• Title/Summary/Keyword: high cell density culture

Search Result 176, Processing Time 0.032 seconds

Continuous Hydrogen Production by Heterotrophic Growth of Citrobacter amalonaticus Y19 in Trickle Bed Reactor (Citrobacter amalonaticus Y19의 영양종속 성장을 이용한 Trickle Bed Reactor에서의 연속적인 수소생산)

  • Park, Ji-Young;Lee, Tae-Ho;Oh, You-Kwan;Kim, Jun-Rae;Seol, Eun-Hee;Jung, Gyoo-Yeol;Kim, Mi-Sun;Park, Sung-Hoon
    • KSBB Journal
    • /
    • v.20 no.6
    • /
    • pp.458-463
    • /
    • 2005
  • [ $H_2$ ] from CO and water was continuously produced in a trickle bed reactor(TBR) using Citrobacter amalonaticus Y19. When the strain C. was cultivated in a stirred-tank reactor under a chemoheterotrophic and aerobic condition, the high final cell concentration of 13 g/L was obtained at 10 hr. When the culture was switched to an anaerobic condition with the continuous supply of gaseous CO, CO-dependent hydrogenase was fully induced and its hydrogen production activity approached 16 mmol/g cell/hr in 60 hr. The fully induced C. amalonaticus Y19 cells were circulated through a TBR packed with polyurethane foam, and the TBR was operated for more than 20 days for $H_2$ production. As gas retention time decreased or inlet CO partial pressure increased, $H_2$ production rate increased but the conversion from CO to $H_2$ decreased. The maximum $H_2$ production rate obtained was 16 mmol/L/hr at the gas retention time of 25 min and the CO inlet partial pressure of 0.4 atm. The high $H_2$ production rate was attributed to the high cell density in the liquid phase circulating the TBR as well as the high surface area of polyurethane foam used as packing material of the TBR.

Chemical signalling within the rumen microbiome

  • Katie Lawther;Fernanda Godoy Santos;Linda B Oyama;Sharon A Huws
    • Animal Bioscience
    • /
    • v.37 no.2_spc
    • /
    • pp.337-345
    • /
    • 2024
  • Ruminants possess a specialized four-compartment forestomach, consisting of the reticulum, rumen, omasum, and abomasum. The rumen, the primary fermentative chamber, harbours a dynamic ecosystem comprising bacteria, protozoa, fungi, archaea, and bacteriophages. These microorganisms engage in diverse ecological interactions within the rumen microbiome, primarily benefiting the host animal by deriving energy from plant material breakdown. These interactions encompass symbiosis, such as mutualism and commensalism, as well as parasitism, predation, and competition. These ecological interactions are dependent on many factors, including the production of diverse molecules, such as those involved in quorum sensing (QS). QS is a density-dependent signalling mechanism involving the release of autoinducer (AIs) compounds, when cell density increases AIs bind to receptors causing the altered expression of certain genes. These AIs are classified as mainly being N-acyl-homoserine lactones (AHL; commonly used by Gram-negative bacteria) or autoinducer-2 based systems (AI-2; used by Gram-positive and Gram-negative bacteria); although other less common AI systems exist. Most of our understanding of QS at a gene-level comes from pure culture in vitro studies using bacterial pathogens, with much being unknown on a commensal bacterial and ecosystem level, especially in the context of the rumen microbiome. A small number of studies have explored QS in the rumen using 'omic' technologies, revealing a prevalence of AI-2 QS systems among rumen bacteria. Nevertheless, the implications of these signalling systems on gene regulation, rumen ecology, and ruminant characteristics are largely uncharted territory. Metatranscriptome data tracking the colonization of perennial ryegrass by rumen microbes suggest that these chemicals may influence transitions in bacterial diversity during colonization. The likelihood of undiscovered chemicals within the rumen microbial arsenal is high, with the identified chemicals representing only the tip of the iceberg. A comprehensive grasp of rumen microbial chemical signalling is crucial for addressing the challenges of food security and climate targets.

Control of dissolved Oxygen Concentration and Specific Growth Rate in Fed-batch Fermentation (유가식 생물반응기에서의 용존산소농도 및 비성장속도의 제어)

  • Kim, Chang-Gyeom;Lee, Tae-Ho;Lee, Seung-Cheol;Chang, Yong-Keun;Chang, Ho-Nam
    • Microbiology and Biotechnology Letters
    • /
    • v.21 no.4
    • /
    • pp.354-365
    • /
    • 1993
  • A novel control method with automatic tuning of PID controller parameters has been developed for efficient regulation of dissolved oxygen concentration in fed-batch fermentations of Escherichia coli. Agitation speed and oxygen partial pressure in the inlet gas stream were chosen to be the manipulated variables. A heuristic reasoning allowed improved tuning decisions from the supervision of control performance indices and it coule obviate the needs for process assumptions or disturbance patterns. The control input consisted of feedback and feedforword parts. The feedback part was determined by PID control and the feedforward part is determined from the feed rate. The proportional gain was updated on-line by a set of heuristics rules based on the supervision of three performance indices. These indices were output error covariance, the average value of output error, and input covariance, which were calculated on-line using a moving window. The integral and derivative time constants were determined from the period of output response. The specific growth rate was maintained at a low level to avoid acetic acid accumulation and thus to achieve a high cell density. The specific growthe rate was estimated from the carbon dioxide evolution rate. In fed-batch fermentation, the simutaneous control of dissolved oxygen concentration (at 0.2; fraction of saturated value) and specific growth rate (at 0.25$hr^{-1}$) was satisfactory for the entire culture period in spite of the changes in the feed rate and the switching of control input.

  • PDF

Characterization of a Ligninase Producing Strain, Serratia marcescens HY-5 isolated from Sympetrum dopressiusculum (고추좀잠자리 (Sympetrum depressiusculum)로부터 분리한 리그닌 분해균주, Serratia marcescens HY-5의 특성)

  • Kim, Ki-Duck;Park, Doo-Sang;Shin, Dong-Ha;Han, Bo-Na;Oh, Hyun-Woo;Youn, Young-Nam;Park, Ho-Yong
    • Korean journal of applied entomology
    • /
    • v.45 no.3 s.144
    • /
    • pp.301-307
    • /
    • 2006
  • A ligniolytic bacterial strain was isolated from the digestive tract of a red dragonfly, Sympetrum dopressiusculum. It was identified as a Serratia marcescens HY-5 by 16S rDNA sequence analysis and physiological and biochemical analysis. The isolated strain showed proportional increase of ligninolytic activity to the cell growth in the culture media which include lignin compounds. It showed about 25-45% decomposition of lignin compound by 48 hr incubation especially, showed effective decomposition of monomer lignin compounds, vanillin and guaiacol, and a dimer, dealkaline lignin. PCR amplification of 16S rDNA followed by denaturing gradient gel electrophoresis analysis showed high density of S. marcescens HY-5 in the gut of the S. depressiusculum at both gut samples which collected at different site.

Enhanced Chondrogenic Differentiation of Human Adipose-derived Stem Cells with Inverse Opal Scaffolds (역오팔 구조 지지체를 이용한 인간 지방 유래 줄기 세포의 연골 분화 촉진)

  • Bhang, Suk Ho;Yu, Taekyung
    • Korean Chemical Engineering Research
    • /
    • v.51 no.6
    • /
    • pp.727-732
    • /
    • 2013
  • In this report, we present an inverse opal scaffold that can enhance the chondrogenic differentiation of human adipose-derived stem cells (hADSCs) without drug, gene, or cytokine supplement. Inverse opal scaffolds based on poly(D,L-lactide-co-glycolide) were formed with uniform $200{\mu}m$ pores. Due to uniform pore sizes and well-controlled interconnectivity of inverse opal scaffold, hADSCs were allowed to distribute homogeneously throughout the scaffolds. As a result, high cell density culture with scaffold was possible. Since the hADSCs cultured in inverse opal scaffolds were subjected to limited supplies of oxygen and nutrients, these cells were naturally preconditioned to a hypoxic environment that stimulated the up-regulation of hypoxia-inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$). As a result, apoptotic activity of hADSCs until 3 weeks after initial cell seeding was significantly reduced and chondrogenic differentiation related molecular signal cascades were up regulated (transforming growth factor-beta, phosphorylated AKT, and phosphorylated p38 expression). In contrast, hADSCs cultured with small and non-uniform porous scaffolds showed significantly increased apoptotic activity with decreased chondrogenic differentiation. Taken together, inverse opal scaffold could potentially be used as an effective tool for improving chondrogenesis using stem cells.

EFFECTS OF HYDROQUINONE ON NEOPLASTIC TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (Hydroquinone이 인체 상피세포의 발암화에 미치는 영향)

  • Sohn, Jung-Hee;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • v.32 no.3
    • /
    • pp.218-228
    • /
    • 2010
  • Components of dental resin-based restorative materials are reported to leach from the filling materials even after polymerization. Hydroquinone (HQ) is one of the major monomers used in the dental resin and is known as a carcinogen. Thus, carcinogenic risk of HQ leaching from the dental resin becomes a public health concern. The present study attempted to examine the carcinogenic potentials of HQ on the human epithelial cell, which is the target cell origin of the most of oral cancers. Cytotoxicity of HQ was observed above 50${\mu}M$ as measured by LDH assay, indicating a relatively low toxicity of this substance in human epithelial cells. The parameters of neoplastic cellular transformation such as cell saturation density, soft agar colony formation and cell aggregation were analyzed to examine the carcinogenic potential of HQ. The study showed that 2-week exposure of HQ showed the tendency of increase in the saturation density and the significant enhancement of soft agar colony formation at the highest dose, 50 ${\mu}M$ only. It is suggested that HQ has a weak potential of carcinogenicity. When cells were treated with HQ and TPA, a well-known tumor promoter, the parameters of neoplastic cellular transformation was significantly increased. This result indicates that the potential risk of carcinogenicity from HQ is largely dependent upon the presence of promoter. Exposure of 50 ${\mu}M$ HQ increased the time-dependent apoptosis as measured by the ELISA kit. This concentration coincides with a dose of neoplastic transformation, indicating a possible link between apoptosis and HQ-induced cellular transformation. Hydroquinone generated Reactive Oxygen Species (ROS) which was evidenced by the treatment of antioxidants such as trolox and N-acetyl cysteine and the GSH depleting agent, BSO. Antioxidants blocked the generation of ROS and the GSH depleting agent, BSO dramatically increased the ROS production. Since HQ is known to increase ROS production thru activation of transcriptional factor such as c-Myb and Pim-1, it is speculated that ROS generation by HQ plays a role in the activation of oncogene, which may lead to neoplastic transformation. In addition, ROS is involved in the alteration of signal transduction, which regulates the apoptosis in many cellular systems. Thus, ROS-mediated apoptosis may be involved in the HQ-induced carcinogenic processes. Protein kinase C (PKC) is known to play pivotal roles in neoplastic transformation of cells and its high expression is often found in a variety of types of tumors including oral cancer. PKC translocation of PKC-${\alpha}$ was observed following HQ exposure. Altered signaling system may also play a role in the transformation process. Taken together, HQ leached from the dental resin does not pose a significant threat as a cancer causing agent, but its carcinogenic potential can be significantly elevated in the presence of promoter. The mechanism of HQ-induced carcinogenesis involved ROS generation, apoptosis and altered signaling pathway. The present study will provide a valuable data to estimate the potential risk of HQ as a carcinogen and understand mechanism of HQ-induced carcinogenesis in human epithelial cells.

Effects of Oils and Dispersant on the Red Tide Organism Cochlodinium Polykrikoides (적조생물 Cochlodinium Polykrikoides에 대한 유류 및 유처리제의 영향)

  • Lee, Sam-Geun;Cho, Eun-Seob;Lim, Wol-Ae;Lee, Young-Sik
    • Journal of Environmental Science International
    • /
    • v.16 no.7
    • /
    • pp.799-804
    • /
    • 2007
  • Oil spill caused severe effects on the marine fauna and flora due to direct contact of organisms with the oil and even in regions not directly affected by the spill. This study was conducted to understand the effects of the oil spill accidents and the use of dispersant on the red tide of Cochlodinium polykrikoides. Crude oil produced in Kuwait, bunker-C, kerosene and diesel oil, and a chemical dispersant produced in Korea, were added with a series of 10 ppb to 100 ppm in the f/2-Si medium at $20^{\circ}C$ under a photon flux from cool white fluorescent tubes of $100\;mol\;m^{-2}\;s^{-1}$ in a 14: 10 h L:D cycle for the culture of C. polykrikoides. In low concentrations of ${\leq}$ 1 ppm of examined oils no impact on the growth of C. polykrikoides was recorded, while in high concentration of ${\geq}$ 10 ppm, cell density was significantly decreased with the range of 10 to 80% in comparison with the control. The growth of C. polykrikoides after the addition of the dispersant and the mixtures combined with oils and a dispersant of ${\geq}$ 10 ppm appeared to decrease, whereas the growth of C. polykrikoides exposed to ${\leq}$ 100 ppb showed little serious impact. However, almost all the C. polykrikoides cells were died regardless of a dispersant and combined mixtures within a few days after the addition of high concentrations.

Environmental resistance of indigenous Saccharomyces cerevisiae with tolerance to potassium metabisulfite at the microbial succession stage of fermenting Campbell Early grape (캠벨얼리 와인발효 중 효모 천이단계에서 분리된 아황산 내성 토착형 효모의 환경내성)

  • Kim, Mi-Sun;Hong, Young-Ah;Yeo, Soo-Hwan;Baek, Seong-Yeol;Yun, Hye-Ju;Rhee, Chang-Ho;Kim, Kwan-Pil;Park, Heui-Dong
    • Food Science and Preservation
    • /
    • v.20 no.6
    • /
    • pp.886-893
    • /
    • 2013
  • Several indigenous sulfite-resistant yeasts were isolated at the microbial succession stage of yeast flora during spontaneous fermentation of Campbell Early grapes using a YPD plate that contained 200 mg/L or 500 mg/L potassium metabisulfite. When they were applied to the wine fermentation using the Campbell Early grape and apple juices, strains S13 and D8 showed strong alcohol fermentation and good flavor production. They were identified as Saccharomyces cerevisiae in the phylogenetic analysis based on their ITS 1-5.8S-ITS II DNA sequences. The two yeast strains grew to a high cell density in the YPD media supplemented with 40%(w/v) glucose. They also grew rapidly in the YPD media at $40^{\circ}C$. While strain S13 showed some differences in cell density at the two temperatures, no marked difference was observed during the culture of strain D8. The strains grew relatively well at pH 5.0 and 9.0 compared with pH 7.0, which was the optimum pH for their growth. Especially, strain S13 cultivated in the YPD media at pH 9.0 grew to 93% of the growth of strain D8, which was obtained at pH 7.0.

EFFECTS OF SIGNAL TRANSDUCTION PATHWAY IN THE RAS-INDUCED CELLULAR TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (인체 상피세포에서 ras-종양유전자의 발암화가 신호 전달 기작에 미치는 영향)

  • Jang, Do-Geun;Byeon, Ki-Jeong;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
    • /
    • v.26 no.3
    • /
    • pp.254-261
    • /
    • 2000
  • The present study has attempted to look into the mechanism of ras-induced carcinogenesis in a human epithelial cell system. Human epithelial cells immortalized with Ad12-SV40 hybrid virus were used to assess carcinogenic potential of the ras-oncogene. Cells transfected with pSV2-ras showed characteristics of cellular transformation. The transformation parameters such as cell density, soft-agar colony formation, and cell aggregation were significantly increased in the cells expressing ras oncoprotein. In addition, the duration required for the appearance of foci was shortened in the ras-transfected cells. Consistent with other reports, our results demonstrated an evidence that the ras-oncogene induced the cellular transformation of human epithelial cell system. When a high concentration of glucocorticoid was added into the media, transformation process was accelerated. It is speculated that glucocorticoid may provide an advantageous environment for the proliferation of the transformed cells. The induction of the intracellular free calcium concentrations following agonist treatment was significantly lower in the transformed cells than in the control cells. These effects were more manifested in the presence of extracellular cacium, indicating that the transformation process may alter the influx pathway of extracellular calcium. The induction of $IP_3$ following agonist treatment was also lower in the transformed cells than in the control cells. Thus, it is suggested that phospholipase C-coupled pathway was down-regulated in the process of the ras-induced transformation. While the levels of $TGF-{\beta}_1$ and PAI-2 mRNAs were decreased, the level of fibronectin mRNA was increased. The results indicate that mechanism of the ras-induced transformation may be associated with the altered expressions of growth regulatory factors. The present study demonstrates an evidence that the ras-induced cellular transformation may be associated with alteration of signal transduction and growth regulatory factors. The study will contribute to improve the understanding of molecular mechanism of epithelium-derived cancers including oral cancer.

  • PDF

Transformation Conditions and Ampicillin-resistant Expression of E. coli Ts-mutant (온도감수성 대장균의 형질전환조건 및 Ampicillin 내성의 표현)

  • JIN Deuk Hee;HONG Yong Ki
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.20 no.1
    • /
    • pp.57-62
    • /
    • 1987
  • The transformational conditions of plasmids $pPL-\lambda$ and pAS 1 are as follows in E. coli, Ts-mutant M 5248 strain at $30^{\circ}C$. When the culture time was 2.5 hours of mid logarithemic phase, the cell concentration was $4.5\times10^7\;cells/ml$, the optical density was equal to 0.45 at 590 nm wave length, the transformational frequencies of plasmid$pPL-\lambda$ and pAS 1 had the highest values as $2\times10^{-6}\;and\;1.5\times10^{-6}\;and\;1.5\times10^{-6}$ and respectively. For $9\times10^6$ competent cells in $200{\mu}l$, the transformational frequency was as high as $4.4\times10^{-6}$ at 510 ng plasmid concentration. The competent cells treated with the mixture of calcium chloride and thymidine twice rates of transformation than those treated with calcium chloride. The ampicillin resistance of transformants was expressed in LB broth after 2 hours at $30^{\circ}C$.

  • PDF