• Title/Summary/Keyword: optimal pathway

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Analysis of Influence of Environmental Conditions on Ganoderic Acid Content: in Ganoderma lucidum Using Orthogonal Design

  • Li Na;Liu Xiao Hua;Zhou Jie;Li Yu Xiang;Zhao Ming Wen
    • Journal of Microbiology and Biotechnology
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    • v.16 no.12
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    • pp.1940-1946
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    • 2006
  • The influence of environmental conditions on the ganoderic acid (GA) content in the fungus Ganoderma lucidum was investigated using a one-factor-at-a-time design and orthogonal design. Among the various medium components examined, sucrose, soybean powder or peptone, ferrous sulfate, and pH 6.0 were the most suitable carbon source (factor A), nitrogen source (factor B), mineral source (factor C), and initial pH (factor D), respectively, for the GA content in the one-factor-at-a-time design. According to the orthogonal design, the order of effect for the four factors on the GA content was A>C>D>B. The best level of factor A was $A_2$ (sucrose) with a value of +0.34 mg/100 mg DW. The optimal treatment combination was $A_2B_1C_3D_1$ with which the GA content reached up to 2.63$\pm$0.011 mg/100 mg DW. The interactions between the mineral ion and the nitrogen source, and the mineral ion and the pH were both highly significant (P<0.01). The highest interaction effect was ($B_2{\times}D_2$) with a value of +0.19 mg/100 mg DW, which was higher than the level effect value for $B_2$ (peptone) and D$_2$ (pH 5.0). Therefore, the results proved that interactions between factors cannot be ignored. The results also indicated the importance of the interactions between the factors, which may help to understand the metabolic pathway leading to triterpene biosynthesis and the expression and regulation of the key enzymes involved.

Purification and Properties of a Novel ${\beta}$-Glucosidase, Hydrolyzing Ginsenoside Rb1 to CK, from Paecilomyces Bainier

  • Yan, Qin;Zhou, Xin-Wen;Zhou, Wei;Li, Xing-Wei;Feng, Mei-Qing;Zhou, Pei
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1081-1089
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    • 2008
  • A novel ginsenoside-hydrolyzing ${\beta}$-glucosidase was purified from Paecilomyces Bainier sp. 229 by a combination of Q-Sepharose FF, phenyl-Sepharose CL-4B, and CHT ceramic hydroxyapatite column chromatography. The purified enzyme was a monomeric protein with a molecular mass estimated to be 115 kDa. The optimal enzyme activity was observed at pH 3.5 and $60^{\circ}C$. It was highly stable within pH 3-9 and at temperatures lower than $55^{\circ}C$. The enzyme was specific to ${\beta}$-glucoside. The order of enzyme activities against different types of ${\beta}$-glucosidic linkages was ${\beta}$-(1-6)>${\beta}$-(1-2)>${\beta}$-(1-4). The enzyme converted ginsenoside Rb1 to CK specifically and efficiently. An 84.3% amount of ginsenoside Rb1, with an initial concentration of 2 mM, was converted into CK in 24 h by the enzyme at $45^{\circ}C$ and pH 3.5. The hydrolysis pathway of ginsenoside Rb1 by the enzyme was $Rb1{\to}Rd{\to}F2{\to}CK$. Five tryptic peptide fragments of the enzyme were identified by a newly developed de novo sequencing method of post-source decay (PSD) matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. By comparing the five identified peptide sequences with the NCBI database, this purified ${\beta}$-glucosidase proves to be a new protein that has not been reported before.

Gamma Knife Radiosurgery for Juxtasellar Tumors (터어키안 주변종양에 대한 감마나이프 방사선 수술)

  • Chang, Jong Hee;Chang, Jin Woo;Park, Yong Gou;Chung, Sang Sup
    • Journal of Korean Neurosurgical Society
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    • v.29 no.10
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    • pp.1345-1351
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    • 2000
  • Objective : Around the sellar area, there are many important structures. But, the optimal radiation dosage for minimal toxicity to surrounding neural tissue has not been firmly established. The purpose of this study is to evaluate the radiosurgical outcome of juxtasellar tumors and to investigate the relationship between radiation dosage and toxicity to neural tissue. Method : Between May 1992 and June 2000, we treated 65 juxtasellar tumors by using the Leksell Gamma Knife. Among them, 52 patients who could be followed more than 1 year were included in this study. The radiosurgical dosage to the optic pathway, cavernous sinus, Meckel's cave, hypothalamus, pituitary gland and stalk, and brain stem was analyzed and correlated with clinical outcome. The mean follow-up period was 33.5 months(range 12.2- 99.0 months). Result : The clinical response rate was 69.2%. The volume response rate was 61.0% and the radiologic control rate was 92.7%. There were 4 complications(7.7%) of 2 trigeminal neuropathy, 1 abducens nerve palsy, and 1 trigeminal and transient abducens nerve palsy. The optic apparatus appeared to tolerate doses greater than 10Gy. The risk of cranial nerve complications in cavernous sinus seemed to be related to doses of more than 16Gy. In 3 of 4 patients who received more than 16Gy to cavernous sinus, the abducens or trigeminal neuropathy occurred. Also, one patient who received more than 15Gy to the Meckel's cave, trigeminal neuropathy developed. The hypothalamus, pituitary gland and stalk, and brain stem were relatively tolerable to radiation. Conclusion : Gamma Knife radiosurgery seems to be an effective method to control the growth of juxtasellar tumors. To avoid injury to surrounding important neural tissue, careful dose planning and further study for radiation toxicity to neural tissue were needed.

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Gene Cloning, Expression, and Characterization of a New Carboxylesterase from Serratia sp. SES-01: Comparison with Escherichia coli BioHe Enzyme

  • Kwon, Min-A;Kim, Hyun-Suk;Oh, Joon-Young;Song, Bong-Keun;Song, Jae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.147-154
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    • 2009
  • The carboxylesterase-encoding gene(bioHs) of a newly isolated strain, Serratia sp. SES-01, was cloned from the genomic DNA library by detecting formation of transparent halo around the colony on LB-tributyrin agar plates. The amino acid sequence of BioHs was highly similar to the members of the BioH enzyme family involved in the biotin biosynthetic pathway; it showed the highest similarity(91%) with that of Serratia proteamaculans. To compare BioHs with other BioH enzymes, the relatively well-known bioHe gene of E. coli was cloned with PCR. After we achieved high-level expression of soluble BioHs and BioHe through the exploration of different culture conditions, the purified BioHs and BioHe enzymes were characterized in terms of specificity, activity, and stability. BioHe was generally more robust to a change in temperature and pH and an addition of organic solvents than BioHs. The two enzymes exhibited a strong preference for carboxylesterase rather than for thioesterase and were optimal at relatively low temperatures($20-40^{\circ}C$) and alkaline pHs(7.5-9.0). The results in this study strongly suggested that both the BioHs and BioHe enzymes would be potential candidates for use as a carboxylesterase in many industrial applications.

A Variational Inequality-based Walkability Assessment Model for Measuring Improvement Effect of Transit Oriented Development (TOD) (대중교통중심개발(TOD) 개선효과 진단을 위한 변동부등식기반 보행네트워크 평가모형)

  • Sohn, Jhieon
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.36 no.2
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    • pp.259-268
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    • 2016
  • The core strategy of transit oriented development (TOD) is to promote high density mixed land use around railway stations. Case studies in advanced countries show that provision of policies for comprehensive maintenance of pedestrian facilities around railway station spheres is being pursued with efficacy. In spite of the importance placed on integrated pedestrian maintenance, domestic construction of integrated pedestrian infrastructure around railway station spheres lacks direction. Thus, there is a clear need for an evaluation standard that can provide the foundation for judgments on TOD improvement. This research proposes a network model that consolidates the interior of the station as well as its surrounding areas to determine the ease of pedestrian flow for effective TOD evaluation. The model considers the railway station and surrounding areas as an assembled network of pedestrian flow. The path chosen by the pedestrian is defined as the optimal degree of inconvenience, and expands on Wardrop's User Equilibrium (1952). To assess the various circumstances that arise on pedestrian facilities including congestion of the pedestrian pathway, constrained elevator capacity, and wait at the crosswalk, a variational inequality based pedestrian equilibrium distribution model is introduced.

Effect of Phorbol 12-Myristate 13-Acetate on the Differentiation of Adipose-Derived Stromal Cells from Different Subcutaneous Adipose Tissue Depots

  • Song, Jennifer K.;Lee, Chang Hoon;Hwang, So-Min;Joo, Bo Sun;Lee, Sun Young;Jung, Jin Sup
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.4
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    • pp.289-296
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    • 2014
  • Human adipose-tissue-derived stromal cells (hADSCs) are abundant in adipose tissue and can differentiate into multi-lineage cell types, including adipocytes, osteoblasts, and chondrocytes. In order to define the optimal harvest site of adipose tissue harvest site, we solated hADSCs from different subcutaneous sites (upper abdomen, lower abdomen, and thigh) and compared their proliferation and potential to differentiate into adipocytes and osteoblasts. In addition, this study examined the effect of phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, on proliferation and differentiation of hADSCs to adipocytes or osteoblasts. hADSCs isolated from different subcutaneous depots have a similar growth rate. Fluorescence-activated cell sorting (FACS) analysis showed that the expression levels of CD73 and CD90 were similar between hADSCs from abdomen and thigh regions. However, the expression of CD105 was lower in hADSCs from the thigh than in those from the abdomen. Although the adipogenic differentiation potential of hADSCs from both tissue regions was similar, the osteogenic differentiation potential of hADSCs from the thigh was greater than that of hADSCs from the abdomen. Phorbol 12-myristate 13-acetate (PMA) treatment increased osteogenic differentiation and suppressed adipogenic differentiation of all hADSCs without affecting their growth rate and the treatment of Go6983, a general inhibitor of protein kinase C (PKC) blocked the PMA effect. These findings indicate that the thigh region might be a suitable source of hADSCs for bone regeneration and that the PKC signaling pathway may be involved in the adipogenic and osteogenic differentiation of hADSCs.

Exploring the Effects of Carbon Sources on the Metabolic Capacity for Shikimic Acid Production in Escherichia coli Using In Silico Metabolic Predictions

  • Ahn, Jung-Oh;Lee, Hong-Weon;Saha, Rajib;Park, Myong-Soo;Jung, Joon-Ki;Lee, Dong-Yup
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1773-1784
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    • 2008
  • Effects of various industrially important carbon sources (glucose, sucrose, xylose, gluconate, and glycerol) on shikimic acid (SA) biosynthesis in Escherichia coli were investigated to gain new insight into the metabolic capability for overproducing SA. At the outset, constraints-based flux analysis using the genome-scale in silico model of E. coli was conducted to quantify the theoretical maximum SA yield. The corresponding flux distributions fueled by different carbon sources under investigation were compared with respect to theoretical yield and energy utilization, thereby identifying the indispensable pathways for achieving optimal SA production on each carbon source. Subsequently, a shikimate-kinase-deficient E. coli mutant was developed by blocking the aromatic amino acid pathway, and the production of SA on various carbon sources was experimentally examined during 51 batch culture. As a result, the highest production rate, 1.92 mmol SA/h, was obtained when glucose was utilized as a carbon source, whereas the efficient SA production from glycerol was obtained with the highest yield, 0.21 mol SA formed per mol carbon atom of carbon source consumed. The current strain can be further improved to satisfy the theoretically achievable SA production that was predicted by in silico analysis.

Cloning, Expression, and Characterization of UDP-glucose Pyrophosphorylase from Sphingomonas chungbukensis DJ77

  • Yoon, Moon-Young;Lee, Kyoung-Jin;Park, Hea-Chul;Park, Sung-Ha;Kim, Sang-Gon;Kim, Sung-Kun;Choi, Jung-Do
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1360-1364
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    • 2009
  • The bacterium Sphingomonas chungbukensis DJ77 produces the extracellular polysaccharide gellan in high yield. Gellan produced by this bacterium is widely used as a gelling agent, and the enzyme UDP-glucose pyrophosphorylase (UGP) is thought to play a key role in the gellan biosynthetic pathway. The UGP gene has been successfully cloned and over-expressed in E. coli. The expressed enzyme was purified with a molecular weight of approximately 32 kDa, as determined by a SDS-polyacrylamide gel, but the enzyme appears as ca. 63 kDa on a native gel, suggesting that the enzyme is present in a homodimer. Kinetic analysis of UDP-glucose for UGP indicates $K_m$ = 1.14 mM and $V_{max}$ = 10.09 mM/min/mg at pH 8.0, which was determined to be the optimal pH for UGP catalytic activity. Amino acid sequence alignment against other bacteria suggests that the UGP contains two conserved domains: An activator binding site and a glucose-1-phosphate binding site. Site-directed mutagenesis of Lys194, located within the glucose-1-phosphate binding site, indicates that substitution of the charge-reversible residue Asp for Lys194 dramatically impairs the UGP activity, supporting the hypothesis that Lys194 plays a critical role in the catalysis.

The Management and Security Plans of a Separated Virtualization Infringement Type Learning Database Using VM (Virtual Machine) (VM(Virtual Machine) 을 이용한 분리된 가상화 침해유형 학습 데이터베이스 관리와 보안방안)

  • Seo, Woo-Seok;Jun, Moon-Seog
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.36 no.8B
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    • pp.947-953
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    • 2011
  • These days, a consistent and fatal attack attribute toward a database has proportionally evolved in the similar development form to that of security policy. Because of access control-based defensive techniques regarding information created in closed networks and attacks on a limited access pathway, cases of infringement of many systems and databases based on accumulated and learned attack patterns from the past are increasing. Therefore, the paper aims to separate attack information by its types based on a virtual infringement pattern system loaded with dualistic VM in order to ensure stability to limited certification and authority to access, to propose a system that blocks infringement through the intensive management of infringement pattern concerning attack networks, and to improve the mechanism for implementing a test that defends the final database, the optimal defensive techniques, and the security policies, through research.

A Study on the Evaluation and Improvement of Student Convenient Facilities at University Campuses, based on Universal Design Concept - Focused on the university campuses in Texas, U.S.A. - (유니버설디자인 개념에 의거한 대학내 학생편의시설 평가 및 개선방향에 대한 연구 - 미국 텍사스주를 중심으로 -)

  • Kim, Won-Pil
    • Journal of the Korean Institute of Educational Facilities
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    • v.21 no.3
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    • pp.19-28
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    • 2014
  • Student halls and their convenient facilities have been a focal point for various student activities at university campus. It has been for most of the student a place of unique memories and of attachment often associated with those good and bad school days. However, it is questionable whether these facilities are supportive and accessible for all of the students and other users including the handicapped. Therefore, based on the concept of UD(universal design) which was widely applied to U.S. institutions, this study intended to evaluate student hall facilities in U.S. and to provide an improvement direction for Korean UD application. For that purpose, four universities were selected for an in-depth analysis and 76 survey indices were utilized based on previous research. It was found that (1) design consideration without braille sign for VIP(visually impaired person) in student buildings can be differently approached with electronic devices; (2) the best demonstration of UD in student buildings can be seen in spacious flat pathway, easy access through ramp and wide entry area, necessary for people in wheel-chairs, but used by all, implying an increase of the ratio of public space; (3) one of the good UD features is an attractive physical environment rather than institutional appearance, in which they ultimately will support and completely adaptable at optimal levels by everyone; (4) consistent maintenance and management maximize the potential of UD principles and minimize physical limitations.