• Applied Biological Chemistry
• /
• v.46 no.4
• /
• pp.353-360
• /
• 2003

For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells $(2{\times}10^6\;cell/ml)$ were infected four times at four-day-intervals. His-tag containing MAChE was purified using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures $(20-50^{\circ}C)$ under experimental conditions. As an extraction solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).

• Applied Biological Chemistry
• /
• v.46 no.2
• /
• pp.79-83
• /
• 2003

Elicitins, proteinaceous elicitors secreted from Oomycetes fungi (Phytophthora spp. and Pythium spp.), have been known as inducer of hypersensitive response (HR) in incompatible interactions between plant and pathogens. Five elicitins among many Korean Phytophthora species caused the reactions of distal HR in radish, chinese cabbage and some hot pepper cultivars, but not in cucumber and tomato. Because the isolation of elicitin from Phytophthora cambivora hasn't been reported yet, we have purified a cambivorein, a new member of the elicitin family, from the culture filtrate of Phytophtilora cambivora (KACC 40160) by using FPLC (Fast Protein Liquid Chromatography, AKTA) with sepharose S and Sephacryl HR columns. We confirmed that it induces necrosis activities in some hot pepper cultivars and its molecular weight is about 10 KDa by Tricine-SDS-PAGE. Comparison of amino acid sequences of its N-terminal ends also informed the identification of Iysine at the 13th position, which is characteristic of a kind of basic elicitin isoform $({\beta}-isoform)$. It Also showed that our elicitin is not identical with N-terminal sequences of many elicitins reported from Phytophthora spp..

• Journal of Food Hygiene and Safety
• /
• v.30 no.1
• /
• pp.74-80
• /
• 2015

In this study, the microbial control effect after treatment of washing solutions (tap water, electrolyzed water, chlorine dioxide solution) and quality changes were investigated when Kwamegi is produced by means of the cold air drying. The initial moisture rate of the sample was 56.62% before drying. At the beginning of drying period, moisture rate sharply fell down but as the experiment progressed, moisture reducing rate was smoothly decreased. The color difference of Kwamegi before drying was 42.40, but it was reduced depending on the increment of drying temperature and time. TBA value of the initial sample was 0.219, rancidity were increased continuously when drying progressed. Total amino acid content was showed the highest value at $25^{\circ}C$ for 36h and the lowest at $40^{\circ}C$ for 12h. From the fatty acids analysis, major fatty acids were consisted of the 14:0, 16:0 and 18:1 (18.1520.96%, 28.0632.51% and 17.0619.81%, respectively). The microbial control effect was biggest when Kwamegi was washed with chlorine dioxide 100ppm for 60s. The microbe of the Kwamegi, Pseudomonas sp. and Pseudomonas putida were identified.

• Korean Journal of Food Science and Technology
• /
• v.36 no.1
• /
• pp.152-157
• /
• 2004

Gluten was extracted from domestic wheat flour using UTH buffer (4 M urea in 0.1 M Tris-HCl, pH 8.6) and validated by SDS-PAGE analysis for production of wheat flour products with reduced gluten content.. Anti-gluten polyclonal antibody was made by administering extracted gluten fraction on animal model. Anti-gluten serum titer of extracted gluten fraction was evaluated by ELISA, and that of antibody titer according to administration period. Anti-gluten sera were used for ELISA and immunoblot analysis before and after hydrolysis of gluten fraction at optimal pH and temperature condition for each protease. Gluten fraction separated by SDS-PAGE showed several bands covering 75 to 10 kDa, in which anti-gluten sera were 25, 34, and 45 kDa. Enzyme hydrolysis of gluten fraction revealed protein band sizes to be lower than 15 kDa. Content of pretense from bovine pancreas (b.p. protease) for gluten hydrolysis was estimated as 1 mg in 10 mL gluten fraction extracted for 4 hr.

• Korean Journal of Food Science and Technology
• /
• v.39 no.3
• /
• pp.276-282
• /
• 2007

In this study, attempts were made to develop a garlic juice extraction method that would result in minimum changes in quality. Protopectinase and a mutienzyme containing cellulase, pectinesterase, ${\beta}-glucanase$, etc. were applied to garlic residue after first extraction, and the yields of garlic juice and the flavor component changes of the juices were investigated. Enzyme concentrations of 0.04, 0.08, and 0.12% which were based on pulp weight before extraction were added and allowed to hydrolyze for 30, 60, 90 and 120 min,. respectively. Increase in the garlic juice yield was observed according to the amount of enzyme added and the reaction time until reaching a maximum point. When 0.12% protopectinase was applied to the garlic residue for 90 min, the yield increased by 13.8%. Under the same conditions, when multienzyme was applied to the garlic residue, the yield increased by 14.5%, which was considered the maximum. The flavor compounds decreased when compared with the total GC peak area of garlic juice prepared without enzymes(control). The volatile flavor compounds in garlic juice prepared with 0.12% protopectinase for 60 min decreased by 6%. The free sugars profile of the garlic juice prepared with 0.12% protopectinase for 60 min was similar to that of the control. The type of enzyme used did not affect the free amino acid profile of the garlic juice. These results indicate that the optimum conditions for extraction of garlic juice are hydrolyzing the residue with 0.12% protopectinase for 60 min, extracting garlic juice from the hydrolyzed reside, and then combining the extracted juice with the first extraction.

• Asian-Australasian Journal of Animal Sciences
• /
• v.14 no.6
• /
• pp.821-830
• /
• 2001

To study the effects of partial replacement of soybean meal (SBM) with palm kernel meal (PKM) and copra meal (CM) on growth performance, nutrient digestibility and carcass characteristics in finishing pigs, a total of 150 crossbred pigs (Landrace$\times$Duroc$\times$Yorkshire; average $52.11{\pm}1.08kg$ body weight) were alloted to five treatments, in a randomized block design. The treatments included 1) Control: without PKM or CM, 2) PKM2: 2% of palm kernel meal, 3) PKM4: 4% palm kernel meal, 4) CM2: 2% of copra meal, 5) CM4: 4% of copra meal. During the early finishing period (52~74 kg), growth performance was better in CM diets than in PKM diets or control diet, and in overall period (74~100 was lower (p<0.05) in PKM4 diet than the other diets. Nutrient digestibilies of PKM or CM substituted diets showed the tendency to be lower than those of control diet. In the early finishing period, total amino acid digestibilities of PKM and CM diets had the tendency to be lower than control diet, and in the late finishing period, they were lower (p<0.05) than control diet. Carcass length was longer (p<0.05) in the pigs fed 2% CM than in the pigs fed 4% PKM diet, but other carcass characteristics were not different among treatments. Although the dietary C14:0 content affected (p<0.05) on the C14:0 content in the carcass, the inclusion of PKM or CM in the diet did not affect the total saturated fatty acids and unsaturated fatty acids in the backfat of finishing pigs. Although it was not significant, supplementation of CM at the 2% and 4% of control group tended to decrease feed cost per kg weight gain by 2.89 to 1.42%, respectively. In conclusion, copra meal can be a valuable source of protein in the diet for finishing pigs and may replace other protein sources in pig diets to a considerable extent.

• Asian-Australasian Journal of Animal Sciences
• /
• v.28 no.11
• /
• pp.1614-1623
• /
• 2015

The objectives of the current experiment were to study the response of the growth performance of early finishing gilts to different net energy (NE) concentrations in diets, and to compare the NE values of diets between calculated NE values and measured NE values using French and Dutch CVB (Centraal Veevoederbureau; Central Bureau for Livestock Feeding) NE systems. In a metabolism trail, the NE concentrations in five diets used for the growth trial were determined based on digestible nutrient concentrations, digestible energy, and metabolizable energy using a replicated $5{\times}5$ Latin square design with 10 barrows (initial body weight [BW], $39.2{\pm}2.2kg$). In a growth trial, a total of 60 early finishing gilts (Landrace${\times}$Yorkshire; initial BW, $47.7{\pm}3.5kg$) were allotted to five dietary treatments of 8.0, 9.0, 10.0, 11.0, and 12.0 MJ NE/kg (calculated, as-is basis) with 12 replicate pens and one pig per pen in a 42-d feeding experiment. The NE and amino acid (AA) concentrations in all diets were calculated based on the values from NRC (2012). Ratios between standardized ileal digestible AA and NE concentrations in all diets were closely maintained. Pigs were allowed ad libitum access to feed and water. Results indicated that calculated NE concentrations in diets (i.e., five dietary treatments) were close to measured NE concentrations using French NE system in diets. The final BW was increased (linear and quadratic, p<0.05) with increasing NE concentrations in diets. Furthermore, average daily gain (ADG) was increased (linear and quadratic, p<0.01) with increasing NE concentrations in diets. There was a quadratic relationship (p<0.01) between average daily feed intake and NE concentrations in diets. Feed efficiency (G:F) was also increased (linear, p<0.01) as NE concentrations in diets were increased. The NE intake per BW gain (kcal NE/kg of BWG) was increased (linear, p<0.01) with increasing NE concentrations in diets that were predicted from both French and Dutch CVB NE systems. Linear regression indicated that predictability of daily NE intake from the BW of pigs was very low for both French ($R^2$, 0.366) and Dutch CVB ($R^2$, 0.374) NE systems. In conclusion, increasing NE concentrations in diets increase BW, ADG, G:F, and NE intake per BW gain of early finishing gilts. The BW of early finishing gilts is not a good sole variable for the prediction of daily NE intake.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.9
• /
• pp.1433-1441
• /
• 2015

Pollution resulting from the discharge of textile dyes into water systems has become a major global concern. Because peroxidases are known for their ability to decolorize and detoxify textile dyes, the peroxidase activity of Vitreoscilla hemoglobin (VHb) has recently been studied. It is found that VHb and variants of this enzyme show great promise for enzymatic decolorization of dyes and may play a role in achieving their successful removal from industrial wastewater. The level of VHb peroxidase activity correlates with two amino acid residues present within the conserved distal pocket, at positions 53 and 54. In this work, sitedirected mutagenesis of these residues was performed and resulted in improved VHb peroxidase activity. The double mutant, Q53H/P54C, shows the highest dye decolorization and removal efficiency, with 70% removal efficiency within 5 min. UV spectral studies of Q53H/P54C reveals a more compact structure and an altered porphyrin environment (λ_Soret = 413 nm) relative to that of wild-type VHb (λ_Soret = 406), and differential scanning calorimetry data indicate that the VHb variant protein structure is more stable. In addition, circular dichroism spectroscopic studies indicate that this variant's increased protein structural stability is due to an increase in helical structure, as deduced from the melting temperature, which is higher than 90℃. Therefore, the VHb variant Q53H/P54C shows promise as an excellent peroxidase, with excellent dye decolorization activity and a more stable structure than wild-type VHb under high-temperature conditions.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.2
• /
• pp.199-209
• /
• 2018

Volatile organic compounds (VOCs) are increasingly been recognized as the chemical mediators of mold interactions, shaping their community dynamics, growth, and metabolism. Herein, we selectively examined the time-correlated (0 D-11 D, where D = incubation days) effects of intraspecies VOC-mediated interactions (VMI) on Aspergillus oryzae KCCM 60345 (S1), following co-cultivation with partner strain A. oryzae KACC 44967 (S2), in a specially designed twin plate assembly. The comparative evaluation of $S1_{VMI}$ (S1 subjected to VMI with S2) and its control ($S1_{Con}$) showed a notable disparity in their radial growth ($S1_{VMI}$ < $S1_{Con}$) at 5 D, protease activity ($S1_{VMI}$ > $S1_{Con}$) at 3-5 D, amylase activity ($S1_{VMI}$ < $S1_{Con}$) at 3-5 D, and antioxidant levels ($S1_{VMI}$ > $S1_{Con}$) at 3 D. Furthermore, we observed a distinct clustering pattern for gas chromatography-time of flight-mass spectrometry datasets from 5 D extracts of $S1_{VMI}$ and $S1_{Con}$ in principle component analysis (PC1: 30.85%; PC2: 10.31%) and partial least squares discriminant analysis (PLS-DA) (PLS1: 30.77; PLS2: 10.15%). Overall, 43 significantly discriminant metabolites were determined for engendering the metabolic variance based on the PLS-DA model (VIP > 0.7, p < 0.05). In general, a marked disparity in the relative abundance of amino acids ($S1_{VMI}$ > $S1_{Con}$) at 5 D, organic acids ($S1_{VMI}$ > $S1_{Con}$) at 5 D, and kojic acid ($S1_{VMI}$ < $S1_{Con}$) at 5-7 D were observed. Examining the headspace VOCs shared between S1 and S2 in the twin plate for 5 D incubated samples, we observed the relatively higher abundance of C-8 VOCs (1-octen-3-ol, (5Z)-octa-1,5-dien-3-ol, 3-octanone, 1-octen-3-ol acetate) having known semiochemical functions. The present study potentially illuminates the effects of VMI on commercially important A. oryzae's growth and biochemical phenotypes with subtle details of altered metabolomes.

• The Korean Journal of Quaternary Research
• /
• v.17 no.2
• /
• pp.15-15
• /
• 2003

In South Korea, marine terraces have been well developed along the eastern coastal zone, and previous researches on the marine terraces have also been focused on to this coastal zone. The marine terraces of the eastern coast of South Korea had been classified into three terrace groups, that is, the higher, middle, and lower surface ones, according to the heights of marine terraces by previous studies(Oh, 1981 ;Chang, 1987 ;Yoon et. al, 1999, 2003 ; Hwang and Yoon, 1996 etc.). Recently, however, it tends to classify the marine terraces based on the concept of geomorphic surface units(Lee, 1987 ; Kim, 1990 ; Choi, S. 2003; Choi S. et. al 2003a,b, etc). For example, it was proposed that the marine terrace surfaces of Eupcheon coast of the southeastern coastal area of Korea could be classified into 16 geomorphic surfaces, i.e., Eupcheon 1terrace(former shoreline height of 160m), 2(153m), 3(140m), 4(130m), 5(124m), 6(115m), 7(100m), 8(92m), 9(82m), 10(71m), 11(62m), 12(53m), 13(43m), 14(35m), 15(18m) and 16(10m) surfaces, in descending order, according to the former shoreline heights(Choi, S, 2003 ; Choi, S. et. al, 2003a,b). Among these terraces, Eupcheon 1, 2, 4, 5 and 7 surfaces had not been reported in previous works. Among the above mentioned marine terraces, Eupcheon 15 terrace, the most widely and continuously distributed marine terrace have been identified as marine terrace of the Last Interglacial culmination period(oxygen isotope stage 5e) which was based on amino acid dates(124∼125ka BP) and geomorphological features such as red soil, pollen analysis, fossil cryogenic structures and crossing terrace concept. Eupoheon 15 terrace surfaces have also been proposed as the key surface for the identification and correlation of the so-called '5e' marine terrace in the eastern coast of South Korea. This terrace was reconfirmed as the Last Interglacial culmination period, which was based on the identification of Ata tephra, one of the wide-spread marker tephra which indicates the Last Interglacial culmination period in Japan by Sasaki et. al(2002). It was thought that marine terraces of the eastern coast of South Korea had been formed by the steady-state uplifting during the Quaternary glacio-eustatic sea level changes(Choi, 1997). The uprift rate of 10cm/1,000years had been proposed in the eastern coast of South Korea based on the former shoreline altitude(18m) of the above Eupcheon 15 terrace. Therefore, it can be estimated that Eupcheon 1 terrace had been formed in the early Pleistocene from the above uprift rate. The OSL dating for the samples of Eupcheon 7, 9, 13, 15 and 16 terraces and identification of marker tephra in the terrace deposits are in progress. It is expected that more elaborate chronology on themarine terraces of the eastern coast of South Korea could be established by these absolute dates and marker-tephra.