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Characterization of a Novel Thermostable Oligopeptidase from Geobacillus thermoleovorans DSM 15325

  • Jasilionis, Andrius;Kuisiene, Nomeda
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1070-1083
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    • 2015
  • A gene (GT-SM3B) encoding a thermostable secreted oligoendopeptidase (GT-SM3B) was cloned from the thermophile Geobacillus thermoleovorans DSM 15325. GT-SM3B is 1,857 bp in length and encodes a single-domain protein of 618 amino acids with a 23-residue signal peptide having a calculated mass of 67.7 kDa after signal cleavage. The deduced amino acid sequence of GT-SM3B contains a conservative zinc metallopeptidase motif (His400-Glu401-X-XHis404). The described oligopeptidase belongs to the M3B subfamily of metallopeptidases and displays the highest amino acid sequence identity (40.3%) to the oligopeptidase PepFBa from mesophilic Bacillus amyloliquefaciens 23-7A among the characterized oligopeptidases. Secretory production of GT-SM3B was used, exploiting successful oligopeptidase signal peptide recognition by Escherichia coli BL21 (DE3). The recombinant enzyme was purified from the culture fluid. Homodimerization of GT-SM3B was determined by SDS-PAGE. Both the homodimer and monomer were catalytically active within a pH range of 5.0–8.0, at pH 7.3 and 40℃, showing the Km, Vmax, and kcat values for carbobenzoxy-Gly-Pro-Gly-Gly-Pro-Ala-OH peptidolysis to be 2.17 ± 0.04 × 10-6 M, 2.65 ± 0.03 × 10-3 µM/min, and 5.99 ± 0.07 s-1, respectively. Peptidase remained stable at a broad pH range of 5.0–8.0. GT-SM3B was thermoactive, demonstrating 84% and 64% of maximum activity at 50℃ and 60℃, respectively. The recombinant oligopeptidase is one of the most thermostable M3B peptidase, retaining 71% residual activity after incubation at 60℃ for 1 h. GT-SM3B was shown to hydrolyze a collagenous peptide mixture derived from various types of collagen, but less preferentially than synthetic hexapeptide. This study is the first report on an extracellular thermostable metallo-oligopeptidase.

The Crystal and Molecular Struture of Cholesteryl Isobutyrate

  • Kim, Mi-Hye;Park, Young-Ja;Ahn, Choong-Tai
    • Bulletin of the Korean Chemical Society
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    • v.10 no.2
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    • pp.177-184
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    • 1989
  • The structure of cholesteryl isobutyrate, $(CH_3)_2CHCOOC_{27}H_{45}$, was determined by single crystal X-ray diffraction methods. Cholesteryl isobutyrate crystallized monoclinic space group $P2_1$, with a = 15.115 (8)${\AA}$, b = 9.636 (5)${\AA}$, c = 20.224 (9)${\AA}$, ${\beta}$ = 93.15 (5)$^{\circ}$, z = 4, $D_c = 1.03 g/cm^3 $and Dm= 1.04 g/$cm^3$. The intensity data were measured for the 3417 reflections, within $sin{\theta}/{\lambda} = 0.59{\AA}^{-1}$, using an automatic four-circle diffractometer and graphite monochromated Mo-$K_{\alpha}$ radiation. The structure was solved by fragment search Patterson methods and direct methods and refined by full-matrix least-squares methods. The final R factor was 0.129 for 2984 observed reflections. The two symmetry-independent molecules (A) and (B) are almost fully extended. The molecules are in antiparallel array forming monolayers with thickness $d_{100}$ = 15.2${\AA}$, and molecular long axes are nearly parallel to the [$\bar{1}$01] directions. The two distinct molecules form separate stacks with almost the same orientations, but with differing degrees of steroid overlap. Thers is a close packing of cholesteryl groups within the monolayers. The packing type is similar to those of cholesteryl hexanoate and cholesteryl oleate.

A STUDY OF BONE RESORPTION AT THE POSTERIOR BORDER OF DISTAL SEGMENT AFTER BILATERAL SAGITTAL SPLIT RAMUS OSTEOTOMY OF MANIBULAR PROGNATHIC PATIENTS (하악전돌환자의 하악지시상분할골절단술 후의 원심골편의 후방경계의 골흡수에 대한 연구)

  • Lee, Yong-In;Lim, Jong-Hwan;Noh, Kwang-Seob;Hong, Jong-Rak;Jun, Ju-Hong;Kim, Chang-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.1
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    • pp.55-59
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    • 2005
  • Purpose : This study is to predict the changes of the distal segment by investing the resorption of the excessive distal segment regarding the amount of setback after BSSRO. Material and Methods : 20 patients with Mandible prognathism treated by Obwegeser-Dal Pont method during the years 2000 to 2002 were selected for this study. Group A consisted of 5 males & 5 females with a setback amount of 10mm and above(Mean $10.80mm{\pm}1.03$, n=20) and Group B consisted of 2 males & 8 females with a setback amount of below 10mm.(Mean $6.10mm{\pm}1.10$, n=20) Panorama X-ray was taken at day 1, 1month, 3months, 6months, and 12months after the surgery. Resorption areas of excessive distal segment were measured on these panorama X-rays and compared. Results : There was bone resorption in both groups. Group A showed more bone resorption than Group B. Group B showed slightly higher resorption rate than Group A. However, there was no statistically significant difference between the resorption rates of Group A and Group B. (P>0.05). Conclusion : More bone resorption occurred with a larger amount of setback and about one third of the excessive distal segment underwent resorption, irrespective of the amount of setback.

Increase of Production Ratio of Pre-selected Superior Dairy Female Offspring by Combination of OPU derived Oocytes and X-bearing Semen

  • Kim, Seong-Su;Choi, Byung-Hyun;Lee, Kyeong-Lim;Jin, Jong-In;Suh, Tae-Kwang;Son, Cheol-Ho;Park, Chan-Ho;Shin, Seung-Oh;Han, Kwang-Jin;Lim, Hyun-Tae;Cho, Kyu-Woan;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.30 no.2
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    • pp.73-82
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    • 2015
  • This study was designed to evaluate the possibility of increase through dairy female offspring's ratio by transfer of pre-selected transferrable blastocyst that was produced by pre-selected X-bearing semen with OPU derived oocytes. Elite dairy female cow is demanded strongly compared with male, the so called, farmer wants to produce only an elite female dairy offspring as a candidate female dairy cow for producing milk. In our study, we selected 2 elite dairy bull semen from National Agricultural Cooperative Federation to pre-select X-bearing semen and 5 elite dairy female cows as donor for collecting of OPU derived oocytes. OPU derived embryo production system was carried out an aspiration of immature oocytes from 5 donor cows 2 times per week, total 200 times for 2 to 7 months by an ultrasonographic guided follicular aspiration system and then produced in vitro-produced blastocysts by in vitro maturation, fertilization and culture. Dairy donor semen selected H-319, 320 bull in National Agricultural Cooperative federation was sorted X-bearing semen by flow-cytometer and frozen for using IVF with OPU derived oocytes. Donor cows were selected 5 elite dairy cows from Gyeongju Dairy Cow Community and then disease tests such as 4 kinds of disease before selecting was checked. Oocyte proportion of grade 1 to 3 from total collected oocytes was significantly lower in donor A and B than those in donor C, D and E (82.16 and 70.03% vs. 90.0, 91.78 and 93.57%), respectively (p<0.05). However, number of oocytes per session in donor A, C and E was significantly higher than those in donor B and D ($7.77{\pm}3.26$, $5.85{\pm}2.10$ and $7.03{\pm}2.14$ vs. $4.68{\pm}2.61$ and $5.21{\pm}1.97$ oocytes), but donor A was significantly higher than donor C (p<0.05). Development to blastocyst in donor B, C and E was significantly higher than those in donor A and D (31.0, 25.0 and 25.0% vs. 14.3 and 4.5%), but donor A was not different in donor C and E (p<0.05). Nine out of 10 blastocysts (90.0%) derived from OPU blastocysts were confirmed male embryos that was induced with Y-bearing semen to confirm sex ratio only. Total 96 blastocysts derived from female bearing semen were transferred into synchronized recipients and then confirmed 42 recipients (43.8%) pregnancy rate, 36 offspring (37.5%) and 91.7% female sex ratio (33 female vs. 3 male offspring). Taken together all data, elite dairy female offspring could be produced effectively by in vitro production system between pre-selected x-bearing semen and OPU derived oocytes that would be influential breeder in the breeding of dairy farm to increase effectively elite dairy offspring ratio as well as net income in the dairy farmer.

The Structures of Alditol Acetates (Alditol Acetates의 분자구조)

  • Park, Yeong Ja;Park, Myeong Hui;Sin, Jeong Mi
    • Journal of the Korean Chemical Society
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    • v.34 no.6
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    • pp.517-526
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    • 1990
  • The crystal structures of two alditol acetates, D-glucitol hexaacetate and xylitol pentaacetate, have been determined by diffraction methods with Mo-K$\alpha$radiation, using direct methods for phase determinations. The crystal data are: for D-glucitol hexaacetate, P2$_1$, with a = 10.275 (2), b = 8.363 (1), c = 12.560 (5) $\AA;\beta$ = 95.97 $(2)^{\circ}$, Z = 2; for xylitol pentaacetate, P2$_1$/C with a = 18.126 (1), b = 11.422 (2), c = 8.649 (1) $\AA$, $\beta = 95.03 (1)^{\circ}$, Z = 4. Both molecules have extended zigzag carbon chain conformations which differ from previous studies of the structures of D-glucitol and xylitol and also differ from NMR studies on alditol acetates. The bond lengths and angles are normal, with mean values over both structures of C($sp^3)-C(sp^3): 1.514 (10),\; C(sp^3)-O: 1.444 (6),\; C(sp^2)-O: 1.347 (9),\; C(sp^2)=O: 1.197 (6),\; C(sp^2)-C(sp^3): 1.479(9){\AA},\; C(sp^3)-C(sp^3)-C(sp^3): 114.6 (17),\; O-C(sp^3)-C(sp^3): 109.4 (23),\; C(sp^2)-O-C(sp^3): 117.4 (6),\; O=C(sp^2)-O: 122.6 (6),\; C(sp^3)-C(sp^2)-O: 111.8 (7),\; C(sp^3)-C(sp^2)=O: 125.5 (4)^{\circ}$. The atoms of acetate groups are in coplanar. There are no particularly short intermolecular contacts and the molecules are held together by van der Waals force only.

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Growth and characterization of ZnSe/GaAs epilayer by hot-wall epitaxy method (Hot-Wal Epitaxy 방법에 의한 ZnSe/GaAs 박막 성장과 특성)

  • 정태수;강창훈;유평렬
    • Journal of the Korean Vacuum Society
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    • v.8 no.3B
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    • pp.302-307
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    • 1999
  • We have grown a high quality ZnSe(100) epilayer on the GaAs(100) substrate by hot-wall epitaxy method. The FWHM value from double-crystal x-ray diffraction rocking curve and growth rate of the ZnSe epilayer grown under the optimal growth conditions were 195 arcsec and 0.03 $\mu \textrm m$/min, respectively. The $I_2^U$ and $I_2^L$ peaks, which split by strain due to lattice mismatch between substrate and epilayer, were measured from the photoluminescence experiment. And we found that the residual impurities in ZnSe epilayer were concerned with Al or CI elements from the calculated binding energy of donor impurity.

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On the Design of a WiFi Direct 802.11ac WLAN under a TGn MIMO Multipath Fading Channel

  • Khan, Gul Zameen;Gonzalez, Ruben;Park, Eun-Chan
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.11 no.3
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    • pp.1373-1392
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    • 2017
  • WiFi Direct (WD) is a state of the art technology for a Device-to-Device (D2D) communication in 802.11 networks. The performance of the WD system can be significantly affected by some key factors such as the type of application, specifications of MAC and PHY layer parameters, and surrounding environment etc. It is, therefore, important to develop a system model that takes these factors into account. In this paper, we focus on investigating the design parameters of the PHY layer that could maximize the efficiency of the WD 802.11 system. For this purpose, a basic theoretical model is formulated for a WD network under a 2x2 Multiple In Multiple Out (MIMO) TGn channel B model. The design level parameters such as input symbol rate and antenna spacing, as well as the effects of the environment, are thoroughly examined in terms of path gain, spectral density, outage probability and Packet Error Rate (PER). Thereafter, a novel adaptive algorithm is proposed to choose optimal parameters in accordance with the Quality of Experience (QoE) for a targeted application. The simulation results show that the proposed method outperforms the standard method thereby achieving an optimal performance in an adaptive manner.

Physico-chemical Properties of Bracken(Pteridium aquilinum) Root Starch -1. Morphology and Chemical Properties- 중복기사검색 (고사리(Pteridium aquilinum) 뿌리 전분의 이화학적 특성에 관한 연구 -1. 전분의 일반 성상 및 화학적 특성-)

  • Jo, Jae-Sun
    • Korean Journal of Food Science and Technology
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    • v.10 no.1
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    • pp.57-62
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    • 1978
  • The morphology and chemical properties of bracken (Pteridium aquilinum) root starch were investigated. The starch granules were mainly sphere and cocoon with the diameter of $5-12\mu$. Polarized micrograph indicated that the starch granule had a hilum at the center of granule, showing a crossed-birefringence. X-ray diffraction pattern demonstrated that the granules showed B-type. The density of the starch was 1.49 and the amylose content was 22%. The ferricyanide number and alkali number were 0.292 and 11.03, respectively. Proximate analysis showed that the starch contained 0.52% lipid, 0.63% ash and 150ppm phosphorus of which over 80% were found in the amylopectin fraction. The iodine affinity and molecular weight of amylose were 16.1 and 83,000 respectively. The degree of branching and glucose units per segment of amylopectin were 3.7% and 27, respectively.

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The Association between Performance of Hepatitis B Vaccination and Health Belief Factors among Some Aged Persons (일부 노인의 B형간염 예방접종 이행과 건강신념과의 관련성)

  • Choi, Chun;Park, Jong;Kang, Myung-Guen;Kim, Ki-Soon
    • Korean Journal of Health Education and Promotion
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    • v.23 no.4
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    • pp.89-104
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    • 2006
  • Objectives: This study was done to find factors related with performance of hepatitis B Vaccination among some aged persons through health belief model. Methods: A questionnaire survey was made during September 2004 toward 230 elderly persons using institutions for the elderly of Gwangju City. The relations between subjects characteristics including health belief, mass media contact, hepatitis B experience and performance of hepatitis B vaccination were tested by t test or X2 test. Multiple logistic regression analysis was done to find final significantly related variables. Results: 24.8% of the subjects were vaccinated against hepatitis B. By simple analysis of relation between performance of hepatitis B vaccination and subjects characteristics including health belief, significant variables were chosen as 6 variables including perceived susceptibility, perceived seriousness, perception of benefits, knowledge on hepatitis B, age, experience of hepatitis through family or friend. After adjusting for confounding variables by multiple logistic regression analysis, hepatitis B vaccine performance showed significantly higher rate as the perception of disease seriousness increased(OR: 1.08, 95% CI: $1.03{\sim}1.14$) and in the group contacted with TV or radio information about hepatitis compared with non-contact. The group who experienced hepatitis among family or friends showed significantly higher hepatitis B vaccination performance rate compared with non-experienced. Conclusion: These results suggested that hepatitis B vaccine performance was related with health belief including hepatitis susceptibility, disease seriousness perception, acquisition of information through TV or radio and indirect hepatitis experience from family or friends.

Comparison of the optical properties of pre-colored dental monolithic zirconia ceramics sintered in a conventional furnace versus a microwave oven

  • Kim, Hee-Kyung;Kim, Sung-Hun
    • The Journal of Advanced Prosthodontics
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    • v.9 no.5
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    • pp.394-401
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    • 2017
  • PURPOSE. The purpose of this study was to compare the optical properties of pre-colored dental monolithic zirconia ceramics of various thicknesses sintered in a microwave and those in a conventional furnace. MATERIALS AND METHODS. A2-shade of pre-colored monolithic zirconia ceramic specimens ($22.0mm{\times}22.0mm$) in 3 thickness groups of 0.5, 1.0, and 1.5 mm were divided into 2 subgroups according to the sintering methods (n=9): microwave and conventional sintering. A spectrophotometer was used to obtain CIELab color coordinates, and translucency parameters and CIEDE2000 color differences (${\Delta}E_{00}$) were measured. The relative amount of monoclinic phase ($X_m$) was estimated with x-ray diffraction. The surface topography was analyzed by atomic force microscope and scanning electron microscope. Statistical analyses were conducted with two-way ANOVA (${\alpha}=.05$). RESULTS. There were small interaction effects on CIE $L^*$, $a^*$, and TP between sintering method and thickness (P<.001): $L^*$ (partial eta squared ${{\eta}_p}^2=0.115$), $a^*$ (${{\eta}_p}^2=0.136$), and TP (${{\eta}_p}^2=0.206$), although higher $b^*$ values were noted for microwave sintering regardless of thickness. Color differences between two sintering methods ranged from 0.52 to 0.96 ${\Delta}E_{00}$ units. The $X_m$ values ranged from 7.03% to 9.89% for conventional sintering, and from 7.31% to 9.17% for microwave sintering. The microwave-sintered specimen demonstrated a smoother surface and a more uniform grain structure compared to the conventionally-sintered specimen. CONCLUSION. With reduced processing time, microwave-sintered pre-colored dental monolithic zirconia ceramics can exhibit similar color perception and translucency to those by conventional sintering.