• Title/Summary/Keyword: Detergents

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Cloning, Expression, Purification, and Properties of an Endoglucanase Gene (Glycosyl Hydrolase Family 12) from Aspergillus niger VTCC-F021 in Pichia pastoris

  • Pham, Thi Hoa;Quyen, Dinh Thi;Nghiem, Ngoc Minh;Vu, Thu Doan
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1012-1020
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    • 2011
  • A gene coding for an endoglucanase (EglA), of the glycosyl hydrolase family 12 and derived from Aspergillus niger VTCC-F021, was cloned and sequenced. The cDNA sequence, 717 bp, and its putative endoglucanase, a 238 aa protein with a predicted molecular mass of 26 kDa and a pI of 4.35, exhibited 98.3-98.7% and 98.3-98.6% identities, respectively, with cDNA sequences and their corresponding endoglucanases from Aspergillus niger strains from the GenBank. The cDNA was overexpressed in Pichia pastoris GS115 under the control of an AOX1 promoter with a level of 1.59 U/ml culture supernatant, after 72 h of growth in a YP medium induced with 1% (v/v) of methanol. The molecular mass of the purified EglA, determined by SDS-PAGE, was 33 kDa, with a specific activity of 100.16 and 19.91 U/mg toward 1% (w/v) of ${\beta}$-glucan and CMC, respectively. Optimal enzymatic activity was noted at a temperature of $55^{\circ}C$ and a pH of 5. The recombinant EglA (rEglA) was stable over a temperature range of $30-37^{\circ}C$ and at pH range of 3.5-4.5. Metal ions, detergents, and solvents tested indicated a slightly inhibitory effect on rEglA activity. Kinetic constants ($K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$) determined for rEglA with ${\beta}$-glucan as a substrate were 4.04 mg/ml, 102.04 U/mg, 2,040.82 $min^{-1}$, and 505.05, whereas they were 10.17 mg/ml, 28.99 U/mg, 571.71 $min^{-1}$, and 57.01 with CMC as a substrate, respectively. The results thus indicate that the rEglA obtained in this study is highly specific toward ${\beta}$-glucan. The biochemical properties of rEglA make it highly valuable for downstream biotechnological applications, including potential use as a feed enzyme.

Pulsatility Estimation of a Pulsatile Decellularizing Device for the Fabrication of Organ Scaffold (생체장기용 지지체 제작을 위한 박동형 탈세포화 장치의 박동성 평가)

  • Kim, Dong Sun;Yang, Se-Ran;Park, Sung Min;Choi, Seong Wook
    • Journal of Biomedical Engineering Research
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    • v.38 no.2
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    • pp.62-73
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    • 2017
  • To identify a solution for the restricted availability of healthy lungs and the high risk of immune rejections following organ transplantation, tissue engineering techniques for culturing lungs have been studied by many research groups. The most promising method for culturing lungs is the utilization of a bio-scaffold that was prepared using harvested organs from human donors or other animals by removing their original cells. In this study, a pulsatile perfusion pump was used to alleviate the cell removal effect with the high fluid-dynamic power of the perfusion stream during the decellularization process, while other conventional studies focused on chemical methods to identify efficient detergents. The purpose of this study was to analyze the developed device by using energy equivalent pressure (EEP), which is an indicator of pulsatility, to understand the characteristics of pulsatile energy transmitted according to the load size by using the artificial model and compare it with the measured EEP. The pulsatility of the device can be estimated with the concept of fluid-dynamic energy during a particular constant time period or fluid-dynamic power represented as EEP and EEP increment. Because the measured EEP of perfusion flow during decellularization can be changed by the amount of fluid leakage and the degree of clogging in the capillary vessels, EEP should be measured to determine whether the decellularization is progressing without problems. The decrement of EEP caused by the high perfusion resistance was observed from some experimental results that were obtained with artificial models. EEP can be used to monitor the decellularization process after analyzing the varying EEP according to the amount of load. It was confirmed that the EEP was maintained at a high level in the experiment using the harvested lungs from 12-13-week-old rats. In addition, it was confirmed that the cell removal time was faster than when continuous perfusion was performed. In this study, pulsatile power delivered to the lungs was measured to monitor the process of cell removal, and it serve as the evidence for efficient decellularization.

A Study on the Hazardousness and the TLV in Working Environments of Benzine (벤진의 유해 위험성과 작업환경 노출기준 연구)

  • Kim, Hyeon-Yeong;Lee, Sung-Bae;Han, Jung-Hee;Shin, Jea-Hoon
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.16 no.3
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    • pp.233-244
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    • 2006
  • Of many volatile organic detergents for metals, benzine(CAS No. 8030-30-6), of which the toxicity has not yet been proven, has been used as an alternative of the halide compounds in the consideration of toxic effects, global warming and the destruction of ozone layer. In order to evaluate the effects of the benzine on human body by investigating the subchronic inhalation toxicity, to obtain the basic data for establishing the criteria of exposure in working environments and to classify the hazardousness in compliance with the Industrial Safety and Health Act by evaluating the hazardousness, repeated inhalation exposure test was carried with SD rats. The rats were grouped by 10 females and males each. The repetitive inhalation exposures were carried out at 4 levels of concentration of 0 ppm, 60 ppm, 300 ppm, and 1,500 ppm, for 6 hours a day, 5 days a week, for 13 weeks. The results are described hereunder. 1. No death of the animals of the exposed and controlled groups in the test period. Not any specific clinical symptoms, change in feed intake quantity, abnormality in eye test, or change in activity were observed. 2. In the 300 ppm and 1,500 ppm groups, weight reduction in the female groups and weight increase of liver and kidney in the male groups compared with control group were observed with statistical significance(p<0.05). 3. In the blood test, the HCT increased in the male 300 ppm group and the number of hematocyte increased, MCV and MCH decreased in the male 1,500 ppm group. In the female 1,500 ppm group, the HB decreased and the distribution width of the hematocyte particle size increased. In the blood biochemistry test, the TP in the male 1,500 ppm group and the LDH in the female 1,500 ppm group were increased with statistical significance(p<0.05). 4. Under the test conditions of the present study with SD rats, the NOEL was evaluated to be from 60 ppm to 300 ppm for both male and female groups. By extrapolation, the NOEL for human who work 8 hours a day was evaluated to be from 128 ppm to 640 ppm 5. Since the NOEL evaluated in this study do not exceed 60ppm(0.184 mg/L) the test material does not belong to the classification of the hazardous substance "NOEL${\leq}$0.5mg/L/6hr/90day(rat), for continuous inhalation of 6hours a day for 90 days" nor to the basic hazardous chemical substance class 1(0.2 mg/L/6hr/90day(rat) defined by the GHS which is a criteria of classification and identification of chemical compounds. However, considering the boiling point($30-204^{\circ}C$), flashing point($-40^{\circ}C$), vapor pressure(40 mmHg), and the inflammable range(1.0 - 6.0 %), sufficient care should be taken for handling in the safety aspects including fire or explosion.

Tissue Distribution and Toxicokinetics of 4-Tert-Octylphenol in Rats (4-Tert-Octylphenol의 랫드에서의 조직분포 및 독성동태에 관한 연구)

  • Kang Mi Kyung;Ahn Mee Ryung;Chung Hye Joo;Choi Sun Ok;Choi Hong Serk;Yang Ji Sun;Lee Yong Bok;Yoo Tae Moo;Sohn Soo Jung
    • Toxicological Research
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    • v.20 no.3
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    • pp.195-203
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    • 2004
  • 4-Tert-Octylphenol (OP) is a surfactant additive widely used in the manufacture of a variety of detergents and plastic products. OP can disrupt endocrine function in humans and animals. This study was carried out to obtain toxicokinetic parameters of OP in male Sprague-Dawley (SD) rats. Male rats were administered with OP by single oral application of 200 mg/kg body weight. Blood, urine and tissues samples were taken at several time intervals after administration. Analysis of samples for OP was performed by column-switching high performance liquid chromatography (HPLC). In addition, we exam-ined tissue distribution and accumulation of OP after single oral application of 50, 100, and 200 mg/kg, single intravenous injection of 1, 5 and 10 mg/kg or daily application of 50 mg/kg for 14 consecutive days. After single oral administration of 200 mg/kg, Cmax of 213 $\pm$ 123 ng/ml was reached within the first 1.3 hr (Tmax) in the plasma. AUC was calculated for 1,333$\pm$484 ngㆍhr/ml. The final elimination half-life of plasma was longer than that of urine, but urinary clearance was lower than oral. A very small fraction of OP (Fe < 0.0017%) was excreted in urine within 24 hr. These results indicated that the major excretion route of OP was not urine. The mean maximal tissue distribution of OP was obserbed at 6 hr after treatment and slowly decreased time-dependently. High OP concentrations were detected in fat at 24 hr. The OP in fat was slowly released with longer elimination half-life and lower clearance than that of other tissues. OP was not accumulated in the liver following single oral application but 14-day oral treatments resulted in two-fold accumulation. It was probably due to the saturation of detoxification pathways. On the other hand, the mRNA expression of cytochrome P450 isoforms except CYP2C11 was not affected by OP at any dose. The expression of CYP2C11 mRNA decreased in a dose-dependent manner. This result suggests that OP changes expression of the male-specific cytochrome P450 isoforms in rat liver, and these changes are closely related to the toxic and estrogenic effect of OP.

A Study on Clean Agents for Halon Replacement in the Portable Extinguisher with CO2 as an Expellant Gas (이산화탄소를 가압원으로 하는 할론대체 소화기용 청정소화약제에 대한 연구)

  • Jeong, Keesin
    • Fire Science and Engineering
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    • v.33 no.3
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    • pp.51-55
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    • 2019
  • To prevent ozone depletion caused by CFCs, the replacement of Halon with clean agents has been developed in the fire protection field along with refrigerants, detergents, and foaming agents. The alternatives for Halon 1211 have been developed in the portable fire extinguisher area and HCFC-123 is used widely as a clean fire extinguishing agent. The type of expellant gas is important because their own vapor pressure is low. In this study, HCFC-123, HCFC-124, HFC-125, and Novec-1230 were selected as fire extinguishing agents and CO2, which is expected to improve the fire extinguishing ability, was chosen as the expellant gas. For each agent, experiments changing the agent and CO2 amount were carried out and HCFC-123 showed a good result, as expected. The extinguisher, HCFC-123 of 1.5 kg, showed the same ability to suppress a class A and B fire as the extinguisher, HCFC-123 of 2.5 kg, which is currently sold on the market. According to this result, the expellant gas has a subsidiary fire extinguish effect. This can reduce the amount of HCFC fire extinguishing agent, which is categorized in the phase-out alternatives, and is a more eco-friendly and economical fire extinguisher than the previous one. This study can also help solve the problems of CO2 fire extinguishers for class B and C fires, and can be used to extinguish electric and electron facilities fire, which contains large amounts of class A fire combustibles.

Toxicity study of GC-100X in rats and beagles (랫드와 비글에서 GC-100X 세정제의 독성에 대한 연구)

  • Kang, Kyung-Sun;Cho, Sung-Dae;Ahn, Nam-Shik;Jung, Ji-Won;Yang, Se-Ran;Park, Joon-Suk;Park, Ki-Soo;Hong, In-Sun;Seo, Min-Soo;Jo, Eun-Hye;Nguyen, Ba Tiep;Lee, Yong-Soon
    • Korean Journal of Veterinary Research
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    • v.44 no.1
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    • pp.29-40
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    • 2004
  • Because cleaning products are part of our everyday lives, it is essential that they should not present significant risks to health. However, many petrochemicals in most soaps and detergents can be absorbed through the scalp and skin and, over time, accumulate in the organs and tissues. This accumulation may result in brain, nerve, and liver damage. Therefore, it is interested in developing non-harmful detergent. According to Korea Research Institute of Chemical Technology, GC-100X may be non-harmful and non-corrosive alkaline ionic water (pH 12). It is composed of hydroxyl radicals and supplemented with xylitol. To evaluate influence of GC-100X on rats and beagles, GC-100X was diluted with distilled water (25%, 50%, and 100% solution respectively). Each of diluted GC-100X was daily treated per oral. In body weight analysis, urinary analysis, ophthalmological test and autopsy, we did not find any significance, but in serum biochemical analysis and hematological analysis, we found some significances in middle dose group compared with control group. These significances in serum biochemical analysis and hematological analysis may be not induced by GC-100X, because it was not found to be significant from control group in histopathological examination. Thus, it is concluded that NOEL(No Observed Effect Level) of GC-100X may be higher than all treatment doses used in this study, and GC-100X may be a non-toxic detergent.

Shape Oscillation and Detachment of Droplet on Vibrating Flat Surface (진동하는 평판 위의 액적의 형상 진동 및 제거 조건에 대한 연구)

  • Shin, Young-Sub;Lim, Hee-Chang
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.38 no.4
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    • pp.337-346
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    • 2014
  • This study aimed to understand the mode characteristics of a droplet subject to periodic forced vibration and the detachment of a droplet placed on a plate surface. An surface was coated with Teflon to clearly observe the behavior of a droplet. The contact angle between the droplet and surface and the hysteresis were found to be approximately $115^{\circ}C$ and within $25^{\circ}C$, respectively. The coating process was performed in a clean room that had an environment with a low level of contaminants and impurities such as air dust, detergents, and particles. To predict the resonance frequency of a droplet, theoretical and experimental approaches were applied. Two high-speed cameras were configured to acquire side and top views and thus capture different characteristics of a droplet: the mode shape, the detachment, the separated secondary droplet, and the waggling motion. A comparison of the theoretical and experimental results shows no more than 18 discrepancies when predicting the resonance frequency. These differences seem to be caused by contact line friction, nonlinear wall adhesion, and the uncertainty of the experiment. For lower energy inputs, the contact line of the droplet was pinned and the oscillation pattern was axisymmetric. However, the contact line of the droplet was de-pinned as the oscillation became more vigorous with increased energy input. The size of each lobe at the resonance frequency is somewhat larger than that at the neighboring frequency. A droplet in mode 2, one of the primary mode frequencies, exhibits vertical periodic movement as well as detachment and secondary ejection from the main droplet.

Feasibility as a Laundry Detergent Additive of an Alkaline Protease from Bacillus clausii C5 Transformed by Chromosomal Integration (Chromosomal Integration에 의해 제조한 Bacillus clausii C5 유래의 alkaline protease의 세제 첨가제 응용성)

  • Joo, Han-Seung;Choi, Jang Won
    • KSBB Journal
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    • v.27 no.6
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    • pp.352-360
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    • 2012
  • Bacillus clausii I-52 which produced SDS- and $H_2O_2$-tolerant extracellular alkaline protease (BCAP) was isolated from heavily polluted tidal mud flat of West Sea in Incheon, Korea and stable strain (transformant C5) of B. clausii I-52 harboring another copy of BCAP gene in the chromosome was developed using the chromosome integration vector, pHPS9-fuBCAP. When investigated the production of BCAP using B. clausii transformant C5 through pilot-scale submerged fermentation (500 L) at $37^{\circ}C$ for 30 h with an aeration rate of 1 vvm and agitation rate of 250 rpm, protease yield of approximately 105,700 U/mL was achieved using an optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_4{\cdot}7H_2O$ 0.01%, $FeSO_4{\cdot}7H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). The enzyme stability of BCAP was increased by addition of polyols (10%, v/v) and also, the stabilities of BCAP towards not only the thermal-induced inactivation at $50^{\circ}C$ but also the SDS and $H_2O_2$-induced inactivation at $50^{\circ}C$ were enhanced. Among the polyols examined, the best result was obtained with propylene glycol (10%, v/v). The BCAP supplemented with propylene glycol exhibited extreme stability against not only the detergent components such as ${\alpha}$-orephin sulfonate (AOS) and zeolite but also the commercial detergent preparations. The granulized enzyme of BCAP was prepared with approximately 1,310,000 U/g of granule. Wash performance analysis using EMPA test fabrics revealed that BCAP granule exhibited high efficiency for removal of protein stains in the presence of anionic surfactants as well as bleaching agents. When compared to Savinase 6T$^{(R)}$ and Everlase 6T$^{(R)}$ manufactured by Novozymes, BCAP under this study probably showed similar or higher efficiency for the removal of protein stains. These results suggest that the alkaline protease produced from B. clausii transformant C5 showing high stability against detergents and high wash performance has significant potential and a promising candidate for use as a detergent additive.

Problems and Solutions of Zymography Techniques (자이모그라피 기술의 문제점과 해결)

  • Kang, Dae-Ook;Choi, Nack-Shick
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1408-1414
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    • 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.

Identifying Characteristics of Incidents at Hazardous Material Facilities

  • Kim, Geun-Young;Kim, Sang-Won;Won, Jai-Mu
    • Journal of the Korean Society of Hazard Mitigation
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    • v.9 no.6
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    • pp.51-56
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    • 2009
  • Safety and quality assessment systems are very important in manufacture, storage, transportation, and handling of hazardous materials(hazmat) to prevent hazmat disasters. At present, hazardous materials exist everywhere in our daily lives with various forms of plastics, household products of cleaning and washing detergents, fertilizers or petroleum-related products. However, hazardous materials are dangerous substances when they are released to human or environment. Hazardous materials become very widely used substances in the age of oil-based industrial economy. The Korean Ministry of Environment (KMOE) describes about one hundred thousand types of chemicals are produced and used worldwide. Over four hundred new chemicals are introduced in every year. A crucial question for the Korean hazardous material management may have been raised: Will you be safe from hazardous material incidents? The gas leak disaster at Union Carbide's Bhopal, India in 1984 that made over 6,400 people killed and 30,000 to 40,000 people seriously injured is the representative case for the safety of hazmat. Korea becomes vulnerable to hazmat disaster due to the development of high-tech industry. Thus, the risk assessment system is required to Korea for transferring abandoned hazmat management systems to self-correcting safety systems. This research analyzed characteristics of various hazmat incidents applying statistical analysis methods including frequency analysis or analysis of category data to hazmat incidents for ten years. All of three analyses of category data indicate the significance of causality between hazmat incident site groups and seasons, regional groups, and incident casualty groups.