• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1516-1524
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• 2014

Flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) can utilize a variety of external electron acceptors and also has stricter substrate specificity than any other glucose oxidoreductases, which makes it the ideal diagnostic enzyme in the field of glucose biosensors. A gene coding for a hypothetical protein, similar to glucose oxidase and derived from Aspergillus terreus NIH2624, was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 260,000 U/l in the culture supernatant after fed-batch cultivation for 84 h. After a three-step purification protocol that included isopropanol precipitation, affinity chromatography, and a second isopropanol precipitation, recombinant FAD-GDH was purified with a recovery of 65%. This is the first time that isopropanol precipitation has been used to concentrate a fermentation supernatant and exchange buffers after affinity chromatography purification. The purified FAD-GDH exhibited a broad and diffuse band between 83 and 150 kDa. The recombinant FAD-GDH was stable across a wide pH range (3.5 to 9.0) with maximum activity at pH 7.5 and $55^{\circ}C$. In addition, it displayed very high thermal stability, with a half-life of 82 min at $60^{\circ}C$. These characteristics indicate that FAD-GDH will be useful in the field of glucose biosensors.

• 한국지능시스템학회논문지
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• 제19권5호
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• pp.713-719
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• 2009

본 논문은 비선형 특성을 내재한 물 수요예측을 위하여 기존의 시계열 자기회귀 알고리즘과 다층신경망 학습방법을 결합한 단기 물 수요 예측 알고리즘을 개발하였다. 제시된 방법을 검증하기 위한 사례연구로 2007년도와 2008년도 전북지역의 광역상수도 A정수장에서 취득된 데이터를 활용하여 알고리즘 구축 및 제안 방법의 정확도를 분석하였다. 실험 결과 다중회귀모델은 MAPE가 5.1%, AR모델은 3.8%, 제안된 방법인 AR+MLP 모델은 3.6%로 나타나 성능이 우수한 것으로 나타났다. 따라서 제안된 방법을 사용할 경우 정수장에서 단기 물 수요예측에 유용하게 활용할 수 있음을 보였다.

• 한국인터넷방송통신학회논문지
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• 제22권2호
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• pp.109-114
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• 2022

최근 미세먼지가 심각한 사회문제로 대두되며 이에 대한 대책으로 공기청정기가 각광을 받고 있다. 이에 따라 본 논문에서는 스마트 자율주행 공기청정기 시스템에 관한 연구 개발을 진행하였다. 개발된 스마트 자율주행 공기청정기는 기존 공기청정기의 표준사용면적의 한계를 개선하고 효율적으로 공기 정화 기능을 수행할 수 있다. 또한 스마트 자율주행 공기청정기 사용의 편의성을 위해 모바일 앱(App)과 웹(Web)기반 프로그램을 같이 개발하였다. 앱을 통해 3가지 공기 정화 모드를 간편하게 조작할 수 있고, 웹을 통해 어디서나 공기오염도에 대한 통계 수치를 모니터링할 수 있다. 그리고 시험을 통해 제안된 스마트 자율주행 공기청정기가 기존의 공기청정기보다 더 효율적임을 보였다.

• 한국지반환경공학회 논문집
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• 제13권12호
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• pp.59-66
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• 2012

본 연구에서는 철도정비창 부지 내에 폐기물 및 중금속오염 구간에서 채취한 토양을 대상으로 하였다. 그리고 효율적인 정화공정 설계를 위하여 고농도 오염구간, 저농도 오염구간, 폐주물사 함유 시료를 대상으로 입도분포 및 입도 분포 오염농도 분석을 실시하였다. 하지만 폐콘크리트, 폐목재 등의 건설폐기물, 폐주물사, 소각재 등이 부지 전반에 걸쳐 매립되어 있어 일반토양 오염과 다른 양상을 보이고 있었다. 따라서 일반적인 중금속정화기술로는 오염원이 감소하지 않아 혼합된 폐기물 중에 자성을 띠는 성분을 자력선별을 적용하여 실험한 결과 중금속 오염도는 감소하는 것으로 나타났다.

• 한국환경보건학회지
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• 제31권3호
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• pp.221-226
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• 2005

Isoflavone was extracted with various concentration of aqueous methanol using whole hypocotyls as the starting material. Whole hypocotyls were preferred as the raw material because the residue could be easily removed from the solvent after the extraction process. Extraction yield was almost constant at the methanol concentration of $20-80\%$. Most of the isoflavone was extracted within 1 hr, and the extraction yield remained almost constant thereafter. When the concentration of methanol was $80\%$, the content of total solid was reduced due to the reduced extraction of contaminating protein as the result of protein insolubilization. Among resins tested, Diaion HP-20, Amberlite XAD-16, and Amberlite IRC-50 showed the highest capacity to absorb the compound. Open column chromatography with Diaion HP-20 showed that $80\%$ aqueous ethanol was most efficient as the eluting solvent with final recovery of the phytochemical being more than $95\%$. Maximum adsorption of the phytochemical occurred at the acidic pH 2-4. When the spatial velocity was increased to 15 and more, the degree of adsorption was decreased, whereas below spatial velocity of 15, the adsorption capacity of isoflavone to the resin was almost constant. The purity of the isoflavone purified by column chromatography was $78\%$.

• Journal of Microbiology and Biotechnology
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• 제25권6호
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• pp.887-892
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• 2015

Uricase is an important microbial enzyme that can be used in the clinical treatment of gout, hyperuricemia, and tumor lysis syndrome. A total of 127 clinical isolates of Pseudomonas aeruginosa were tested for uricase production. A Pseudomonas strain named Ps43 showed the highest level of native uricase enzyme expression. The open reading frame of the uricase enzyme was amplified from Ps43 and cloned into the expression vector pRSET-B. Uricase was expressed using E. coli BL21 (DE3). The ORF was sequenced and assigned GenBank Accession No. KJ718888. The nucleotide sequence analysis was identical to the coding sequence of uricase gene puuDof P. aeruginosa PAO1. We report the successful expression of P. aeruginosa uricase in Escherichia coli. E. coli showed an induced protein with a molecular mass of about 58 kDa that was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting. We also established efficient protein purification using the Ni-Sepharose column with activity of the purified enzyme of 2.16 IU and a 2-fold increase in the specific activity of the pure enzyme compared with the crude enzyme.

• 약학회지
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• 제54권4호
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• pp.316-321
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• 2010

Adefovir dipivoxil which was originally developed by Gilead Sciences has been used as treatments of HIV and HBV, especially a therapeutics for HBeAg positive and negative chronic patients. We developed highly efficient purification method using reverse phase column chromatography for mass production and a stable amorphous Adefovir dipivoxil using solid dispersion method. Reverse phase column chromatography led to highly pure product, more than 99.7% by HPLC and can be used for mass production compared with normal column chromatography. Solid dispersion method containing watersoluble polymer and Isomalt showed improved stability of amorphous Adefovir dipivoxil against heat and moisture.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2008년도 제39회 하계학술대회
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• pp.1466-1467
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• 2008

We describe here a novel micro total analysis system for the purification and identification of the affinity-captured proteins. Also we demonstrated the mass analysis of the Carcinoembrionic antigen (CEA) and Alpha femtoprotein which were chosen as the target cancer marker. For MALDI-TOF analyses, the proteins should to be separated from a protein mixture and be concentrated when needed. This procedure usually takes a long time even before protease-digested samples are to be obtained from them. Here, we describe integrated and efficient micro chip for protein purification and digestion for MALDI-TOF analyses. At first, disease protein is purified by passing the micro chamber from a protein mixture or human whole serum and released from the micro affinity beads by thermal heating. Purified protein is then transfer to the hole for trypsin digestion. The final sample is analyzed by MALDI-TOF. All the processes could be finished successfully within one hour, which renders MALDI-TOF analyses of a target protein quite simple.

• BMB Reports
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• 제31권1호
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• pp.77-82
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• 1998

To purify the geranylgeranyl protein transferase type I (GGPT-I) efficiently, a gene expression system using the pGEX-4T-1 vector was constructed. The cal1 gene, encoding the ${\beta}$ subunit of GGPT-I, was subcloned into the pGEX-4T-1 vector and co-transformed into E. coli cells harboring the ram2 gene, the ${\alpha}$ subunit gene of GGPT-I. GGPT-I was highly expressed as a fusion protein with glutathione S-transferase (GST) in E. coli, purified to homogeneity by glutathione-agarose affinity chromatography, and the GST moiety was excised by thrombin treatment. The purified yeast GGPT-I showed a dose-dependent increase in the transferase activity, and its apparent $K_m$ value for an undecapeptide fused with GST (GST-PEP) was $0.66\;{\mu}M$ and the apparent value for geranylgeranyl pyrophosphate (GGPP) was $0.071\;{\mu}M$.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권1호
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• pp.32-37
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• 2006

In this study, we have purified and characterized the membrane bound nitrate reductase obtained from the denitrifying bacteria, Ochrobactrum anthropi SY509, which was isolated from soil samples. O. anthropi SY509 can grow in minimal medium using nitrate as a nitrogen source. We achieved an overall purification rate of 15-fold from the protein extracted from the membrane fraction, with a recovery of approximately 12% of activity. The enzyme exhibited its highest level of activity at pH 5.5, and the activity was increased up to $70^{\circ}C$. Periplasmic and cytochromic proteins, including nitrite and nitrous oxide reductase, were excluded during centrifugation and were verified using enzyme essay. Reduced methyl viologen was determined to be the most efficient electron donor among a variety of anionic and cationic dyestuffs, which could be also used as an electron donor with dimethyl dithionite. The effects of purification and storage conditions on the stability of enzyme were also investigated. The activity of the membranebound nitrate reductase was stably maintained for over 2 weeks in solution. To maintain the stability of enzyme, the cell was disrupted using sonication at low temperatures, and enzyme was extracted by hot water without any surfactant. The purified enzyme was stored in solution with no salt to prevent any significant losses in activity levels.