• 한국수산과학회지
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• 제26권6호
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• pp.529-537
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• 1993

가다랭이 안와유로부터 docosahexaenoic acid(DHA)를 정제하기 위해 기존의 방법들을 적용하여 비교 검토하고, DHA의 효과적인 정제를 위해 조작방법을 개량하였다. 가다랭이 안와조직의 총지질은 $55.4\%$였으며, 이 중 DHA는 $23.7\%$였다. 저온분별결정법과 요소결정법을 적용한 결과 순도에서 각각 약 $46\%$ 및 $61\%$의 DHA가 얻어졌다. 이들 방법들은 순도면에서는 다소 떨어지나, 정제조작이 단순하여 다량의 DHA 분리에 적합하였다. 질산은 수용액법은 상기 2가지 방법에 비하여 순도면에서는 약간 개선되었으나, 회수율이 대단히 낮았다($10\%$ 이하). 질산은 함침 실리카 칼럼 크로마토그래피법은 고순도 DHA의 정제방법으로써 적합하였다(순도 $98\%$ 이상, 회수율 $90\%$ 이상). 결과적으로 저온분별결정법과 질산은 함침 실리카 칼럼 크로마토그래피법을 조합한 개량법(2단계 정제법)이 가다랭이 안와유로부터 고순도 DHA($99.9\%$)의 정제를 위한 가장 효과적인 방법으로 평가되었다.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.231-236
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• 2004

The inexpensive large-scale production of pure PGA (Penicillin G Acylase) has been a commercial goal. PGA has been used as a model enzyme in the development of simple one-step purification methods. In this study, the purification of poly-His tagged PGA protein secreted into the periplasmic space was carried out by using immobilized metal-ion affinity chromatography (IMAC). The PGA gene was obtained from E. coli ATCC 11105. Codons encoding histidines were fused at the C-terminus of the PGA gene by PCR. E. coli JM109 harboring pPGA-HIS6 vector produced active his-tagged acylases in the presence of lac promoter during cultivation at $26^{\circ}C$. The maximum specific activity of the acylase purified by using one-step chromatography after osmotic shock was 38.5 U/mg and was recovered with the yield of 70％. Both 23 kDa ($\alpha$) and 62 kDa ($\beta$) subunits were recovered by using IMAC with just C-terminus tagging of the $\beta$ subunit. The purification of the periplasmic fraction by osmotic shock and that of purified acylase was increased by 2.6-fold and 19-fold, respectively, compared to the crude extract.

• 한국환경과학회지
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• 제21권7호
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• pp.825-835
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• 2012

This study for the development of water purification Artificial floating island maximizing domestic Artificial floating island patent trends and product development, according to the timing of patent registration was analyzed for trends. In addition, domestic invention patent technology Artificial floating island typed according to the purpose and characteristics of domestic patents were Artificial Floating Island. In particular, domestic leisure space with a growing population and the need for securing emerging role as a reservoir of water only in the past, who do appeal as a tourist destination or as an ecological space utilized, and accordingly will transform and the need to secure a hydrophilic, degrade water quality problems using this aquatic environment (water acquisition and hydrophilic), the requirements are a big obstacle is the reality factor. This patented product differentiation strategy through the analysis of the development of technology progressiveness (Field Application) in terms of water quality improvement and maintenance side, and the hydrophilic side scenery, ecological restoration aspects, and applicability to the field and taking into account existing technology economic aspects of distinction were presented and advertised a lot in terms of cost compared to other techniques without the use of highly efficient methodology for building a water purification and also appears identity appeal, wetlands, rivers, etc. can be applied broadly technician widespread deployment and installation time to less simple and more are expected to spread.

• 한국농공학회논문집
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• 제51권5호
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• pp.95-100
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• 2009

The water purification systems have been hardly used for agricultural purpose due to their complicated compositions and high costs for farmers, while only simple filtrations have been applied to irrigation systems in order to prevent the system from clogging problems. This study therefore developed a clean water supplying system, the Filter-Disinfection-Adsorption (FDA) system, especially for greenhouse cultivation of where low quality of water is available. This system has also been produced for providing convenience water to farmers in the areas of no water supply service systems for the purpose of washing their bodies or agricultural machineries after farm work. The FDA system consists of three stages of purification processes with an integral module, including disk and teflon filtrations and Ultraviolet (UV) sterilization processes. Indoor experiments were undertaken with a trial product of the FDA system to test its performance. The operation test of the process was performed as well as the condition check of each item including UV module, filters, control panel, pump, valves, etc. The results shows good performance of each test with no critical problems. The initial and maintenance costs were also analysed with other purification systems. From the comparisons, the FDA system found to be very economical and easy to use.

• BMB Reports
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• 제44권4호
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• pp.279-284
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• 2011

The glutathione S-transferase (GST) system is useful for increasing protein solubility and purifying soluble GST fusion proteins. However, purifying half of the GST fusion proteins is still difficult, because they are virtually insoluble under non-denaturing conditions. To optimize a simple and rapid purification condition for GST-pyruvate kinase muscle 2 (GST-PKM2) protein, we used 1% sarkosyl for lysis and a 1 : 200 ratio of sarkosyl to Triton X-100 (S-T) for purification. We purified the GST-PKM2 protein with a high yield, approximately 5 mg/L culture, which was 33 times higher than that prepared using a conventional method. Notably, the GST-high-temperature requirement A2 (GST-HtrA2) protein, used as a model protein for functional activity, fully maintained its proteolytic activity, even when purified under our S-T condition. This method may be useful to apply to other biologically important proteins that become highly insoluble in the prokaryotic expression system.

• International Journal of Naval Architecture and Ocean Engineering
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• 제7권1호
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• pp.212-225
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• 2015

In order to preserve the quality of fresh water in the artificial lake after the reclamation of an intertidal flat at the mouth of a river, we suggest two novel methods of water purification by using tidal potential energy and an enclosed permeable embankment called an utsuro (Akai et al., 1990) in the reclaimed region. One method uses an inflatable bag on the seabed within an utsuro, while the other uses a moored floating barge out of a dyke. Each case employs a subsea pipe to allow flow between the inside and outside of the utsuro. The change in water level in the utsuro, which is pushed through the pipe by the potential energy outside, caused circulation in the artificial lake. In this paper, we analyzed the inflatable bag and floating barge motion as well as the pipe flow characteristics and drafts as given by a harmonic sea level, and compared the theoretical value with an experimental value with a simple small model basin. The numerical calculation based on theory showed good agreement with experimental values.

• BMB Reports
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• 제40권1호
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• pp.113-118
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• 2007

Tolaasin, a pore-forming peptide toxin, is produced by Pseudomonas tolaasii and causes brown blotch disease of the cultivated mushrooms. P. tolaasii 6264 was isolated from the oyster mushroom damaged by the disease in Korean. In order to isolate tolaasin molecules, the supernatant of bacterial culture was harvested at the stationary phase of growth. Tolaasin was prepared by ammonium sulfate precipitation and three steps of chromatograpies, including a gel permeation and two ion exchange chromatographies. Specific hemolytic activity of tolaasin was increased from 1.7 to 162.0 HU $mg^{-1}$ protein, a 98-fold increase, and the purification yield was 16.3%. Tolaasin preparation obtained at each purification step was analyzed by HPLC and SDS-PAGE. Two major peptides were detected from all chromatographic preparations. Their molecular masses were analyzed by MALDI-TOF mass spectrometry and they were identified as tolaasin I and tolaasin II. These results demonstrate that the method used in this study is simple, time-saving, and successful for the preparation of tolaasin.

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.63-67
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• 2004

A simple procedure for the extraction of the lipolytic enzyme from rice bran has been developed. High activity of lipolytic enzyme was obtained by first defatting the rice bran to remove lipid components with various extraction conditions. Then, after rove cycles of aqueous extraction, rice bran lipolytic enzyme was purified using micro- and ultrafiltration apparatus. Lipolytic enzyme activity was estimated by its hydrolytic action of tributyrin. The result indicated that the standard activity curve of butyric acid showed that the potential rice bran enzyme is a hydrolytic lipase enzyme. In addition, it showed higher lipolytic activity and specific enzyme activity with further purification by micro- and ultrafiltration. The size of rice bran lipase enzyme was identified through 15 ％ SDS-PAGE. The molecular weight of the rice bran lipase enzyme was 41 kDa.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.719-722
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• 2001

Silk proteins from Nephila clavipes are fibrous proteins containing repetitive sequences with both crystalline and amorphous domains. In order to obtain high-level production of silk protein, the synthetic genes had 16 contiguous units of the consensus repeat sequence of the silk protein were expressed in Escherichia coli BL21(DE3) under the strong inducible T7 promoter. For production of recombinant silk protein in large amounts, pH-stat fed-batch cultures were carried out. The recombinant silk protein was produced as soluble forms in E. coli, and the recombinant silk protein content was as high as 11% of the total protein. When cells were induced at $OD_{600}$ of 60, the amount of silk protein produced was 6.49 g/L. After simple purification steps, 9.2 mg of silk protein that was more than 80% pure was obtained from a 50 mL culture, and the recovery yield was 26.3%.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.501-504
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• 2003

A novel prepurification method was developed aiming at increasing yield and purity, also reducing solvent usage for purification of paclitaxel. This method was a simple and efficient procedure, for the isolation and prepurification of paclitaxel from the biomass of Taxus chinensis, consisting of micelle formation, followed by two steps of precipitation. The use of a micelle and precipitation in the prepurification process allows for rapid separation of paclitaxel from interfering compounds and dramatically reduces solvent usage compared to alternative methodologies. This prepurification process serves to minimize the size and complexity of the HPLC operations for paclitaxel purification. This process is readily scalable to a pilot plant and eventually to a production environment where multikilogram quantities of material are expected to be produced. As much as possible, the process has been optimized to minimize solvent usage, complexity, and operating costs.