• Title/Summary/Keyword: aprE

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An Economic Assessment for APR+ Standard Detailed Design Developing Phase (APR+ 표준상세설계 개발단계에서의 경제성 평가)

  • Ha, Gak-Hyeon;Suh, Yong-Pyo;Kim, Man-Won;Kim, Sung-Choon;Park, Sun-Eung
    • Journal of Energy Engineering
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    • v.21 no.3
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    • pp.292-300
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    • 2012
  • KHNP CRI has been developing APR+ nuclear power plant since 2007, which is GEN III+ model with 4,361 MWth capacity. To develop safer and more economical nuclear power plant than APR1400, we studied domestic and foreign nuclear power plants under construction. We also reviewed nuclear power plants which are appropriate for domestic construction in Korea and also for export. Economic assessments were made twice during the second phase of standard detailed design of the plant. The result of the second phase of economic analysis for APR+ standard detailed design showed that APR+ N-th plant was 24.6% more economical than coal-fired 1,000MW power plant, and was evaluated to be competitive enough in global market for construction of the nuclear power plant.

Characterization of AprE176, a Fibrinolytic Enzyme from Bacillus subtilis HK176

  • Jeong, Seon-Ju;Heo, Kyeong;Park, Ji Yeong;Lee, Kang Wook;Park, Jae-Yong;Joo, Sang Hoon;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.89-97
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    • 2015
  • Bacillus subtilis HK176 with high fibrinolytic activity was isolated from cheonggukjang, a Korean fermented soyfood. A gene, aprE176, encoding the major fibrinolytic enzyme was cloned from B. subtilis HK176 and overexpressed in E. coli BL21(DE3) using plasmid pET26b(+). The specific activity of purified AprE176 was 216.8 ± 5.4 plasmin unit/mg protein and the optimum pH and temperature were pH 8.0 and 40℃, respectively. Error-prone PCR was performed for aprE176, and the PCR products were introduced into E. coli BL21(DE3) after ligation with pET26b(+). Mutants showing enhanced fibrinolytic activities were screened first using skim-milk plates and then fibrin plates. Among the mutants, M179 showed the highest activity on a fibrin plate and it had one amino acid substitution (A176T). The specific activity of M179 was 2.2-fold higher than that of the wild-type enzyme, but the catalytic efficiency (kcat/Km) of M179 was not different from the wild-type enzyme owing to reduced substrate affinity. Interestingly, M179 showed increased thermostability. M179 retained 36% of activity after 5 h at 45℃, whereas AprE176 retained only 11%. Molecular modeling analysis suggested that the 176th residue of M179, threonine, was located near the cation-binding site compared with the wild type. This probably caused tight binding of M179 with Ca2+, whichincreased the thermostability of M179.

Cloning of Fibrinolytic Enzyme Gene from Bacillus subtilis Isolated from Cheonggukjang and Its Expression in Protease-deficient Bacillus subtilis Strains

  • Jeong, Seon-Ju;Kwon, Gun-Hee;Chun, Ji-Yeon;Kim, Jong-Sang;Park, Cheon-Seok;Kwon, Dae-Young;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.1018-1023
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    • 2007
  • Bacillus subtilis CH3-5 was isolated from cheonggukjang prepared according to traditional methods. CH3-5 secreted at least four different fibrinolytic proteases (63, 47, 29, and 20 kDa) into the culture medium. A fibrinolytic enzyme gene, aprE2, encoding a 29kDa enzyme was cloned from the genomic DNA of CH3-5, and the DNA sequence determined. aprE2 was overexpressed in heterologous B. subtilis strains deficient in extracellular proteases using a E. coli-Bacillus shuttle vector. A 29 kDa AprE2 band was observed and AprE2 seemed to exhibit higher activities towards fibrin rather than casein.

Characterization of a Fibrinolytic Enzyme Secreted by Bacillus amyloliquefaciens CB1 and Its Gene Cloning

  • Heo, Kyeong;Cho, Kye Man;Lee, Chang Kwon;Kim, Gyoung Min;Shin, Jung-Hye;Kim, Jong Sang;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.974-983
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    • 2013
  • Bacillus amyloliquefaciens CB1 was isolated from cheonggukjang, a Korean fermented soy food. B. amyloliquefaciens CB1 secretes proteases with fibrinolytic activities. A gene homologous to aprE of Bacillus subtilis, aprECB1, was cloned from B. amyloliquefaciens CB1, and DNA sequencing showed that aprECB1 can encode a prepro-type serine protease consisting of 382 amino acids. When aprECB1 was introduced into B. subtilis WB600 using an E. coli-Bacillus shuttle vector, pHY300PLK, transformants showed fibrinolytic activity and produced a 28 kDa protein, the size expected for the mature enzyme. The 28 kDa fibrinolytic enzyme was purified from the culture supernatant of B. subtilis WB600 transformant. AprECB1 was completely inhibited by phenylmethylsulfonyl fluoride and almost completely inhibited by EDTA and EGTA, indicating that it is a serine metalloprotease. AprECB1 exhibited the highest specificity for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide, a known substrate for ${\alpha}$-chymotrypsin. $A{\alpha}$ and $B{\beta}$ chains of fibrinogen were quickly degraded by AprECB1, but the ${\gamma}$-chain was resistant.

Temporal variation of magma chemistry in association with extinction of spreading, the fossil Antarctic-Phoenix Ridge, Drake Passage, Antarctica

  • Choe, Won-Hie;Lee, Jong-Ik;Lee, Mi-Jung;Hur, Soon-Do;Jin, Young-Keun
    • 한국지구과학회:학술대회논문집
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    • 2005.09a
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    • pp.136-141
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    • 2005
  • The K Ar ages, whole rock geochemistry and Sr Nd Pb isotopes have been determined for the submarine basalts dredged from the P2 and P3 segments of the Antarctic-Phoenix Ridge (APR), Drake Passage, Antarctica, for better understanding on temporal variation of magma chemistry in association with extinction of seafloor spreading. The fossilized APR is distant from the known hot spots, and consists of older N-MORB prior to extinction of spreading and younger E-MORB after extinction. The older N-MORB (3.5-6.4 Ma) occur in the southeast flank of the P3 segment (PR3) and the younger E-MORB (1.4-3.1 Ma) comprise a huge seamount at the P3 segment (SPR) and a big volcanic edifice at the P2 segment (PR2). The N-type PR3 basalts have higher Mg#, K/Ba, and CaO/Al2O3 and lower Zr/Y, Sr, and Na8.0 with slight enrichment in incompatible elements and almost flat REE patterns. The E-type SPR and PR2 basalts are highly enriched in incompatible elements and LREE. The extinction of spreading occurring at 3.3 Ma seems to have led to a temporal magma oversupply with E-MORB signatures. Geochemical signatures such as Ba/TiO2, Ba/La, and Sm/La suggest heterogeneity of upper mantle and formation of E-MORB by higher contribution of enriched materials to mantle melting, compared to N-MORB environment. E-MORB magmas beneath the APR seem to have been produced by low melting degree (up to 1% or more) at deeper low-temperature regime, where metasomatized veins consisting of pyroxenites have preferentially participated in the melting. The occurrence of E-MORB at the APR is a good example to better understand what kinds of magmatism would occur in association with extinction of spreading.

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Cloning and Expression of a Fibrinolytic Enzyme Gene, aprECJ1, from Bacillus velezensis CJ1 Isolated from Myeolchi Jeotgal

  • Yoo, Ji Yeon;Yao, Zhuang;Lee, Se Jin;Jeon, Hye Sung;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.289-297
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    • 2021
  • Bacillus velezensis CJ1, showing significant fibrinolytic activity, was isolated from Myeolchi Jeotgal, a popular Korean fermented seafood. When B. velezensis CJ1 was grown on four different culture media, the culture on the Luria-Bertani (LB) broth showed the highest fibrinolytic activity (102.94 mU/μl) at 48 h. LB was also the best medium for growth. SDS-PAGE of culture supernatant showed four major bands, 38, 35, 27, and 22 kDa in size. Fibrin zymography showed four active bands, 50, 47, 40, and 30 kDa in size. A gene homologous to aprE of the Bacillus species was cloned by PCR. DNA sequencing showed that aprECJ1 can encode a protease consisting of 382 amino acids. The translated amino acid sequence of AprECJ1 showed high identity values with those of B. velezensis strains and other Bacillus species. The aprECJ1 gene was introduced into B. subtilis WB600 using an E. coli-Bacillus shuttle vector, pHY300PLK, and overexpressed. A 27 kDa band corresponding to the mature form of AprECJ1 was produced and confirmed by SDS-PAGE and fibrin zymography. B. subtilis WB600 [pHYaprECJ1] showed 1.8-fold higher fibrinolytic activity than B. velezensis CJ1 at 48 h.

Environmental Fatigue Evaluation of APR1000 Reactor Vessel (APR1000 원자로용기의 환경피로 평가)

  • Kim, Jong Min;Kim, Yong Hwan
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.26 no.3
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    • pp.207-212
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    • 2013
  • APR1000(Advanced Power Reactor 1000) was developed to export 1000MW nuclear power plants by adding ADFs(Advanced Design Features) including 60 years design life, local frequency control operation, 0.3g SSE, etc. to OPR1000(Optimized Power Reactor 1000). In this paper, environmental fatigue analyses for the reactor vessel in APR1000 have been performed as per Reg. Guide 1.207. Outlet nozzle, which has a relatively high cumulative usage factor in the reactor vessel was evaluated and a structural integrity is maintained under the reactor coolant environment.

Development of a new CVAP structural analysis methodology of APR1400 reactor internals using scaled model tests

  • Jongsung Moon;Inseong Jin;Doyoung Ko;Kyuhyung Kim
    • Nuclear Engineering and Technology
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    • v.56 no.1
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    • pp.309-316
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    • 2024
  • The U.S. Nuclear Regulatory Commission (NRC) Regulatory Guide (RG) 1.20 provides guidance on the comprehensive vibration assessment program (CVAP) to be performed on reactor internals during preoperational and startup tests. The purpose of the program is to identify loads that could cause vibration in the reactor internals and to ensure that these vibrations do not affect their structural integrity. The structural vibrational analysis program involves creating finite element analysis models of the reactor internals and calculating their structural responses when subjected to vibration loads. The appropriateness of the structural analysis methodology must be demonstrated through benchmarks or any other reasonable means. Although existing structural analysis methodologies have been proven to be appropriate and are widely used, this paper presents the development of an improved new structural analysis methodology for APR1400 reactor internals using scaled model tests.

Isolation of 2 Bacillus Strains with Strong Fibrinolytic Activities from Kimchi

  • Yao, Zhuang;Meng, Yu;Le, Huong Giang;Lee, Se Jin;Jeon, Hye Sung;Yoo, Ji Yeon;Afifah, Diana Nur;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.48 no.4
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    • pp.439-446
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    • 2020
  • Two Bacillus strains, K3 and K208, both demonstrating strong fibrinolytic activities were isolated from Kimchi, a traditional Korean preparation of fermented vegetables. Isolates were subjected to various molecular biology based identification methods including RAPD-PCR and identified as B. subtilis and B. velezensis, respectively. Tryptic soy broth (TSB) was found to best maintain both the growth and the fibrinolytic activity of these strains. Culture supernatants were analyzed by SDS-PAGE and fibrin zymography, and the results indicate that a 40 and 27 kDa band seem to be responsible for the fibrinolytic activities of these two isolates and the 27 kDa band was subsequently identified as the mature form of AprE, the major fibrinolytic enzyme. Thus the aprE genes were cloned and the translated amino acid sequences demonstrated 99.3% identity with each other, and 86.5% identity with BsfA, a fibrinolytic enzyme from B. subtilis ZA400 also isolated from Kimchi, and AprE2, a fibrinolytic enzyme from B. subtilis CH3-5 isolated from Cheonggukjang, a traditional Korean fermented soy. Given this B. subtilis K3 and B. velezensis K208 may be promising starter cultures in the production of fermented foods.

Variation of fibrinolytic enzyme activity produced Bacillus subtilis by gene cloning (유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화)

  • 이홍석;유천권;이철수;강상모
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.14-20
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    • 2000
  • The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity. Bands for the aprN and prtR genes were identified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN), pENC2 (prtR) and pFLA1 (aprN and prtR), respectively. The constructed vector was used for transformation of the strains of B.subtilis J-10 and B. subtilis K-54 and the fribrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolyitic enzyme activity in B. subtilis J-10 by 27.3% and 16%, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B.subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

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