• KSBB Journal
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• 제17권4호
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• pp.365-369
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• 2002

수용성 형태로 존재하는 융합 페리틴을 정제함에 있어서 실리카 분말을 이용한 전처리 공정은 전체 정제 공정효율 증가에 기여하였다. 전처리 공정을 통해 정제된 융합 페리틴의 순도를 높이기 위해 젤 여과 크로마토그래피를 통해 보다 정제된 응합 페리틴을 얻을 수 있었다. 이렇게 정제된 융합 페리틴의 철분결합능력을 분석해본결과 320 moles $F_{H}+F_{L}$ mole로 활성이 우수함을 알 수 있었으며, 분자량 분석을 통해 융합페리틴(40 k dalton)은 trimer와 monomer형태로 존재함을 확인할 수 있었다

• Mycobiology
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• 제37권3호
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• pp.240-242
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• 2009

An improved procedure for preparing PCR cloning vectors was developed. This procedure includes the incorporation of adapters to create XcmI restriction enzyme sites in pBluescript II SK(+) vectors, digestion with XcmI followed by further digestion of the small fragment produced by XcmI digestion with additional enzymes, and purification with PCR purification kits. Using this procedure, PCR cloning vectors with high ligation efficiencies and low blue or false-positive colonies were obtained.

• Bulletin of the Korean Chemical Society
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• 제23권7호
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• pp.985-989
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• 2002

For efficient biopulping process, very active and stable lignase is essential. Laccase is one of the best enzyme in terms of environmentally benign processes, since the enzyme uses oxygen as an oxidant to degrade lignin and produces no hamful prod ucts. We could purify a laccase homogeneously from Cerrena unicolor in a very active state. It shows characteristic absorption feature with blue band at λmax = 604 ㎚. Molecular weight of the enzyme is 57,608 which could be accurately determined by MALDI/TOF MS. The enzyme has 2.8 copper ions per enzyme implying apoenzymes might exist together. The enzyme is active in lignin degradation and the activity increases 4 times in the presence of ABTS as a mediator.

• 한국해양바이오학회지
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• 제2권4호
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• pp.245-249
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• 2007

High quality of purified alginate and fucoidan is required for the medical uses to prevent the unexpected side effects from the contaminated endotoxin present in the materials. We attempted to establish an efficient and fast mass scale production method for the highly purified poly-G alginate from the sea weeds. In addition, lab scale method was established to obtain the fraction of higher purity from the commercially available fucoidan source (about 85% purity).

• KSBB Journal
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• 제8권3호
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• pp.287-294
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• 1993

사람의 뇨로부터 유용한 단백질을 동시에 분리하 기 위한 목적으로 효과적인 통합 분리정제 공정이 고안되었다. Ultrafiltration 방법을 이용한 농축과정 과 pH침전법을 전처리 단계로 사용하였고, 이어서 g gel filtration과 흡착, 이온교환, 친화, 그리고 역상 C column을 선택적으로 혼합한 chromatography를 수행하였다. 이 공정으로 정제한 urokinase, epider­m mal growth factor, albumin은 각각 SDS-poly­a acrylamide gel 전기이동상에서 단일의 단백질 띠로 이동하였고, 자신의 단백질 활성을 유지하고 있었다. 이들 세 가지 목적 단백질의 전체 수율은 각각 48 % %, 17%, 46%로 나타났다.

• 미생물학회지
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• 제23권4호
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• pp.271-281
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• 1985

Bacillus thuringiensis var. thuringiensis produces an extracellular insecticidal thermostable .betha.-exotoxin, which was purified through microfiltering, barium precipitation, charcoal absorption chromatography, ion exchange column chromatography and gel filtration. The exotoxin in each purification step was detedted by thin layer chromatography, high pressure liquid chromatography and paper electrophoresis with efficient results. The exotoxin productivity on time course was checked by spectrophotometric absorbance at 258nm with the result that the exotoxin was initially produced in 6 hour culture and reached maximum value in 36 hour culture. Anti-bacterial effect test on Micrococcus flava was applied as toxicity test. The results showed that frowth inhibition of M. flava could be shown in plate assay of cell free filtered supernatant, alkaline eluant from charcoal and purified exotosin obtained from gel filtration column chromatography on Sephadex G-10 appeared to be 740. Heat stability of the exotoxin was confirmed through autoclaving twice.

• Journal of Microbiology and Biotechnology
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• 제17권11호
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• pp.1751-1757
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• 2007

Elastin-like polypeptides (ELPs) undergo a reversible inverse phase transition upon a change in temperature. This thermally triggered phase transition allows for a simple and rapid means of purifying a fusion protein. Recovery of ELPs-tagged fusion protein was easily achieved by aggregation, triggered either by raising temperature or by adding salt. In this study, levansucrase has been used as a model enzyme in the development of a simple one-step purification method using ELPs. The levansucrase gene cloned from Pseudomonas aurantiaca S-4380 was tagged with various sizes of ELPs to functionally express and optimize the purification of levansucrase. One of two ELPs, ELP[V-20] or ELP[V-40], was fused at the C-terminus of the levansucrase gene. A levansucrase-ELP fusion protein was expressed in Escherichia coli $DH5{\alpha}$ at $37^{\circ}C$ for 18 h. The molecular masses of levansucrase-ELP[V-20] and levansucrase-ELP[V-40] were determined as 56 kDa and 65 kDa, respectively. The phase transition of levansucrase-ELP[V-20] occurred at $20^{\circ}C$ in 50 mM Tris-Cl (pH 8) buffer with 3 M NaCl added, whereas the phase transition temperature ($T_t$) of levansucrase-ELP[V-40] was $17^{\circ}C$ with 2 M NaCl. Levansucrase was successfully purified using the phase transition characteristics of ELPs, with a recovery yield of higher than 80%, as verified by SDS-PAGE. The specific activity was measured spectrophotometrically to be 173 U/mg and 171 U/mg for levansucrase-ELP[V-20] and levansucrase-ELP[V-40], respectively, implying that the ELP-tagging system provides an efficient one-step separation method for protein purification.

• Korean Chemical Engineering Research
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• 제52권4호
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• pp.497-502
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• 2014

바이오매스 유래 타르 및 왁스 성분은 파클리탁셀의 분리정제에 많은 악영향을 미치기 때문에 반드시 제거되어야 한다. 흡착제 처리는 식물세포배양으로부터 유래된 타르 및 왁스 성분을 매우 간단하고 효과적으로 제거할 수 있는 방법이다. 본 연구에서는 전처리 단계에서 상용흡착제 실로퓨트를 이용한 흡착제 처리 공정에서 주요 공정 변수들(온도, 시간, 용매, 건조시료/실로퓨트 비율)을 최적화하였다. 최적의 흡착제 처리 온도, 시간, 용매, 건조시료/실로퓨트 비율은 각각 $30^{\circ}C$, 15분, 메틸렌 클로라이드, 1:1(w/w)이었다. 이러한 결과는 TGA와 HPLC 분석을 통한 유기물의 흡착 양상으로도 확인하였다. 흡착제 처리 단계에서 순도 증가는 미미하였으나 타르 및 왁스 성분의 제거로 인한 후속공정의 공정 가능성과 편리성에 상당히 영향을 미쳤다.

• 상하수도학회지
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• 제33권5호
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• pp.329-339
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• 2019

Recently, the advancement of information and communication technology(ICT) is expanding the connectivity through Internet of Things(IoT), and the media of connection is also expanding from wire/cable transmission to broadband wireless communication, which has significantly improved mobility. This hyperconnectivity has become a key element of the fourth industrial revolution, whereas the supervisory control network of purification plants in korea is operated as a communication network separated from the outside, thereby lagging in terms of connectivity. This is considered the best way to ensure security, and thus there is hardly any consideration of establishing alternatives to operate an efficient and stable communication network. Moreover, security for management of a commercialized communication network and network management solution may be accompanied by immense costs, making it more difficult to make new attempts. Therefore, to improve the conditions for the current supervisory control network of purification plants, this study developed a industrial security L2 switch that supports modbus TCP(Transmission Control Protocol) communication and encryption function of the transmission section. As a result, the communication security performance improved significantly, and the cost for implementing the network management system using Historical Trend and information of HMI(Human Machine Interface) could be reduced by approximately KRW 200 million. The results of this study may be applied to systems for gas, electricity and social safety nets that are infrastructure communication networks that are similar to purification plants.

• Biomolecules & Therapeutics
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• 제1권2호
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• pp.188-195
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• 1993

In order to understand the machanism of action and regulation of ${\beta}$-adrenergic receptor in terms of molecular level, the purification of receptor protein has a fundamental importance. Moreover, species differences among avian, amphibian and mammalian ${\beta}$-adrenergic receptors make it more important to purify mammalian ${\beta}$-adrenergic receptor. Because ${\beta}$-adrenergic receptor is an integral membrane protein, it must be solubilized from the membrane for the purification. The purpose of the present study was to solubilize and characterize the mammalian $\beta$-adrenergic receptor from guinea pig lung in quantities by more efficient and practical method eventually to purify receptor. Guinea pig lung membrane preparation was solubilized by sequential treatment of buffers containing low and high concentration of digitonin which are 0.2 and 1.2% respectively. About 50% of the total receptor pool was released by this double extraction procedure. The $\beta$-adrenoceptors in the digitonin extract were identified using the ${\beta}$-adrenergic antagonist, (-)-[$^3H$]-dihydroalprenolol ([$^3H$]DHA). The solubilized receptor retained all of the essential characteristics of membrane-bound receptor, namely saturability; stereoselectivity; high affinity to ${\beta}$-adrenergic drugs. For the measurement of soluble receptor activity, Sephadex G-50 chromatography method has been widely used. Inspite of its accuracy and wide acceptance, this technique employed troublesome column work which required long time to assay the activity of receptor. We employed another methods to measure receptor activity. When using 0.5% polyethylenimine pretreated GF/B glass fiber filter, filtration technique could be used to measure soluble receptor activity. This technique enabled us to reduce the total amount of time to assay by a factor of 4 as well as to detect soluble receptor. In the present study, we could establish more efficient and practical solubilization method of mammalian $\beta$-adrenergic receptor. The rapidity and high yield of this solubilization scheme, together with the favorable recovery of the receptor activity, are significant steps toward the ultimate purification of the mammalian $\beta$-adrenergic receptor. The result of this study together with more convenient purification method could provide large amount of purified receptor with ease for various research purposes.