• BMB Reports
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• 제29권6호
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• pp.540-544
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• 1996

Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.

• Bulletin of the Korean Chemical Society
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• 제30권11호
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• pp.2647-2650
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• 2009

Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

• 한국미생물·생명공학회지
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• 제18권4호
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• pp.338-343
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• 1990

Streptomyces sp. DMCJ-49 균주의 배양액으로부터 분리된 $\alpha$-amylase 저해물질 MB4-03은 구조상 질소원자와 methyl기를 포함하는 일종의 직선형 Alpha(1-4) 결합으로 연결된 glucose의 변형된 oligomer 형태로서 구성 단당류는 12 정도로 추정된다.

• The Korean Journal of Physiology and Pharmacology
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• 제2권3호
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• pp.385-393
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• 1998

Human neutrophil elastase (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, was purified by Ultrogel AcA54 gel filtration and CM-Sephadex ion exchange chromatography. HNElastase was inhibited by phenylbutazone in a concentration dependent manner up to 0.4 mM, but as the concentration increased, the inhibitory effect gradually diminished. Binding of phenylbutazone to the human neutrophil elastase caused strong Raman shifts at 200, 440, and 1194 $cm^{-1}$. The peak at 1194 $cm^{-1}$ might be evidence of the presence $of\;-N=N-{\Phi}$ radical. The core area of the elastase, according to the visual molecular model of human neutrophil elastase, was structurally stable. A deeply situated active center was at the core area surrounded by hydrophobic amino acids. Directly neighboring the active site was one positively charged atom and two atoms carrying a negative charge, which enabled the enzyme and the drug to form a strong interaction. Phenylbutazone may form a binding, similar to a key & lock system to the atoms carrying opposite charges near the active site of the enzyme molecule. Furthermore, the hydrophobicity of the surrounding amino acid near the active site seemed to enhance the binding strength of phenylbutazone. Binding of phenylbutazone near the active site may cause masking of the active site, preventing the substrate from approaching the active site and inhibiting elastase activity.

• 미생물학회지
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• 제53권2호
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• pp.79-82
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• 2017

먹물버섯 Coprinus comatus를 yeast extract peptone dextrose 액체배지에서 배양하고 laccase의 생성 분비를 확인하였다. 이 때 분비된 laccase를 이온교환수지 방법과 전기영동 분획수집방법으로 순수하게 정제하였다. 정제된 효소단백질의 분자량은 SDS-PAGE로 분석하였을 때 약 67 kDa으로 나타났고 최적산도는 pH 4.3, 최적온도는 $25^{\circ}C$로 확인되었다. 기질 o-tolidine에 대한 Km 값은 0.45 mM, Vmax 값은 0.0132 OD/min/unit로 나타났다. 정제된 laccase를 사용하여 염료 RBBR과 Poly R-478의 탈색을 분석한 결과 배양 12시간 후에 RBBR은 49.3% 탈색되었고 poly R-478은 41.6% 탈색되었다.

• 생약학회지
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• 제24권4호
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• pp.289-295
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• 1993

Polysaccharides fractions from the leaves of Artemisia selengensis$(ASP_1)$ and Artemisia iwayomogi$(ASP_1)$ were extracted and purified by Sephadex gel filtration and DEAE-Sephadex ion exchange chromatographies. Both $ASP_1$ and $AIP_1$ fractions were tested for their effects on the spleen cell culture in vitro. Both $ASP_1$ and $AIP_1$ fractions allow growth of spleen cells up to 3 months in culture, suggesting the immunoregulatory activities of those polysaccharide fractions. The molecular weights of $ASP_1$ and $ASI_1$ fractions were found to be about 2,500 daltons by Sephadex gel filtration chromatography using standard dextrans. Both $ASP_1$ and $AIP_1$ fractions were composed of glucose, xylose and galactose.

• 생약학회지
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• 제24권3호
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• pp.203-212
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• 1993

To find antitumor components in the hot water extract of the cultured mycelia of Ganoderma lucidum, protein-bound polysaccharides were purified and fractionated (Fr. I-V) by DEAE-cellulose ion exchange column chromatography and Sepbarose CL-4B gel filtration. When a dose of 20 mg/kg/day of each was, i.p., injected into ICR mice, the inhibition ratios against the solid form of sarcoma 180 were $64.2{\sim}75.8%$. The antitumor component was examined for immunological activity. It increased the amount of superoxide anion released by induced macrophages in peritoneal cavity to 1.8 times and the count of hemolytic plaque-forming cells (PFC) was increased to 4.4 times as compared with those of the control group. It contained 68.6% polysaccharide which consisted of mannose, glucose, galactose, fucose and xylose and 5.1% protein consisting of 17 amino acids. The contents of hexosamine were 0.78%. The molecular weight of Fr. V that showed the highest antitumor activity was $5.8{\times}10^4$ dalton by Sepharose CL-4B gel filtration. It was named lucidan.

• Journal of Nutrition and Health
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• 제32권2호
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• pp.158-165
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• 1999

Effects of oral taurine supplementation (6g/day) on plasma concentration and urinary of free amino acids were evaluated in healthy female adults. Among twenty five female volunteers(23.6$\pm$0.3 years old) participated in the taurine supplementation program, twenty four subjects successfully completed the two supplementation program. Plasma and urinary levels of free amino acids were determined by using an automated amino acid analyzer based on ion-exchange chromatography. Two weeks of taurine supplementation resulted in a 65% increase in plasma taurine concentration (p<0.001), Changes in fasting plasma amino acid concentrations followed by taurine supplementation were not spectacular, and were all within the normal range for human aldults. Taurine supplementation significantly elevated urinary methionine, asparagine, hydorxyproline and phosphoserine excretions(31~280%), and significantly decreased the urinary excretions of isoleucine, glutamate and serine compared to the values prior to taurine supplementation. For almost every individual amino acids, 24 hr urinary excretion level was significantly correlated to the urinary excretion value expressed as nmol/mg creatinine(p<0.001). A significant negative correlation found between plasma glutamine concentration and urinary glutamine excretion level suggests that the decrease in plasma glutamine concentration might be associated with the enhanced glutamine excretion in urine followed by taurine supplementation.

• 한국식품영양과학회지
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• 제27권5호
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• pp.801-807
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• 1998

Taurine content in Korean foods of plant origin was determined for 118 commonly used food items including cereals, potatoes, pulses, nuts, seeds, vegetables and fruits. Taurine concentration in food sample was analyzed using an automated amino acid analyzer(Biochrom 20, Pharmacia LKB) based on ion-exchange chromatography. Taurine was frequently detected in plant kingdom in much lower concentrations(1/100~1/1000) than those found in marine lifes and mammals. Glutinous rice, glutinous millent and sorghum did not contain taurine, while 0.7~3.9mg taurine/100g dry wt were detected in rice, barley and their products. Potatoes and sweet potatoes contained 0.3~1.2mg taurine/100g wet wt, and seasame seeds, perilla seeds, almonds, walnuts and gigko nuts contained 0.7~3.0mg taurine/100g wt. Taurine concentration was undetectable in most of the pulses, and in a large number of vegetables. Garlic bulbs, eggplants, green peppers, lotus roots, and cabbages have a relatively high level of taurine(around 1mg taurine/100g wet wt) among vegetables. Taurine was absent or found in very low levels(<1mg taurine/100g wet wt) in most of the commonly used fruits.

• 생명과학회지
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• 제5권4호
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• pp.170-180
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• 1995

A carotennnoprotein from the skin of Ascidian(Halocynthia roretzi) was extracted by Triton X-100 and purified by ammonium sulfate fraction, SephadexG-200 charomatography and DEAE-cellulose ion exchange chromatography. The carotenoprotein was redwith broad $\lambda$$_{max}$ between 495, 467 and 318nm. The red carotenoprotein had an approximate molecular weight of 326KDa(gel filtration). SDS-PAGE indicated the presence of two polypeptodes of 84.1KDa and 74.4KDa, with different mobility in polyacrylamide gel electrophoresis. In the presence of denaturing agents such as organic solvent aand extreme pH, the red complex readily disociates to liberate the yellow carotenoid($\lambda$$_{max}$ 452nm) and a colourless apoprotein. The amino acid composition of carotenoprotein were mainly threonine(15.2%), aspartic acid(12.2%), glutamic acid(11.9%) and serine(9.6%), while proline was not found. The carotenoprotein consisted of lipids as structure units. Its major fatty acids composion were C$_{18:1}$, C$_{16:1}$, and C$_{16:0}$. The monounsaturated fatty acids(41.5%) contained abundant content compared to other fatty aacids(polyunsaturated fatty acids 37.4%, saturated fatty acids 20.6%).