP), cells were cultured under the IHP with various resting times. Then the cell adhesive forces were measured quantitatively utilizing a cell detachment test system and immunofluorescent staining was performed using fluorescent microscopy. In the results, immediately after mechanical stimuli (150 minutes after seeding) and one hour later (210 minutes after seeding), the average adhesive force of experimental group 5 (resting time: 15min) compared with that of control group at same culture time was increased significantly (p<0.05). The results indicated that IHP can contribute in improving cell adhesive force and some of time intervals were required for the expression of cell response.

  • PDF

토착미 생물을 이용한 TNT(2,4,6-Trinitrotoluene)의 생물학적분해

  • 배범한;유경민;장윤영;이인숙
    • 한국지하수토양환경학회:학술대회논문집
    • /
    • 한국지하수토양환경학회 2002년도 추계학술발표회
    • /
    • pp.235-238
    • /
    • 2002
  • Microorganisms were isolated from military shooting site. Aerobic batch reactor and resting cell condition experiments were carried out using isolated microorganisms. Experiments were examined at room temperature on shaker and ten-roll mixer. During 10 days of reaction time, TNT was degraded 15.51 ~ 22.47 mg/L from initial concentration(31$\pm$1 mg/L) by aerobic batch reactor. Aerobic resting cell condition experiments were carried out ill phosphate buffer with 58($\pm$1) mg/L TNT at pH of 6.0($\pm$0.2). TNT was degraded 67.8% of initial concentration. The mai or component was found 4-ADNT(4-Aminodinitrotoluene).

  • PDF

인삼추출물이 흥분성세포의 막전압에 미치는 영향 (Effects of Ginseng Extract on Excitable Cell Membrane Potential)

  • 정진모;백광세;남택상;김인교;강두희
    • The Korean Journal of Physiology
    • /
    • 제15권1호
    • /
    • pp.3-8
    • /
    • 1981
  • Studies have been conducted to test the effect of Ginseng alcohol extract on the membrane potentials of frog skeletal muscle. The gastrocnemius muscle was isolated and placed in a chamber containing the Clark-frog Ringer solution. Membrane potentials were recorded using microelectrodes filled with 3 M KCI and muscle was electrically stimulated to obtain action potential. Changes in both the action potential and the resting membrane potential were observed after adding an appropriate amount of Ginseng alcohol extract in the perfusing Ringer solution. The results obtained from 346 muscle cells are summarized as follows : 1) The average resting membrane potential of the normal frog gastrocnemius muscle cell was -92.8 mV and the peak of the action potential reached at 29.8 mV. 2) Both the resting membrane potential and the peak of the action potential decreased by Ginseng alcohol extract, the effect being proportional to the dose of Ginseng alcohol extract. 3) The resting membrane potential and the peak of the action potential continuously decreased until about 40 min after Ginseng addition and leveled off thereafter. The potentials recovered to its original value after Ginseng was washed out. 4) The resting membrane potential was more sensitive to the Ginseng alcohol extract than was the action potential. These results strongly suggest that Ginseng alcohol extract increases both the $Na^+$ and $K^+$ permeability in the skeletal muscle cell membrane.

  • PDF

Proteomic Analysis of Resting and Activated Human $CD8^+$ T Cells

  • Koo Jung-Hui;Chae Wook-Jun;Choi Je-Min;Nam Hyung-Wook;Morio Tomohiro;Kim Yu-Sam;Jang Yang-Soo;Choi Kwan-Yong;Yang Jung-Jin;Lee Sang-Kyou
    • Journal of Microbiology and Biotechnology
    • /
    • 제16권6호
    • /
    • pp.911-920
    • /
    • 2006
  • [ $CD8^+$ ] T Iymphocytes with the cytotoxic activity and capability to release various cytokines are the major players in immune responses against viral infection and cancer. To identify the proteins specific to resting or activated human CD8$^+$ T cells, human CD8$^+$ T cells were activated with anti-CD3+anti-CD28 mAb in the presence of IL-2. The solubilized proteins from resting and activated human CD8$^+$ T cells were separated by high-resolution two-dimensional polyacrylamide gel electrophoresis, and their proteomes were analyzed. Proteomic analysis of resting and activated T cells resulted in identification of 35 proteins with the altered expression. Mass spectrometry coupled with Profound and SWISS-PROT database analysis revealed that these identified proteins are to be functionally associated with cell proliferation, metabolic pathways, antigen presentation, and intracellular signal transduction pathways. We also identified six unknown proteins predicted from genomic DNA sequences specific to resting or activated CD8$^+$ T cells. Protein network studies and functional characterization of these novel proteins may provide new insight into the signaling transduction pathway of CD8$^+$ T cell activation.

Candida sp. SY16의 휴식세포를 이용한 생물계면활성제 Mannosylerythritol Lipid의 생산 (Production of a Biosurfactant Mannosylerythritol Lipid by Resting Cell of Candida sp. SY16.)

  • 김희식;전종운;최우영;오희목;이기형;권태종;윤병대
    • 한국미생물·생명공학회지
    • /
    • 제30권2호
    • /
    • pp.167-171
    • /
    • 2002
  • Candida sp. SY16의 휴식세포를 이용하여 탄소원만 포함되 있는 증류수로부터 많은 양의 생물계면활성제(mannosylerythritol lipid: MEL)를 생산할 수 있었다. 플라스크를 이용한 생산반응의 경우, 20 g/l의 휴식세포를 75g/1의 soybean oil이 포함되있고, pH는 4~5범위에서 반응할 때, 가장 높은 생산수율을 얻을 수 있었으며, 높은 농도($PO_4$-P 농도 198 $\mu\textrm{g}$/ml 이상)의 인은 MEL 생산을 저해하는 것으로 나타났다. 최적화된 조건으로 jar fermentor를 이용하여 휴식세포를 반응한 경우 생물계면활성제를 120 h 반응 후, 58 g/l로 얻을 수 있었으며, 그 생산성은 성장세포를 이용한 회분식 발효의 경우보다 높았고, 배양시간도 단축시킬 수 있었다.

Differences in Their Proliferation and Differentiation between B-1 and B-2 Cell

  • Yeo, Seung-Geun;Cha, Chang-Il;Park, Dong-Choon
    • IMMUNE NETWORK
    • /
    • 제6권1호
    • /
    • pp.1-5
    • /
    • 2006
  • Background: B cell subset has been divided into B-1 cells and B-2 cells. B-1 cells are found most prominently in the peritoneal cavity, as well as constituting a small pro portion of splenic B cells and they are larger and less dense than B-2 cells in morphology. This study was designed to compare the differences in their proliferation and differentiation between B-1 and B-2 cell. Methods: We obtained sorted B-1 cells from peritoneal fluid and B-2 cells from spleens of mice. Secreted IgM was measured by enzyme-linked immunosorbent assay. Entering of S phase in response to LPS-stimuli was measured by proliferative assay. Cell cycle analysis by propidium iodide was performed. p21 expression was assessed by real time PCR. Results: Cell proliferation and cell cycle progression in B-1 and B-2 cells, which did not occur in the absence of LPS, required LPS stimulation. After LPS stimulation, B-1 and B-2 cells were shifted to Sand G2/M phases. p21 expression by resting B-1 cells was higher than that of resting B-2 cells. Conclusion: B-1 cells differ from conventional B-2 cells in proliferation, differentiation and cell cycle.