To improve the growth of mouse hybridoma 2c3.1 secreting anti-Hepatitis B surface antigen monoclonal antibody (anti-HBsAg MAb), we had constructed an enriched medium and observed the effects of fetal bovine serum and serum-free supplements including human serum albumin, 'insulin and transferrin', and monoethanolamine. For further enhancement of growth, the concentrations of two major energy sources, glucose and glutamine, were strengthened with various ratios in the enriched medium. Maximum cell growth and monoclonal antibody production obtained in various ratios of glucose/glutamine with an inoculation concentration of 2$\times$105 cells/ml were 0.73$\times$106-4.62$\times$106 cells/ml and 65.1-422.6 $\mu\textrm{g}$/ml, respectively. Glutamine was round to be a major energy source and a limiting nutrient in comparison to glucose for 2c3.1 cell cultivation in enriched media with low serum.
Fourteen different taxa of dictyostelid cellular slime molds were recovered from the alpine and subalpine zone in Mt. Paektu. In subalpine zone, six species were recovered : Dictyostelium minutum, D. brefeldianum, D. crassicaule, D. capitatum, Polysphondylium solitarium and P. pallidum. One of these species which were isolated from the soils of Larix olgensis community exhibited several distinctive features which differed from the published species. This species was designated as a new species, Polysphondylium solitarium Kang et Chang, sp. nov. When cultivated at 22~24$^{\circ}C$ on low-nutrient agar media with Escherichia coli, Polysphondylium solitarium is distinguishable from other species by the following combination of features: (ⅰ) the sorocarps with vinaceous pigmentation; (ⅱ) pseudoplasmodia radial. usually centralized and rarely subdivided; (ⅲ) the various number of whorls; (ⅳ) the spores with unconsolidated and nonconspicuous polar granules. Also, we confirmed this new species by analyzing ribosomal DNA (ITS1, ITS2 and 5.8s DNA) sequences of P. solitarium and P. violaceum.
It was discovered that ammonium phosphate in the medium played an important role in both growing mycelium and producing exopolysaccharides (EPS) from G. lucidum. In lower concentration levels of ammonium phosphate (0-3 g/l), an improved mycelial growth was observed by maintaining more filamentous morphology than in high concentrations (5-11 g/l). In addition, it was confirmed by comparing the factual dimension and frequency of the area regarding the mycelial pellets. This must be attributed to limitations of nutrient transfer by maintaining filamentous mycelium during the cultivation in a low ammonium phosphate containing medium. On the other hand, the best EPS production was observed in medium with the absence or low concentration of ammonium phosphate. The shear stress of the culture broth was greatly affected by the shear rate, as compared with that of the culture broth with high ammonium phosphate concentration. The rheological characteristics of the fermentation broth and filtrate worked well according to the Herschel-Bulkley model. It was also found that the morphological changes of the mycelium resulting from the ammonium phosphate concentration directly affected the rheological characteristics of the system and resulted in reversely affecting the EPS production levels. Based on these results, it can be concluded that delicate regulation of the ammonium phosphate concentration in the culture media should be provided in order to obtain optimal mycelial growth and/or EPS production.
Kim, Sun-Ae;Lee, Yu-Mi;Oh, Se-Wook;Gwak, Hyo-Sun;Hwang, In-Gyun;Kang, Dong-Hyun;Woo, Gun-Jo;Rhee, Min-Suk
Food Science of Animal Resources
/
v.29
no.6
/
pp.702-708
/
2009
We investigated biofilm formation in various media, growth in low pH, and the hemolytic activity of 14 strains of Cronobacter spp. (Enterobacter sakazakii) isolated from a variety of foods including powdered infant formula (n=75), infant cereal (n=100), honey (n=30), and other infant foods (n=100) in Korea. The Cronobacter spp. adhered and formed biofilms on polyethylene, and a greater extent of biofilm was observed in nutrient-rich media. No clear difference in biofilm-forming ability was noted among the media constituents and the pattern of biofilm formation was strain-dependent. Seven strains out of 14 strains (50%) grew at pH 4.1, indicating that the acid resistance of these Cronobacter spp. isolated in Korea was relatively low. Hemolytic activity was not observed in any of the strains. This study provides basic information for the physiological and biochemical characteristics of Cronobacter spp. isolated from a variety of infant foods in Korea.
Journal of Korean Society for Atmospheric Environment
/
v.19
no.6
/
pp.689-699
/
2003
This study was to examine characteristics of treating toluene vapor, which gets to be problematic due to its harmful carcinogenicity and mass generation from various sources, through a biological treatment facility which is environment-friendly and adopts a high-efficient and low-cost clean technology. In order to identify whether Alnus Firma Fruit (AFF) can be used as a media for a bioreactor, its utility and basic operating factors, a study was conducted on pressure drop, supply of nutrient substances and retention time which are operating factors of a biofilter, and eliminating characteristics were compared between AFF and the conventional biological activatedcarbon (BAC) widely used as filter media. In the case of AFF, the initial microbial deposits was 2.3${\times}$10$^{7}$ CFU/g dry AFF, which represents the initial microbial density higher than the case of BAC showing 5.5${\times}$10$^{6}$ CFU/g dry BAC And it took about 2 weeks to acclimate until its eliminating rate got to be increased over 90%. As a result of comparing pressure loss taking place with the lapse of time between BAC and AFF, after 130 days passed at SV 25h$^{-1}$ , BAC showed that its eliminating efficiency had a tendency to drop greatly due to a great pressure loss (0.53\longrightarrow54.7 mm$H_2O$/m) caused by an excess of biomass as accumulated. On the other hand. AFF showed that the pressure drop was 0.53 mm$H_2O$/m, about 2 times as much as the initial pressure loss of 0.4 mm$H_2O$/m, which represents no great change in the pressure loss, and its eliminating efficiency was also shown to be continuously high. Therefore, when AFF was used as a filler for a biological treatment facility, a biological filter enabling improvement of the purifying efficiency to be promoted could be provided, and moreover, the pressure loss was so small that the filler replacement cycle or the back flushing cycle could be extended. So, even in terms of the operating cost, it was identified to be an economical filler When an inorganic material was used as a filler, the biofilters performance acted sensitively on whether nutrient substances were supplied or not. In the case of AFF with low adsorptivity, addition of ethyl-alcohol increased the solubility of toluene, and consequently, biodegradation got to be actively made by microbes, and thus, its eliminating rate could be increased. As the flow velocity and the inflow concentration got to be more increased, its eliminating rate got to be lower, and particularly, an increase in the flow velocity made its eliminating rate drop more greatly than an increase in the concentration.
Proceedings of the Korean Society of Crop Science Conference
/
2017.06a
/
pp.196-196
/
2017
Oilseed crop Camelina (Camelina sativa L.) is a suitable for biodiesel production that has high adaptability under low-nutrient condition like marginal land and requires low-input cost for cultivation. Enhanced abiotic stress tolerance of Camelina is very important for oil production under the wide range of different climate. CsRCI2s (Rare Cold Inducible 2) are related proteins in various abiotic stresses that predicted to localized at plasma membrane (PM) and endoplasmic reticulum (ER). These proteins are consist of eight-family that can be divided into tail (CsRCI2D/E/F/G) and no-tail (CsRCI2A/B/E/H) type of C-terminal. However, it is still less understood the function of C-terminal tail. In this study, CsRCI2D/H genes were cloned through gateway cloning system that used pCB302-3 as destination vector. And we used agrobacterium-mediated transformation system for generation of overexpression (OX) transformants. Overexpression of target gene was confirmed using RT-PCR and segregation ratio on selection media. We analyzed physiological response in media and soil under abiotic stresses using CsRCI2D and CsRCI2H overexpression plant. To compare abiotic stresses tolerance, wild type and CsRCI2D/H OX line seeds were sown on agar plate treated with various NaCl and mannitol concentration for 7 days. In the test of growth rate under abiotic stress on media, CsRCI2H OX line showed similar to NaCl and mannitol stress. In the other hand, CsRCI2D OX line showed to be improved stress tolerance that especially increased in 200mM NaCl but was similar on mannitol media. In greenhouse, WT and CsRCI2D/H OX lines for physiological analysis and productivity under abiotic stresses were treated 100, 150, 200mM NaCl. Then it was measured various parameters such as leaf width and length, plant height, total seed weight, flower number, seed number. CsRCI2H OX line in greenhouse did not show any changes in physiological parameters but CsRCI2D OX line was improved both physiological response and productivity under NaCl stress. Among physiological parameters of CsRCI2D OX line under NaCl stress, leaf length and width were observed shorter than WT but it were slightly longer than WT in 200mM NaCl stress. Furthermore, total seed weight of CsRCI2D OX line under stress displayed to decrease than WT in normal condition, but it was gradually raised with increasing NaCl stress then more than WT relatively. These results suggested CsRCI2D might be contribute to improve abiotic stress tolerance. However, function of CsRCI2H is need to more detail study. In conclusion, overexpression of CsRCI2s family can generate various environmental stress tolerance plant and may improve crop productivity for bio-energy production.
In this study the removal possibility of nutrients of T-P, NH3-N, NO3-N and T-N is examined through a positive experimental study using submerged biofilter of media packing channel method. From the analysis of nutrients removal efficiency for each run of the collected sample following results are obtained. Firstly the result of N/P surveying for inflow shows serious value that excess the limit value of 20 as the values are in the range of 12.0~42.7 and the average is 25.73. Secondly the highest concentration of the incoming NH3-N reaches double of the standard since the concentrations of NH3-N and NO3-N for inflow shows 0.06mg/$\ell$ and 2.5~3.8mg/$\ell$ respectively and the average removal rate which passed the submerged biofilter adopted in this study is a satisfactory level. Next the average removal rate of T-P of 51.5% shows the possiblity of entrophication removal since the removal rate of T-P of 66.8~68.8% in relative low temperature period of RUN 1~2 appeared higher than in RUN 3~6 and T-N shows relatively poor result with the average removal rate of 34.1% And it is known that the bigger BOD/P and BOD/N are the more removal rate increases from the examination result of the relation between BOD/P and BOD/N and the treatment water T-P and T-N to decide the relation with the concentration of organic matters and though that the appropriate proportion is necessary for effective removal of nitrogen and phsophorus.
We isolated and identified culturable Arctic bacteria that had inhabited soils around the Korean Arctic Research Station Dasan located at Ny-Alsund, Svalbard, Norway $(79^{\circ}N,\;12^{\circ}E)$. The collected soils were diluted in distilled water; the diluted soil-water was spread on 3M petri-films at Dasan Station. The petri-films were transported to the laboratory at KORDI, and cultured at $4^{\circ}C$. Colonies grown on the petri-films were subsequently cultured on nutrient agar plates at $4^{\circ}C$ every 7 days. The pure colonies were inoculated into nutrient liquid media, genomic DNA was extracted, and phylogenetic analysis was performed on the basis of 165 rDNA sequences. A total of 227 strains of bacteria were isolated. Among them, 16S rDNA sequences of 185 strains were identical with those of known strains isolated in this study, and 42 strains were finally identified. Phylogenetic analysis using 16S rDNA indicated that the 30 strains belonged to Pseudomonas, 7 strains to Arthrobacter, two strains to Flavobacterium, and the remaining to Achromobacter, Pedobacter, and Psychrobacter. Among the 42 strains, 14 bacteria produced protease: they were 6 strains of Pseudomonax, 4 strains of Arthrobater, an Achromobacter strain, 2 strains of Flavobacterium, and a Pedohacter strain. We expect these Arctic bacteria can be used for screening to develop new industrial enzymes that are active at low temperatures.
Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$$vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$$carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$$Mosaic$$Virus$ and the gray mold disease caused by $Botrytis$$cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$$manteilii$ B001 and $Bacillus$$cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.
Addition of carbon and nitrogen source to an alcohol distillery wastewater was tried to increase the cell concentration of a b baker's yeast cultivated in that wastewater. Carbon was found to be primary limiting nutrient and nitrogen secondary limiting o one. Glucose addition increased the cell concentration 1.3 times higher than no addition, and both glucose and $(NH_4)_2S0_4$ a addition did 5.8 times. A fed-batch cultivation by the automatic addition of glucose and ammonium was executed. Added g glu$\infty$se was automatically controlled to low concentration by a method using DO as control parameter. Ammonium was a automatically added as NH40H used as pH $\infty$ntrol agent after initiating glucose addition. By this simple cultivation method t the cell concentration $\infty$내d be efficiently increased from 2.6g/L to 12.0g/L, and maximum specific growth rate and biomass y yield to glu$\infty$se were $0.18hr^{-1}$ and about 0.54g/g respectively. By increasing cell concentration, COD of the wastewater m media could be additionally reduced by about 22%.
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