The prevalence of food allergies was investigated using questionnaires with 300 subjects whose ages ranged from 19 to 24 years old and the causative food allergens was analyzed using immunological analysis with serum of the subjects who answered that they have/had food allergy. The questionnaire showed that 11.33% of subjects have/had experience of food hypersensitivity, where the main causative foods were fish, beef, chicken, milk, egg, and pork in order. The meat allergy shared 4.65% (2.33% for beef, 1.66% for chicken, 0.66% for pork) in the prevalence of food allergies. The causative beef allergens were investigated with the serum of 6 subjects who have had beef allergy. Western blots were carried out with the serum of P6 subject who showed a positive reaction to beef extract in ELISA. The two specific bands were detected in beef extract on the PVDF membrane, and no band was detected in extracts of pork and chicken. A calculation of the distance of migration by SDS-PAGE enabled the molecular masses of the two bands to be estimated as 67kDa and 31kDa, respectively. The 67kDa was revealed as bovine serum albumin (BSA) which is one of the important beef allergens as reported previously though an analysis of the N-terminal amino acid sequence. However we could not identify the sequence of 31kDa, probably because they comprised several subunits and were modified proteins such as glycoprotein that were unlikely to be easily degraded by the Edman method. The 31kDa band were dyed with the PAS (periodic acid-schiff reagent), suggesting that it might be a glycoprotein. These results suggested that the 31kDa might be considered as a novel potential beef allergen which is not reported previously, although further studies are needed.
The effect of gamma irradiation of microbiological and physicochemical properties of raw ingredients (thirteen kinds of protein sources) for the mixed feed were investigated. The total aerobic bacteria counts in the samples was ranged from $10^2$ to $10^7$cells/g. After 5 to 7 kGy irradiation, the total count was reduced by 3 to 4 log cycles. Coliforms were detected in seven samples and the range was from $10^4$ to $10^6$cells/g. Enteric pathogens were found only in fish meal and poultry byproduct as $10^2$ to $10^6$cells/g. They were sensitive to radiation and completely sterilized by 3 to 5 kGy irradiation. Total fungi count was ranged from $10^1$ to $10^4$cells/g in all samples. They were osmophiles such as Aspergillus, Penicillium, Cladosporium, Aureobasidium and Rhizopus and were destroyed by 3 to 7 kGy irradiation. Five species of potential mycotoxin producers including Aspergillus flavus were also identified. Physicochemical properties, such as total amino acid content, TBA value, POV and color difference showed that irradiation with optimum does was less detrimental than autoclaving.
Journal of The Korean Society of Grassland and Forage Science
/
v.35
no.2
/
pp.137-144
/
2015
This study was carried out to investigate the growth characteristics, yield, and chemical compositions of corn hybrids for silage according to the drainage depths in a lowland paddy field. The experimental design was arranged in a randomized block design with four treatments and three replications. The drainage depths of the four treatments were 0 cm, 20 cm, 30 cm and 50 cm. Tip-filling degree, ear length, and ear circle were higher in the 30 cm and 50 cm treatments compared with the other treatments (p < 0.05); but the plant length, leaf length, leaf width, ear height, stem diameter, and stem hardness were not significantly different (p < 0.05). The brix was higher in the order of 20 cm > 0 cm > 50 cm > 30 cm treatment. The fresh yield, dry matter yield, and TDN yield increased as the drainage depth increased (p < 0.05). The crude-protein and crude-ash content were highest in the 50 cm treatment (p < 0.05), but the crude-fat, NDF, ADF, and crude-fiber levels were not significantly different (p < 0.05). The total mineral content was higher in the order of 0 cm > 30 cm > 20 cm > 50 cm treatment (p < 0.05). Fructose, glucose, and sucrose were highest in the 20 cm treatment. The total amino-acid (EAA + NEAA) content was the highest in the 50 cm treatment. Based on these results, greater drainage depths increased the yield and feed value of silage corn.
Park, Ju-Yeon;Ahn, Jeung-Yeub;Hong, Hee-Ok;Hahn, Young-Sook
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.
Arylsulfatase catalyzes the release of SO$\sub$4//sup2/- from sulfate esters of simple phenols. Arylsolfatase occurs widely in animal tissues and in microorganisms including soil bacteria. Its widespread distribution suggests that it has a rather fundamental function and environmental meaning. It has been shown previously that arylsulfatase of Klebsiella was purified and characterized. A condition of arylsulfatase synthesis was tested with several strains of Serratia. Serratia marcescens could not utilize some sugars, such as xylose, rhamnose, glucosamine and arabinose hut glucose and mannitol as a sole carbon source. However, arylsulfatase synthesis was repressed by glucose but not by mannitol. The enzyme synthesis was repressed ob inorganic sulfate and methionine, and this repression was relieved by addition of tyramine. Arylsulfatase of S. marcescen was purified by fractionation with ammonium sulfate and followed by chromatographies on DEAE-Cellulose CM-Cellulose, and DEAE-Sephadex A-25. The molecular weight of arylsulfatase was determined to be 46,000 by SDS-Gel electrophoresis and 49,000 by Sephadex G-100 column chromatography. The enzyme showed some different properties with that of K. aerogenes. The activity was maximum at pH 6.8. The Km and Vmax values for p-nitrophenyl sulfate were 2.5${\times}$10$\^$-4/ M and 20 nmoles/min/mg protein, respectively. The enzyme showed high activities toward phenyl sulfate, ο-and p-nitro phenyl sulfates, and p-nitrocatechol sulfate. The inhibition of enzyme was strongly affected by hydroxylamine, inorganic fluoride, sulfide and phosphate, but by inorganic sulfate. Like Klebsiella arylsulfatase, tyramine, octopamine, and dopamine gave signifcant inhibitory effect.
For the effective utilization of the coastal fish resources in Korea, an investigation on the optimum processing conditions and the quality of a textured fish protein concentrate similar to the texture of animal meat has been carried out with the fish meat of Alaska pollack and mackerel. A noodle shaped product was prepared with the fish meat paste after the adjustment of pH and salt content. The product was soaked in $96\%$ ethyl alcohol to produce textured fish protein concentrate and then dried. The processing conditions were estimated with the rehydration capacity of the textured fish protein concentrate(FFC). The quality of the final product was evaluated with chemical composition, sensory test and texture measurement. The optimum pH and salt content of the fish meat for the processing of meaty textured FPC were 7.5 and $1.0\%$ respectively. The most effective soaking conditions were as follows:soaking time, 40 min. ; temperature of alcohol, 5 to $20^{\circ}C$;amount of alcohol, 4 times the weight of tile fish meat paste, number of soaking in alcohol, 4 times. The alcohol remaining in meaty textured FPC could be removed effectively by forced air drying. The yield and the contents of protein and lipid in the meaty textured FPC from Alaska pollack were $19.9\%\;84.3\%\;and\;0.5\%$ and those from mackerel were $29.8\%,\;78.1\%\;and\;3.6\%$ respectively. The content of essential amino acid in the meaty textured FPC from Alaska pollack and mackerel was not inferior to that of beef, textured soybean protein and FAO pattern. Beef meat can be substituted with the meaty textured FPC up to $50\%$ in processing meat balls withoutanysignificantlossinthetaste, ordor and texture.
Seo, Yong Bae;Choi, Seong Seok;Nam, Soo-Wan;Kim, Gun-Do
Microbiology and Biotechnology Letters
/
v.45
no.3
/
pp.226-235
/
2017
Carotenoids such as phytoene, lycopene, and ${\beta}-carotene$ are used as food colorants, animal feed supplements, and for human nutrition and cosmetic purposes. Previously, we reported the isolation of a novel marine bacterium, Kocuria gwangalliensis, which produces a pink-orange pigment. Phytoene desaturase (CrtI), encoded by the gene crtI, catalyzes lycopene formation from phytoene and is an essential enzyme in the early steps of carotenoid biosynthesis. CrtI is one of the key enzymes regulating carotenoid biosynthesis and has been implicated as a rate-limiting enzyme of the pathway in various carotenoid synthesizing organisms. Here, we report the cloning of the crtI gene responsible for lycopene biosynthesis from K. gwangalliensis. The gene consisted of 1,584 bases encoding 527 amino acid residues. The nucleotide sequence of the crtI gene was compared with that of other species, including Kocuria rhizophila and Myxococcus xanthus, and was found to be well conserved during evolution. An expression plasmid containing the crtI gene was constructed (pCcrt1), and Escherichia coli cells were transformed with this plasmid to produce a recombinant protein of approximately 57 kDa, corresponding to the molecular weight of phytoene desaturase. Lycopene biosynthesis was confirmed when the plasmid pCcrtI was co-transformed into E. coli containing the plasmid pRScrtEB carrying the crtE and crtB genes required for lycopene biosynthesis. The results from this study will provide valuable information on the primary structure of K. gwangalliensis CrtI at the molecular level.
Kim, Soon-Hag;Choi, Cheol-Young;Hwang, Joo-Yeon;Kim, Young-Youl;Park, Chan;Oh, Berm-Seok;Kimm, Ku-Chan;Scott A. Gahr;Sohn, Young-Chang
Journal of Aquaculture
/
v.17
no.1
/
pp.1-7
/
2004
A rainbow trout uncoupling protein 3 (UCP3) cDNA clone, encoding a 310 amino acid protein, was cloned and sequenced from a liver cDNA library. Two different splice variants designated UCP3-vl and UCP3-v2, were identified through liver cDNA library screening using rainbow trout UCP3 cDNA clone as a probe. UCP3-vl has 3 insertions in the UCP3 cDNA: the first insertion (133 bp), the second (141 bp), and the third (370 bp) were located 126 bp, 334 bp and 532 bp downstream from the start codon, respectively. UCP3-v2 contained a single insertion, identical in sequence and location to the second insertion of UCP3-vl. UCP3, a mitochondrial protein, functions to modulate the efficiency of oxidative phosphorylation. UCP3 has been detected from heart, testis, spinal cord, eye, retina, colon, muscle, brown adipose tissue and white adipose tissue in mammalian animals. Human and rodent UCP3s are highly expressed in skeletal muscle and brown adipose tissue, while they show weak expression of UCP3 in heart and white adipose tissue. In contrast to mammalian studies, RT-PCR and Southern blot analysis of the rainbow trout demonstrated that UCP3 is strongly expressed in liver and heart. UCP3, UCP3-vl, and UCP3-v2 all contain an Ava III short interspersed element (SINE), located in the 3'untraslated region (UTR). PCR using primers from the Ava III SINE and the UCP3 3'UTR region indicates that the UCP3 cDNA is structurally conserved among salmonids and that these primers may be useful for salmonid species genotyping.
Proceedings of the Botanical Society of Korea Conference
/
2002.04a
/
pp.62-72
/
2002
This study has investigated the biosynthesis and function of the heavy metal binding peptides, the phytochelatins, in plants. PCs are synthesised enzymatically from glutathione by the enzyme PC synthase in the presence of heavy metal ions. Using Arabidopsis thaliana as a model organism cadmium-sensitive, phytochelatin-deficient mutants have been isolated and characterised in previous studies. The cadl mutants have wildtype levels of glutathione, are PC deficient and lack PC synthase activity. Thus, the CADl gene has been proposed to encode PC synthase. The CADl gene was isolated by a positional cloning strategy The gene was mapped and a candidate identified. Each of four cadl mutants had a single base pair change in the candidate gene and the cadmium-sensitive, cadl phenotype was complemented by the candidate gene. This demonstrated the CADl gene had been cloned. A homologous gene in the fission yeast, Schizosaccharomyces pombe was identified through database searches. A targeted-deletion mutation of this gene was constructed and the mutant, like cadl mutants of Arabidopsis, was cadmium-sensitive and PC-deficient. A comparison of the redicted amino acid sequences reveals a highly conserved N-terminal region Presumed to be the catalytic domain and a variable C-terminal region containing multiple Cys residues proposed to be involved in activation of the enzyme by metal ions. Similar genes were also identified in animal species. The Arabidopsis CADl/AtPCSl and S. pombe SpbPCS genes were expressed in E. coli and were shown to be sufficient for glutathione-dependent, heavy metal activate PC synthesis in vitro, thus demonstrating these genes encode PC synthase enzymes. Using RT-PCR, AtPCSl expression appeared to be independent of Cd exposure. However, at higher levels of Cd exposure a AtPCSl-CUS reporter gene construct appeared to be more highly expressed. Using the reporter gene construct, AtPCSl was expressed most tissues. Expression appeared to be greater in younger tissues and same higher levels of expression was observed in some regions, including carpels and the base of siliques. AtPCS2 was a functional gene encoding an active PC synthase. However, its Pattern of expression and the phenotype of a mutant (or antisense line) have not been determined. Assuming the gene is functional then it has clearly been maintained through evolution and must provide some selective advantage. This implies that, at least in some cells or tissue, it is likely to be the dominant PC synthase expressed. This remains to be determined
Central analgesic effect of capsaicin was assessed by the tail flick reflex (TFR) test, using male Sprague-Dawley rats under anesthesia with pentobarbital sodium (induction with 40 mg/kg and maintenance with $4{\sim}8\;mg/kg/hr$). Level of norepinephrine in the spinal cord was also measured. Capsaicin, $35{\sim}150\;{\mu}g$, was injected intrathecally, and the TFR latency was measured before, 10, 30, and 60 minutes after the drug administration. TFR latency was increased 100% or more immediately by intrathecal capsaicin, from 2.9 seconds to the maximum of 7.0 seconds at 10 minute after the drug; P<0.01. The increase in TFR latency was maintained during the course of experiment of 2 hours. Concomitant reduction of NE content in the spinal cord was observed; from 16 ng/mg protein to 7 ng/mg protein. On the other hand, subcutaneous injection of capsaicin of 50 mg/kg did not change the TFR latency although the NE content reduced similarly to the case of intrathecal injection. Pretreatment of the animal with 0.5 mg/kg of MK-801 reversed the increase of TFR latency and NE reduction induced by intrathecal capsaicin. These results suggest that capsaicin causes analgesia at the spinal cord level by activating the excitatory amino acid-NE-dorsal horn interneurons axis of the descending inhibitory pain modulation pathway.
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