• Bulletin of the Korean Chemical Society
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• 제35권9호
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• pp.2665-2668
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• 2014

Aptamers are oligonucleotides or peptide molecules that are able to bind to their specific target molecules with high affinity via molecular recognition. In this study, we present development of aptamer-based precipitation assays (or simply aptamoprecipitation) for His-tagged proteins and thrombin to compare their purification efficiency with other conventional affinity precipitation methods. A crosslinking method was employed to immobilize thiol-functionalized aptamers onto the surface of polystyrene resins, enabling them to specifically bind to His-tag and to thrombin, respectively. The resulting aptamer-functionalized resins were successfully applied via a one-step experiment to purification of His-tagged proteins from complex E. coli and to thrombin extraction, exhibiting superior or at least comparable purification results to the conventional immobilized metal affinity precipitation or immunoprecipitation.

• 대한본초학회지
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• 제22권4호
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• pp.21-28
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• 2007

Objectives : Kyenegum has been popularly used long as the digestive. The purpose of this study was to investigate the purification and characteristics of protease obtained from Kyenegum crude enzyme. Methods : Kyenegum protease was purified by precipitation with ammonium sulfate followed by SP-Spharose ion exchange chromatography. The molecular weight of Kyenegum protease was estimated by SDS-PAGE electrophoresis. Results : Kyenegum protease was 3,087 units/mg protein specific activity, 14.5 purification fold and 9.8 % recovery. The molecular weight of protease was estimated to be 18 kDa. The isoelectric point was pI 8.97 and values of Km and Vmax of its were 48 mg/mL and 2 units/min, respectively. Conclusion : The result suggests that the protease obtained from Kyenegum has excellent stability of temperature, acid and collagen substrate specificity.

• 한국미생물·생명공학회지
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• 제46권2호
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• pp.111-113
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• 2018

In this study, purification of cold-adapted protease from Janthinobacterium sp. was investigated. First, using gradient precipitation, protease was confirmed to be deposited in the 30-80% range of ammonium sulfate. Next, DEAE-Sepharose column was used for the binding of the protease under various conditions. The optimal binding condition was found to be pH 8.5 and flow rate of 30 ml/h. Under the optimal condition, the protease was purified with 29% recovery yield. This result can be useful for the purification of other cold-adapted protein.

• Journal of Microbiology
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• 제35권2호
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• pp.97-102
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• 1997

Cysteine desulfhydrase (EC 4.4.1.1.) was purified from the culture supernatant of Streptomyces albidoflavus SMF301 by hydroxyapatite, gel filtration and Resource Q ion-exchange chromatography with a purification fold of six identical subunits. The enzyme was stabilized by dithiothreitol and pyridoxal 5'-phosphate during the purification procedures. The optimum pH and temperature were pH 8.6 and 35$^{\circ}C$, respectively. The N-terminal amino acid sequence was identified as A-P-L-P-T-A-D-V-R-S-D-P-G-Y-R-E-W-L-G-E-A-V. The purified cystein desulfhydrase had a high substrate specificity toward cysteine, and exhibited no cystahionine $\gamma$-lyase activity. The $K_m$ value for cysteine was determined to be 0.37 mM.

• 한국콘크리트학회:학술대회논문집
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• 한국콘크리트학회 2006년도 춘계 학술발표회 논문집(II)
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• pp.649-652
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• 2006

This paper describe the performance of seawater purification, to which living organisms can adapt, and the physical properties of porous concrete with continuous void. Although conventional concrete has been regarded as a destroyer of nature, seawater and air can pass freely through concrete when it is made porous by forming continuous void. This not only enables plants to vegetables, but also makes it possible for microscopic animals and plants, including bacteria, to attach to and inhabit uneven surface as well as internal voids when the concrete is provided in a natural seawater area or seawater side area. As a result, porous concrete using recycled aggregate improved the performance of seawater purification. In this study, The performance of seawater purification of porous concrete using recycled aggregate analyzed by T-P, T-N.

• 한국미생물·생명공학회지
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• 제28권5호
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• pp.251-257
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• 2000

Serratia marcescens purine nucleoside phosphorylase (PNP) was purfied to homogeneity by streptomycin sulfate treatment, Sephacry HR S-200 gel filtration chromatography and AMP-agarose affinity chromatography. The specific activity of the enzyme was increased 49-fold during purification with an overall yield of 7.0%. The molecular weight was 168kD as estimated by Sephadex G-150 gel filtration chromatography. The S. marcescens enzyme was composed of six identical subunits with subunit molecular weight of 28kD, as estimated by SDS-PAGE. The Km values of S. marcescens enzyme for inosine and deoxyinsoine were 0.38 and 1.20 mM, respectively. The ph optimum was near 8.0, and the enzyme was relatively heat-stable protein. The enzyme was inactivated com-pletely by 0.5 mM of $Cu^{ 2+}$.

• 한국미생물·생명공학회지
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• 제21권5호
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• pp.446-452
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• 1993

The cholesterol oxidase produced from Bacillus sphaericus was purified and characterized. Through a series of purification procedures including DEAE-Toyoperal 650C, Sephadex G-200 and DEAE-Sephadex A-50 column chromatography, the purified enzyme was shown to have a specific activity of 0.179 units/mg protein having 31.8 fold purification and final yield of 12%. The molecular weight of the enzyme was estimated to be 47kDa and 47.tkDa by Sephadex G-200 chromatography and SDS-PAGE. The optimum temperature and pH for the enzyme were 30C and 6.0, respectively. The activity of the purified cholesterol oxidase was inhibited by Fe2+ and Hg+.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 1996년도 추계학술대회 논문집
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• pp.313-317
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• 1996

This work deals with description of gas purifing system to product high pure helium gas using low temperature absorption. The system controls temperature of heaters, open/close of solenoid valves and levels of liquid nitrogen to purify a raw gas and continuously products purified gas with perfoming alternatively purification and regeneration. We develop the monitoring and control program to monitor the gas purification process on real-time and control the process time with checking the impurities in purified gas. From the result of system operation, the developed monitoring and control system continuously products high pure helium gas with reducing impurities in raw gas to permitted limits(less than 0.01 ~ 0.05 ppm)

• 한국식품과학회지
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• 제48권6호
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• pp.542-547
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• 2016

The thermal properties of ramie leaf ${\beta}$-amylase (RBA) were examined to develop a novel process for enzyme purification. The thermostability of RBA extract prepared from ramie leaf powder was examined at various temperatures. RBA activity decreased slightly, whereas other carbohydrate-active enzymes, such as $\small{D}$-enzyme, were rapidly inactivated during 30 min incubation at $60^{\circ}C$. When the heat-treated extract was incubated with various substrates, maltose was produced exclusively as the major product, whereas the untreated crude extract produced maltose and other maltooligosaccharides. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, fewer protein bands were observed for the heat-treated extract than the untreated extract, indicating that the thermostable RBA was partially purified and other thermolabile enzymes were eliminated. Thus, the treatment of the RBA extract at $60^{\circ}C$ for 30 min resulted in 5.4-fold purification with a recovery yield of 90%.

• 한국식물병리학회지
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• 제14권6호
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• pp.559-562
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• 1998

Odontoglossum ringspot virus (ORSV) was finally purified from ORSV-infected orchid plants by diethylaminoethyl (DEAE) cellulose anion exchange column chromatography. The virus was reliably eluted by potassium chloride at the concentration from 0.1 M to 0.13 M. Partial purification was done by solubilization with Triton X-100 (allkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG; MW 8,000). The finally purified ORSV represented one distinct homogeneous band and the molecular weight of its capsid protein was about 17,500 Dalton in electrophoretic analysis. Electron microscopy showed not only intact particles ranged from 280 nm to 340 nm in length, but also segmented particles that final 140 nm to 220 nm and even disks. Enzyme-linked immunosorbent assay (ELISA) showed that final yield was 12 mg/100 g of the infected leaves. Bioassay demonstrated that the purified ORSV had the normal infectivity to orchid plants and Nicotiana glutionsa. Based on these data, anion exchange column chromatography could be efficiently applied to the purification of ORSV and other viruses similar to ORSV.