• Biotechnology and Bioprocess Engineering:BBE
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• 제9권3호
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• pp.184-190
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• 2004

The influences of impeller types on morphology and protein expression were investigated in a submerged culture of Aspergillus oryzae. The impeller types strongly affected mycelial morphology and protein production in batch and fed-batch fermentations. Cells that were cultured by propeller agitation grew in the form of a pellet, whereas cells that were cultured by turbine agitation grew in a freely dispersed-hyphal manner and in a clumped form. Pellet-grown cells showed high levels of protein production for both the intracellularly heterologous protein (${\beta}$-glucuronidase) and the extracellularly homologous protein (${\alpha}$-amylase). The feeding mode of the carbon source also influenced the morphological distribution and protein expression in fed-batch fermentation of A. oryzae. Pulsed-feeding mainly showed high protein expression and homogeneous distribution of pellet whereas continuous feeding resulted in less protein expression and heterogeneous distribution with pellet and dispersed-hyphae. The pellet growth with propeller agitation paralleling with the pulsed-feeding of carbon source showed a high level of protein production in the submerged fed-batch fermentation of recombinant A. oryzae.

• Interdisciplinary Bio Central
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• 제1권2호
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• pp.8.1-8.6
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• 2009

Introduction: It is a challenge to design a protein score function which stabilizes the native structures of many proteins simultaneously. The coarse-grained description of proteins to construct the pairwise-contact score function usually ignores the backbone directionality of protein structures. We propose a new two-body score function which stabilizes all native states of 1,006 proteins simultaneously. This two-body score function differs from the usual pairwise-contact functions in that it considers two adjacent amino acids at two ends of each peptide bond with the backbone directionality from the N-terminal to the C-terminal. The score is a corresponding propensity for a directional alignment of two adjacent amino acids with their local environments. Results and Discussion: We show that the construction of a directional adjacency-score function was achieved using 1,006 training proteins with the sequence homology less than 30%, which include all representatives of different protein classes. After parameterizing the local environments of amino acids into 9 categories depending on three secondary structures and three kinds of hydrophobicity of amino acids, the 32,400 adjacency-scores of amino acids could be determined by the perceptron learning and the protein threading. These could stabilize simultaneously all native folds of 1,006 training proteins. When these parameters are tested on the new distinct 382 proteins with the sequence homology less than 90%, 371 (97.1%) proteins could recognize their native folds. We also showed using these parameters that the retro sequence of the SH3 domain, the B domain of Staphylococcal protein A, and the B1 domain of Streptococcal protein G could not be stabilized to fold, which agrees with the experimental evidence.

• 농업과학연구
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• 제37권2호
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• pp.245-248
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• 2010

This study was conducted to investigate effect of changes in target fat and protein contents in milk on feed cost using a simulation modeling approach based on the 2001 dairy NRC. Two simulations were done; simulation I had a limitation (up to 20%), but simulation II had no limitation for the use of cottonseed hull in a diet. Using commonly used feed ingredients in Korea, we formulated least cost diets that meet nutrient requirement of a lactating dairy cow producing 36 kg of milk with combinations of 0.1% decrease or 0.1% increase in target milk fat or protein, respectively, from the national average milk fat (4.0%) and milk protein (3.1%). The contents of alfalfa and corn in a least-cost diet were decreased and those of tall fescue, whole cottonseed and rapeseed meal were increased with decreasing fat and/or increasing protein in milk. Scenarios that decreased target milk fat percentage from 4.0% to 3.9% reduced feed cost by 2 won per kg. Due to decrease in feed intake, daily feed cost was even more reduced (136 won per head) by decreasing target milk fat percentage. Increase in target milk protein percentage from 3.1% to 3.2% reduced feed cost by 6 won per kg. Among scenarios simulated, the least feed cost was obtained in scenario aimed for 3.9% fat and 3.2% of protein in milk. We conclude that a feeding practice for increasing milk protein percentage does not directly increase feed cost. In addition, feeding practices that increase protein content in milk is expected to improve economic life-span and reproductive performance of dairy cows.

• BMB Reports
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• 제43권3호
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• pp.193-198
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• 2010

In this study, we report the identification and characterization of a novel C2H2 zinc finger protein, ZNF552, from a human embryonic heart cDNA library. ZNF552 is composed of three exons and two introns and maps to chromosome 19q13.43. The cDNA of ZNF552 is 2.3 kb, encoding 407 amino acids with an amino-terminal KRAB domain and seven carboxyl-terminal C2H2 zinc finger motifs in the nucleus and cytoplasm. Northern blotting analysis indicated that a 2.3 kb transcript specific for ZNF552 was expressed in liver, lung, spleen, testis and kidney, especially with a higher level in the lung and testis in human adult tissues. Reporter gene assays showed that ZNF552 was a transcriptional repressor, and overexpression of ZNF552 in the COS-7 cells inhibited the transcriptional activities of AP-1 and SRE, which could be relieved through RNAi analysis. Deletion studies showed that the KRAB domain of ZNF552 may be involved in this inhibition.

• Journal of Korean Neurosurgical Society
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• 제67권4호
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• pp.451-457
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• 2024

Objective : Exploring protein requirements for critically ill patients has become prominent. On the other hand, considering the significant impact of coma therapy and targeted temperature management (TTM) on the brain as well as systemic metabolisms, protein requirements may plausibly be changed by treatment application. However, there is currently no research on protein requirements following the application of these treatments. Therefore, the aim of this study is to elucidate changes in patients' protein requirements during the application of TTM and coma therapy. Methods : This study is a retrospective analysis of prospectively collected data from March 2019 to May 2022. Among the patients admitted to the intensive care unit, those receiving coma therapy and TTM were included. The patient's treatment period was divided into two phases (phase 1, application and maintenance of coma therapy and TTM; phase 2, tapering and cessation of treatment). In assessing protein requirements, the urine urea nitrogen (UUN) method was employed to estimate the nitrogen balance, offering insight into protein utilization within the body. The patient's protein requirement for each phase was defined as the amount of protein required to achieve a nitrogen balance within ±5, based on the 24-hour collection of UUN. Changes in protein requirements between phases were analyzed. Results : Out of 195 patients, 107 patients with a total of 214 UUN values were included. The mean protein requirement for the entire treatment period was 1.84±0.62 g/kg/day, which is higher than the generally recommended protein supply of 1.2 g/kg/day. As the treatment was tapered, there was a statistically significant increase in the protein requirement from 1.49±0.42 to 2.18±0.60 in phase 2 (p<0.001). Conclusion : Our study revealed a total average protein requirement of 1.84±0.62 g during the treatment period, which falls within the upper range of the preexisting guidelines. Nevertheless, a notable deviation emerged when analyzing the treatment application period separately. Hence, it is recommended to incorporate considerations for the type and timing of treatment, extending beyond the current guideline, which solely accounts for the severity by disease.

• 농업과학연구
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• 제45권2호
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• pp.248-257
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• 2018

Overexpression of AtCYP78A7, a gene encoding a cytochrome P450 protein, has been reported to improve tolerance to drought stress in genetically modified (GM) rice (Oryza sativa L.). The aim of this study was to evaluate the potential allergenicity and acute oral toxicity of the AtCYP78A7 protein expressed in GM rice. Bioinformatics analysis of the amino acid sequence of AtCYP78A7 did not identify any similarities with any known allergens or toxins. It showed that no known allergen had more than a 35% amino acid sequence homology with the AtCYP78A7 protein over an 80 amino acid window or more than 8 consecutive identical amino acids. The gene encoding the AtCYP78A7 protein was cloned in the pGEX-4T-1 vector and expressed in E. coli. Then, the AtCYP78A7 protein was purified and analyzed for acute oral toxicity. The AtCYP78A7 protein was fed at a dose of 2,000 mg/kg body weight in mice, and the changes in mortalities, clinical findings, and body weight were monitored for 14 days after the dosing. Necropsy was carried out on day 14. The protein did not cause any adverse effects when it was orally administered to mice at 2000 mg/kg body weight. These results indicate that the AtCYP78A7 protein expressed in GM rice would not be a potential allergen or toxin.

• Toxicological Research
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• 제12권2호
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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.

• Journal of Nutrition and Health
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• 제6권3호
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• pp.1-8
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• 1973

Metabolic responses to the protein-free, high-carbohydrate diet and subsequent food restriction on the same diet at the level of 50% and 75% has been studied on the adult albino rats. The energy source was either corn starch or sugar. In experiment I, adult male rats weighing $509{\pm}8g$ were divided into two groups 10 rats each. Rats fed on the stock diet served as a control. Rats of restriction group received a protein free diet until they reduced their weight down to 400g and continue on a protein-calorie restriction diet until they reduced their weight down to 300g. In experiment II, 28 adult male rats and the same numbers of female rats weighing $329{\pm}5g$ and $223{\pm}4g$ respectively were divided into four groups, 7 males and females in each. Rats fed on a stock diet were sacrificed at the point when others started a protein free diet. These were served as the control. The protein free group received a protein free diet ad libitum for 4 weeks. The 50% restriction group and 75% restriction group were fed on a protein free diet coupled with food restriction at levels of 50% and 75% respectively for 3 weeks. In the result of this study: 1. The rate of body weight changes was similar between the males and the females. Feeding protein free diet ad lib. initiated a rapid weight lost of approximately 25% and protein free diet coupled with food restriction showed 37-43% reduction of their initial weight. 2. There was no significant differences in the value of the N concentration in liver, spleen, brain and muscle between controls and experimental groups. 3. Rats fed on protein free diet showed 1/10 value of the control in the nitrogen excretion in urine. However female showed less N excretion than male. 4. Observing blood picture, the effects of protein depletion and calorie restriction were not appeared any remarkable changes. 5. There was no sign of fatty liver which might result from protein depletion and calorie restriction. 6. Following semi-starvation, FAO and HMP-DH total enzyme activity was reduced, but activity per unit weight was relatively stable.

• Journal of Nutrition and Health
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• 제18권2호
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• pp.98-114
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• 1985

In the previous studies, one of our author had observed the obligatory nitrogen losses through urine and feces in young Korean male adults, and utilization of whole egg protein in Korean young adults with ordinary intake of energy as a basic study for estimating protein requirements of Korean. This study has tried to determine the protein requirement and utilization when subjects ate the Korean mixed diet at the energy level of 45 kcal per Kg body weight per day. Six healthy Korean young female adults, college students, aged $20{\sim}25$ years old, and $45{\sim}61Kg$ of body weight participated as the study subjects. They were given isocaloric diets with four different protein levels for five days each successively. These diets contained protein levels of 0.45, 0.60, 0.75 and 0.90g per Kg of body weight per day, respectively. It was attempted to observe energy and protein intake, urinary and fecal nitrogen losses, true digestibility of protein, net protein utilization of protein in the body over last two days of period of eating each diet and nitrogen balances were calculated. The body weight change and hematological observation were also performed. The results obtained were summarized as follows ; 1) True digestibility of protein of Korean mixed diets ranged from 83.1% to 86.5% and the average of which was $84.7{\pm}1.7%$. 2) Net protein utilization rate of Korean mixed diets was range of 49% to 55% and the average of which was $52{\pm}3%.$ 3) The body weight of subjects were slightly increased and the values of hemoglobin and hematocrit of the blood were also little increased during the experimental periods of twenty days. 4) Protein requirements of Korean mixed diet with the energy level of 45 kcal/Kg body weight per day of Korean young female adults were estimated as 1.00g/Kg/day on this experimental condition.

• IMMUNE NETWORK
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• 제3권1호
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• pp.16-22
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• 2003

Background: The S100A2 gene, also known as S100L or CaN19, encodes a protein comprised of 99-amino acids, is a member of the calcium-binding proteins of EF-hand family. According to a recent study, this gene was over-expressed in several early and malignant carcinomas compared to normal tissues. To elucidate the role of S100A2 protein in the process during carcinogenesis, production of monoclonal antibody specific to the protein is essential. Methods: First, cDNA sequence coding for ORF region of human S100A2 gene was amplified and cloned into an expression vector to produce GST fusion protein. Recombinant S100A2 protein and subsequently, monoclonal antibody to the protein were produced. The specificity of anti-S100A2 monoclonal antibody was confirmed by immunoblot analysis of cross reactivity to other recombinant proteins of S100A family (GST-S100A1, GST-S100A4 and GST-S100A6). To confirm the relation of S100A2 to cervical carcinogenesis, S100A2 protein in early cervical carcinoma tissue was immunostained using the monoclonal antibody. Results: GST-S100A2 recombinant protein was purified by affinity chromatography and then fusion protein was cleaved and S100A2 protein was isolated. The monoclonal antibody (KK0723; Korean patent pending #2001-30294) to the protein was produced and the antibody did not react with other members of EF-hand family proteins such as S100A1, S100A4 and S100A6. Conclusion: These data suggest that anti-S100A2 monoclonal antibody produced in this study can be very useful for the early detection of cervical carcinoma and elucidation of mechanism during the early cervical carcinogenesis.