• Journal of Life Science
• /
• v.25 no.4
• /
• pp.385-389
• /
• 2015

In this study, it is reported that a large polyprotein can be stoichiometrically cleaved by the use of caspase-3-dependent proteolysis. Previously, it has been shown that the proteolytic IETD motif was partially processed when treated with caspase-3, while the DEVD motif was completely cleaved. The cleavage efficiency of the DEVD-based substrate was approximately 2.0 times higher than that of the IETD substrate, in response to caspase-3. Based on this, 3 protein genes of interest were genetically linked to each other by adding two proteolytic cleavage sequences, DEVD and IETD, for caspase-3. Particularly, glutathione-S transferase (GST), maltose binding protein (MBP), and red fluorescent protein (RFP) were chosen as model proteins due to the variation in their size. The expressed polyprotein was purified by immobilized metal ion affinity chromatography (IMAC) via a hexa-histidine tag at the C-terminal end, showing 93 kDa of a chimeric GST:MBP:RFP fusion protein. In response to caspase-3, cleavage products, such as MBP:RFP (68 kDa), MBP (42 kDa), RFP (26 kDa), and GST (25 kDa), were separated from a large precursor GST:MBP:RFP (93 kDa) via SDS-PAGE. The results obtained from this study indicate that a multi-protein can be stoichiometrically produced from a large poly-protein by using proteolytic recognition motifs, such as DEVD and IETD tetra-peptides, for caspase-3.

• Toxicological Research
• /
• v.35 no.2
• /
• pp.119-129
• /
• 2019

As the use of cosmetics has greatly increased in a daily life, safety issues with cosmetic ingredients have drawn an attention. Drometrizole [2-(2'-hydroxy-5'-methylphenyl)benzotriazole] is categorized as a sunscreen ingredient and is used in cosmetics and non-cosmetics as a UV light absorber. No significant toxicity has been observed in acute oral, inhalation, or dermal toxicity studies. In a 13-week oral toxicity study in beagle dogs, No observed adverse effect level (NOAEL) was determined as 31.75 mg/kg bw/day in males and 34.6 mg/kg bw/day in females, based on increased serum alanine aminotransferase activity. Although drometrizole was negative for skin sensitization in two Magnusson-Kligman maximization tests in guinea pigs, there were two case reports of consumers presenting with allergic contact dermatitis. Drometrizole showed no teratogenicity in reproductive and developmental toxicity studies in which rats and mice were treated for 6 to 15 days of the gestation period. Ames tests showed that drometrizole was not mutagenic. A long-term carcinogenicity study using mice and rats showed no significant carcinogenic effect. A nail product containing 0.03% drometrizole was nonirritating, non-sensitizing and non-photosensitizing in a test with 147 human subjects. For risk assessment, the NOAEL chosen was 31.75 mg/kg bw/day in a 13-week oral toxicity study. Systemic exposure dosages were 0.27228 mg/kg bw/day and 1.90598 mg/kg bw/day for 1% and 7% drometrizole in cosmetics, respectively. Risk characterization studies demonstrated that when cosmetic products contain 1.0% of drometrizole, the margin of safety was greater than 100. Based on the risk assessment data, the MFDS revised the regulatory concentration of drometrizole from 7% to 1% in 2015. Under current regulation, drometrizole is considered to be safe for use in cosmetics. If new toxicological data are obtained in the future, the risk assessment should be carried out to update the appropriate guidelines.

• Korean Journal of Veterinary Service
• /
• v.44 no.4
• /
• pp.283-290
• /
• 2021

This is a study on the improvement of the chemical treatment method of the livestock carcass treatment newly introduced in the livestock infectious disease prevention method in order to improve the problems of the existing burial-centered carcass treatment method when a livestock infectious disease occurs. It was conducted to establish detailed treatment standards for the chemical treatment method of pig carcasses based on the results of proof of the absence of infectious diseases in pigs. After inoculating pig carcasses with 10 pathogens (6 viruses [FMDV, ASFV, CSFV, PCV2, PRRSV, PEDV] and 4 bacteria [Lawsonia intracellularis, Clostridium perfringens type C, E. coli, Salmonella Typhimurium]) It was treated at 90℃ for 5 hours in a potassium hydroxide (KOH) liquid solution corresponding to 15% of the body weight. This method liquefies all cadaveric components and inactivates all inoculated pathogens. Based on these results, it was possible to prove that chemical treatment of pig carcasses is effective in killing pathogens and is a safe method without the risk of disease transmission. Although there are problems to be solved in the processing and operation of the chemical treatment products of livestock carcasses, the chemical treatment method of livestock carcasses can be suggested as an alternative to the current domestic burial-centered livestock carcass treatment method, preventing environmental pollution, and contributing to public health.

• Journal of the Korean Wood Science and Technology
• /
• v.39 no.3
• /
• pp.221-229
• /
• 2011

Due to the increase of oil price and the environmental issue such as the emission of volatile organic compounds, the necessity for developing alternative resins of petroleum-based adhesive resins, which have extensively been used for the manufacture of wood-based products, has been speculation since the early 1990. In our study, rapeseed flour (RSF), which is the by-product of bio-diesel produced from rapeseed, were hydrolyzed by enzymes. As a crosslinking agents of the RSF hydrolyzates, phenol-formaldehyde prepolymers (PF) were prepared. The RSF hydrolyzates and PF were mixed to complete the formulation of RSF-based adhesive resins, and the resins were applied to make the laminated veneer lumber (LVL). The physical and mechanical properties of the LVL were measured to examine whether RSF can be used as raw materials of adhesive resins for the fabrication of LVL or not. The average moisture content and soaking delamination rate of the LVL bonded with RSF-based adhesive resins exceeded the minimum requirement of KS standard. Moreover, thermal analysis of the RSF-based resins showed similar tendencies except for the RSF-based adhesive resins formulated with pectinase-hydrolyzed RSF. The bending strengths of the LVL were higher than that of the LVL made with commercial PF resins. These results showed the potential of RSF as a raw material of alternative adhesives for the production of LVL. Further works on the optimal conditions of RSF hydrolysis and spreading characteristics for RSF-based adhesive resins is required to improve the adhesive performance of RSF-based resins.

• Korean Journal of Weed Science
• /
• v.30 no.4
• /
• pp.340-360
• /
• 2010

The depletion of fossil fuels, ecological problems associated with $CO_2$ emissions climate change, growing world population, and future energy supplies are forcing the development of alternative resources for energy (heat and electricity), transport fuels and chemicals: the replacement of fossil resources with $CO_2$ neutral biomass. Several options exist to cover energy supplies of the future, including solar, wind, and water power; however, chemical carbon source can get from biomass only. When used in combination with environmental friend production and processing technology, the use of biomass can be seen as a sustainable alternative to conventional chemical feedstocks. The biorefinery concept is analogous to today's petroleum refinery, which produce multiple fuels and chemical products from petroleum. A biorefinery is a facility that integrates biomass conversion processes and equipment to produce fuels, power, and value-added chemicals from biomass. Biorefinery is the co-production of a spectrum of bio-based products (food, feed, materials, and chemicals) and energy (fuels, power, and heat) from biomass [definition IEA Bioenergy Task 42]. By producing multiple products, a biorefinery takes advantage of the various components in biomass and their intermediates therefore maximizing the value derived from the biomass feedstocks. A biorefinery could, for example, produce one or several low-volume, but high-value, chemical or nutraceutical products and a low-value, but high-volume liquid transportation fuel such as biodiesel or bioethanol. Future biorefinery may play a major role in producing chemicals and materials as a bridge between agriculture and chemistry that are traditionally produced from petroleum. Industrial biotechnology is expected to significantly complement or replace the current petroleum-based industry and to play an important role.

• Applied Biological Chemistry
• /
• v.48 no.2
• /
• pp.132-139
• /
• 2005

The major obstacle in the popularization of Chungkookjang is the short shelf-life of $2{\sim}3$ months and some problems concerning storage including the growth of molds even in the products even within shelf-life. To solve these problems we conducted a research to improve its storage by using the vacuumed packaging and sanitary method through seed culture, innoculation and sterilization. For the optimization of storage time, temperature and sterilization temperature, we measured viable cell numbers of bacteria and fungi, amount of gas outbreak and contents of amino type nitrogen and monitored these experimental results by response surface methodology of SAS program, so that we could observe the quality changes of Chungkookjang during shelf-life. Especially fungi, which are the biggest troublemaker in Chungkookjang shelf-life, couldn't be detected from the generally and vacuum-packed samples; also, viable cell numbers were highly influenced by sterilization temperature and in vacuum-packed samples. In the case of vacuum-packed samples, amount of gas outbreak was highly influenced by sterilization temperature of its storage conditions and it was higher in generally packed samples as compared to vacuum-packed samples even at any storage conditions. The changes of pH in generally and vacuum-packed samples were highly influenced by the storage temperature. As the temperatures of storage and sterilization were higher and the storage time was longer, so the amount of gas outbreak was accordingly lower. These results showed that amino type nitrogen contents in generally and vacuum-packed samples were systematically influenced by the temperature, storage time and sterilization temperature. Also the result showed that the change of amino type nitrogen contents during storage was less in vacuum-packed samples than in general ones. Based on the above results, we can produce Chungkookjang products with extended shelf-life of as far as 6 months without any quality change using sanitary manufacturing method, vacuumed packaging condition, sterilization in $70^{\circ}C$ for 60 minutes and storage under $10^{\circ}C$ during shelf-life. According to this research, we have the possibility to greatly increase the goods value of Chungkookjang by developing the manufacture processing and packaging.

• Journal of Plant Biotechnology
• /
• v.44 no.1
• /
• pp.97-106
• /
• 2017

Living modified crops are genetically modified living organisms and are widely used in biotechnical research and desired goods. As the reliance on LM products, concerns about safety of LMOs have been continuously increased in South Korea. We established the detection methods for unintentional released LMOs in environmental conditions. To detect six LM event genes of 1 canola, 1 maize and 4 soybeans, PCR conditions were based upon consideration of the Joint Research Centre information. Genomic DNAs were isolated from LM samples and PCR analysis were performed using each event-specific primer pair. Event-specific genes of all events were efficiently recognized by our methods. To investigate the insertion site of LM genes in each genome, we verified PCR product sequence by DNA sequencing. These results suggest that the LM event-specific gene amplification can be efficiently developed. In addition, our detection method is fit for monitoring and post-management of LM crops in the environment.

• Journal of Microbiology and Biotechnology
• /
• v.27 no.4
• /
• pp.739-746
• /
• 2017

As alternatives to antibiotics in livestocks, probiotics have been used, although most of them in the form of liquid or semisolid formulations, which show low cell viability after oral administration. Therefore, suitable dry dosage forms should be developed for livestocks to protect probiotics against the low pH in the stomach such that the products have higher probiotics survivability. Here, in order to develop a dry dosage forms of probiotics for poultry, we used hydroxypropyl methylcellulose phthalate 55 (HPMCP 55) as a tablet-forming matrix to develop probiotics in a tablet form for poultry. Here, we made three different kinds of probiotics-loaded tablet under different compression forces and investigated their characteristics based on their survivability, morphology, disintegration time, and kinetics in simulated gastrointestinal fluid. The results indicated that the probiotics formulated in the tablets displayed higher survival rates in acidic gastric conditions than probiotics in solution. Rapid release of the probiotics from the tablets occurred in simulated intestinal fluid because of fast swelling of the tablets in neutral pH. As a matrix of tablet, HPMCP 55 provided good viability of probiotics after 6 months under refrigeration. Moreover, after oral administration of probiotics-loaded tablets to chicken, more viable probiotics were observed, than with solution type, through several digestive areas of chicken by the tablets.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.6
• /
• pp.880-886
• /
• 2007

Ensiled or oven-dried green tea by-products (GTB) were evaluated in goats for their nutritive potential as protein feedstuffs based on in vitro and in vivo digestibility. To elucidate the effects of tea tannin on in vitro digestibility, polyethylene glycol (PEG) was used as a tannin binding agent. Both ensiled and dried GTB contained 31.9 to 32.6% of crude protein (CP) on a dry matter (DM) basis. Phenolics and tannins in soybean meal and alfalfa hay were low or not detected, but they were high in both ensiled and dried GTB (7.3-10.1% DM as total extractable tannins). In vitro protein digestibility in the rumen ranked: soybean meal>alfalfa hay cube>ensiled GTB = dried GTB. The protein digestibility post-ruminally of these feedstuffs showed a similar trend to that in the rumen, but the digestibility of ensiled GTB was significantly higher than that of dried GTB. Addition of PEG improved the in vitro protein digestibility of both kinds of GTB in the rumen and post-ruminally, indicating that tannins suppressed the potential protein digestibility of GTB. The increased protein digestibility by PEG addition was not significantly different between ensiled and dried GTB in the rumen, but the percentage increment of ensiled GTB was higher than dried GTB post-ruminally. In the in vivo digestibility trial, ensiled and dried GTB were offered to goats as partial substitutes for soybean meal and alfalfa hay cubes. Offering both GTB to goats as 5-10% on a DM basis did not affect nutrient digestibility, ruminal pH, volatile fatty acids, and ammonia concentration. However, the eating time of the GTB-incorporated diet was longer than that of the basal diet. It took 1.4 and 1.6 times longer than the control diet, to eat the diet completely when GTB silage was offered at 5 and 10% levels, respectively, of the total diet. These results show that ensiled and dried GTB are useful as partial substitutes for soybean meal and alfalfa hay cubes for goats with respect to nutritive value. Because of lessened palatability, it is recommended that GTB be incorporated into the diet at 5% on a DM basis.

• Journal of Life Science
• /
• v.29 no.12
• /
• pp.1408-1414
• /
• 2019

Enzymes are widely used in industrial applications such as detergents, food, feed production, pharmaceuticals and medical applications and major contributors to clean industrial products and processes. To screen, identify, and characterize the enzymes the zymography techniques are routinely used. The zymography technique is a simple, sensitive, and quantifiable technique that is widely used to detect functional enzymes following electrophoretic separation in sodium dodecyl sulfate (SDS)-polyacrylamide gels. The method is a versatile two-stage technique involving protein separation by electrophoresis followed by the detection of enzyme activity in polyacrylamide gels under non-reducing conditions. It is based on SDS-polyacrylamide gel (PAG) copolymerization with substrates, which are degraded by the hydrolytic enzymes restored in enzyme reaction buffer after the electrophoretic separation. Any kind of biological sample can be applied and analyzed on zymography, including culture supernatants of microbes, plants extracts, blood, tissue culture fluids, enzymes in foods extracts and metaproteome. The advantage of zymography is that it is possible to directly detect the protein with activity on the electrophoretic gel as well as confirm the activity at the nanogram level. Thus, this zymography technology can be applied in various fields. However, these advantages are rather disadvantageous and can often lead to experimental errors. In this review, the advantages, disadvantages, and problem solving of zymography technique are described.