• Title/Summary/Keyword: specific yield

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Isolation and Characterization of Dextrans Produced by Leuconostoc sp. strain JYY4 from Fermented Kimchi

  • Gu, Ji-Joong;Ha, Yoo-Jin;Yoo, Sun-Kyun
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.4
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    • pp.758-766
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    • 2015
  • Dextran is a generic term for a bacterial exopolysaccharide synthesized from sucrose and composed of chains of D-glucose units connected by ${\alpha}$-1,6-linkages by using dextransucrases. Dextran could be used as vicosifying, stabilizing, emulsifying, gelling, bulking, dietary fiber, prebiotics, and water holding agents. We isolated new strain capable of producing dextran from Korean traditional kimchi and identified as Leuconostoc sp. strain JYY4. Batch fermentation was conducted in bioreactor with a working volume of 3 L. The media was MMY and 15% (w/v) sucrose. Mineral medium consisted of $3.0g\;KH_2PO_4$, $0.01g\;FeSO_4$, $H_2O$, $0.01g\;MnSO_4$, $4H_2O$, $0.2g\;MgSO_4\;7H_2O$, 0.01 g NaCl, $0.05g\;CaCl_2$ per 1 liter deionized water. The pH of media was initially adjusted to 6.0. The inoculation rate was 1.0% (v/v) of the working volume. Temperature was maintained at $28^{\circ}C$. The agitation rate was 100 rpm. The production pattern of dextran was associated with the cell growth. After 24 hr dextran reached its highest concentration of 59.4 g/L. The sucrose was consumed completely after 40 hr. Growth reached stationery phase when sucrose became limiting, regardless of the presence of fructose or mannitol. When the specific growth rate was 0.54 hr-1, utilization averaged 5.8 g/L-hr. The yield and productivity of dextran were 80% and 2.0 g/L-hr, respectively. Dextrans produced by were separated to two different size by an alcohol fraction method. The size of high molecular weight dextran (45% alcohol, v/v), less soluble dextran, was between MW 500,000 and 2,000,000. Soluble dextran (55% alcohol, v/v) was between 70,000 and 150,000. The molecular weight average of total dextran (70% alcohol, v/v) was between 150,000 to 500,000. The enzymatic hydrolyzates of total dextran of ATCC 13146 showed branched dextrans by Penicillium dextranase contained of glucose, isomaltose, isomaltotriose, and isomaltooligosaccharides greater than DP4 (degree of polymerization) that had branch points. Compounds greater than DP4 were branched isomaltooligosaacharides. Hydrolysates by the Lipomyces dextranase produced the same composition of oligosaccharides as those by Penicillin dextranase.

Purification and Characterization of Degradative Enzyme of Dental Plaque from Streptomyces sp. Y9343 (Streptomyces sp. Y9343이 生産하는 齒面細菌膜 分解酵素의 精製와 特性)

  • Kim, Seong-Joo;Han, Hong-Keun;Yoon, Jeong-Weon
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.9-18
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    • 1996
  • Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.

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Development of Thinning Effect Analysis Model (TEAM) Using Individual-Tree Distance-Independent Growth Model of Pinus koraiensis Stands (잣나무 임분의 개체목 거리독립생장모델을 이용한 간벌효과 분석모델 개발)

  • Kwon, Soonduk;Kim, Seonyoung;Chung, Joosang;Kim, Hyung-Ho
    • Journal of Korean Society of Forest Science
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    • v.96 no.6
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    • pp.742-749
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    • 2007
  • The objective of this study was to develop thinning effect analysis model (TEAM) using individual-tree distance-independent growth model of Pinus koraiensis Stands. The TEAM was designed to analyze thinning effects associated with such thinning prescriptions as the number, timing, intensity, and method of thinnings. To testing TEAM application, stand growth effects were compared with seven scenarios according to thinning prescription plan. In the results, it was possible to estimate the number of trees, height, volume with diameter (DBH) class of individual trees, and average diameter growth, height growth, the number of trees and volume growth per ha of stands. The result of sensitivity analysis on one Pinus koraiensis stand, it was not sure to expect the much more volume at the rotation age by stand density control applying thinning prescription. In the case of thinning, total yield volume has much more $40{\sim}75m^3$ per ha, within 5 cm in average diameter growth and within 1 m in average height growth than thats of non-thinning over increasing stand age. TEAM, as decision making support system, can be used for selecting the thinning prescription trial and determining one of some thinning prescription plan in different site specific stand environments.

Optimum Conditions for Growing Gem-quality Colorless Cubic Zirconia (보석용 무색 큐빅 저코니아의 최적 육성 조건)

  • 김원사;유영문;신현숙
    • Journal of the Mineralogical Society of Korea
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    • v.14 no.2
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    • pp.99-110
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    • 2001
  • A gem-quality yttria-stabilized zirconium oxide crystals were synthesized by the skull-melting method, using the RF electrical apparatus. Principal raw materials used were $ZrO_2$and 25 wt.% $Y_2O_3$as stabilizer and 0.03~0.05 wt.% $Nd_2O_3$decolorizing agent were added to it. The single crystals were approximately 20$\times$63 mm in size with chemical composition $Zr_{0.73}$ $Y_{0.27}$ $O_{1.87}$ . The crystals are isotropic with no appreciable anisotropism under a polarizing microscope. Their refractive indices are in the range of 2.15~2.18, specific gravity 5.85, Mohs' hardness 8~8.5, and reflectivity 13.47%. The zirconia crystals were confirmed to have cubic structure with Face-centered lattice(Z=4), space group Fm3m ($CaF_2$-type structure) and unit cell parameters are a=5.157 $\AA$. The optimal growing conditions for yttria-stabilized zirconia are 50 kW, 2.94 MHz in power and to use a crucible with 105 mm $\times$ 135 mm in size. When the lowering speed of the crucible was set 16mm/hr gave the best yield, 42%. Since the refractive index(2.15~2.18) of cubic zirconia is smaller than that of diamond, the angle between crown and pavilion should be fashioned to make it smaller than $40.5^{\circ}$ to show the maximum brilliancy and fire.

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Industrial-Scale Production of High-Purity Antihemophilic Factor IX from Human Plasma (사람 혈장으로부터 고순도 혈액응고 제9인자의 산업적 생산)

  • Kang, Yong;Choi, Yong-Woon;Sung, Hark-Mo;Sohn, Ki-Whan;Shin, Jeong-Sup;Kim, In-Seop
    • KSBB Journal
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    • v.23 no.1
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    • pp.37-43
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    • 2008
  • The use of antihemophilic factor IX complex has been associated with a variety of thrombotic complications, the major cause of which was the contamination of thrombogenic proteins such as vitamin K-dependent clotting factors II, VII, and X. In order to produce a commercial factor IX (GreenNine VF) free from thrombogenic potential, industrial-scale production process for high-purity factor IX from human plasma has been developed. The purification process contains cryo-precipitation, DEAE-sephadex A-50 anion-exchange chromatography, DEAE-toyopearl 650M anion-exchange column chromatography, heparin-sepharose 6FF affinity column chromatography, and CM-sepharose FF cation-exchange column chromatography. Also the process includes two viral inactivation and removal procedures, solvent/detergent treatment and nanofiltration using Viresolve NFP filter. The purification yield was 35.4%. The specific activity in the purified concentrate was 190.8 IU/mg which exceeded that in the factor IX complex (FacNine) by a factor of 48. The activities of factor II, VII, and X were not detected in GreenNine VF. SDS-PAGE analysis showed that GreenNine VF had the highest purity in comparison with commercially available high purity factor IX concentrates, Mononine, Octanyne, Berinin HS, and Immunine STIM plus 600. One batch size of the production was 2,400 vials of 250 IU product or 1,200 vials of 500 IU product from 1,600 L cryo-poor plasma.

Comparison of Enzymatic Activity and Cleavage Characteristics of Trypsin Immobilized by Covalent Conjugation and Affinity Interaction (공유결합과 친화력결합에 의한 고정화 Trypsin의 효소역가와 절단특성 비교)

  • Jang, Dae-Ho;Seong, Gi-Hun;Lee, Eun-Kyu
    • KSBB Journal
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    • v.21 no.4
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    • pp.279-285
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    • 2006
  • We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.

Thermal Acid Hydrolysis Pretreatment, Enzymatic Saccharification and Ethanol Fermentation from Red Seaweed, Gracilaria verrucosa (꼬시래기 홍조류로부터 열산가수분해, 효소당화 및 에탄올 발효)

  • Ra, Chae Hun;Choi, Jin Gyu;Kang, Chang-Han;Sunwoo, In Yung;Jeong, Gwi-Taek;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.9-15
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    • 2015
  • The seaweed, Gracilaria verrucosa, was fermented to produce bioethanol. Optimal pretreatment conditions were determined to be 12% (w/v) seaweed slurry and 270 mM sulfuric acid at 121℃ for 60 min. After thermal acid hydrolysis, enzymatic saccharification was carried out with 16 U/ml of mixed enzymes using Viscozyme L and Celluclast 1.5 L to G. verrucosa hydrolysates. A total monosaccharide concentration of 50.4 g/l, representing 84.2% conversion of 60 g/l total carbohydrate from 120 g dw/l G. verrucosa slurry was obtained by thermal acid hydrolysis and enzymatic saccharification. G. verrucosa hydrolysate was used as the substrate for ethanol production by separate hydrolysis and fermentation (SHF). Ethanol production by Candida lusitaniae ATCC 42720 acclimated to high-galactose concentrations was 22.0 g/l with ethanol yield (YEtOH) of 0.43. Acclimated yeast to high concentrations of specific sugar could utilize mixed sugars, resulting in higher ethanol yields in the seaweed hydrolysates medium.

Purification and Characterization of a Fibrinolytic Enzyme Produced by Bacillus amyloliquefaciens HC188 (Bacillus amyloliquefaciens HC188이 생산하는 혈전분해 효소의 정제 및 특성)

  • Shin, So Hee;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.33-43
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    • 2013
  • A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

Evaluation of the Characteristics of THM Formation by Chlorination in Extracted Humic Acid from Nakdong River (낙동강 원수에서 추출한 Humic Acid에서의 염소처리에 의한 THM 생성 특성 평가)

  • Son, Hee-Jong;Roh, Jae-Soon;Bae, Sang-Dae;Choi, Young-Ik;Jung, Chul-Woo
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.4
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    • pp.412-418
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    • 2007
  • The purpose of this study was to determine the several factors for affecting chlorine disinfection by-products(DBPs) characteristics by reacting chlorine and organic matters in the aquatic phase. The results of this research yield the following specific conclusions: The concentration of trihalomethanes(THMs) was increased with increasing dissolved organic carbon(DOC), and a trend of THMs formation was parabolic with increasing organic matters. Formations of THMs increased straightly for the first 4 hours and the amounts of producted THMs for the 30 minutes were up to $25\sim43%$ in the entire experiment periods(168 hours). When keeping up the concentration of organic matters at constant and changing that of bromide, the quantity of formed THMs did not show distinguished difference with the reaction times. THMs were gradually increased at $4^{\circ}C$ even though a reaction phase was parabolic formation(PF) phase. However, THMs were increased rapidly in the instantaneous formation(IF) phase and then became slowdown in the PF phase between $20\sim35^{\circ}C$. THMs were gradually increased although entering in the PF phase at pH 5. However, THMswere increased rapidly in the IF phase and then became slowdown in the PF phase at pH 7 and pH 9, and these treads were much more clear at pH 9 than at pH 7.

Development of Carbonization Technology and Application of Unutilized Wood Wastes(I) -Carbonization and It's Properties of Thinned Trees- (미이용 목질폐잔재의 탄화 이용개발(I) -수종의 간벌재 탄화와 탄화물의 특성-)

  • Kim, Byung-Ro;Kong, Seog-Woo
    • Journal of the Korean Wood Science and Technology
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    • v.27 no.2
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    • pp.70-77
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    • 1999
  • Objective of this research is to obtain fundamental data of carbonized wood wastes for soil condition, de-ordorization, absorption of water, carrier for microbial activity, and purifying agent for water quality of river. The carbonization technique and the properties of carbonized wood wastes(thinned trees) are analyzed. Proximate analysis shows the thinned wood contains 0.22-0.73% ash, 77-80% volatile matter, and 10-14% fixed carbon. The charcoal yield decreases and the shrinkage rate increases as the carbonization temperature and time increase. The charcoal yields of Larix leptolepis, Pinus rigida and Pinus densiflora are high, whereas those of Pinus koraiensis and Quercus variabilis are low. The shrinkage rate by carbonization has same trend as water removal of wood. The specific gravity after the carbonization decreases about 50% comparing to green wood. The charcoal has 0.89-4.08% ash, 6.31-13.79% volatile matter, and 73.9-83.5% fixed carbon. As the carbonization temperature and time increase, pH of charcoal increases. When the carbonization temperature is $400^{\circ}C$, pH is about 7.5. When the temperature is between 600 to $800^{\circ}C$, pH is about 10 with small difference. The water-retention capacity is not affected by the carbonization temperature and time. The water-retention capacity within 24hr is about 2.5 - 3times of sample weight, and the equivalent moisture content becomes 2-10% after 24 hr.

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