A $\alpha$-l, 4-D-glucan maltohydrolase $(\beta$-amylase), secreted by the mesophilic aerobic bacterium Bacillus polymyxa No.26, was purified and characterized. The enzyme production was increased after a logarithmic phase of bacterial growth and paralleled with the onset of bacterial sporulation. By applying anion exchange chromatography and gel filtration the enzyme was purified 16.7-fold and had a specific activity of 285.7 units/mg. Two enzyme activities were eluted on a column of DEAE-Sephadex chromatography, and they were designated as E-I for a major enzyme peak and E-II for a minor peak. Of them, E-I enzyme peak was further purified by using gel chromatography. The molecular mass of this enzyme was determined to be 64, 000 daltons and consisted of a single subunit, showing an isoelectric point of 8.9. The enzyme was able to attack specifically the $\alpha$-l, 4-glycosidic linkages in soluble starch and caused its complete hydrolysis to maltose and $\beta$-limited dextrin. This amylolytic enzyme displayed a temperature optimum at $45^\circ{C}$ and a pH optimum at 7.0. The amino acid composition of the purified enzyme was quite similar to the other bacterial $\beta$-amylases reported. Surprisingly, the purified enzyme from this aerobe only exhibited hydrolytic activity on soluble starch, not on starch granules. The degradation of from starch by $\beta$-amylase was greatly stimulated by pullulanase addition. These results differentiated from other $\beta$-amylases reported. Based on a previous result that showed the enzyme system involves in effective degradation of raw starch granules, this result strongly suggested that the purified enzyme (E-I) can be a synergistic part of starch granule-digestion and E-II plays a crucial role in digestion of starch granules.
Park, Woo-Kyu;Kong, Jae-Yang;Kim, Hyun-Jung;Lee, Dong-Ha;Lim, Hong;Cheon, Hyae-Gyeong
Biomolecules & Therapeutics
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제10권1호
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pp.19-24
/
2002
The effects of a newly synthesized $H^+/K^+$ ATPase inhibitor,1-(2-methyl-4-methoxypheny)-4-[(3-hy-droxypropyl)amino] -6-methyl-2,3-dihydropyrrolo (3,2-c) quinoline (DBM-819) , on the central nervous system, isolated smooth muscle, cardiovascular and digestive systems and renal function were investigated in various experimental animals. Oral administration of DBM-819 had no effect on the central nervous system except body temperature of mice slightly decreased at doses of 15 and 50 mg/kg. DBM-819 produced a moderate analgesic effect in acetic acid-induced writhing test in mice at 50 mg/kg (p.o.). In conscious rats, DBM-819 (15 and 50 mg/kg, p.o.) showed a slight increase in blood pressure and a small decrease in heart rate. DBM-819 had an significant effect on agonist-induced contraction of guinea pig ileum at $1.5{\times}10^{-5}g/ml.$ No significant effect of DBM-819 (5 and 15 mg/kg, i.p) on urinary volume or urinary excretion of $Na^+,\;K^+$ and Cl- was observed in rats. DBM-819 had no significant effect on intestinal transport of a semisolid meal in mice at 15 and 50 mg/kg (p.o.). These findings suggest that DBM-819 exerts no significant pharmacological effects on the central nervous system and renal function at 15 mg/kg (p.o.), but produces some effects on the smooth muscle and circulatory system.
Applications of liposomes as a drug carrier for the oral delivery of poorly-absorbable macromolecular drugs have been limited, because of their instability in gastrointestinal environments including pH, bile salts, and digestive enzymes. Two polysaccharides, dextran(DX) and pullulan(PL), were introduced to the preformed liposomes in order to enhance the stability. Palmitoyl derivatives of polysaccharides, palmitoyldextran(PalDX) and palmitoylpullulan(PalPL), were synthesizd and introduced to the liposomes during preparation for the same purpose of stability. The effects of these polysaccharides coating were evaluated basically by physical properties of particle size distribution and optical microscopy, then compared with uncoated liposomes by the observations of both in vitro stability and in vovo absorption characteristics. The geometric mean diameters of polysaccharide-coated liposomes were greater than that of uncoated liposome, showing the outermost polysaccharide-coated layer under the optical microscopy. In vitro stabilities of uncoated or polysaccharides-coated liposomes were measured by turbidity changes in various pH buffer solutions containing sodium choleate as bile salts. While uncoated liposome was very sensitive to bile salts, polysaccharides-coated liposomes were stable in relatively higher concentrations of sodium choleate, giving the results of better stability of PalDX- and PalPL-coated liposomes than that of DX- and PL-coated liposomes. After liposomal encapsulation of acyclovir(ACV), an antiviral agent as a model drug, it has been administered orally to rats as dose of ACV 40 mg/kg. Plasma concentrations of ACV were assayed by HPLC and analyzed by model-independent pharmacokinetics. Pharmacokinetic parameters of Cmax, tmax, and [AUC] have been compared.
Park, Jong-Moon;Han, Na-Young;Kim, Hokeun;Hwang, Injae;Kim, Jae Bum;Hahm, Ki-Baik;Lee, Sang-Won;Lee, Hookeun
Mass Spectrometry Letters
/
제5권1호
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pp.16-23
/
2014
Liquid chromatography based mass spectrometry (LC-MS) is a key technology for analyzing highly complex and dynamic proteome samples. With highly accurate and sensitive LC-MS analysis of complex proteome samples, efficient data processing is another critical issue to obtain more information from LC-MS data. A typical proteomic data processing starts with protein database search engine which assigns peptide sequences to MS/MS spectra and finds proteins. Although several search engines, such as SEQUEST and MASCOT, have been widely used, there is no unique standard way to interpret MS/MS spectra of peptides. Each search engine has pros and cons depending on types of mass spectrometers and physicochemical properties of peptides. In this study, we describe a novel data process pipeline which identifies more peptides and proteins by correcting precursor ion mass numbers and unifying multi search engines results. The pipeline utilizes two open-source software, iPE-MMR for mass number correction, and iProphet to combine several search results. The integrated pipeline identified 25% more proteins in mouse epididymal adipose tissue compared with the conventional method. Also the pipeline was validated using control and colitis induced colon tissue. The results of the present study shows that the integrated pipeline can efficiently identify increased number of proteins compared to the conventional method which can be a breakthrough in identification of a potential biomarker candidate.
Ponciri fructus, the unripe fruits of Poncirus trifoliata, are widely used in oriental traditional medicine as a remedy for inflammation, gastritis, emesis, digestive ulcers, allergy, and dysentery. To study the anti-wrinkle effects of Ponciri fructus extract (PFE) containing flavanone glycosides, PFE was fermented with Ganoderma lucidum mycelia and its biological activities were investigated. In Ponciri fructus extracts fermented with G. lucidum (G-PFE), polyphenol content was $1,021.00{\pm}0.50{\mu}g/mL$ and flavonoid content was $589.41{\pm}0.21{\mu}g/mL$. G-PFE was found to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals and superoxide anion radical by a dose dependent manner, respectively. G-PFE showed higher antioxidant activity than that of PFE. In addition, the photoprotective properties of G-PFE was tested in human dermal fibroblasts (HDF) exposed to UVA radiation. G-PFE inhibited the activity of matrix metalloproteinase-1 (MMP-1) and showed a dose dependent decrease in the expression level of MMP-1. G-PFE also increased collagen biosynthesis in HDF. These results demonstrate that G-PFE could be useful as a potential cosmetic ingredient for anti-wrinkle.
There are about 2,000 species of termite in the World, but one species of termite inhabits in the southern part of Korean peninsula. Termites are social insects that live in colonies. The colonies are composed of king, queen, soldiers and workers. Termite food consists of cellulose obtained from wood. Protozoa in their digestive tracts convert the cellulose into usable food. Korean termite is a subspecies of Reticulitermes speratus Kolbe, Rhinotermitidae. It's subterranean termite and the scientic name is R. speratus kyushensis Morimoto. The subterranean termite must nest in the soil in order to survival, and infest soft-wood which contact with the soil near the nest. There are several ways which subterranean termite infestations can be noticed. Atcertain times of the year during daylight hours, king and queen termites emerge from the colonies. The propose of these flight is to establish new colonies. The termite is a decomposer of biological ecosystem, but an invader in the preservation of cultural properties as like wood buildings. There are serveral control methods for the prevention of wood building from termite's damages. Those are biological control, ecological control, physical control and chemical control. Ecological and Physical control are the best methods in the new constructing wood-building. Fumigation which is a method of chemical control, is the best method for the building damaged by the termite. After the fumigation, we have totake wood & soil treatments for the building and nearby in order not to be reinvaded by the termites.
Noni has been used for medicinal purposes for more than 2,000 years in South Pacific Polynesia, China and India, and has been heavily ingested as an extract for its excellent antioxidant and anti-inflammatory effects. However, a recent study found that the noni extract causes digestive disorders, kidney problems, and liver diseases, which made it necessary to use it for other purposes than as an extract. In this study, we want to evaluate the potential of noni as an anti-oxidant, anti-inflammatory and anti-wrinkling agent. Methods: Noni was freeze-dried, extracted in water, and concentrated. Skin cells were treated with the noni extract for 24 hrs and then were exposed to UVB (55 mJ/cm2). After 48 hrs of incubation, pro-inflammatory cytokine, elastase, MMP-1 and type-1 procollagen levels were measured by ELISA. Results: To find out the antioxidant effect of the noni extract, the DPPH and ABTS radical scavenging activity experiments were conducted and the noni extract showed 97.0 % and 92.0 % antioxidant efficacy at 200 ㎍/mL respectively. The noni extract (50 and 100 ㎍/mL) decreased IL-6 and TNF-α in RAW 264.7 cells induced by LPS in a concentration-dependent manner. In the RT-PCR experiment involving NO production, the noni extract (50 and 100 ㎍/mL) inhibited NO production by strongly inhibiting iNOS mRNA expression, and also inhibited the elevation of MMP-1 and elastases caused by UVB irradiation by 25.0 % and 7.0 % respectively. In addition, type-1 procollagen was elevated by 20.0 % by the noni extract treatment in HaCaT cells. Conclusion: The noni extract has photoprotective ability by reducing proinflammatory mediators, elastase and MMP-1 production, and elevation of collagen synthesis. Our findings suggest that the noni extract might be a good natural substance to protect against UVB-induced premature skin aging.
Non-alcoholic fatty liver disease (NAFLD), especially including non-alcoholic steatohepatitis (NASH) is one of the common diseases with 25% of prevalence globally, but there is no thera-peutic access available. Amomum villosum var. xanthioides (Wall. ex Baker) T.L.Wu & S.J.Chen (AX), which is a medicinal herb and traditionally used for treating digestive tract disorders in Asia countries. We aimed to examine pharmacological effects of ethyl acetate fraction of AX (AXEF) against ER stress-induced NASH mice model using C57/BL6J male mice by tunicamycin (TM, 2 mg/kg) injection focusing on the oxidative stress. Mice were orally administrated AXEF (12.5, 25, or 50 mg/kg), silymarin (50 mg/kg) or distilled water daily for 5 days, and outcomes for fatty liver, inflammation, and oxidative stress were measured in serum or liver tissue levels. AXEF drastically attenuated hepatic ER stress-induced NASH which were evidenced by decreases of li-pid droplet accumulations, serum liver enzymes, hepatic inflammations, and cell death signals in the hepatic tissue or serum levels. Interestingly, AXEF showed potent antioxidant effects by quenching of reactive oxidative stress and its final product of lipid peroxide in the hepatic tissue, specifically increase of metallothionein (MT). To confirm underlying actions of AXEF, we ob-served that AXEF increase MT1gene promoter activities in the physiological levels. Collectively, AXEF showed antioxidant properties on TM-induced ER stress of NASH by enhancement of MTs.
Objectives : The aim of this study is to find the therapeutic meaning of the Portulacea in herbal medication. Methods : About the origin, the component, the processing the drug, the properties and tastes of drugs, the meridian tropism, the effects, the treating disease, the contraindication and the method of administration, I have researched thirty literatures to mention the Portulacea in time sequence. Results : 1. The Portulacea belongs to the Portulacaceae herbs and it consists of noradrenaline, potassium sometimes containing small amounts of dopamine, dopa, malic acid, citric acid, glutamine acid, asparagin acid, alanine, cane sugar, grape sugar, fruit sugar. 2. The processing of drug is wash of water clealy, and remove the foreign substance. Then the drug is cutting to use. The method of burnt to ashes is used, too. 3. The properties and tastes of drugs is acid, cold, nontoxic. The meridian tropism is mainly liver and the large intestine meridian. 4. From old times, Portulacea has come into general use to treat eczema, the rose erysipelas, an acne, hemorrhoids, discharge from the womb etc. because it is effective on neutralizing poison, reduce a swelling, a tumor, an abscess and stopping of bleeding 5. Portulacea must be stoped When person have a weak digestive organ becase it is cold herba. And don't take use with Fish and shellfish. 6. Portulacea is useful method to external care. To use the herba, pulverize amount of property and then apply to the the affected part Conclusions : This study showed that the Portulacea is useful herb to treat of skin disease and useful method to external care.
This study evaluated the quality of commercial extruded pellet (EP) diet of five companies (A, B, C, D and E) for olive flounder Paralichtys olivaceus by biochemical analyses, physical properties and growth performance. The proximate analyses of five EP diets showed $3.2-10.0\%$ of moisture, $49.3-55.5\%$ of crude protein, $4.6-14.7\%$ of crude lipid, $7.0-13.8\%$ of crude ash, $0.7-10.5\%$ of crude fiber, $10.0-27.3\%$ of itrogen free extract (NFE), 304.3-395.4kcal/100g of digestible energy (DE) and 6.1-7.1 of calorie/protein ratio (C/P). Peroxide value (POV) was highest in diet D (47.4 meq/kg) as compared to other diets which in the range of 4.0-11.7 meq/kg. Total amino acid contents were ranged from 46.54 to $55.46\%$ with the highest content in diet B and the lowest content in diet C. Essential amino acid of diet C was lowest $(7.43\%)$ as compared to other diets which in the range of $19.43-20.30\%$. Saturated fatty acid was higher in diet A $(37.65\%)$ followed by diet B $(36.32\%)$, diet E$(34.39\%)$, diet C$(30.95\%)$ and diet D$(30.10\%)$. EPA+DHA were highest in diet E$(30.78\%)$ and lowest in diet C$(15.48\%)$. The floating rate after 6 hours on the sea water was highest in diet C$(100\%)$ followed by diet B$(40\%)$ and A$(10\%)$. However, diets D and E were completely settled down after 1 and 2 hours, respectively. The range of relative expansion rate was $27.2-49.3\%$ for all diets and all reached the peak at 2-3 hours. The water absorption rate of diets C and D was lowest, and diet E was highest at 1 hour after deposition of sea water. Growth rate was higher in diet B$(22.3\%)$ and E$(21.3\%)$. Feed efficiency was higher in diet A$(109.7\%)$ and E$(105.3\%)$ and was significantly lowest in diet D$(80.7\%)$. The protein efficiency ratio was highest in diet E (2.72) and lowest in diet D (1.76). These results suggest that there is a necessity for improvement of nutrients balance and feed physical properties to fulfill the nutrient requirements and digestive characteristics of fishes in commercial EP diets.
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