• Title/Summary/Keyword: cell production

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Overexpression of S-Adenosylmethionine Synthetase in Recombinant Chlamydomonas for Enhanced Lipid Production

  • Jeong Hyeon Kim;Joon Woo Ahn;Eun-Jeong Park;Jong-il Choi
    • Journal of Microbiology and Biotechnology
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    • v.33 no.3
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    • pp.310-318
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    • 2023
  • Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to ithstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.

Effect of Increased Glutamate Availability on L-Ornithine Production in Corynebacterium glutamicum

  • Hwang, Joong-Hee;Hwang, Gui-Hye;Cho, Jae-Yong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.704-710
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    • 2008
  • Glutamate availability in the argF-argR-proB${\Delta}$ strain of Corynebacterium glutamicum was increased by addition of glutamate to the cell or inactivation of the phosphoenolpyruvate carboxykinase activity and simultaneous overexpression of the pyruvate carboxylase activity to assess its effect on L-ornithine production. When glutamate was increased in an L-ornithine-producing strain, the production of L-ornithine was not changed. This unexpected result indicated that the intracellular concentration and supply of glutamate is not a rate-limiting step for the L-ornithine production in an L-ornithine-producing strain of C. glutamicum. In contrast, overexpression of the L-ornithine biosynthesis genes (argCJBD) resulted in approximately 30% increase of L-ornithine production, from 12.73 to 16.49 mg/g (dry cell weight). These results implied that downstream reactions converting glutamate to L-ornithine, but not the availability of glutamate, is the rate-limiting step for elevating L-ornithine production in the argF-argR-proB${\Delta}$ strain of C. glutamicum.

연속배양을 통한 L-prolinc 발효공정의 최적화 연구

  • Yu, Ji-Myeong;Choe, Sun-Yong;Jang, Hyeong-Uk;An, Jeong-O;Jo, Yeong-Il;Lee, Hong-Won;Jeong, Jun-Gi
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.426-429
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    • 2001
  • The continuous production of L -proline was studied using L-histidine auxotrophic mutant of Corynebacterium acetoacidophilum under various substrate limited conditions. Among the $NH_4\;^+$ $PO_4\;^3$ and L -histidine limited conditions, the highest production of L -proline was observed under the L-histidine limited condition. Under $NH_4\;^+$ and $PO_4\;^3$ limited conditions, no or poor L-proline production was observed, respectively. For the kinetic parameters under L -histidine limitation the specific rate of L -proline production was increased with dilution rate upto $0.1hr^{-1}$ but decreased above $0.1hr^{-1}$. The volumetric rate of L -proline production was showed similar pattern with specific rate. The dried cell weight was gradually increased according to decrease the dilution rate. Specific rate of glucose consumption was proportionally increased with dilution rate. The results of continuous culture (higher production of L-proline at dilution rate $0.1hr^{-1}$) will be used in fed-batch culture for the control of cell growth rate and mass production of L-proline.

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Effect of Osmotic Pressure of Salts on Growth of Torula sp. and Erythritol Production (염의 삼투압이 Torula sp.의 증식과 Erythritol 생산에 미치는 영향)

  • 김경아;노봉수;김상용;오덕근
    • Microbiology and Biotechnology Letters
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    • v.27 no.2
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    • pp.91-95
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    • 1999
  • To investigate the effect of salts on the production of erythritol by Torula sp., cells were grown on the media containing various concentrations of KCl or NaCl. Cell growth and glucose consumption rates decreased when KCl or NaCl concentration increased from 0.0 to 0.5M. The production of erythritol, however, was maximal at 0.3M aCl. The erythritol concentration of 54.3g/l in the medium containing 0.3M NaCl and 200g/l glucose was obtained after 120h. The production of erythritol decreased in cultures above 0.3M NaCl or 0.4M KCl due to the inhibition of cell growth. To elucidate the effect of salts more quantitatively, KCl and NaCl concentrations were converted to osmotic pressure. As the osmotic pressure increased, the yield of erythritol from glucose increased regardless of the kinds of salts and the yield of erythritol was approximately 49% at the osmolality of 2.4Os/kg. When the osmotic pressure increased to 2.5Os/kg, the specific growth rate of cells decreased but the production rate of erythritol increased. For the effective production of erythritol, osmotic pressure must be adjusted not to inhibit markedly the growth rate of cells and to stimulate the production rate of erythritol by supplementing salt.

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Somatic Cell Counts in Milk of Buffaloes Administered Oxytocin During Early Lactation

  • Prasad, Jyotsna;Singh, Mahendra
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.5
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    • pp.684-692
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    • 2001
  • To find out the effect of oxytocin on somatic cell count and milk production, 12 primiparous and multiparous Murrah buffaloes were selected, immediately after the parturition, from the Institute's buffalo herd. These were divided into two groups of 6 each. Buffaloes of group I did not receive oxytocin injection (control); whereas, buffaloes of group II were administered oxytocin during early lactation (av. 42.50 days). The oxytocin injection was given in doses of 2.5 IU i.m. before the start of milking, to let down the milk, for a period of 5 days. Samples of milk from individual buffaloes were collected for 5 days before (Period I), during (Period II) and after (Period III) from both the group of buffaloes. Milk samples of A. M. and P. M. milking were composited in proposition to milk yields for analysis of milk constituents. Normal values of somatic cell counts in group I of buffaloes varied from 0.54 to $0.75{\times}10^{5}cells/ml$. Mean cytoplasmic particles and epithelial cells varied from 3.68 to $7.19{\times}10^{5}cells/ml$ and 0.13 to $0.54{\times}10^{5}cells/ml$. On percentage basis the epithelial and the total leucocyte count were 60 and 40. Total leucocyte count, in the study varied from 0.17 to $0.69{\times}10^{5}cells/ml$. The differential cell count of milk indicated presence of lymphocytes (16.50 to $61.16{\times}1000$), neutrophil (0.00 to $2.00{\times}1000$) and monocyte (0.00 to $18.16{\times}1000$). Somatic cell count (p<0.01) and epithelial cells (p<0.05) varied between buffaloes and between periods of study. Total leucocyte counts of milk were also significantly varied between periods (p<0.05). The change in fat, lactose, chloride, EC and NEFA concentrations during different periods of study, were highly significant, indicated diurnal variations in different buffaloes during different days of experiment. Administration of oxytocin resulted in increase in somatic cell counts of milk (p<0.01) due to the increases in total leucocyte count (p<0.01) during the treatment period. The differential cell count indicated that oxytocin administration increased lymphocyte number significantly (p<0.01). However, secretion of neutrophil, monocyte and cytoplasmic particles were not affected by oxytocin. Eosinophil and basophil cell, though present in few samples, remain unaffected by oxytocin administration. There was no effect of oxytocin on milk production, composition, pH, EC and NEFA concentration.

Correlation between Redox Potential and State Variables in Batch Cultures for Ornithine Production. (오르니틴 생산을 위한 회분식 배양에서 산화환원전위와 상태변수들간의 상관관계)

  • 이태호;나정걸;장용근;정봉현
    • Microbiology and Biotechnology Letters
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    • v.26 no.2
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    • pp.167-172
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    • 1998
  • In batch cultures of Brevibacterium ketoglutamicum for the L-ornithine production in which the pH and dissolved oxygen concentration were regulated constant, the profiles of redox potential were observed in parallel with the profiles of state variables such as cell, glucose, and ornithine concentrations. It was found that the redox potential had a close relationship with cell concentration and was also affected by ornithine concentration. The effects of ornithine and glucose on redox potential were examined in a separate series of experiments. Based on the experimental results, a correlation of redox potential to glucose, cell and ornithine concentrations has been proposed. The proposed correlation can be used for on-line estimation of ornithine concentration from on-line data of redox potential, glucose concentration, and cell concentration.

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Inhibitory Effect of Mixture of Ethanol Extracts in Agastachis Herba and Pueraria Radix on the Proliferation and $PGE_2$ Production of HT-29 Human Colon Cancer Cell Line (곽향과 갈근 복합제제의 대장암 세포주 HT-29 증식 저해효과 및 $PGE_2$ 생성 억제효과)

  • Lee, Seung-Youn;Kim, Hee-Seok;Kim, Jeoung-Ok;Hwang, Sung-Wan;Hwang, Sung-Yeoun
    • Korean Journal of Pharmacognosy
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    • v.37 no.4 s.147
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    • pp.283-289
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    • 2006
  • Ethanol extracts of the whole herb of Agastachis Herba (A) and of Pueraria Radix (P) alone and of their mixture (A+P) downregulated the cell growth, cyclooxygenase-2 (COX-2) expression, prostaglandin $E_2\;(PGE_2)$, and cGMP production. A, P, and A + P inhibited the cell growth of HT-29 colon cancer cells in a concentration- and time-dependent manner but not the growth of normal colon cell, CCD-112CoN. In addition, they markedly inhibited the productions of $PGE_2$ and cGMP as well as the mRNA expression of COX-2. These data suggest that non-toxic concentration of A, P, and A + P have a significant effect on the in vitro growth of HT-29 cells, specifically through the inhibition of the $PGE_2$ production via COX-2.

Present Status and Prospects of in vitro Production of Secondary Metabolites from Plant sin China

  • Chen, Xian-Ya;Xu, Zhi-Hong
    • Proceedings of the Botanical Society of Korea Conference
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    • 1995.06a
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    • pp.40-56
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    • 1995
  • During the past two decades, China has seen her great progress in plant biotechnology. Since the Chinese market of herb medicine is huge, while the plant resources are shrinking, particular emphasis has been placed in plant tissue and cell cultures of medicinal plants, this includes fast propagation, protoplast isolation and regeneration, cell suspension cultures and large scale fermentation. To optimize culture conditions for producing secondary compounds in vitro, various media, additives and elicitors have been tested. Successful examples of large scale culture for the secondary metabolite biosynthesis are quite limited : Lithospermum ery throrhizon and Arnebia euchroma for shikonin derivatives, Panax ginseng, P. notoginseng, P. quinquefolium for saponins, and a few other medicinal plants. Recent development of genetic transformation systems of plant cells offered a new approach to in vitro production of secondary compounds. Hairy root induction and cultures, by using Ri-plasmid, have been reported from a number of medicinal plant species, such as Artemisia annua that produces little artemisinin in normal cultured cells, and from Glycyrrhiza uralensis. In the coming five years, Chinese scientists will continue their work on large scale cell cultures of a few of selected plant species, including Taxus spp. and A. annua, for the production of secondary metabolites with medicinal interests, one or two groups of scientists will be engaged in molecular cloning of the key enzymes in plant secondary metabolism.

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Single Cell Protein Production by a Yeast Utilizing Alcoholic Waste Fluid of Sweet Potatoes (고구마 주정폐액을 자화하는 효모에 의한 SCP 생산)

  • Chung, Dong-Ok;Jung, Ji-Heun
    • Applied Biological Chemistry
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    • v.30 no.3
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    • pp.258-263
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    • 1987
  • The waste gained from alcohol production with sweet potato was considered as a potential substrate for the production of single cell protein (SCP). The high content in organic materials, simplicity to filtrate and a suitable pH for the growth of yeasts were indicated as a good substrate. A yeast utilizing this substrate was isolated from the compost ground and identified as Candida boidinii. The strain was the highest in assimilation of this alcoholic waste and the yield was maximized at pH 5.0, $30^{\circ}C$ after 72 hrs incubation. The dry cell weight and crude protein content at optimal conditions were 1.02g/100ml and 54.5%, respectively. The growth of the yeast was stimulated with the addition of 0.2% urea, 0.1% $K_2HPO_4$, and 0.02% $MgSO_4{\cdot}7H_2O$.

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Antioxidant potential of buffalo and cow milk Cheddar cheeses to tackle human colon adenocarcinoma (Caco-2) cells

  • Huma, Nuzhat;Rafiq, Saima;Sameen, Aysha;Pasha, Imran;Khan, Muhammad Issa
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.2
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    • pp.287-292
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    • 2018
  • Objective: The aim of present study was to assess the anti-oxidant potential of water-soluble peptides (WSPs) extract derived from buffalo and cow milk Cheddar cheeses at different stages of ripening. Methods: The antioxidant potential of WSPs extract was assessed through 2,2'-azinobis-3-ethylbenzothiazoline-6sulfonic acid (ABTS)-radical scavenging activity. In addition, impact of WSPs extract on cell viability and production of reactive oxygen species (ROS) in human colon adenocarcinoma Caco-2 (tert-butylhydroperoxide-induced) cell lines was also evaluated. Results: The ABTS-radical scavenging activity increased progressively with ripening period and dose-dependently in both cheeses. However, peptide extract from buffalo milk Cheddar cheese demonstrated relatively higher activity due to higher contents of water-soluble nitrogen. Intracellular ROS production in Caco-2 cells decreased significantly (p<0.05) till 150th day of cheese ripening and remained constant thereafter. Additionally, dose-dependent response of WSPs extract on antioxidant activity was noticed in the Caco-2 cell line. Conclusion: On the basis of current in vitro study, the Cheddar cheese WSPs extract can protect intestinal epithelium against oxidative stress due to their antioxidant activity.