• Membrane Journal
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• v.26 no.4
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• pp.266-272
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• 2016

The separation and purification of lysozyme enzyme from chicken egg white (CEW) solution was studied using the dead-end filtration. The optimum operation conditions of the dead-end filtration reveal that the maximum value of permselectivity of lysozyme to the other proteins of ovalbumin and conalbumin in the CEW solution was tested with change of molecular weight cut-off (MWCO) of ultrafitration membrane and pH of the CEW solution. The optimum operation conditions for the efficient permeable separation of lysozyme from the CEW solution are that the membrane MWCO is 30 kD and the pH of CEW solution is 11. At this optimum operation conditions, the maximum value of permselectivity of lysozyme to total proteins in CEW solution is about 83.

• BMB Reports
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• v.31 no.4
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• pp.413-417
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• 1998

Transgenic mice containing a bovine ${\beta}-Casein/Human$ lysozyme fusion gene (pBZ) were generated in order to produce human lysozyme in their milk. The expression vector was a quadripartite fusion consisting of a 2 kb upstream DNA of the bovine ${\beta}-casein$ gene, human lysozyme gene, intron II of the rabbit ${\beta}-globin$ gene, and the polyadenylation/termination signals of SV40 DNA. Fertilized mouse zygotes were microinjected with pBZ, then transferred into the oviduct of foster mothers. Out of 20 mice born, 11 survived until postweaning and three were identified as positivetransgenic by Southern blot analysis (one male and two females). The founder mice were mated to BCFl mice to produce transgenic progeny. It was confirmed by RT-PCR and Northern blot analyses that the transgene was specifically expressed in the mammary gland of the founder mice. Furthermore, the artificial introns within the transgenic RNA was proven to be correctly spliced out as judged by RT-PCR analysis. These results indicated that transgenic mice generated in this study properly expressed the human lysozyme RNA in their mammary gland.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.288-292
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• 2006

Through statistically designed experiments, lysis agents were optimized to effectively disrupt bacterial cells in a microfluidic device. Most surfactants caused the efficient lysis of Gram-positive microbes, but not of Gram-negative bacteria. A Plackett-Burman design was used to select the components that increase the efficiency of the lysis of the Gram-negative bacteria Escherichia coli. Using this experimental design, both lysozyme and benzalkonium chloride were shown to significantly increase the cell lysis efficiency, and ATP was extracted in proportion to the lysis efficiency. Benzalkonium chloride affected the cell membrane physically, while lysozyme destroyed the cell wall, and the amount of ATP extracted increased through the synergistic interaction of these two components. The two-factor response-surface design method was used to determine the optimum concentrations of lysozyme and benzalkonium chloride, which were found to be 202 and 99 ppm, respectively. The lysis effect was further verified by microscopic observations in the microchannels. These results indicate that Gram-negative cells can be lysed efficiently in a microfluidic device, thereby allowing the rapid detection of bacterial cells using a bioluminescence-based assay of the released ATP.

• Journal of Life Science
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• v.27 no.10
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• pp.1145-1151
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• 2017

In this study, we report on a simple, efficient method for obtaining penicillin G amidase (PGA) from recombinant Escherichia coli using a formulation mixed with detergent and lysozyme. Research was conducted on the extraction efficiency of PGA from the periplasmic space in cells in terms of the type of detergent, detergent concentration, pH, reaction time, and temperature of permeabilization. The extraction yield of PGA in the formulated surfactant/lysozyme treatment was increased by approximately (55-65 U/ml) in comparison with that in the single surfactant treatment. The released PGA solution was concentrated and exchanged with buffer using an ultrafiltration (U/F) system. The yields of diatomite filtration, membrane filtration (M/F), and U/F were 69.7%, 93.8%, and 77.3%, respectively. A total of 212 KU of PGA was recovered. At the 25-L culture scale, the overall yield of extraction using the mixed surfactant/lysozyme method was 49.2%. The specific activity of extracted PGA was 11 U/mg in protein. The concentrated PGA solution was immobilized on microporous silica beads without further purification of PGA. The total immobilization yield of PGA on the resin was 48.7%, while the enzyme activity was 101 U/g. The immobilized PGA was successfully used to produce 6-APA from penicillin G. Our results indicated that a simple extraction method from periplasmic space in E. coli may be used for the commercial scale production of ${\beta}-lactam$ antibiotics using immobilized PGA.

• Journal of Animal Science and Technology
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• v.47 no.6
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• pp.1033-1040
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• 2005

Protein function of ovotransferrin with various pH and temperature, and its antimicrobial characteristics were determined. Foaming ability of ovotransferrin was high in alkali condition (pH 11), then diminished as time follows. In acidic condition (pH 3.0), very little amount of foam was produced and disappeared promptly in 30min. However, neutral condition (pH 7.0) was revealed as the best area for foam production and foam stability of ovotransferrin. Temperature effect on foam stability of ovotransferrin showed that the highest foam was produced at 60℃. Ovotransferrin was shown weak antimicrobial activity against E. Coli, S .typhi, P. aerug and Candida albicans at dose of 12.5mg/ml and 25mg/ml. Anti-microbial effect of ovotransferrin with either lysozyme or albumine on pathogenic bacteria and fungi shows that the most effective dose was 25mg/ml, especially on S. typhi and C. albicans.

• Fisheries and Aquatic Sciences
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• v.11 no.4
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• pp.183-189
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• 2008

In this study, tests were conducted to investigate the effects of three dietary growth hormones, administered in various amounts, on the growth performance and lysozyme activity in juvenile olive flounder, Paralichthys olivaceus. Three dietary growth hormones, recombinant human growth hormone (rHGH), recombinant bovine somatotropin A (rBST A) and recombinant bovine somatotropin B (rBST B) were tested at three different supplemental levels (10, 20 or 40 mg/kg body weight per week) by a $3{\times}3$ factorial design and a complete randomized design in comparison to a control group. Fish were fed one of the ten experimental diets (control, $rHGH_{10}$, $rHGH_{20}$, $rHGH_{40}$, rBST $A_{10}$, rBST $A_{20}$, rBST $A_{40}$, rBST $B_{10}$, rBST $B_{20}$ and rBST $B_{40}$) for 6 weeks and afterward were analyzed for growth performance by measuring weight gain (WG), feed efficiency (FE), specific growth rate (SGR) and protein efficiency ratio (PER). Based on the factorial design analysis, fish fed rHGH diets demonstrated significantly higher growth performance than fish fed rBST A or rBST B diets. However there were no significant differences in WG, FE, SGR and PER between fish fed rBST A and rBST B diets. Neither hormone level nor the interaction between the different hormones and their various levels had a significant effect on WG, FE, SGR, PER, lysozyme activity or whole-body proximate composition. A complete randomized design analysis confirmed fish fed $rHGH_{10}$, $rHGH_{20}$, $rHGH_{40}$, rBST $A_{10}$, rBST $A_{20}$, rBST $A_{40}$, rBST $B_{20}$ and rBST $B_{40}$ diets for 6 weeks showed higher WG than fish fed the control diet (P<0.05). A higher FE was observed in fish fed $rHGH_{10}$, $rHGH_{20}$, $rHGH_{40}$, rBST $A_{20}$ and rBST $A_{40}$ diets in comparison to fish fed the control diet. Fish fed all graded rHGH, rBST A and rBST B supplemented diets showed a higher SGR than fish fed the control diet. Regarding PER, fish fed $rHGH_{10}$, $rHGH_{20}$, $rHGH_{40}$, rBST $A_{10}$, rBST $A_{20}$, rBST $A_{40}$ and rBST $B_{20}$ diets were higher than fish fed the control diet. Furthermore, the lysozyme activity of fish fed a diet of $rHGH_{20}$ was significantly higher than that of fish fed any other diet. The results measuring the growth and development of the fish clearly suggest the biopotency of dietary rHGH could be higher than those of both dietary rBST A and rBST B. Further implied is the probability that within the range of 10 to 40 mg/kg BW/week the dietary growth hormones could accelerate growth performance, and that 20 mg rHGH/kg BW/week could possibly enhance lysozyme activity in juvenile olive flounder, Paralichthys olivaceus.

• 한국유가공학회:학술대회논문집
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• 1997.05a
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• pp.41-61
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• 1997

젖산균의 유전자 연구를 촉진하기 위해 간단하고 신속한 plasmid DNA의 분리방법과 electro-poration을 이용하여 vector plasmid의 간단하고 신속한 전이방법을 얻기 위해 젖산균의 형질전환에 영향하는 요인에 대하여 연구하였으며 연구결과는 다음과 같다. 1. O'Sullivan과 Klaenhammer의 방법을 개선하여 젖산균 plasmid DNA의 분리에 좋은 결과를 얻을 수 있는 신속하고 쉬운 방법을 고안하였으며, genomic DNA 분리에 이용되는 guanidium thio-cyanate 처리방법을 plasmid의 분리에 적용할 수 있었다. 2. L. casei, L. acidophilus. L. delbruekii var. bulgaricus. L. brevis와 L. plantarum 균주에서 plasmid를 확인하였으며, 돼지 분에서 분리된 L. lactis ssp. lactis. L. fermentum과 L. plantarum에서도 plasmid를 분리 확인하였다. 3. Lactococci의 plasmid분리는 lactobacilli와는 달리 mutanolysin의 처리없이도 잘 되었으며, L. lactis ssp. lactis와 Ent. faecalis에서 plasmid를 확인하였다. 4. E. coli plasimd 분리에 이용되는 MPS membrane filter 방법으로 젖산균 plasmid pLZ12의 분리가 가능하였으나, 세포파편이 filter를 막아 사용에 어려움이 있는 것으로 확인되었다. 5. Plasmid 분리없이 electroporation을 이용한 세포 대 세포 전이법으로 간편하고 빠르게 E. coli DH5${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12를 electroporation시 낮은 전압에서 형질전환 효율이 비교적 좋았으며, 배양 시기를 달리하여 전이시켰을 때 대수생장 말기의 세포가 형질전환 효율이 좋았다. 12. L. casei 102S세포를 각각 10% glycerol, EB, 2차 증류수 등에 녹여 electroporation을 실시하였을 때 각각 $3.8{\times}10^3$, $5.0{\times}10^2$,1.5${\times}10^2$cfu의 형질전환 효율을 보였으며, 1.0mM HEPES, TE buffer를 사용하였을 때에는 전이가 이루어지지 않았다. 13. Plasmid pLZ12의 농도를 달리하여 electroporation을 하였을 때 형질전환 효율이 농도에 비례하여 증가하였다. 14. L. casei 102S에 대수생장 말기의 세포를 채취하여 10% glycerol, 200 Ohms, 25 ${\mu}$FD, 10kV/cm로 plasmid pLZ12를 electroporation할 때 최대 형질전환 효율인 3.8${\times}$10$^{3}$cfu를 얻었으며, lysozyme 처리가 다른 젖산균과는 달리 형질전환 효율을 증가시키지 못하였다. 15. L. casei 102S 세포를 10% glycerol과 EB에 녹여 -20$^{\circ}C$에서 냉동시킨 다음 1일과 7일 후의 세포를 electroporation한 결과 냉동시 세포에 손상을 주는 것으로 인식되었다.

• Journal of Food Hygiene and Safety
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• v.22 no.3
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• pp.151-158
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• 2007

Among 68 strains of lactic acid bacteria (LAB) isolated from Dongchimi, a strain K11 was selected due to its bactericidal activity against Escherichia coli O157 The strain K11 was identified as Lactobacillus plantarum, based on physiological and biochemical characteristics. In the late exponential phase, La. plantarum K11 showed maximum bacteriocin activity (12,800 BU/mL) and maintained until the early stationary phase. The bacteriocin activity was completely inactivated by all the proteolytic enzymes such as pepsin, protease, proteinase K, papain, chymotrypsin, and trypsin, but the activity was not affected by catalase, a-amylase, lysozyme, and lipase, suggesting proteinaceous nature of the bacteriocin. Additionally, this activity was not affected in the pH range from 3.0 to 9.0 and under storage conditions like 30 days at -20,4, or $25^{\circ}C$. Although the bacteriocin activity was absolutely lost after 15 min treatment at 121, it was relatively stable at $70^{\circ}C$ for 60 min or $100^{\circ}C$ for 30 min. The activity was disappeared by treatment with acetone, benzene, ethanol, or methanol, but it was not affected by treatment with chloroform or hexane. The antibacterial activity of the bacteriocin was good against some LAB including Lactobacillus spp., Enterococcus spp., and Streptococcus spp., but not against food-borne pathogens such as Bacillus spp., Listeria spp., and Staphylococcus spp. as well as yeasts and molds. Especially, some intestinal bacteria such as Enterobacter aerogenes and E. coli were significantly affected by the bacteriocin of La, plantarum K11. Furthermore, the addition of 640 BU/mL resulted in the complete clearance of E. coli O157 after 10 hr.

• Proceedings of the Korea Society of Information Technology Applications Conference
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• 2006.06a
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• pp.185-207
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• 2006

산업단지는 지난 30년간 한국 산업의 성장을 이끌어 온 발전모형으로서 존재하여 왔으나, 최근 지식에 기초한 혁신창출형 경제체제가 국가 및 지역사회의 경쟁력을 위한 핵심요소로 부각되면서 '효율성' 측면에서 그 의미가 크게 퇴색되었다. 이를 위한 대안으로서 '클러스터'가 대두되어 다양한 분석연구가 수행되고 있으며, 정부와 지방자치단체들은 이를 바탕으로 각자의 특색에 맞는 클러스터 조성 정책을 펼치고 있다. 그러나, 기존의 연구들은 클러스터의 종류 및 발전단계에 관한 프레임워크 제시 등의 이론적 수준에 국한되어 있거나, 지역사례 연구를 통한 성공요인분석(CFS) 및 단순한 정책방향 제시 수준에 머물러 있는 한계를 보이고 있다. 본 연구는 '클러스터'에 관한 선행연구를 분석해 보고, 클러스터의 중요한 판단기준이 되는 군집도와 네트워크 연계 정도를 기준으로 한 '$2{\times}2$ 클러스터 전략격자모형'을 효과적인 클러스터 구축전략 수립을 위한 이론적 틀로서 제시하였다. 또한, 분석틀에 실질적인 사례로서 '충북지역의 SW산업'을 전략격자모형에 대응시켜 분석함으로써 전략격자의 유용성을 제시하였다. 이를 위해, 충북지역의 SW 공급업체와 수요업체를 대상으로 설문조사를 실시, 분석한 후 그 결과를 전략격자모형에 대응시켰다. 그 결과, 충북지역의 SW산업은 아직 산업단지 수준에 있는 것으로 분석되었고 충북의 SW산업의 충북 내의 수요만으로는 더 큰 성장이 어려운 것으로 분석, 지역 내에서의 수요창출을 목표로 하는 '단일 클러스터' 구축보다는 지역적 제약을 벗어난 '매가 클러스터'의 구축으로 지역 내외에서의 수요창출이 가능한 클러스터의 구축을 그 대안으로 제시하였다.${\alpha}$에 E. coli Jm109의 plasmid pBX19, pBR322를 전이시켰다. 6. L. lactis ssp. lactis 균주에 lysozyme 처리시 30${\sim}$80%의 생존율을 보였으며, 대부분의 L. acidophilus 균주의 경우 약 70%의 생존율을 보였다. L. casei 102S의 경우는 45분간 처리 시에도 100%의 생존율을 보였다. 8. L. lactis ssp. lactis 균주에 pLZ12를 6.0kV에서 전이시킨 결과 12.5kV에서보다 형질전환 효율이 훨씬 높았으며 lysozyme 처리에 의해 형질전환 효율이 증가되었다. 9. L. acidophilus 균주에 pLZ12를 전이시 6.0kV에서는 전이가 모두 이루어졌으나, 12.5kV에서는 L. acidophilus WIESBY와 NCFM에서 전이가 이루어지지 않았으며, lysozyme 처리 후 pLZ12를 전이시켰을 때 12kV보다 6.0kV에서 형질전환 효율이 증가되었다. 10. Gene Pulser와 Progenitor II를 사용하여 pLZ12를 L. lactis ssp. lactis 균주에 전이하였을 때 Gene Pulser에 비해 Progenitor II의 형질전환 효율이 현저히 떨어졌다. L. acidophilus HY7008과 HY7001은 두 기기 모두 형질전환이 이루어졌으나, L. acidophilus WEISBY와 NCFM은 Progeni-tor II에서 전이가 일어나지 않았으며, Gene Pulser에서 전이균주를 얻어 두 electroporator간에 형질전환 효율의 차이를 보였다. 11. L. casei 102S에 pLZ12

• Fisheries and Aquatic Sciences
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• v.20 no.6
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• pp.11.1-11.8
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• 2017

Juvenile black sea bream, Acanthopagrus schlegelii, were exposed to waterborne zinc (Zn) at concentrations of 0, 200, and $400{\mu}g/L$, at temperatures of 18 or $26^{\circ}C$ for 4 weeks. Superoxide dismutase (SOD) activities in the liver and gill of A. schlegelii significantly increased following exposure to waterborne Zn. Significant reduction in glutathione S-transferase (GST) activity in the liver and gill was observed following exposure to waterborne Zn. Glutathione (GSH) concentrations in the liver and gill also significantly decreased following exposure to waterborne Zn. Phagocytosis and lysozyme in the plasma and kidney were significantly increased following exposure to waterborne Zn. High water temperature increased alterations in the antioxidant and immune responses. The results of the present study suggest that waterborne Zn induced significant alterations in oxidative stress, increased immune responses and high temperature that trigger Zn toxicity.