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Establishment of the expression system of human HtrA2 in the zebrafish (Zebrafish 동물모델에서 human HtrA2의 expression system 정립에 관한 연구)

  • Cho, Sung-Won;Park, Hyo-Jin;Kim, Goo-Young;Nam, Min-Kyung;Kim, Ho-Young;Ko, In-Ho;Kim, Cheol-Hee;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.4
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    • pp.571-578
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    • 2006
  • HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

Polymorphisms and Allele Distribution of Novel Indel Markers in Jeju Black Cattle, Hanwoo and Imported Cattle Breeds (제주흑우, 한우 및 수입 소 품종에서 새로운 indel 마커의 다형성과 대립인자 분포)

  • Han, Sang-Hyun;Kim, Jae-Hwan;Cho, In-Cheol;Cho, Sang-Rae;Cho, Won-Mo;Kim, Sang-Geum;Kim, Yoo-Kyung;Kang, Yong-Jun;Park, Yong-Sang;Kim, Young-Hoon;Park, Se-Phil;Kim, Eun-Young;Lee, Sung-Soo;Ko, Moon-Suck
    • Journal of Life Science
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    • v.22 no.12
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    • pp.1644-1650
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    • 2012
  • The aim of this study was to screen the polymorphisms and distribution of each genotype of insertion/ deletion (indel) markers which were found in a preliminary comparative study of bovine genomic sequence databases. Comparative bioinformatic analyses were first performed between the nucleotide sequences of Bovine Genome Project and those of expressed sequence tag (EST) database, and a total of fifty-one species of indel markers were screened. Of these, forty-two indel markers were evaluated, and nine informative indel markers were ultimately selected for population analysis. Nucleotide sequences of each marker were re-sequenced and their polymorphic patterns were typed in six cattle breeds: Holstein, Angus, Charolais, Hereford, and two Korean native cattle breeds (Hanwoo and Jeju Black cattle). Cattle breeds tested in this study showed polymorphic patterns in eight indel markers but not in the Indel-15 marker in Charolais and Holstein. The results of analysis for Jeju Black cattle (JBC) population indicated an observed heterozygosity (Ho) that was highest in HW_G1 (0.600) and the lowest in Indel_29 (0.274). The PIC value was the highest in HW_G4 (0.373) and lowest in Indel_6 (0.305). These polymorphic indel markers will be useful in supplying genetic information for parentage tests and traceability and to develop a molecular breeding system for improvement of animal production in cattle breeds as well as in the JBC population.

A STUDY ON FRACTURAL BEHAVIOR OF DENTIN-RESIN INTERFACE (상아질-복합레진 접착계면의 파괴거동에 대한 연구)

  • Ryu, Gil-Joo;Choi, Gi-Woon;Park, Sang-Jin;Choi, Kyung-Kyu
    • Restorative Dentistry and Endodontics
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    • v.32 no.3
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    • pp.208-221
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    • 2007
  • The fracture toughness test is believed as a clinically relevant method for assessing the fracture resistance of the dentinal restoratives. The objectives of this study were to measure the fracture toughness $(K_{1C})$ and microtensile bond strength of dentin-resin composite interface and compare their relationship for their use in evaluation of the integrity of the dentin-resin bond. A minimum of six short-rod specimens for fracture toughness test and fifteen specimens for microtensile bond strength test was fabricated for each group of materials used. After all specimens storing for 24 hours in distilled water at $37^{\circ}C$, they were tensile-loaded with an EZ tester universal testing machin. Statistical analysis was performed using ANOVA and Tukey's test at the 95% confidence level, Pearson's coefficient was used to verify the correlation between the mean of fracture toughness and microtensile bond strength. FE-SEM was employed on fractured surface to describe the crack propagation. Fracture toughness value of Clearfil SE Bond (SE) was the highest, followed by Adper Single Bond 2 (SB), OptiBond Solo (OB), ONE-STEP PLUS (0S), ScotchBond Multi-purpose (SM) and there was significant difference between SE and other 4 groups (p < 0.05). There were, however, no significant difference among SB, OB, OS, SM (p > 0.05). Microtensile bond strength of SE was the highest, followed by SB, OB, SM, OS and OS only showed significant lower value (p < 0.05). There was no correlation between fracture toughness and microtensile bond strength values. FE-SEM examination revealed that dentin bonding agent showed different film thickness and different failure pattern according to the film thickness. From the limited results of this study, it was noted that there was statistically no correlation between K1C and ${\mu}TBS$. We can conclude that for obtaining the reliability of bond strength test of dentin bonding agent, we must pay more attention to the test procedure and its profound scrutiny.

CHANGES IN μ-TBS TO PULP CHAMBER DENTIN AFTER THE APPLICATION OF NAOCL & REVERSAL EFFECT BY USING SODIUM ASCORBATE (NaOCl의 적용 후 치수강 상아질에 대한 결합강도의 변화와 Sodium Ascorbate에 의한 환원 효과)

  • Kwon, Su-Mi;Kim, Tae-Gun;Yu, Mi-Kyung;Lee, Kwang-Won
    • Restorative Dentistry and Endodontics
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    • v.34 no.6
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    • pp.515-525
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    • 2009
  • Clinical suggestion for the limitation of application time of NaOCl solution is needed to avoid large reductions in resin-dentin bond strength. The aim of this study was to measure the change of ${\mu}$-tensile bond strength after the various application time of 5.25% NaOCl solution to pulp chamber dentin in endodontic access cavity, and to evaluate the effect of 10% sodium ascorbate application for 10 min on bond strength after the treatment of 5.25% NaOCl solution. In this experiment, there were no statistical differences(p > 0.05) in bond strengths between upper chamber dentin and lower chamber dentin. NaOCl-treated group for 20 min did not show any significant decrease(p > 0.05) in bond strength than non-treated control group. In contrast to that, bond strengths of NaOCl-treated groups for 40 & 80 min were significantly lower(p < 0.05) than that of non-treated control group. 10% sodium ascorbate retreated group for 10 min after 5.25% NaOCl application for 40 min to chamber dentin showed the recovery of bond strength significantly. However, the bond strength of sodium ascorbate retreated group after 5.25% NaOCl application for 80 min was still significantly lower(p < 0.05) compared to the non-treated control group, which means the reductions in resin-dentin bond strength were not fully reversed. On the contrary, sodium ascorbate retreated group after 5.25% NaOCl application for 5 min showed significantly higher(p < 0.05) bond strength compared to the control group, which demonstrates its superior recovery effect. In SEM exminations of specimens retreated with 10% sodium ascorbate after NaOCl application for 40 & 80 min showed that resin tags were formed clearly and densely, but weakly in density and homogeneity of individual resin tag compared to the control specimen.

Case Study on the Enterprise Microblog Usage: Focusing on Knowledge Management Strategy (기업용 마이크로블로그의 사용행태에 대한 사례연구: 지식경영전략을 중심으로)

  • Kang, Min Su;Park, Arum;Lee, Kyoung-Jun
    • Journal of Intelligence and Information Systems
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    • v.21 no.1
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    • pp.47-63
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    • 2015
  • As knowledge is paid attention as a new production factor that generates added value, studies continue to apply knowledge management to business environment. In addition, as ICT (Information Communication Technology) was engrafted in business environment, it leads to increasing task efficiency and productivity of individual workers. Accordingly, the way that a business achieves its goal has changed to one in which its individual members are willing to take part in the organization and share information to create new values (Han, 2003) and studies for the system and service to support such transition are carrying out. Of late, a new concept called 'Enterprise 2.0' newly appears. It is the extension of Wen 2.0 and its technology, which focus on participation, sharing and openness, to the work environment of a business (Jung, 2013). Enterprise 2.0 is being used as a collaborative tool to prop up individual creativity and group brain power by combining Web 2.0 technologies such as blog, Wiki, RSS and tag with business software (McAfee, 2006). As Tweeter gets popular, Enterprise Microblog (EMB), which is an example of Enterprise 2.0 for business, has been developed as equivalent to Tweeter in business circle and SaaS (Software as a Service) such as Yammer was introduced The studies of EMB mainly focus on demonstrating its usability in terms of intra-firm communication and knowledge management. However existing studies lean too much towards large-sized companies and certain departments, rather than a company as a whole. Therefore, few studies have been conducted on small and medium-sized companies that have difficulty preparing separate resources and supplying exclusive workforce to introduce knowledge management. In this respect, the present study placed its analytic focus on small-sized companies actually equipped with EMB to know how they use it. And, based on the findings, this study examined their knowledge management strategies for EMB from the point of codification and personalization. Hypothesis -"as a company grows, it shifts EMB strategy from codification to personalization'- was established on the basis of reviewing precedent studies and literature. To demonstrate the hypothesis, this study analyzed the usage of EMB by small companies that have used it from foundation. For case study, the duration of the use was divided into 2 spans and longitudinal analysis was employed to examine the contents of the blogs. Using the key findings of the analysis, this study is aimed to propose practical implications for the operation of knowledge management of small-sized company and the suitable application of knowledge management system for operation Knowledge Management Strategy can be classified by codification strategy and personalization strategy (Hansen et. al., 1999), and how to manage the two strategies were always studied. Also, current studies regarding the knowledge management strategy were targeted mostly for major companies, resulting in lack of studies in how it can be applied on SMEs. This research, with the knowledge management strategy suited for SMEs, sets an Enterprise Microblog (EMB), and with the EMB applied on SMEs' Knowledge Management Strategy, it is reviewed on the perspective of SMEs' Codification and Personalization Strategies. Through the advanced research regarding Knowledge Management Strategy and EMB, the hypothesis is set that "Depending on the development of the company, the main application of EMB alters from Codification Strategy to Personalization Strategy". To check the hypothesis, SME that have used the EMB called 'Yammer' was analyzed from the date of their foundation until today. The case study has implemented longitudinal analysis which divides the period when the EMBs were used into three stages and analyzes the contents. As the result of the study, this suggests a substantial implication regarding the application of Knowledge Management Strategy and its Knowledge Management System that is suitable for SME.

In vivo Radioprotective Effects of Basic Fibroblast Growth Factor in C3H Mice (Basic Fibroblast Growth Factor (bFGF)의 방사선보호작용에 대한 실험적 연구)

  • Kim, Yeon-Shil;Yoon, Sei-Chul
    • Radiation Oncology Journal
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    • v.20 no.3
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    • pp.253-263
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    • 2002
  • Purpose : In order to understand in vivo radiation damage modifying of bFGF on jejunal mucosa, bone marrow and the effect of bFGF on the growth of transplanted mouse sarcoma 180 tumor in mice. Materials and Methods : Mice were treated with $6\;{\mu}g$ of bFGF at 24 hours and 4 hours before exposing to 600 cGy, 800 cGy and 1,000 cGy total body irradiation (TBI), and then exposed to 3,000 cGy local radiation therapy on the tumor bearing thigh. Survival and tumor growth curve were plotted in radiation alone group and combined group of bFGF and irradiation (RT). Histologic examination was performed in another experimental group. Experimental groups consisted of normal control, tumor control, RT (radiation therapy) alone, $6\;{\mu}g$ bFGF alone, combined group of $3\;{\mu}g$ bFGF and irradiation (RT), combined group of $6\;{\mu}g$ bFGF and irradiation (RT). Histologic examination was peformed with H-E staining in marrow, jejunal mucosa, lung and sarcoma 180 bearing tumor. Radiation induced apoptosis was determined in each group with the DNA terminal transferase nick-end labeling method ($ApopTag^{\circledR}$ S7100-kit, Intergen Co.) Results : The results were as follows 1) $6\;{\mu}g$ bFGF given before TBI significantly improved the survival of lethally irradiated mice. bFGF would protect against lethal bone marrow syndrome. 2) $6\;{\mu}g$ bFGF treated group showed a significant higher crypt depth and microvilli length than RT alone group (p<0.05). 3) The bone marrow of bFGF treated group showed less hypocellularity than radiation alone group on day 7 and 14 after TBI (p<0.05), and this protective effect was more evident in $6\;{\mu}g$ bFGF treated group than that of $3\;{\mu}g$ bFGF treated group. 4) bFGF protected against early radiation induced apoptosis in intestinal crypt cell but might have had no antiapoptotic effect in bone marrow stem cell and pulmonary endothelial cells. 5) There was no significant differences in tumor growth rate between tumor control and bFGF alone groups (p>0.05). 6) There were no significant differences in histopathologic findings of lung and mouse sarcoma 180 tumor between radiation alone group and bFGF treated group. Conclusions : Our results suggest that bFGF protects small bowel and bone marrow from acute radiation damage without promoting the inoculated tumor growth in C3H mice. Improved recovery of early responding normal tissue and reduced number of radiation induced apoptosis may be possible mechanism of radioprotective effect of bFGF.

Clinical Feature and Dietary Pattern of Infantile Constipation Under Two Years of Age (2세 이하 영유아 변비의 임상 양상과 식이 방식)

  • Kim, Hye-Jin;Moon, Jin Soo;Hwang, Jong Hee;Jang, Hyun Oh;Nam, Seung Yeun;Kim, Dong Wook;Lee, Chong Guk
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.1
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    • pp.31-38
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    • 2006
  • Purpose: Infantile constipation is one of the most common problems in pediatric gastrointestinal outpatient clinic. We planed to show the clinical feature of infantile constipation and explore the possible relationship between diets and symptoms of constipation. Methods: We analyzed the medical records and telephone questionnaire about infants under 2 years of age with constipation, who visited outpatient clinic of Department of Pediatrics, Ilsan Paik Hospital during the time from March 2002 to February 2005. Data including the symptoms and signs of constipation, diet history, and past-medical history were analyzed. Results: Total 96 infants, 40 male and 56 female, were enrolled in this study. The mean age was 9 months. Twenty-three infants were exclusively breast milk fed (BMF), 20 infants were cow's milk fed (CMF) and 53 infants had a history of mixed feeding with cow's milk and breast milk (MMF). Patients showed painful defecation (95.8%), abdominal distension (53.1%), palpable rectal stool (35.1%), hard stool (30.2%), blood-tinged stool (29.2%) and anal fissure (16.7%). Patients with exclusive cow milk feeding had prominent clinical features, such as anal fissure (p=0.03), hematochezia (p=0.04) and palpable rectal stool (p=0.025). Patients who had a history of larger intake of liquid food had a tendency to get anal skin tag (p<0.05). Conclusion: Exclusive breast milk feeding seemed important to avoid constipation with clinical significance. To educate caregivers in appropriate way of the weaning food may help the infants with constipation.

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The Need for Paradigm Shift in Semantic Similarity and Semantic Relatedness : From Cognitive Semantics Perspective (의미간의 유사도 연구의 패러다임 변화의 필요성-인지 의미론적 관점에서의 고찰)

  • Choi, Youngseok;Park, Jinsoo
    • Journal of Intelligence and Information Systems
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    • v.19 no.1
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    • pp.111-123
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    • 2013
  • Semantic similarity/relatedness measure between two concepts plays an important role in research on system integration and database integration. Moreover, current research on keyword recommendation or tag clustering strongly depends on this kind of semantic measure. For this reason, many researchers in various fields including computer science and computational linguistics have tried to improve methods to calculating semantic similarity/relatedness measure. This study of similarity between concepts is meant to discover how a computational process can model the action of a human to determine the relationship between two concepts. Most research on calculating semantic similarity usually uses ready-made reference knowledge such as semantic network and dictionary to measure concept similarity. The topological method is used to calculated relatedness or similarity between concepts based on various forms of a semantic network including a hierarchical taxonomy. This approach assumes that the semantic network reflects the human knowledge well. The nodes in a network represent concepts, and way to measure the conceptual similarity between two nodes are also regarded as ways to determine the conceptual similarity of two words(i.e,. two nodes in a network). Topological method can be categorized as node-based or edge-based, which are also called the information content approach and the conceptual distance approach, respectively. The node-based approach is used to calculate similarity between concepts based on how much information the two concepts share in terms of a semantic network or taxonomy while edge-based approach estimates the distance between the nodes that correspond to the concepts being compared. Both of two approaches have assumed that the semantic network is static. That means topological approach has not considered the change of semantic relation between concepts in semantic network. However, as information communication technologies make advantage in sharing knowledge among people, semantic relation between concepts in semantic network may change. To explain the change in semantic relation, we adopt the cognitive semantics. The basic assumption of cognitive semantics is that humans judge the semantic relation based on their cognition and understanding of concepts. This cognition and understanding is called 'World Knowledge.' World knowledge can be categorized as personal knowledge and cultural knowledge. Personal knowledge means the knowledge from personal experience. Everyone can have different Personal Knowledge of same concept. Cultural Knowledge is the knowledge shared by people who are living in the same culture or using the same language. People in the same culture have common understanding of specific concepts. Cultural knowledge can be the starting point of discussion about the change of semantic relation. If the culture shared by people changes for some reasons, the human's cultural knowledge may also change. Today's society and culture are changing at a past face, and the change of cultural knowledge is not negligible issues in the research on semantic relationship between concepts. In this paper, we propose the future directions of research on semantic similarity. In other words, we discuss that how the research on semantic similarity can reflect the change of semantic relation caused by the change of cultural knowledge. We suggest three direction of future research on semantic similarity. First, the research should include the versioning and update methodology for semantic network. Second, semantic network which is dynamically generated can be used for the calculation of semantic similarity between concepts. If the researcher can develop the methodology to extract the semantic network from given knowledge base in real time, this approach can solve many problems related to the change of semantic relation. Third, the statistical approach based on corpus analysis can be an alternative for the method using semantic network. We believe that these proposed research direction can be the milestone of the research on semantic relation.

A Case Study on Universal Dependency Tagsets (다국어 범용 의존관계 주석체계(Universal Dependencies) 적용 연구 - 한국어와 일본어의 비교를 중심으로)

  • Han, Jiyoon;Lee, Jin;Lee, Chanyoung;Kim, Hansaem
    • Cross-Cultural Studies
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    • v.53
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    • pp.163-192
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    • 2018
  • The purpose of this paper was to examine universal dependency UD application cases of Korean and Japanese with similar morphological characteristics. In addition, UD application and improvement methods of Korean were examined through comparative analysis. Korean and Japanese are very well developed due to their agglutinative characteristics. Therefore, there are many difficulties to apply UD which is built around English refraction. We examined the application of UPOS and DEPREL as components of UD with discussions. In UPOS, we looked at category problem related to narrative such as AUX, ADJ, and VERB, We examined how to handle units. In relation to the DEPREL annotation system, we discussed how to reflect syntactic problem from the basic unit annotation of syntax tags. We investigated problems of case and aux arising from the problem of setting dominant position from Korean and Japanese as the dominant language. We also investigated problems of annotation of parallel structure and setting of annotation basic unit. Among various relation annotation tags, case and aux are discussed because they show the most noticeable difference in distribution when comparing annotation tag application patterns with Korean. The case is related to both Korean and Japanese surveys. Aux is a secondary verb in Korean and an auxiliary verb in Japanese. As a result of examining specific annotation patterns, it was found that Japanese aux not only assigned auxiliary clauses, but also auxiliary elements to add the grammatical meaning to the verb and form corresponding to the end of Korean. In UD annotation of Japanese, the basic unit of morphological analysis is defined as a unit of basic syntactic annotation in Japanese UD annotation. Thus, when using information, it is necessary to consider how to use morphological analysis unit as information of dependency annotation in Korean.

Expression and Purification of Three Lipases (LipAD1, LipAD2, and LipAD3) and a Lipase Chaperone (LipBD) from Acinetobacter schindleri DYL129 (Acinetobacter schindleri DYL129 유래의 3개 lipases와 chaperone의 발현과 정제)

  • Kim, Sun-Hee;Lee, Yong-Suk;Jeong, Hae-Rin;Pyeon, Hyo-Min;You, Ju-Soon;Choi, Yong-Lark
    • Journal of Life Science
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    • v.29 no.4
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    • pp.492-498
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    • 2019
  • Previously, three kinds of lipases, lipAD1, lipAD2, and lipAD3, and lipase chaperone, lipBD, of Acinetobacter schindleri DYL129 isolated from soil sample were reported. In this report, three lipase and lipase chaperone were cloned into the pET32a(+) or pGEX-6P-1 vectors for protein expression in Escherichia coli, and named as pETLAD1, pETLAD2, pETLAD3 and pETLBD or pGEXLAD1, pGEXLAD 2, pGEXLAD3 and pGEXLBD, respectively. Protein expression rate was higher in pET system than in pGEX system. Although LipAD1 and LipAD2 were produced as inclusion bodies, their expression levels were high. So LipAD1 and LipAD2 could be solubilized in 8 M urea buffer and purified. LipAD3 and LipBD were overexpressed in soluble form and purified. Those proteins were purified by His-tag affinity chromatography connected in AKTA prime system. The activities of the purified lipases were demonstrated with 1% tributyrin agar plate. After purification, molecular mass was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis. LipAD1 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl butyrate, LipAD2 showed high activity toward ${\rho}$-nitrophenyl acetate and ${\rho}$-nitrophenyl myristate, and LipAD3 showed high activity toward ${\rho}$-nitrophenyl acetate, ${\rho}$-nitrophenyl butyrate, and ${\rho}$-nitrophenyl miristate, respectively. Three lipases, LipAD1, LipAD2, and LipAD3, showed optimal reaction at $50^{\circ}C$ using ${\rho}$-nitrophenyl butyrate, as substrate.