• Journal of Life Science
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• v.7 no.2
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• pp.127-133
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• 1997

The phytase(myo-inositol hexkisphosphate phosphohydrolase ; EC 3.1.3.8) activity was observed only in the homogenate of intestinal mucosa, though the activity of alkaline phisphatase was measurable in various organs. In addition, no protein bands were detected in any other organs on immunoblotting using the anti-90kDa phytase antiserum. Thses results suggest that phytase is specifically present in small intestinal mucosa, and that hydrolysis of phytic acid(inositol-hexakisphosphate) can be allotted for a physiological role of the intestine-specific enzyme. The activities of phytase was increased during development of rat. The 70kDa phytase appeared just after birth, but the 90kDa phytase was not observed until adult period, suggesting that the 90kDa phytase was synthesized in response to weanling.

• Journal of fish pathology
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• v.21 no.3
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• pp.247-257
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• 2008

To understand the activity of non-specific defence factors in cultured Sebastes, the antibacterial effect of the serum, skin mucus and homogenate of various organs from cultured oblong rockfish (Sebastes oblongus) and rockfish(Sebastes schlegeli) against pathogenic bacteria, Aeromonas hydrophila, Edwardsiella tarda, Vibrio anguillarum, and Streptococcus sp. was compared with that of flounder(Paralichthys olivaceus) and seabass(Leteolabrax japonicus). And the activities of proteolytic enzyme, chitinolytic enzyme and haemolycin as non-specific defence factor were investigated on the oblong rockfish and rockfish. Samples from oblong rockfish showed the highest antibacterial activity by lysoplate assay on agar plate mixed with pathogens, followed in descending order by rockfish, seabass, and flounder. Turbidimetric assay was carried to evaluate the lysozyme activity of fish samples against lyophilized cells of Micrococcus lysodeiktikus. The serum, kidney, liver, stomach, intestine and eyeball of oblong rockfish and the mucus and gill of rockfish appeared to have the highest lysozyme activity among the fish strains investigated. All samples except skin mucus, liver, and eyeball of oblong rockfish and rockfish showed proteolytic enzyme activity. Chitinolytic enzyme activity was showed in random sampling and haemolytic activity was remarkable in oblong rockfish. Therefore, Sebastes strain was proved to have effective defense mechanisms based on the antibacterial activities, and lysozyme, proteolytic enzyme, chitinolytic enzyme, and haemolycin were considered to act as the non-specific defence factor of Sebastes.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.232.2-232.2
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• 2002

Corticosteroids have been used most frequently for inflammatory bowel disease. They are well absorbed and only a limited fraction of the dose is delivered to the inflammatory site in the colon. To reduce side effects by the systemic absorption. colon-specific delivery is highly desirable. We designed dexamethasone 21-sulfate sodium (DS) as a cOlon-specific prodrug of dexamethasone (D) expecting that it might be stable and non absorbable in the upper intestine and dissociate in the colon by the sulfatase, an enzyme solely found in the colon. (omitted)

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.6
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• pp.1152-1159
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• 1998

In order to develop more rapid and reproducible analysis of biotin known as vitamin H, attempts were made to establish the condition for enzyme linked immunosorbent assay(ELISA) compared with traditional microbiological assay(MBA). Antibiotin and antiserum were obtained from the immunized rabbits injected with emulsion of biotin KLH conjugate and Freund's adjuvant. The antiserum showed cross reactivity on biocytin, a derivative of biotin, which is converted to biotin in intestine, at the rate of 177%(median inhibitory concentration(IC50)=12.58ppb), but not on other derivatives such as desthiobiotin, diaminobiotin and 2 imino biotin. Specific antibody for biotin was purified from the antiserum through protein A column and desalting column. The conditions of competitive direct ELISA (cdELISA) were established. Detection range of biotin concentration by cdELISA was 0.01∼300ng/ ml(ppb). In the spike test with milk, fruit flake and pine carrot juice, the correlation coefficient between two methods of MBA and ELISA was reliably consistent at the value of r=0.992. But detection of biotin by microbiological assay(MBA) was rather restricted in range and nonspecific. Detection range of biotin by MBA was 0.1∼0.5ng/ml(ppb). It showed cross reactivities on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. In conclusion, ELISA revealed a significant improvement compared with MBA for the biotin detection in terms of sensitivity, detection range and cross reactivity. In addition, a variety of samples could be analyzed rapidly and conveniently at one time by using ELISA. These results strongly suggest that the ELISA is very promising for the practical application to detect biotin contents in a wide range of food stuffs.

• BMB Reports
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• v.37 no.6
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• pp.684-690
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• 2004

Heparin lyase I was purified to homogeneity from Bacteroides stercoris HJ-15 isolated from human intestine, by a combination of DEAE-Sepharose, gel-filtration, hydroxyapatite, and CM-Sephadex C-50 column chromatography. This enzyme preferred heparin to heparan sulfate, but was inactive at cleaving acharan sulfate. The apparent molecular mass of heparin lyase I was estimated as 48,000 daltons by SDS-PAGE and its isoelectric point was determined as 9.0 by IEF. The purified enzyme required 500 mM NaCl in the reaction mixture for maximal activity and the optimal activity was obtained at pH 7.0 and $50^{\circ}C$. It was rather stable within the range of 25 to $50^{\circ}C$ but lost activity rapidly above $50^{\circ}C$. The enzyme was activated by $Co^{2+}$ or EDTA and stabilized by dithiothreitol. The kinetic constants, $K_m$ and $V_{max}$ for heparin were $1.3{\times}10^{-5}\;M$ and $8.8\;{\mu}mol/min{\cdot}mg$. The purified heparin lyase I was an eliminase that acted best on porcine intestinal heparin, and to a lesser extent on porcine intestinal mucosa heparan sulfate. It was inactive in the cleavage of N-desulfated heparin and acharan sulfate. In conclusion, heparin lyase I from Bacteroides stercoris was specific to heparin rather than heparan sulfate and its biochemical properties showed a substrate specificity similar to that of Flavobacterial heparin lyase I.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1324-1329
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• 2008

This study was conducted to evaluate the potential of the transformed Lactobacillus reuteri Pg4 (T-Pg4) harboring the ${\beta}$-glucanase gene as a poultry probiotic. The probiotic properties of the T-Pg4 strain were evaluated in vitro by their adherence capability and acid and bile salt tolerance, and were evaluated in vivo by their survival and adhesion in the gastrointestinal tract (GIT) of specific-pathogen-free (SPF) chickens. The results showed that the T-Pg4 strain exhibited resistance to acidic conditions and contact with bile salt, and adhered efficiently to the crop and intestinal epithelial cells of chickens in vitro. The T-Pg4 strain also could survive and colonize the gastrointestinal epithelium of the experimental SPF chickens in vivo. In addition, radial enzyme diffusion was used to demonstrate that the Lactobacillus spp. randomly isolated from the GIT of the SPF chickens fed T-Pg4 possessed ${\beta}$-glucanase secretion capability. These findings have demonstrated that the transformed L. reuteri Pg4 survives transit through the stomach and intestine, and may secrete ${\beta}$-glucanase in the chicken GIT. Therefore, it is suggested that this organism could be used as a multifunctional poultry probiotic.

• Journal of Nutrition and Health
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• v.19 no.6
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• pp.363-373
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• 1986

This study was devised to elucidate whether soused shrimp exhibits a digestive action on boiled pork meats. and the mechanism by which sousing with a high concentration of sodium chloride preserves nutrients in foods for a prolonged pe\ulcornerriod. Protease was isolated from soused shrimp using a combination of ammonium sulfate fractionation. DEAE - cellulose ion exchange chromatography and gel filtra\ulcornertion. The isolated protease had specific activity of 1.560 units. 210 purification fo\ulcornerld with an yield of 38%. Its optimum pH and temperature were 8.0 and $43^{\circ}C$ respectively. The molecular weight of the enzyme was 35.000. The Km value of the enzyme for casein was 1.6 x $10^{-6}$ M The e=yme required the presence of cu\ulcornerpric ion to exhibit its full activity. Eighty eight percent of the enzyme activity was in\ulcornerhibited by 3.5M NaCI showing a reversibly linear decrease of the enzyme activity as NaCI concentration increased. The nature of the inhibition by NaCl was rever\ulcornersible and noncompetitive. The protease activity in soused shrimp was well preser\ulcornerved with the elapse of time at least in part due to NaCI induced suppression of autodigestion. The enzyme was denatured by acid easily. i.e. 1% of the original activity remained after staying at pH 2 for 10 minutes. which is within the norm\ulcorneral range of pH of the human stomach. Soused shrimp was observed to be one of those containing the highest protease activity compared with the other soused foo\ulcornerds such as soused oyster. squid. clam. and Pollack intestine with respect to spec\ulcornerific activities of dialized 1:4 whole homogenates(w/v) in 5 mM sodium phospha\ulcornerte - 2.4 mM j3 - mercaptoethanol buffer. pH 8.0. Casein and boiled meats including pork, beef, and chicken appeared to be the good substrates for the protease. Casein was the best. Therefore. the ingestion of boiled meats including pork together with soused sh\ulcornerrimp would help digestion of boiled pork in human not only by increasing appe\ulcornertite also by the direct proteolytic digestion of boiled meats by soused shrimp to\ulcorner some extent. And a high concentration of sodium chloride inhibited the protease activity reversibly in a remarkable degree, which ensued in a significant retardat\ulcornerion of autodigestion of protein in foods by proteases, and hereby contributed to the preservation of foods for an extended period.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1162-1167
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• 2004

The present study was designed to define whether dietary conjugated linoleic acid (CLA) could affect antioxidant enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and glutathione S transferase (GST), and the level of malondialdehyde (MDA), a marker of lipid peroxidation, in the small intestine and liver from broiler chickens. A total of twenty-four 3 wk-old male broiler chickens were assigned to three dietary treatments (1.5% corn oil, 0.75% corn oil plus 0.75% CLA, and 1.5% CLA, isocalorically), and fed a grower-finisher diet from 22 to 35 days. In the small intestinal mucosae, the specific activities of SOD, GSH-Px, CAT, and GST, and the level of MDA were not substantially influenced by dietary CLA. In the liver, the specific activities of SOD, GSH-Px, and GST, and the level of MDA were also unaffected by dietary CLA at the level of either 0.75% or 1.5% compared with corn oil at the level of 1.5%. However, the broiler chickens fed the diet containing 1.5% CLA resulted in a significant increase in peroxisomal CAT activity and a marked decrease in total lipid and non-esterified fatty acids (NEFA) from liver tissues compared with those fed the diet containing 1.5% corn oil. In conclusion, ability of CLA to increase hepatic CAT activity suggest that dietary CLA may affect, at least in part, antioxidant defense system as well as lipid metabolism in the liver of broiler chickens.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.118-125
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• 1999

Porcine Proliferative Enteropathy(PPE) is an infectious enteric disease and a major cause of economic loss in swine industry due to weight loss, poor growth and sudden death in growing and finishing pigs at 6 to 20 weeks of age. PPE has been diagnosed by clinical signs, syndrom and lesions in the intestine in Korea. However, the diagnostic method had several problems in the detection of infected or carrier pigs. Therefore, in this study, we established the polymerase chain reaction(PCR) which was a fast, specific and sensitive method for identification of Lawsonia intracellularis (L intracellularis). We designed and synthesized primer on the 16S rDNA and p78 gene encoding L intracellularis. Specificity of the method was confirmed by comparison of the PCR results using other enteric bacteria and the study has shown that PCR method was sensitive to detect 1ng of genomic DNA as a template. Identity of the PCR products was confirmed by comparison of pattern of restriction endonuclease analysis with restriction enzyme Hae III and Pst I. Also, the PCR method was applicable to the naturally affected pigs with PPE. Based on the results from this study, the PCR method could be used as a fast and specific diagnostic tool for PPE.

• Journal of Animal Science and Technology
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• v.48 no.1
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• pp.77-90
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• 2006

The purpose of this study was to isolate cellulolytic and hemicellulolytic anaerobic bacteria inhabiting from gut of ruminants and investigate their hydrolytic enzyme activities. Extracellular CMCase activities of H-strains isolated from the rumen of a Holstein dairy cow were higher than those of D- and DC- strains from the rumen and large intestine of Korean spotted deer. Most isolated bacteria utilized more efficiently Dehority's artificial medium containing starch, glucose and cellobiose (DAS) than those in Dehority's artificial medium containing cellulose only (DAC). The results of biochemical reactions and sugar fermentation indicated that the isolated bacteria belong to one of bacterial strains of Peptostreptococcus spp., Bifidobacterium spp., Prevotela ruminicola/buccae, Clostridium beijer/butyricum and Streptococcus intermedis which are not highly cellulolytic. Activities of Avicelase, xylanase, β-D-glucosidase, α-L-arabinofuranosidase and β-xylosidase of the isolated anaerobic bacteria in DAS were higher than those in DAC. In conclusion, the results indicated the higher enzyme activities of the isolated strains cultured in DAS medium were mainly caused by their specific carbohydrate utilization for enzyme production and growth rate. The highly cellulolytic bacteria were not isolated in the present experiment. Thus further research is required to investigate characteristics of gut bacteria from Korean spotted deer.