• Title/Summary/Keyword: optimum properties

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Cloning of a Novel vpr Gene Encoding a Minor Fibrinolytic Enzyme from Bacillus subtilis SJ4 and the Properties of Vpr

  • Yao, Zhuang;Meng, Yu;Le, Huong Giang;Lee, Se Jin;Jeon, Hye Sung;Yoo, Ji Yeon;Kim, Hyun-Jin;Kim, Jeong Hwan
    • Journal of Microbiology and Biotechnology
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    • v.30 no.11
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    • pp.1720-1728
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    • 2020
  • We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

Optimization of Extraction Conditions for Functional Components of Roasted Pleurotus eryngii by Microwave-Assisted Extraction (볶음 새송이버섯 기능성분의 마이크로웨이브 추출조건 최적화)

  • Lee, Myung-Hee;Yoon, Sung-Ran;Jo, Deok-Jo;Kim, Hyun-Ku;Lee, Gee-Dong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1062-1069
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    • 2007
  • Response surface methodology was employed to optimize extraction conditions for finding the maximizing the functional properties of roasted Pleurotus eryngii. Based on the central composite design, independent variables were ethanol concentration ($0{\sim}100%$), extraction time ($1{\sim}9$ min) and microwave power ($25{\sim}125$ W). Soluble solid content, electron donating ability and nitrite-scavenging ability were mainly affected by ethanol concentration, but ACE inhibition activity was largely affected by extraction time. The optimum ranges of extraction conditions resulting from superimposing the response surface were predicted to be ethanol concentration ($25{\sim}50%$), extraction time ($3{\sim}9$ min) and microwave power ($80{\sim}125$ W). Total protein and total phenolic compound content of optimal extracts were 45.80 mg/g and 7.42 mg/g, respectively. In phenolic compounds of roasted Pleurotus eryngii extracts, protocatechuic acid was the highest concentration at 1226.32 ${\mu}g/g$, followed by salicylic acid, catechin, p-hydroxybenzoic acid, caffeic acid, coumaric acid and hesperidin.

Journal of Ginseng ResearchHighly regioselective biotransformation of ginsenoside Rb2 into compound Y and compound K by β-glycosidase purified from Armillaria mellea mycelia

  • Kim, Min-Ji;Upadhyaya, Jitendra;Yoon, Min-Sun;Ryu, Nam Soo;Song, Young Eun;Park, Hee-Won;Kim, Young-Hoi;Kim, Myung-Kon
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.504-511
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    • 2018
  • Background: The biological activities of ginseng saponins (ginsenosides) are associated with type, number, and position of sugar moieties linked to aglycone skeletons. Deglycosylated minor ginsenosides are known to be more biologically active than major ginsenosides. Accordingly, the deglycosylation of major ginsenosides can provide the multibioactive effects of ginsenosides. The purpose of this study was to transform ginsenoside Rb2, one of the protopanaxadiol-type major ginsenosides, into minor ginsenosides using ${\beta}$-glycosidase (BG-1) purified from Armillaria mellea mycelium. Methods: Ginsenoside Rb2 was hydrolyzed by using BG-1; the hydrolytic properties of Rb2 by BG-1 were also characterized. In addition, the influence of reaction conditions such as reaction time, pH, and temperature, and transformation pathways of Rb2, Rd, F2, compound O (C-O), and C-Y by treatment with BG-1 were investigated. Results: BG-1 first hydrolyzes 3-O-outer ${\beta}$-$\text\tiny{D}$-glucoside of Rb2, then 3-O-${\beta}$-$\text\tiny{D}$-glucoside of C-O into C-Y. C-Y was gradually converted into C-K with a prolonged reaction time, but the pathway of Rb2 ${\rightarrow}$ Rd ${\rightarrow}$ F2 ${\rightarrow}$ C-K was not observed. The optimum reaction conditions for C-Y and C-K formation from Rb2 by BG-1 were pH 4.0-4.5, temperature $45-60^{\circ}C$, and reaction time 72-96 h. Conclusion: ${\beta}$-Glycosidase purified from A. mellea mycelium can be efficiently used to transform Rb2 into C-Y and C-K. To our best knowledge, this is the first result of transformation from Rb2 into C-Y and C-K by basidiomycete mushroom enzyme.

Characterization of geopolymer made of municipal solid waste incineration ash slag (도시쓰레기 소각재 슬래그로 제조된 지오폴리머의 특성)

  • Kim, Yongsung;Kang, Seunggu
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.24 no.1
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    • pp.15-20
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    • 2014
  • In this research, the geopolymer was fabricated using municipal solid waste incineration ash (denoted as MSWIA) slag and alkali activator, NaOH and its properties were analyzed. Particularly, the effects of NaOH molarity, particle size of MSWIA, and liquid/solids ratio on the compressive strength of geopolymers were investigated. The compressive strength of geopolymers fabricated increased with finer grain size of MSWIA, and optimum value of the liquid/solids ratio was identified as 0.13. As the molarity of the NaOH increased, the compressive strength of geopolymers was increased. Even more the 20 M of NaOH, but the strength was not increased. The calcium aluminum silicate and calcium aluminum silicate hydrate zeolites were generated in the geopolymer fabricated with more than 20 M of NaOH, with some unreacted silica and unknown crystals remained. The highest compressive strength, 163 MPa, of geopolymer was appeared at conditions of curing temperature $70^{\circ}C$, and 20 M of NaOH, indicating that the high concentration of NaOH accelerates the geopolymer reaction and dense microstructure. The high-strength geopolymer fabricated in the present study is expected to contribute significantly to develop the field of cement alternative substances and to improve the recycling rate of MSWIA slag.

A Study on the Thermal Crack Control of the In-Ground LNG Storage Tank as Super Massive Structures (지하식 LNG 저장탱크 구조물의 온도균열 제어에 관한 연구)

  • Kwon, Yeong-Ho
    • Journal of the Korea Concrete Institute
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    • v.23 no.6
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    • pp.773-780
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    • 2011
  • In this study, thermal stress analysis are carried out considering material properties, curing condition, ambient temperature, and casting date of the mass concrete placed in bottom slab and side wall of the in-ground type LNG tank as a super massive structure. Also, based on the numerical results, cracking possibility is predicted and counter measures to prevent the cracking are proposed. For the tasks, two optimum mix proportions were selected. From the results of the thermal stress analysis, the through crack index of 1.2 was satisfied for separately caste concrete lots except for the bottom slab caste in 2 separate sequences. For the double caste bottom slab, it is necessary introduce counter measures such as pre-cooling prior to the site construction. Also, another crack preventive measure is to lower the initial casting temperature by $25^{\circ}C$ or less to satisfy 1.2 through crack index criterion. In the $1^{st}$ and $2^{nd}$ caste bottom slab, the surface crack index was over 1.2. Therefore, the surface cracks can be controlled by implementing the curing conditions proposed in this study. Since the side wall's surface crack index was over 1.0, it is safe to assume that the counter preventive measures can control width and number of cracks.

Utilizability of Waste Concrete Powder as a Material for Soil Pavement (흙도로포장용 재료로서 폐콘크리트 미분말의 활용성 연구)

  • Kim, Yong-Jic;Choi, Yun-Wang;Kim, Young-Jin
    • Journal of the Korean Recycled Construction Resources Institute
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    • v.3 no.3
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    • pp.277-282
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    • 2015
  • This study is conducted to utilize waste concrete powder (WCP) made as a by-product manufacturing high quality recycled aggregate. The blaine fineness of the used waste concrete powder was $928cm^2/g$. As the main characteristic of waste concrete powder, it showed an angular type similar to cement, but hydrated products were attached on the surface of particles. In addition, the size of the particles of waste concrete powder was larger than OPC and in terms of chemical components it had higher $SiO_2$ contents. For using WCP in soil cement-based pavement, the qualities, physical and chemical properties, of WCP should be researched. In the first step, the specified compressive strength of mortar for two types of clay sand soil and clay soil respectively was experimented to be 15 MPa and then optimum mixing ratio of chemical solidification agent were decided in the range of 1.5 - 3.0% in the replacement with cement weight content. In the second step, based on the prior experimental results, recycling possibility of WCP in soil cement-based pavement was studied. In the result of experiment the mixing ratio of WCP were 5, 10, 15 and 20% in the replacement with soil weight and the compressive strength of mortar was somewhat decreased according to the increase of the mixing ratio of WCP.

Optimization of Extraction Conditions for Extracts from Cucurbita moschata Duch. by Response Surface Methodology (반응표면분석에 의한 늙은 호박 추출물의 추출조건 최적화)

  • Lee, Hye-Jin;Do, Jeong-Ryong;Kwon, Joong-Ho;Kim, Hyun-Ku
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.3
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    • pp.449-454
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    • 2010
  • Extraction characteristics of pumpkin (Cucurbita moschata Duch.) and the functional properties of the corresponding extracts were monitored by the response surface methodology (RSM). The maximum electron donating ability was found as 24.57% at the conditions of 33.13 watt microwave power, 53.67% ethanol concentration and 3.76 min extraction time. The maximum tyrosinase inhibition was 96.59% at 56.21 watt, 68.02% and 7.97 min. The SOD-like activity was 45.57% at 36.00 watt, 71.51% and 5.28 min. The total polyphenol content was 85.02% at 127.39 watt, 76.18% and 2.68 min. Based on superimposition of four dimensional RSM with respect to electron donating ability, tyrosinase inhibition, SOD-like activity and total polyphenol contents obtained under the various extraction conditions, the optimum ranges of extraction conditions were found to be microwave power of 72~144 watt, ethanol concentration of 0~38% and extraction time of 6~9 min.

Degradation Assessment of Forest Trails in Mt. Jiri Area of Gyeongnam Province (경남권역 지리산 둘레길 훼손 실태 분석)

  • Lim, Hong-Geun;Park, Jae-Hyeon
    • Journal of Korean Society of Forest Science
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    • v.102 no.2
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    • pp.255-263
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    • 2013
  • This study was carried out to evaluate degradation status of forest trails by analysis of soil physical and chemical properties in Mt. Jiri area of Gyeongnam province. Soil texture was loam in the forest area and sandy loam in the forest trails. Soil bulk density was significantly higher (P<0.05) in the forest trails (1.15 g/$cm^3$) than in the forest area (1.00 g/$cm^3$). The rates of pore space were lower in the forest trails (56.6%) than in the forest area (62.4%). Soil moisture content was significantly different (P>0.05) between the forest trails (13.3%) and the forest area (11.3%) Soil strength was higher at 5 cm of soil depth than at 10 cm of soil depth. It indicates that soil compaction by visitors could be affected at 5 cm of soil depth. Soil strength over 30 cm of soil depth was not significantly different between the forest trails and the forest areas. The content of organic matter, total nitrogen, available phosphorus, and exchangeable cations were lower in the forest trails compared with the optimum content of forest soils because of soil erosion with increasing visitors. These results indicate that it needs a counterplan to protect forest trails from overcrowded visitors.

Exploration of Optimum Retention of Antibacterial Agents in Functional Packaging Paper (항균 포장원지내 항균소재의 최적 정착법 탐색)

  • Kim, Chul-Hwan;Kim, Jae-Ok;Jung, Jun-Ho;Cho, Sung-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.298-305
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    • 2005
  • Antimicrobial packaging paper was prepared with a powder-type botanical antimicrobial agent from grapefruit seed extract (BAAG) and zeolite according to TAPPI standard method. The functional fillers containing BAAG fixed to CaCO$_3$ and zeolite were well retained in the fiber network by a retention aid such as cationic polyacrylamide, which was ascertained by the ash contents of paper and the SEM microphotographs. With addition of the functional fillers to paper, tensile strength and burst strength of the paper decreased by interference of the functional fillers with interfiber bonding but bending stiffness and tear strength increased by improved elastic modulus of paper and delayed transfer of tearing energy. Finally, it was confirmed that the antimicrobial packaging paper might be able to be used to make packaging bags and corrugated containers due to the minor deterioration of strength properties.

Extraction of Carbohydrates and Minerals from Laminaria Using Organic Acid (다시마 뿌리로부터 유기산을 이용한 다당과 미네랄 추출)

  • Chun, Ji Yeon;Han, Cha Seong;Lee, Jung Shik;Kim, Young Suk;Park, Kwon Pil
    • Korean Chemical Engineering Research
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    • v.50 no.2
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    • pp.238-243
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    • 2012
  • Laminaria roots have not been practically used in Korea. In this study, the extraction process of carbohydrates and minerals from Laminaria roots was investigated and the properties of extracted components were measured. Hydrochloric acid generally used in carbohydrate extraction from seaweeds in order to obtain high extraction yield. But in this work, to utilize extracted components as a functional food material, organic acids such as citric acid were used. Organic acid as extraction solvent has low extraction yield compared to strong acids. Therefore optimum condition for maximum yield was investigated in carbohydrate extraction from Laminaria roots using organic acid. We measured the extraction yields of carbohydrate with variation of extraction temperature, extraction time, concentration of organic acid and particle size of samples. The extraction yield increased as the particle size decreased and temperature became high. The extraction yield was 19.0 wt% after 4.0 hours extraction with 0.2 wt% citric acid at $100^{\circ}C$. Potassium concentration was high compared other minerals in extraction solution, that is, the ratio of K/Na was about 3.0. Fucoidan from Laminaria roots had same carbohydrate composition and lower molecular weight compared that of Undaria pinnatifida.