• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.3
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• pp.235-239
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• 2006

Due to recent advances in genome sequencing, there has been a dramatic increase in the quantity of genetic information, which has lead to an even greater demand for a faster, more parallel expression system. Therefore, interest in cell-free protein synthesis, as an alternative method for high-throughput gene expression, has been revived. In contrast to in vivo gene expression methods, cell-free protein synthesis provides a completely open system for direct access to the reaction conditions. We have developed an efficient cell-free protein synthesis system by optimizing the energy source and S30 extract. Under the optimized conditions, approximately $650{\mu}g/mL$ of protein was produced after 2h of incubation, with the developed system further modified for the efficient expression of PCR-amplified DNA. When the concentrations of DNA, magnesium, and amino acids were optimized for the production of PCR-based cell-free protein synthesis, the protein yield was comparable to that from the plasmid template.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.21-21
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• 2011

A series of Quasi-Elastic Neutron Scattering (QENS) experiments helps us to understand the single-particle (hydrogen atom) dynamics of a globular protein and its hydration water and strong coupling between them. We also performed Molecular Dynamics (MD) simulations on a realistic model of the hydrated hen-egg Lysozyme powder having two proteins in the periodic box. We found the existence of a Fragile-to-Strong dynamic Crossover (FSC) phenomenon in hydration water around a protein occurring at TL=$225{\pm}5K$ by analyzing Intermediate Scattering Function (ISF). On lowering of the temperature toward FSC, the structure of hydration water makes a transition from predominantly the High Density Liquid (HDL) form, a more fluid state, to predominantly the Low Density Liquid (LDL) form, a less fluid state, derived from the existence of a liquid?liquid critical point at an elevated pressure. We showed experimentally and confirmed theoretically that this sudden switch in the mobility of the hydration water around a protein triggers the dynamic transition (so-called glass transition) of the protein, at a temperature TD=220 K. Mean Square Displacement (MSD) is the important factor to show that the FSC is the key to the strong coupling between a protein and its hydration water by suggesting TL${\fallingdotseq}$TD. MD simulations with TIP4P force field for water were performed to understand hydration level dependency of the FSC temperature. We added water molecules to increase hydration level of the protein hydration water, from 0.30, 0.45, 0.60 and 1.00 (1.00 is the bulk water). These confirm the existence of the FSC and the hydration level dependence of the FSC temperature: FSC temperature is decreased upon increasing hydration level. We compared the hydration water around Lysozyme, B-DNA and RNA. Similarity among those suggests that the FSC and this coupling be universal for globular proteins, biopolymers.

• Journal of Aquaculture
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• v.19 no.4
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• pp.254-260
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• 2006

A 6-week feeding trial was conducted to determine apparent protein and phosphorus digestibilities in order to evaluate nine different dietary protein sources in growing olive flounder, Paralichthys olivaceus. Nine diets containing fish meal analog ($BAIFA-M^{TM}$), white fish meal (WFM), flounder muscle powder (FMP), squid liver powder (SLP), leather meal (LM), soybean meal (SM), corn gluten meal (CGM), poultry by-product (PBP) or egg albumin(EA) were prepared by mixing a basal diet (BD) with one of nine test ingredients at the ratio of 7 to 3. Apparent protein digestibilities of FMP, SLP, WFM, SM, CGM, LM, PBP, $BAIFA-M^{TM}$, and EA were 94%, 92%, 86%, 82%, 75%, 72%, 72%, 71%, and 30%, respectively. Apparent phosphorus digestibilities of FMP, SLP, $BAIFA-M^{TM}$, LM, WFM, PBP, CGM, EA and SM were 77%, 72%, 65%, 55%, 54%, 50%, 20%, 20%, and 17%, respectively. Weight gain of fish fed FMP ($323^a$) was significantly higher than those of fish fed the other diets, and those of fish fed basal diet ($302^b$), SLP ($305^b$) and WFM ($308^b$) diet were significantly higher than those of fish fed SM ($274^c$), $BAIFA-M^{TM}\;(268^{cd}),\;PBP\;(261^{de}),\;LM\;(251^e),\;CGM\;(254^e)$ and EA ($181^f$). Based on the results of apparent protein digestibilities, apparent phosphorus digestibilities and weight gain, SLP, SM and $BAIFA-M^{TM}$ could be one of good protein source to replace fish meal in flounder diets.

• Journal of Korean Ophthalmic Optics Society
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• v.10 no.2
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• pp.91-97
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• 2005

We investigated the question whether protein removing activities of enzyme cleaner - protein remover for soft contact lens - are associated with the material of soft contact lens as well as action time, temperature and pH of enzyme solution. We used a subtilisin cleaner as protein remover and estimated the protein amount remained on soft contact lens after using the subtilisin cleaner under the different conditions. The remained protein in soft contact lens was greatly decreased until treatment for 60min, but no significant differences were found from 60min to 24hr. The cleaning effect of the enzymatic treatment in the range of $15{\sim}30^{\circ}C$ was constant. however, there was a significant decline of the protein removing effect at $10^{\circ}C$ and less. The pH of the solution was also important for the efficacy of the enzymatic treatment. The activity of the enzyme cleaner was highest in pH 8.0 and significantly decreased a pH below 7. The pH dependence was found to be related to the conformational change of subtilisin. Furthermore, significant differences in the protein deposit removing efficacy of the subtilisin cleaner were found among the soft contact lens materials.

• Bulletin of the Korean Chemical Society
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• v.30 no.12
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• pp.2999-3005
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• 2009

Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product, E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwichtype binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used for the cell lysis. Temperature also affected the stability of the E7 protein, and we found that the E7 protein was stabilized at 4$^{\circ}C$ for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.8
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• pp.1135-1142
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• 2017

Objective: The objective of this study was to evaluate effects of heat treatment and soybean oil inclusion on protein oxidation of soy protein isolate (SPI) and of oxidized protein on redox status of broilers at an early age. Methods: SPI mixed with soybean oil (SPIO) heated at $100^{\circ}C$ for 8 h was used to evaluate protein oxidation of SPI. A total of two hundred and sixteen 1-day-old Arbor Acres chicks were divided into 3 groups with 6 replicates of 12 birds, receiving basal diet (CON), heat-oxidized SPI diet (HSPI) or mixture of SPI and 2% soybean oil diet (HSPIO) for 21 d, respectively. Results: Increased protein carbonyl, decreased protein sulfhydryl of SPI were observed as heating time increased in all treatments (p<0.05). Addition of 2% soybean oil increased protein carbonyl of SPI at 8 h heating (p<0.05). Dietary HSPI and HSPIO decreased the average daily gain of broilers as compared with the CON (p<0.05). Broilers fed HSPI and HSPIO exhibited decreased glutathione (GSH) in serum, catalase activity and total sulfhydryl in liver and increased malondialdehyde (MDA) and protein carbonyl in serum, advanced oxidation protein products (AOPPs) in liver and protein carbonyl in jejunal mucosa as compared with that of the CON (p<0.05). Additionally, broilers receiving HSPIO showed decreased glutathione peroxidase activity (GSH-Px) in serum, GSH and hydroxyl radical scavenging capacity in liver, GSH-Px activity in duodenal mucosa, GSH-Px activity and superoxide anion radical scavenging capacity in jejunal mucosa and increased AOPPs in serum, MDA and protein carbonyl in liver, MDA and AOPPs in jejunal mucosa (p<0.05). Conclusion: Protein oxidation of SPI can be induced by heat and soybean oil and oxidized protein resulted in redox imbalance in broilers at an early age.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.53 no.4
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• pp.577-582
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• 2020

This study was conducted to investigate the replacement of fish meal (FM) with a plant and animal protein mixture (wheat gluten, soy protein concentrate, tankage meal and poultry by-product meal) in the diets of juvenile olive flounder Paralichthys olivaceus. The basal diet was formulated to contain 65% FM (Con). Four other experimental diets were formulated with alternative proteins replacing 20%, 30%, 40% and 50% of FM (FM20, FM30, FM40 and FM50, respectively). Taurine and betaine were added to the FM replacement diets. Triplicate groups of fish (mean±SD, 5.41±0.01) were fed the diets to apparent satiation for 15 weeks. After the feeding trial, no significant differences were found between any dietary groups in growth performance, feed utilization, survival, hematological parameters or whole-body composition. This result indicates that a proper mixture of the four protein sources with taurine and betaine supplements can be used as FM replacement to reduce FM levels from 65% to 32.5% in juvenile olive flounder diets.

• Journal of Sericultural and Entomological Science
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• v.34 no.1
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• pp.30-34
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• 1992

Isolation, purification and molecular properties of adult specific protein (ASP) were investigated in the silkworm, Bombyx mori. ASP was purified from adult haemolymph by gel filtration on Sephadex G-100 and ion exchange chromatography on CM52. The preparation was shown to be homogeneous by native-PAGE and immunoelectrophoresis. The purified protein was consisted of two different subunits with molecular weights of 19.5kDa and 17.5kDa, and contained no sugars and no lipids.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1568-1572
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• 2002

A 152 day trial was conducted to see the effect of feeding naturally fermented wheat straw (FWS) with either energy, protein or energy protein supplements on the growth of buffalo calves. Twenty four male buffalo calves (10-12 months old) divided in 6 equal groups were individually offered FWS as sole roughage along with either conventional concentrate mixture (conc), maize grains (M), solvent extracted mustard cake (DMC), M-DMC mixture (50:50), deoiled rice bran (DRB) or uromol bran mixture (UBM) in 70:30 ratio. The digestibility of nutrients, nitrogen retention and nutritive value was maximum in FWS:UBM followed by FWS:DMC and FWS:Conc groups. Almost, all the blood parameters were observed well within the normal range except that of blood urea (FWS:UBM) and creatinine (FWS:DMC and FWS:DRB). The dietary combination in which FWS was supplemented with only conventional protein supplement like DMC proved to be highly efficient as far as live weight gain was concerned. FWS supplemented with energy-protein combination i.e. MDMC could also be used as complete feed for growing calves in comparison to conventional feeding system.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.30 no.6
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• pp.948-955
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• 1997

Immunomagnetic separation of virus coupled with .reverse transcription-polymerase chain reaction (IMS-PCR) was performed with infectious hematopoietic necrosis virus (IHNV). A DNA fragment of expected size was synthesized in the RT-PCR with total RNA extracted from IHNV inoculated CHSE-214. In a SDS-PAGE analysis, a protein band of over 70kDa was detected from non-infected cells and cells inoculated with IHNV and infectious pancreatic necrosis virus (IPNV). This protein was detected in the Western blot analysis probably because of non-specific reaction to monoclonal antibody against IHNV nucleocapsid protein. In the immunomagnetic separation, magnetic beads coated with monoclonal antibody against the IHNV nucleocapsid protein was incubated with supernatant from IHNV inoculated CHSE-214 cells. During this process, the non-specifically reacting protein could be removed by washing the magnetic bead with PBS in the presence of an external magnetic field, and viral proteins were detected from the remaining, cleaned magnetic beads. It was necessary to extract viral RNA from the captured virus particles before RT-PCR, and no DNA product was detected when the captured virus was only heated 5 min at $95^{\circ}C$. A PCR-product of expected size was synthesized from IMS-PCR with magnetic beads double coated either by goat anti-mouse IgG antibody -monoclonal antibody or streptavidin - biotin conjugated monoclonal antibody.