• Microbiology and Biotechnology Letters
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• v.14 no.6
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• pp.455-460
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• 1986

Dane particle was prepared from the plasma of chronic HBsAg carrier with high levels of HBsAg activity. DNA extracted front Dane particle core after a DNA polymerase reaction with $\alpha$-($^{32}$P) dNTP, was identified as HBV DNA by liquid scintillation counter and agarose gel electrophoresis-G.M. counting. To produce Hepatitis B surface antigen for use as a vaccine against Hepatitis B virus infection, yeast strains harboring recombinant plasmid with Apase promoter was used. Recombinant plasmid was construced from pHBV 130 and pAN 82, transformed into E coli, and then transferred into yeast strains. HBsAg was produced by derepression in Burkholder minimal medium with controlled inorganic phosphate concentration. The kinetics of HBsAg production was also investigated. Total HBsAg activity increased rapidly between 3 and 6 hours after transfer to phosphate-free medium and reached a maximum at around 9th hour. The transfer into phosphate-free medium after 6 hours in high phosphate cell growth medium gave maximum activity.

• Clinical and Experimental Reproductive Medicine
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• v.26 no.1
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• pp.75-81
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• 1999

The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.

• Journal of Life Science
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• v.11 no.2
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• pp.63-69
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• 2001

bacterium having high abilities to solubilize in-organic phosphate was isolated from cultivated soils. The strain was identified as Aeromonas hydrophila DA33, based on the physiological and biochemical properties. The optimum temperature and initial pH to solubilize insoluble phosphate in sucrose minimal medium were 3$0^{\circ}C$ and pH 5.0, respectively. In these conditions, phosphate-solubilizing activities of the strain against two types of insoluble phosphate were quantitatively determined. When glucose was used for carborn source, the strain had a marked mineral phospahte solubilizing activity. Inorganic phospahte solubilization was directly related to the pH drop by the strain. Analysis of the culture medium confirmed the production of gluconic acid as the main organic acid released by Aeromonas hydrophila DA33.

• The Korean Journal of Zoology
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• v.7 no.2
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• pp.25-36
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• 1964

The present paper deals with the comparative study on phylogenic difference in the patterns of energy metabolism of brain slices of several vertebrate species by measuring oxygen consumptionwith glucose-6-phosphate, glucose-1-phosphate, glyceraldehyde-3-phosphate or glutamate as respiratory substrate employing Warburg's manometric method, by determination of the utilization rate of glucose using glucose-1-C14 by analyzing patterns of free amino acid distribution , and by histochemical determination using glucose-1-C14 by analyzing patterns of free amino acid distribution acid distribution , and by histochemical determination of glycogen contents. 1. Glucose enhances the oxygen consumption of brain slices of animals belinging to reptile, aves and mammalia while it shows a tendency to decrease that of animals belonging to pisces and amphibia. 2. Glucose-6--phosphate increase oxygen consumption more than glucose in every species examined, while glucose-1-phosphate and glyceraldehyde-3-phosphate increase that of Rana nigromaculata only . In general m, it appears that phosphosugars are more effective as a respiratory substrate to those species which have less endogenous respiration than to those having larger endogenous respiration. 3. Similar patterns of free amino acid distribution and the relative amount are found among the species and in every species examined glutamic acid is detected in the larges amount . ${\gamma}$-Amino butyric acid, glycine, alanine and aspartic acid are found in every species. 4. Ophicephalus showed less oxygen consumption than endogenous respiration when glutamate was added to the medium. When sodium fluoride was added, the oxygen consumption was some what increased . Such phenomenon wasnot found in the frog. 5. The result of histochemcial analysis of the brain showed that glycogen was abundantly present in the fish , amphibia , and especially in the reptile and that no distinctive grains of glycogen were found in the bird and mammal . From these facts, it may be supposed that anaerobic glycolysis as energy source dominates in fish and amphibia and aerobic respiration through the oxidation of glucose dominates in bird and mamal , the reptile occupying transitional position between these two categories. The way of obtaining energy for brain activity by the oxidation of glucose supplied from the circulating blood is seemed to be first acquired by reptile and the function is completed both in aves and mammal.

• Journal of Life Science
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• v.6 no.3
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• pp.204-212
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• 1996

The aim of this study was to evaluate the effects of phosphate, aimno acid, and BSA on in vitro development of mammalian embryos. In vitro-matured and -fertilized(IVM/IVF) bovine embryos were cultured in simple, chemically defined, protein-free medium(mTLP-PVA0. When the phosphate concentration of mTLP-PVA supplemented with 19 amino acid were adjusted to 0.0, 0.10, 0.35, 1.05 and 2.10mM by the concentration of sodium phoshpate, there were no significant different in development ability of IVM/IVF bovine embryos cultured in the medium containing from 0.00 to 1.05mM phosphate until 48 hours post-insemination, However, proportion of embryos developing to $$8-cell and morula at 96 and 144 hours post- insemination, respectively, was significantly increased in the medium with o.35 mM phosphate(p<0.05). There was significant difference between O.10(18%)-0.35(24%)mM phosphate and 1.05(13%)-2.10(1%)mM phosphate in supporting development to blastocyst(p<0.05). When IVM/IVF bovine embryos were cultured in the medium supplemented with 19 amino acids, significant different was observed in the proporton of embryos reaching $$8-cell(49-50%), morula(38-40%) and blastocyst (29-32%) stages at 96, 144, and 192 hours post-insemination, respectively(p<0.05). Glutamine alone had no benefit on embryo development. When BSA was added to mTLP-PVA with 0.35mM phosphate, glutamine and 19 amino acids at 8, 48, 120 hours post-insemination, BSA significantly enhanced the development ability ofb embryos reaching $$2-cell (74-77%), $$8-cell (49-53%), morula(43-47%), and blastocyst(38-42%) stages at 48, 96, 144, and 192 hours post-insemination, respectively, regardless of the time of BSA addition.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.551-563
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• 1996

This study investigated the properties of primary cultured proximal tubule cells in hormonally defined(insulin, transferrin, and hydrocortisone), serum-free medium or 10% serum-supplemented medium. The growth rate of the primary cultured proximal tubule cells was lower in the hormonally defined, serum-free medium than in the 10% serum- supplemented medium(p < 0.05), while the activities of brush border marker enzymes, alkaline phosphatase(AP), leucine aminopeptidase(LAP), and y-glutamyl transpeptidase(${\gamma}$-GTP) were increased(p < 0.05). The activities of these enzymes, however, decreased with the lapse of incubation time to 50-70% after 6 days culture compared to those of the freshly-prepared proximal tubules. The enzymatic activities of the primary cultured proximal tubul cells on 6, 9, 12, and 15 days of culture were significantly increased in the hormonally defined, serum-free medium compared to the 10% serum-supplemented medium(p < 0.05). The functional differentiation of the primary culture was examined by observing multicellular domes of the confluent monolayer, which is indicative of transepithelial solute transport. The dome formation by the proximal tubule cultures occurred at a higher frequency in the hormonally defined, serum-free medium than in the 10% serum-supplemented medium(p < 0.05). Upon electron microscopic examination, an increased density of the brush border was observed in the hormonally defined, serum-free medium compared to the cells grown in 10% serum-supplemented medium. The activities of $Na^+$glucose cotransporter($^{14}C$-a-MG uptake), $Na^+$phosphate cotransportere($^{32}P$ uptake) and $Na^+$ transporter($^{22}Na^+$ uptake) in the brush border membrane, and of $Na^+/K^+$-ATPase($^{86}Rb$ uptake) in the basolateral membrane were significantly stimulated in the hormonally defined, serum-free medium than in 10% serum-supplemented medium(p < 0.05). In conclusion, the primary cultured proximal tubule cells grown in the hormonally defined, serum-free medium demonstrated a slower growth rate, but the functions of cell were enhanced.

• Applied Biological Chemistry
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• v.40 no.4
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• pp.329-333
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• 1997

Phosphate solubilizing microorganisms (1,000 bacteria and 200 fungi) were isolated from soil around Kyungnam and Kyungbook regions using potato dextrose agar-calcium phosphate medium. A fungus with the greatest phosphate solubilizing activity was selected and identified to Penicillium sp. GL-101, based on the morphological characteristics of conidiophore and conidia; flask shape of phialide, simple branching type of conidiophore, and columnar shape of conidial head, in malt extract agar and potato dextrose agar media. The optimum temperature and initial pH to solubilize rock phosphate in potato dextrose broth-rock phosphate medium were $25^{\circ}C$ and pH 7.5, respectively. In these optimum conditions, phosphate solubilizing activities of Penicillium sp. GL-101 against four types of insoluble phosphate: tricalcium-phosphate, aluminium phosphate, hydroxyapatite and rock phosphate, were quantitatively determined. As results, this fungus highly discharged free phosphates to the culture broth with the concentrations of 1,152 ppm against tricalcium-phosphate, 565 ppm against rock phosphate, 292 ppm against aluminium phosphate, and 217 ppm against hydroxyapatite, respectively.

• Environmental Engineering Research
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• v.25 no.3
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• pp.393-399
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• 2020

The use of different types of wastewater (WW) for the cultivation of microalgae and cyanobacteria during recent decades has provided important economic and environmental benefits. However, direct use of WW can lead to growth inhibition and biomass contamination. In the present study, we separated the key WW nutrients, namely nitrate and phosphate, by adsorption and regeneration and used the resulting regenerated water to cultivate the cyanobacterium Spirulina platensis. The adsorbent was granular γ-alumina derived from waste aluminum cans. This procedure recovered 19.9% of nitrate and 23.7% of phosphate from WW. The cyanobacterial cultures efficiently assimilated the nutrients from the medium prepared using regenerated WW, and the growth and nutrient uptake were similar to those in a synthetic medium. In addition, imposing nutrient limitations to increase carbohydrate productivity was easily achieved using regenerated wastewater nutrients, without requiring additional dilution or complex processing. In acute toxicity tests, the harvested biomass in a regenerated medium had similar toxicity levels compared to the biomass obtained from a synthetic medium. The proposed method of using regenerated WW to produce contamination-free biomass has broad potential applications.

• Food Science of Animal Resources
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• v.38 no.1
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• pp.26-42
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• 2018

Today incorporation of natural ingredients as inorganic phosphate replacers has come into prominence as a novel research topic due to health concerns about phosphates. In this study, we aimed to investigate the quality of emulsified chicken meatballs produced with Jerusalem artichoke powder (JAP), either alone or in combination with sodium carbonate (SC) as sodium tripolyphosphate (STPP) replacers. The results showed that naturally dried JAP showed favorable technological properties in terms of water-oil binding and gelling. Emulsion batters formulated with JAP-SC mixture showed lower jelly and fat separation, higher water-holding capacity and higher emulsion stability than control samples with STPP. In final product, incorporation of JAP-SC mixture increased moisture and reduced lipid and energy values, and kept the pH value similar to control. Added JAP lead to increments in $b^*$ values whereas decreases $L^*$ values. Cook yield was similar to control in phosphate-free samples formulated with JAP-SC mix. Either low or medium ratios of JAP in combination with SC managed to protect most of the sensory parameters, while sensory scores tend to decrease in samples containing high levels of JAP. Addition of JAP to formulations presented samples that have equivalent behavior to phosphates in terms of lipid oxidation. In conclusion, our study confirms that utilization of JAP in combination with SC had promising effects as phosphate replacers by presenting natural solutions and providing equivalent quality to standard phosphate containing products.

• The Korean Journal of Mycology
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• v.13 no.4
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• pp.235-241
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• 1985

The present study was designed to investigate biosynthetic patterns of polysaccharides under catabolic repression and derepression in Saccharomyces uvarum. Correlation coefficients between polysaccharide synthesis and polyphosphate accumulation were examined, according to the culture phase and under various phosphate concentrations (free, limited, sufficient). During catabolic derepression, biosynthesis of glycogen was enhanced. rapidly and highly in the cells grown on minimal medium, compared with those grown on the complete medium. Acid soluble glycogen type was the main component of total glycogen and alkali soluble glycogen was synthesized in small amount, after 24 hr culture, at the time of almost exhaustion of sugar in the medium. Total glycogen was accumulated highly in proportion to the amount of phosphate added to the medium. It could be postulated that type 'C' isoenzyme among ALPase was directly or indirectly correlated with the glucan synthesis. Mannan synthesis indicated maximal amount at the early exponential phase and stationary phase, and also acid soluble sugars at the stationary phase. Correlation coefficient between the mannan synthesis and poly-p-'C' accumulation, and also between mannan synthesis and phospholipid content indicated 0.866 and 0.726, respectively.