• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.92-102
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• 2006

Abundant proteins often cause problems in proteome study. Glutelin family proteins (hereafter referred to glutelin) are present in rice proteome sample as over-whelming constituents with very high abundance. In order to increase the number of identified proteins in rice proteome study, we developed a newly improved method for sample preparation through the removal of glutelin. When the protein samples from rice seed were extracted by the conventional trichloroacetic acid (TCA) extraction method, glutelin accounts for about 60% of total rice seed proteins in SDS gels. Using our new water extraction method, glutelin consists of only about 10% of total proteins. After analyzing on a two-dimensional gel electrophoresis (2-DE), 937 protein spots were detected using the conventional TCA extraction method. On the other hand, 1240 proteins could be seen using the new water extraction method. The selectivity for non-glutelin and less abundant protein by the water extraction method was also confirmed by ESI-Q/TOF mass spectrometry analysis. Thus, the new water extraction method developed here can be efficiently used to study the proteome analysis of rice storage seed.

• KSBB Journal
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• v.16 no.2
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• pp.146-152
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• 2001

To avoid the intrinsic problem of aggregation associated with the traditional solution-phase refolding process, we propose a solid-phase refolding method integrated with expanded bed adsorption chromatography. The model protein used was a fusion protein of recombinant human growth hormone and a glutathione S transferase fragment. It was demonstrated that the EBA-mediated refolding technique could simultaneously remove cellular debris and directly renature the fusion protein inclusion bodies in the cell homogenate with much higher yields and less agregation. To demonstrate the applicability of the method, we successfully tested the three representative types of starting materials, i. e., rhGH monomer, washed inclusion bodies, and the E. coli homogenate. This direct and simplified refolding process could also reduce the number of renaturation steps required and allow refolding at a higher concentration, at approximately 2 mg fusion protein per ml of resin. To the best of our knowledge, it is the first approach that has combined the solid-phase refolding method with expanded bed chromatography.

• Proceedings of the KAIS Fall Conference
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• 2012.05a
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• pp.239-242
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• 2012

효모 (Saccharomyces Cervisiae)는 단일 세포의 형태로 존재하는 진핵 세포로써 동물세포와 유사한 기작으로 분비성 단백질을 생성한다. 따라서 박테리아와 달리 효모를 이용하면 당단백질이나 이황결합을 포함하는 분비성 단백질을 경제적으로 대량 합성할 수 있다. 효모의 필수 단백질 중 하나인 단백질 이황이성질화 효소는 소포체에 위치하며 분비성 단백질에 구조적으로 안정한 이황결합을 제공하는 효소이다. 본 연구는 단백질 이황이성질화 효소 (protein disulfide isomerase)가 지니고 있는 두 개의 활성도메인 중 분비성 단백질들의 합성 효율에 직접적으로 관여하는 부위를 찾는 연구이다. 효모 유전체로부터 단백질 이황이성질화 효소의 유전자 (PDI1)을 제거하고 효소의 변이 유전자를 주입한 후 효모의 성장 속도를 측정하였다. 또한 효모의 대표적 분비성 단백질을 각 변이 효소를 지니는 효모에 과발현시켜 합성 및 이황결합 형성 효율을 측정하였다. 단백질 이황이성질화 효소내 두 개의 활성 부위 중 아미노 말단쪽에 위치한 a 도메인에 있는 활성 부위가 분비성 단백질의 활성에 중요한 역할을 한다는 것을 알 수 있었다. 이 결과는 이황결합이나 당을 포함하는 외래 단백질의 고효율 합성을 위한 새로운 효모종 개발에 중요한 정보를 제공할 것으로 기대 된다.

• Proceedings of the Korea Contents Association Conference
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• 2011.05a
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• pp.339-340
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• 2011

본 연구는 N말단 아미노산 종류에 따라 단백질의 수명(half-life)이 결정된다는 N-end rule을 기반으로 단백질 수명을 예측해주는 프로그램인 protparam의 결과와 bleach-chase를 이용한 실험 데이터를 비교 분석하였다. 단백질 수명을 결정하는 여러 요인들을 고려하지 않고 한 가지 요인만을 반영한 protparam의 결과는 실제 측정값과 현격한 차이를 나타낸다. 특히 실제 단백질은 NME(N-terminal Methionine Excision) 현상이 일어나는데 이를 고려하지 않고 유전체에서 번역한 그대로의 아미노산 서열을 가지고 단백질 수명을 계산하는 한계를 가지고 있다. 이에 본 연구에서 N말단 아미노산을 순차적으로 제거하여 N-end rule을 적용한 결과도 실험 데이터와 일치하지 않는 결과를 보여주고 있음을 확인하였다. 따라서 현재 사용되고 있는 단백질 수명 예측 프로그램은 이런 문제점을 가지고 있기 때문에 새로운 예측 알고리즘의 개발이 요구된다.

• Journal of Korean Society of Environmental Engineers
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• v.29 no.4
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• pp.460-465
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• 2007

In this study, the removal characteristic of ammonia nitrogen and behavior of nitrogen was investigated using Leclercia adecarboxylata, which was derived from the culture contaminated by ammonia nitrogen of high concentration. The method of ammonia nitrogen removal was not biological nitrification and denitrification but elimination of nutrient salt with internal synthesis of microorganisms which use ammonia nitrogen as substrate. L. adecarboxylata(one of ammonia synthesis microorganisms) was highly activated and showed the most high removal efficiency in free salt condition but the removal efficiency decreased badly in salt concentration of more than 4%. About 80 mg/L of $NH_3-N$ was mostly removed within 20 hours and 500 mg/L of $NH_3-N$ showed less then removal efficiency of 50% because carbon source was not enough. However, ammonium nitrogen concentration was decreased again when the carbon source was inserted additionally thus, ammonium nitrogen removal efficiency by L. adecarboxylata, was related to amount of carbon source. pH decreased from 8.0 to 6.36 according to growth of L. adecarboxylata. Concentration of nitrite nitrogen and nitrate nitrogen did not increase and TKN concentration showed no variation while ammonia nitrogen was removed by L. adecarboxylata. In addition to, when content of protein in organic nitrogen was measured, protein was not detected at the beginning of microorganism synthesis but protein of 193.1 mg/L was detected after 48 hours. Hence, ammonium nitrogen was not decomposed as nitrate nitrogen and nitrite nitrogen but synthesized by L. adecarboxylata, which has excellent ability of nitrogen synthesis and can threat ammonia nitrogen of high concentration in wastewater.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.38 no.5
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• pp.377-382
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• 1993

Effects of leaf and pod removal on changes in dry weight and on the contents of soluble sugar, starch, protein and oil in leaves and seeds of soybean [Glycine max(L.) Merr.] cultivar ‘Hwangkeumkong’ were measured at the research farm of Korea University in 1992. The upper 40% and lower 60% of leaves and pods were subjected to treatments at the growth stage of beginning pod(R3). Upper leaf-lower pod removal showed the highest leaf and the lowest seed dry weights. Soluble sugar content was no different among treatments in leaves and seeds. The highest starch content was found in leaves of upper leaf-lower pod removal. Protein content was higher in lower leaves than upper leaves and the lowest in seeds of lower leaf-upper pod removal which had the highest oil content in leaves and seeds. These results apparently indicated that photoassimilates were mobilized from upper leaves to lower seeds, and protein sources were moved from lower to upper parts but weak in remobilization from leaves for the long distance translocation during the reproductive growth period.

• Journal of Conservation Science
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• v.12 no.1 s.15
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• pp.83-91
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• 2003

The paste used for conserving and mounting has much to do with the life of cultural properties of paintings. Previous studies on the pastes dealt with elimination of protein from wheat flour as protein has been known to do harm to paste and investigated the effects of starch and protein content on adhesive strength and conservativeness. We found that protein content inversely affected adhesive strength while adhesive strength and viscosity of paste were proportionally related to starch concentration of the paste. The adhesives with more protein showed less conservativeness. Types and number of microorganisms were found to increase as protein percentage increased. All these results points that the higher content of protein in adhesive formula support higher microbial growth with reduced adhesive strength, but higher flexibility. The optimized paste of conservation treatment was wheat starch paste with not only minimize viscosity but also maximized adhesive strength.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.257-262
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• 2004

ErmSF is one of the ERM proteins which transfer the methyl group to A2058 in 23S rRNA to confer the resis­tance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism. Unlike other ERM pro­teins, ErmSF contains long N-terminal end region (NTER), of which $25\%$ is composed of arginine that is known to interact with RNA well. Gradual deletion of NTER leaded us to the point where mutant protein lost much of activity in vivo. Overexpressed and purified mutant protein showed much reduced activity in vitro: $2\%$ activity relative to that of wild type protein. This fact suggests that this amino acid interact with RNA close to meth­ylatable adenine to locate it at an active site properly.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.39 no.6
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• pp.594-601
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• 1994

Effects of translocation on seed yield and carbon-nitrogen compounds at five leaf-pod ratios of soybean [Glycine max. (L.)Merr. ] culti bars, 'Paldalkong', 'Baekunkong', and 'Danyeobkong' were measured. The upper 50 and lower 50% of leaves and pods were subjected to treatments at growth stage R3. Three soybean cultivars showed the similar trends on changes in dry matter accumulation and on the contents of soluble sugar, starch and protein in seeds among the treatments. Mean stem dry weight was increased with upper leaf-lower pod and lower leaf-upper pod removals, and decreased with upper leaf and lower leaf removals. Leaf dry weight was appeared higher at the upper leaves among the treatments. Seed numbers and dry weights were decreased with leaf and leaf-pod removals, and were higher in lower part of the plants. Soluble sugar and starch contents in seeds were also showed slightly higher in lower part. Protein content of seeds was decreased in upper part with upper leaf removal and in lower part with lower leaf removal, however, that of the upper seeds was the highest with lower leaf removal. The results of this study are assumed that carbon and nitrogen compounds were translocated opposite directions and protein source was weak in remobilization for the long distance transport during the reproductive growth period of soybean plants.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.10a
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• pp.179-180
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• 2001

두부는 대두의 염용성 단백질을 추출, 응고시켜 만든, 동양의 식생활에는육류 단백질을 대체하는 가장 중요한 단백질 급원 식품이다. 두부는 다른 두류가공품에 비해 인체에서의 이용효율이 높고, 제조 공정 중 소화에 저해가 되는 조섬유질 등이 제거되어 소화율도높다. 또한 영양가가 우수하며, 다른 곡류에 많이 결핍되어 있는 필수 아미노산이 내재해 있을 뿐 아니라, 가격 면에서도 저렴한 등 다양한 장점을 지니고있다. (중략)