The optima] conditions for the formation, the regeneration. and the spheroplast fusion of Flavimonas aryz/habitans spheroplasts were investigated. Cells were transformed to spherop]asts effectively by treatment of 0.5% volume (v/v) of 0.] M EDTA and ]00 flg/ml lysozyme at $37^{\circ}C$ for 30 min without shaking. Magnesium chloride and calcium chloride were effective on the stabilization of spheroplasts. and 20 mM calcium chloride in the rich regeneration medium improve the yield of regenerants as much as 3.5-fo]d. Addition of 0.8% bovine serium albumine (BSA) in dilution buffer for spheroplast formation improved the stabilization of spheroplasts over extended periods (4-6 hr) at room temperature. and thus increased the yield of recombinants to 4.5-fold. The spheroplast formation frequency and regeneration frequency of F aryzihabitans strain was 90.10% and 3.800/." respectively. The first regenerated cell of F. aryzihabitans spheroplasts were appeared 6 hours after plating. By I I hours after plating, 80% of spheroplasts were regenerated on thc rich regeneration medium containing 0.5 M sucrose. The intraspeci11c spheroplast fusion of F urvz/habitans was carried out and the properties of obtained fusants were investigated. Formation of fusion products was effective when the Flav/munas spheroplast mixture was treated with 40%(w/v) PEG6000 and 20 mM CaCl, for 10 min at room temperature. and thc formation of frequency of recombinants were $2.0{\times}10^{-5}~3.6{\times}10^{-5}$. All tested recombinant clones were very stable on further propagation.
Kim, Hoe-Geun;Song, Myeon-Gyu;Kim, Dong-U;Lee, Sang-Yul
Proceedings of the Korean Institute of Surface Engineering Conference
/
2018.06a
/
pp.62-62
/
2018
Pt is still considered as one of the most active electrocatalysts for ORR in alkaline fuel cells. However, the high cost and scarcity of Pt hamper the widespread commercialization of fuel cells. As a strong candidate for the replacement of Pt catalyst, silver (Ag) has been extensively studied due to its high activity, abundance, and low cost. Ag is more stable than Pt in the pH range of 8~14 as the equilibrium potential of Ag/Ag+ being ${\approx}200mV$ higher than that of Pt/PtO. However, Ag is the overall catalytic activity of Ag for oxygen reduction reaction(ORR) is still not comparable to Pt catalyst since the surface Ag atoms are approximately 10 times less active than Pt atoms. Therefore, further enhancement in the ORR activity of Ag catalysts is necessary to be competitive with current cutting-edge Pt-based catalysts. We demonstrate the architectural design of Ag catalysts, synthesized using plasma discharge in liquid phase, for enhanced ORR kinetics in alkaline media. An attractive feature of this work is that the plasma status controlled via electric-field could form the Ag nanowires or dendrites without any chemical agents. The plasma reactor was made of a Teflon vessel with an inner diameter of 80 mm and a height of 80 mm, where a pair of tungsten(W) electrodes with a diameter of 2 mm was placed horizontally. The stock solutions were made by dissolving the 5-mM AgNO3 in DI water. For the synthesis of Agnanowires, the electricfield of 3.6kVcm-1 in a 200-ml AgNO3 aqueous solution was applied across the electrodes using a bipolar pulsed power supply(Kurita, Seisakusyo Co. Ltd). The repetition rate and pulse width were fixed at 30kHz and 2.0 us, respectively. The plasma discharge was carried out for a fixed reaction time of 60 min. In case of Ag nanodendrites, the electric field of 32kVcm-1 in a 200-ml AgNO3 aqueous solution was applied and other conditions were identical to the plasma discharge in water in terms of electrode configuration, repetition rate and discharge time. Using SEM and STEM, morphology of Ag nanowires and dendrites were investigated. With 3.6 kV/cm, Ag nanowire was obtained, while Ag dendrite was constructed with 32 kV/cm. The average diameter and legth of Ag nanowireses were 50 nm and 3.5 um, and thoes values of Ag dendrites were 40 nm and 3.0 um. As a results of XPS analysis, the surface defects in the Ag nanowires facilitated O2 incorporation into the surface region via the interaction between the oxygen and the electron cloud of the adjacent Ag atoms. The catalytic activity of Ag for oxygen reduction reaction(ORR) showed that the catalytic ORR activity of Ag nanowires are much better than Ag nanodendrites, and electron transfer number of Ag nanowires is similar to that of Pt (${\approx}4$).
An apatite drain was constructed on September 30, 1994 at the Green Valley Abandoned Coal Mine site near Terre Haute in west central Indiana. The primary objective of this experiment is to evaluate the long-term ability of the apatite drain to mitigate acid mine drainage (AMD) under field conditions. The drain 9 m long, 3.3 m wide, and 0.75 m deep, contain 95 rum to No. 30 mesh-size apatite ore (francolite) and receive AMD seepage from reclaimed gob piles, and designed according to the laboratory testing. The apatite drain was covered with limestone riprap and filter fabric to protect the drainage system from stormwater and siltation. The drain consists of about 50 metric tons of apatite ore obtained from a phosphate mine in Florida. A gabion structure was constructed downstream of the apatite drain to create a settling pond to collect precipitates. Apatite effectively removed iron up to 4,200 mg/l, aluminum up to 830 mg/l and sulfate up to 13,430 mg/l. The pH was nearly constant for the influent and effluent, ranging between 3.1 and 4.3. Flow rate measured at the gabion structure ranged from 3 to 4.5 l/m. Precipitates of iron and aluminum phosphate (yellow and white suspendid solids) continued to accumulate in the settling pond.
Polygalacturonase(PG) and pectinesterase(PE) were extracted from Chinese cabbage and physicochemical properties of the enzymes were characterized. The preheating conditions for maximum retention of Kimchi texture were also studied. The activity of PE was highest at $50^{\circ}C$ and at 0.02M $CaCl_2$ but decreased in 0.2M $CaCl_2$, PG exhibited maximum activity at $65^{\circ}C$ with 0.3mM $CaCl_2$ but was inhibited by $CaCl_2$ at 0.5mM. Both of the enzymes, however, exhibited the maximum activity with 0.25M NaCl. Optimum preheating treatment was determined for minimum PG activity and maximum PE activity. Thus a maximum crispness and firmness was obtained with preheating in 0.05M $CaCl_2$ solution at $50^{\circ}C$ for 1.5hr results indicated that PE activity and calcium ion were very effective in preserving firmness.
Park, In-Cheol;Kim, Gwang-Su;Park, Myeong-Jin;Lee, Seung-Hun;Hong, Seok-Il;Choe, Tae-Bu
KSBB Journal
/
v.14
no.4
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pp.460-464
/
1999
Methicillin-resistant Staphyloccus aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes a problem of nosnocomial infection since the third generation of cephalosporines has been introduced in the 1980s. As antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions such as, pH, temperature, and time, etc., it is difficult to decide in the case of borderline- or low-level of MRSA. Therefore it would be necessary to develope a new method based on the molecular biological technique to overcome these problems. In this study, we extracted DNA from S. aureus and performed polymerase chain reaction (PCR) to amplify mec A gene, encoding penicillin-binding protein 2' (PBP-2'), which is known to confer bacteria resistance to the bacteriostatic action of methicillin. The results were compares with those of minimal inhibitory concentration (MIC) test. When MIC test with oxacillin was performed on the 120 isolates of S. aureus from each patient's specimens, 64 of them were MRSA and 56 of them were methicillin-sensitive Staphylococcus aureus (MSSA). In pus specimen, more precisely, 61.9% (26/42) of MRSA was detected, and 44.2% (19/43), 60% (9/15) and 50% (10/20) of MRSA were detected in sputum, body fluid, and other specimen respectively. When 40 isolates of MRSA and MSSA were tested by PCR method and compares with the results of MIC method, different results were obtained from 1 isolate of MRSA (2.5%) and in 2 isolates of MSSA (5%) suggesting that PCR method should be performed at the same time for more accurate clinical test of MRSA.
We have investigated water quality of the coastal saline groundwaters utilized for fish farms in Jeju Island. The water quality investigation included the spatial observations for 75 fish farms during March-May, 1994 and the hi-monthly observations for both coastal saline groundwaters and seawaters at four fish farms from August 1994 to December 1995. Water temperature of the saline groundwaters ranged from 16 to 18$^{\circ}C$ over the study period. Salinity of the saline groundwaters varied between 20.60 ppt and 34.02 ppt, slightly lower than that of the coastal seawaters(26.47~34.53 ppt). This salinity variation must be associated with local precipitation conditions in Jeju Island. The oxygen saturation for most saline groundwater samples was lower than 80%, ranging from 24.7 to 89.8%. The COD and pH values for the saline groundwaters were similar to those for the coastal seawaters. The concentrations of DIP for the saline groundwaters varied between 0.021 mg/L and 0.121 mg/ L, and seasonal variation of DIP in the saline groundwater ranged from 0.014 to 0.077 mg/L, which were higher than that of the coastal seawaters(0.000~0.015 mg/L). Nitrate in the saline groundwaters accounted for more than 90% of the DIM. The maximum concentrations of ammonia, nitrite, nitrate and DIN in the saline groundwaters were 0.085, 0.012, 2.294 and 2.309 mg/L, respectively. These concentrations of the saline groundwaters were considerably lower than those affected culture organisms. Overall, the saline groundwaters utilized for fish farms in Jeju Island appear to maintain good waterquality for fish farms.
Aronia (Black chokeberry, Aronia melanocarpa) belonging to the Rosaceae family, is native to eastern North America. Aronia contain high levels of flavonoids, mostly anthocyanins and proanthocyanidins, which are known as condensed tannins. The dominant proanthocyanidins in aronia are (-)-epicatechin and (+)-catechin. The concentration of proanthocyanidins in aronia is higher than in other berries, however due to the astringent taste it is not desirable for consumption. Therefore, the purpose of this study is to evaluate the effect of aronia on the reduction in tannins by yeast isolated from regional Jeotgal. We isolated strains of yeast with high ${\beta}$-glucosidase activity from Jeotgal, with the MTY2 strains exhibiting a reduction in final tannin concentration according to thin layer chromatography (TLC) analysis. MTY2 was confirmed as Kazachstania servazzii using an 18S rDNA sequence and named as K. servazzii MTY2. K. servazzii MTY2 showed most significant growth when K. servazzii MTY2 was cultured in a solution of 10% (w/v) glucose, 3% (w/v) tryptone and 0.1% (w/v) sodium chloride. According to the high performance liquid chromatography (HPLC) analysis, the (+) - catechin peak is present, but (-) - epicatechin peak was reduced at culture condition added with 10% glucose in medium.
The lignocellulytic enzymes including a-amylase (EC 3.2.1.1), lignin peroxidase (EC 1.11.1.14), laccase (EC 1.10.3.2), xylanase (EC 3.2.1.8), ${\beta}$-xylosidase (EC 3.2.1.37), ${\beta}$-glucosidase (EC 3.2.1.21) and cellulase (EC 3.2.1.4) were extracted from spent mushroom compost (SMC) of Pleurotus eryngii. Different extraction buffers and conditions were tested for optimal recovery of the enzymes. The optimum extraction was shaking incubation (200 rpm) for 2 h at $4^{\circ}C$. ${\alpha}$-Amylase was extracted with the productivity range from 1.20 to 1.6 Unit/SMC g. Cellulase was recovered with the productivity range from 2.10 to 2.80 U/gf. ${\beta}$-glucosidase and ${\beta}$-xylosidase productivities showed lowest recovery producing 0.1 U/g and 0.02 U/g, respectively. The P. eryngii SMCs collected from three different mushroom farms showed different recovery on laccase and xylanse, cellulase. Furthermore, the water extracted SMC was compared to commercial enzymes for its industrial application in decolorization and cellulase activity.
The germination characteristics of Diplachne fusca (L.) P. Beauv. seeds were investigated under different seed storage conditions to find out reliable system for maintaining the seeds with high and uniform germination rate and thus for possible use of the seeds in herbicide screening with a continuous seed supply. When the seeds were stored under wet-low temperature($4^{\circ}C$) condition, the germination rate was 88% after 4-week-storage. The germination rate slightly declined after the storage for longer than 3 months. Dry seeds stored at room temperature exhibited very low germination rate. The wet-low temperature treatment was effective for inducing the germination of the seeds which had been stored under dry-room temperature condition for 4 months. The germination rate was 70% after 2-week- storage under the wet-low temperature condition. The germination rate was much higher under an alternate temperature condition than under a continuous temperature condition. The optimum temperature was 35/$25^{\circ}C$(14/10hrs). The seeds had a capability to germinate under NaCl-treated condition even at a concentration of 1.0%, whereas the germination of Echinochloa crus-galli seeds was completely inhibited by 0.5% NaCl. This result indicates that D. fusca has an advantage over E. crus-galli to survive in reclaimed 1and from the sea.
Although microorganisms are, in fact, the most diverse and abundant type of organism on Earth, the ecological functions of microbial populations remains poorly understood. A variety of bacteria including marine Vibrios encounter numerous ecological challenges, such as UV light, predation, competition, and seasonal variations in seawater including pH, salinity, nutrient levels, temperature and so forth. In order to survive and proliferate under variable conditions, they have to develop elaborate means of communication to meet the challenges to which they are exposed. In bacteria, a range of biological functions have recently been found to be regulated by a population density-dependent cell-cell signaling mechanism known as quorum-sensing (QS). In other words, bacterial cells sense population density by monitoring the presence of self-produced extracellular autoinducers (AI). N-acylhomoserine lactone (AHL)-dependent quorum-sensing was first discovered in two luminescent marine bacteria, Vibrio fischeri and Vibrio harveyi. The LuxI/R system of V. fischeriis the paradigm of Gram-negative quorum-sensing systems. At high population density, the accumulated signalstrigger the expression of target genes and thereby initiate a new set of biological activities. Several QS systems have been identified so far. Among them, an AHL-dependent QS system has been found to control biofilm formation in several bacterial species, including Pseudomonas aeruginosa, Aeromonas hydrophila, Burkholderia cepacia, and Serratia liquefaciens. Bacterial biofilm is a structured community of bacterial cells enclosed in a self-produced polymeric matrix that adheres to an inert or living surface. Extracellular signal molecules have been implicated in biofilm formation. Agrobacterium tumefaciens strain NT1(traR, tra::lacZ749) and Chromobacterium violaceum strain CV026 are used as biosensors to detect AHL signals. Quorum sensing in lactic acid bacteria involves peptides that are directly sensed by membrane-located histidine kinases, after which the signal is transmitted to an intracellular regulator. In the nisin autoregulation process in Lactococcus lactis, the NisK protein acts as the sensor for nisin, and NisR protein as the response regulator activatingthe transcription of target genes. For control over growth and survival in bacterial communities, various strategies need to be developed by which receptors of the signal molecules are interfered with or the synthesis and release of the molecules is controlled. However, much is still unknown about the metabolic processes involved in such signal transduction and whether or not various foods and food ingredients may affect communication between spoilage or pathogenic bacteria. In five to ten years, we will be able to discover new signal molecules, some of which may have applications in food preservation to inhibit the growth of pathogens on foods.
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