• Title/Summary/Keyword: E protein

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Investigation of the Relationship between Protein, Message and Inducer Concentrations in Recombinant E. coli Cells

  • Jorgensen, Lene;Connor J. Thomas;Brian K. Oneill;Anton P.J. Middelbeg
    • Journal of Microbiology and Biotechnology
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    • v.7 no.1
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    • pp.21-24
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    • 1997
  • Chloramphenicol acetyl transferase (CAT) protein and mRNA levels in E. coli were determined following induction of a tac::cat construct by isopropyl-${\beta}$-thiogalactopyranoside (IPTG). High cat mRNA levels did not directly reflect CAT protein levels, in either shakeflask experiments or fermentations. Furthermore, concentrations of IPTG resulting in the highest levels of expression of cat mRNA, were different to those resulting in highest levels of CAT protein. The data suggest that high transcriptional activities lead to limitations at the translational level.

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Direct characterization of E2-dependent target specificity and processivity using an artificial p27-linker-E2 ubiquitination system

  • Ryu, Kyoung-Seok;Choi, Yun-Seok;Ko, Jun-Sang;Kim, Seong-Ock;Kim, Hyun-Jung;Cheong, Hae-Kap;Jeon, Young-Ho;Choi, Byong-Seok;Cheong, Chae-Joon
    • BMB Reports
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    • v.41 no.12
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    • pp.852-857
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    • 2008
  • Little attention has been paid to the specificity between E2 and the target protein during ubiquitination, although RING-E3 induces a potential intra-molecular reaction by mediating the direct transfer of ubiquitin from E2 to the target protein. We have constructed artificial E2 fusion proteins in which a target protein (p27) is tethered to one of six E2s via a flexible linker. Interestingly, only three E2s (UbcH5b, hHR6b, and Cdc34) are able to ubiquitinate p27 via an intra-molecular reaction in this system. Although the first ubiquitination of p27 (p27-Ub) by Cdc34 is less efficient than that of UbcH5b and hHR6b, the additional ubiquitin attachment to p27-Ub by Cdc34 is highly efficient. The E2 core of Cdc34 provides specificity to p27, and the residues 184-196 are required for possessive ubiquitination by Cdc34. We demonstrate direct E2 specificity for p27 and also show that differential ubiquitin linkages can be dependent on E2 alone.

RNA Helicase activity of SecA protein of Escherichia coli

  • Park, Sukyung;Kim, Hyoungman
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.24-24
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    • 1996
  • SecA protein of E. coli is essential for the translocation of various precursor proteins across the plasma membrane. Along with it, SecA protein interacts with precursor proteins, SecY/E, SecB and is an ATPase which has multiple ATP binding sites. There is little known about the regulation mechanism of the protein translocation machinery. (omitted)

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Developing a Protein-chip for Depigmenting Agents Screening (미백제 스크리닝용 단백질칩의 개발)

  • Kim, Eun-Ki;Kwak, Eun-Young;Han, Jung-Sun;Lee, Hyang-Bok;Shin, Jung-Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.13-16
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    • 2005
  • For the high-throughput-screening system (HTS) of depigmenting agents using a protein chip, effects of oligonucleotide-inhibitor sequence on the binding of Mitf protein to E box of MC1R was investigated. The sequence of oligonucletide-inhibitor affected the binding of the target DNA to Mitf, depending on the location of the sequence variation in the inhibitor nucleotide. The oligonucletide-inhibitor that changed the CATGTG sequence didn't show enough inhibition of the target DNA to Mitf, whereas significant inhibition was observed when the sequence outside the CATGTG was changed. This result indicated that CATCTG is crucial sequence for the binding of Mitf to I-box which initiates the transcription of pigmenting genes.

Enzymatic Properties of a Fusion Protein between Human Cytochrome P450 1A1 and Rat NADPH-P450 Reductase Expressed in Escherichia Coli (대장균에서 발현된 인간 Cytochrome P450 1A1과 Rat NADPH-P450 Reductase와의 Fusion Protein의 효소 특성 연구)

  • 천영진;정태천;이현걸;한상섭;노정구
    • Toxicological Research
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    • v.12 no.2
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    • pp.155-161
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    • 1996
  • The enzymatic properties for NADPH-P450 reductase domain of a fusion protein between human cytochrome P450 1A1 and rat NADPH-P450 reductase expressed in Escherichia coli were investigated. The fusion plasmid pCW/1A1OR-expressed E. coli membrane showed high NADPH-cytochrome c reductase activity ($830.1\pm 85.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$), while pCW control vector and P 450 1A1 expression vector pCW/1A1 showed relatively quite low activity ($4.35\pm 0.49, 3.27\pm 0.50 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively). The kinetic curves for NADPH-cytochrome c reductase followed typical Michaelis-Menten kinetics. The $K_{max}$ and $V_{max}$ for NADPH-dependent reductase activity were $8.24\pm 2.61\mu $and $817.9\pm 60.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively, whereas those for cytochrome c-dependent reductase activity were $19.97\pm 2.86\mu M$ and $1303.5\pm 67.1 nmol\cdot min^{-1}\cdot mg protein^{-1}$. The reductase activities were also compared with those of rat, porcine and human liver microsomes. The activity of pCW/ 1A1OR-expressed E. coli membrane was 15.2-fold higher than that of rat liver microsome. Treatment with benzo(a)pyrene, 7-ethoxyresorufin and $\alpha$-naphthofiavone which are known as specific substrates or inhibitor for human P450 1A1 increased NADPH-cytochrome c reductase activity of fusion protein in E. coli membrane dose-dependently. These results demonstrate that the membrane topology of fused enzyme may be important for activity of its NADPH-P450 reductase domain.

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Effects of Heat Treatment History of Skim Milk Powder on the Quality Characteristics of RTD Coffee with High Protein (탈지분유의 열처리 공정이 고단백질 RTD 커피의 품질 특성에 미치는 영향)

  • Ko, Bong Soo;Lee, Seok Ryong;Han, Sung Hee
    • Culinary science and hospitality research
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    • v.24 no.1
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    • pp.24-30
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    • 2018
  • This study investigated the quality of characteristics of high-protein RTD coffee using domestic and imported skim milk powder with different heat treatment. Skim milk powders (A, B) had high-heat treatment, C had medium-heat, and D and E had low-heat treatment. The transmittance of A and B were higher than that of C, and that of C were higher than that of D and E (p<0.05). The precipitate attached on bottom of container of RTD coffee using A and B were 2.993~3.053% and higher than those (0.753~0.803%) of RTD coffee using C, D and E (p<0.05), but there was no difference between those of RTD coffee using C, D and E (p<0.05). The centrifuged precipitate of RTD coffee using A and B were 1.987~2.040% and higher than those (0.820~0.830%) of RTD coffee using C, D and E (p<0.05), but there was no difference between those of RTD coffee using C, D and E (p<0.05). The proximate composition of precipitate attached on bottom of container of RTD coffee using A, which showed the highest amount of precipitate, showed 65.7% of carbohydrate, 19.0% of protein, 4.8% of fat and 4.8% of ash in dry basis, that of RTD coffee being 72.7%, 15.1%, 7.9% and 4.3% in dry basis respectively. Protein and fat content of precipitate were lower and protein and ash content were higher than those of RTD coffee. But seeing that the most increased portion was protein, precipitation of RTD coffee appears to be attributed to heat-denatured proteins.

Rheological Studies of the Fish Protein upon the Thermal Processing (열처리 공정에 따른 생선단백질의 물성 연구)

  • Kang, Byung-Sun;Kim, Byung-Yong;Lee, Jae-Kwun
    • Korean Journal of Food Science and Technology
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    • v.26 no.2
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    • pp.103-109
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    • 1994
  • Changes in the rheological properties and the linear viscoelasticity of fish protein gel upon the thermal processing were studied by using mathematical models with stress-relaxation data. The linear viscoelasticity of surimi gel was observed in the range of the true strain $0.105{\sim}0.693$ and cross-head speed $50{\sim}250\;mm/min$ applied in this study. The results of the generalized Maxwell analysis showed that the magnitudes of elastic elements $(E,\;E_e)$ were increased, but the viscous element $({\eta}) $was decreased, as the cross-head speeds and strain levels were increased. Compared to the protein gel heated directly at $90^{\circ}C$ without preheating, the protein gel pretreated at $4^{\circ}C$ and $40^{\circ}C$ showed the higher elastic modulus, but showed different trends in the viscous component, depending on the rheological model applied. Thus, the approaching methods and curve fitting of two mathematical models of stress-relaxation to describe the viscoelastic properties of fish protein gel were discussed.

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Orthokinetic Stability of $\beta$-Lactoglubulin-Stabilized Emulsions : Effects of Protein Heat Treatment and Surfactant Addition

  • Hong, Soon-Taek
    • Preventive Nutrition and Food Science
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    • v.3 no.2
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    • pp.133-142
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    • 1998
  • Effects of protein heat treatment and surfactant additionoo the orthokindetic stability of $\beta$-lactoglobulin-stabilized emulsions have been investigated under turbulent flow conditions. In studies on protein-stabilized emulsions, samples which had been subjected to heat treatment(i.e. the protein solution orthe emulsion) have been found to be more prone to orthokinetic coalescene than the untreated ones. The emulsions stabilized with protein heated above the denaturation temperature(i.e. 7$0^{\circ}C$) showed the bigger initial average droplet size, which resulted in an increased orthokinetic coalescenece rate. The storage of the protein-stabilized emulsion at high temperature prior to the shearing experiment also made the emulsion less stable in the shear field. Interestingly. the addition of DATEM has been found to produce a substantial increase in orthokinetic stability of the heat-denatured protein-stabilized emulsion system, although Tween 20 is the opposite case.

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Effects of the Dietary Protein and Energy Levels on Growth in Fat Cod (Hexagrammos otakii Jordan et Starks) (사료의 단백질 및 에너지 함량이 쥐노래미 성장에 미치는 영향)

  • LEE Jong Kwan;LEE Sang-Min
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.4
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    • pp.464-473
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    • 1996
  • Two-month feeding experiment was conducted to investigate the optimum dietary protein level and energy to protein ratio in fat cod (Hexagrammos otakii Jordan et STARKS). The fish averaging 29 g were fed with one of the isocaloric diets containing 30, 40, 50 or $60\%$ of protein, or with one of the isoproteic diets containing 9, 10, 11 or 12 of available energy/protein (E/P) ratio. Weight gain and feed efficiency increased significantly with dietary protein level up to $50\%$, then decreased with $60\%$ protein diet (P<0.05). Daily protein intake increased significantly with dietary protein level, whereas protein efficiency ratio decreased with dietary protein level (P<0.05). Second order polynomial regression analyses of percent weight gain and daily protein intake may indicate that the adequate dietary protein level is $45\%$ and daily protein requirement per 100g fish is 1.5g for maximal growth. Weight gain, feed efficiency and protein efficiency from fish led the diet containing 12 of E/P ratio were significantly higher than those from fish fed the other diets (P<0.05). Daily feed or protein intake from fish fed the diet containing 12 of E/P ratio was significantly lower than those from fish fed the other diets (P<0.05). Daily lipid intake increased significantly with dietary E/P ratio (P<0.05).

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A Study on the Change of Thiamine and Riboflavin Value in the Organs of Rats (장기내(臟器內) Thiamine 과 Riboflavin 함량(含量) 변동(變動)에 대(對)한 연구(硏究) - 전아(餞餓), 고당질식(高糖質食), 고조백질식(高蚤白質食) 및 Vitamin E 첨가(添加) 유식(裕食)에 의(依)한 영향(影響)에 대(對)하여 -)

  • Lee, Ky-Dong
    • Journal of Nutrition and Health
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    • v.1 no.2
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    • pp.93-105
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    • 1968
  • The variations of both thiamine and riboflavin value in the organs, viz. liver, small intestine, spleen and kidney of the rats were measured for observing some metabolic changes in the animals during fasting and feeding different quality of diets without V-E supplement. The animal used for the experiment was adult female ablino rat from a pure strain, weighing 225-280g. The animals were divided into 6 groups; the control group, the high carbohydrate diet group, the high carbohydrate diet with V-E group, the high protein diet group, the high protein diet with V-E group, and fasting group. The result obtained are summarized as follows; 1. The thiamine contents in the liver were once increased during early stage of starvation compared with the control group, thereafter they were decreased on the 8 days fasting while the contents in the small intestine and spleen were decreased during 1 to 8 days fasting. 2. The riboflavin contents in the liver and kidney were increased during starvation and the content in the small intestine was no marked change compared with control group. 3. The thiamine contents in the liver and small intestine during feeding the high carbohydrate with V-E supplement diet group were lower than that of the diet without V-E group and the content in the spleen was increased by feeding V-E enriched high carbohydrate diet. 4. The thiamine contents in the liver, small intestine and spleen during feeding the V-E supplemented diets were lower than that of the non-supplemented one's. 5. The riboflavin contents in the liver, small intestine, and kidney were increased during feeding the high carbohydrate diet compared to the control group, and they were decreased during feeding the V-E enriched high carbohydrate diet. 6. The riboflavin contents in each organ were increased during feeding the high protein diet compared to the control group, and they were much increased during 20 to 30 days of feeding the V-E supplemented high protein diet. 7. Therefore, as the above results showed, the variation of thiamine value in the each organs were not markedly changed during feeding different quality of the diets. The thiamine and riboflavin contents in the each organ in the V-E enriched high carbohydrate diet group were lower than without V-E supplemented one's The riboflavin contents in each organ were increased during feeding the high protein diet compared with the control group and the centents were increased during 20 to 30 days of the feeding V-E enriched high protein diet.

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