• Title/Summary/Keyword: Liquid medium

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Increased Production of an Alkaline Protease from Bacillus clausii I-52 by Chromosomal Integration (Bacillus clausii I-52의 Chromosomal Integration에 의한 Alkaline Protease의 생산성 향상)

  • Joo, Han-Seung;Park, Dong-Chul;Choi, Jang-Won
    • Journal of agriculture & life science
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    • v.46 no.1
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    • pp.163-176
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    • 2012
  • TTo increase productivity of a strong extracellular alkaline protease (BCAP), stable strains of Bacillus clausii I-52 carrying another copy of BCAP gene in the chromosome were developed. Integrative vector, pHPS9-fuBCAP carrying BCAP promoter, ribosome binding site, signal sequence and active protease gene was constructed and transferred into B. clausii I-52, and integration of the constructed plasmid into chromosome was identified by PCR. An investigation was carried out on BCAP production by B. clausii I-52 and transformant C5 showing the highest relative activity of alkaline protease using submerged fermentation. Maximum enzyme activity was produced when cells were grown under the submerged fermentation conditions at $37^{\circ}C$ for 48 h with an aeration rate of 1 vvm and agitation rate of 650 rpm in a optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_47H_2O$ 0.01%, $FeSO_47H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). A protease yield of approximately 134,670U/ml was achieved using an optimized media, which show an increase of approximately 1.6-fold compared to that of non-transformant (83,960 U/ml). When the stability of transformant C5 was examined, the integrated plasmid pHPS9-fuBCAP was detected in the transformant after cultivation for 8 days, suggesting that it maintained stably in the chromosomal DNA of transformant C5.

Dispersion Polymerization of Acrylate Monomers in Supercritical $CO_2$ using GMA-functionalized Reactive Surfactant (초임계 이산화탄소에서 Glycidyl methacrylate 반응성 계면활성제를 이용한 아크릴레이트의 분산중합)

  • Park, Kyung-Kyu;Kang, Chang-Min;Lee, Sang-Ho
    • Elastomers and Composites
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    • v.45 no.4
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    • pp.256-262
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    • 2010
  • Dispersion polymerization of methyl acrylate, ethyl acrylate, butyl acrylate, and glycidyl methacrylate were performed in supercritical $CO_2$ at $80\;^{\circ}C$ and 346 bar. Glycidyl methacrylate linked poly(dimethylsiloxane) (GMS-PDMS) surfactant, which was prepared by linking glycidyl methacrylate to monoglycidyl ether terminated PDMS with amino-propyltriethoxysilane, was used as surfactant for the dispersion polymerization in $CO_2$. The yield of the poly(alkyl acrylate) polymers, synthesized in $CO_2$ medium, decreased as the alkyl tail of the acrylate monomers increased. Poly(glycidyl methacrylate) and poly(methyl acrylate) were produced in bead form whereas poly(ethyl acrylate) and poly(butyl acrylate) were viscous liquid. The poly(glycidyl methacrylate) particles had a number average diameter of 2.45 ${\mu}m$ and monodisperse distribution. The poly(methyl acrylate) had a number average diameter of 0.52 ${\mu}m$ and the particle size distribution was bimodal. The glass transition temperatures ($T_g$) of the poly(glycidyl methacrylate) and the poly(alkyl acrylate) products were 4~9 K higher than the $T_g$ of the corresponding acrylate polymers synthesized in conventional processes.

Usability Evaluation of Gel Pad Materials for Superficial Structures used in Musculoskeletal Ultrasonography (근골격계 초음파검사에 사용되는 표재성 검사용 젤 패드 물질의 유용성 평가)

  • Han, Mi-Ran;Kwak, Jong-Gil;Song, Jong-Nam;Jang, Seong-Ju;Han, Jae-Bok
    • Journal of the Korean Society of Radiology
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    • v.15 no.6
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    • pp.899-909
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    • 2021
  • In Musculoskeletal ultrasound examination, a linear transducer (12-5 MHz) with a contact surface length of 6 cm, which is commonly distributed in medical institutions, does not contact with bent fingers and wrist depressions only by using a conventional liquid ultrasound gel, which causes image loss. In order to solve this image loss, the use of commercially available Water gel pad is increasing. However, it is a disposable medical product. Repeated use is prohibited and expensive. In this study, in order to reduce the economic burden caused by the use of the commercialized Water gel Pad, we searched for water-soluble food thickener powder that can be easily purchased around our lives and manufactured Gel pads. Among them, kappa-Carrageenan Gel Pad, iota-Carrageenan Gel Pad, and Agar-Agar Gel Pad satisfied the evaluation criteria without artifacts in image evaluation using the N-365 Multipurpose Phantom. Also, the price to purchase the food thickener powder was about 38 times cheaper than the existing commercialized Water gel pad. As a result, if you use the food thickener powder such as kappa-Carrageenan, iota-Carrageenan and Agar-Agar to make solidified elastic Gel pad without a cover instead of the commercially available expensive disposable Water gel pad, the convenience and usefulness of use are improved. It is judged that the manufactured Gel pads can contribute well to the role of efficient ultrasound propagation medium.

An empirical model of air bubble size for the application to air masker (에어마스커의 기포크기 추정 경험적 모델)

  • Park, Cheolsoo;Jeong, So Won;Kim, Gun Do;Park, Youngha;Moon, Ilsung;Yim, Geuntae
    • The Journal of the Acoustical Society of Korea
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    • v.40 no.4
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    • pp.320-329
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    • 2021
  • In this paper, an empirical model of air bubble size to be applied to an air masker for reduction of underwater radiation noise is presented. The proposed model improves the divergence problem under the low-speed flow condition of the existing model derived using Rayleigh's jet instability model and simple continuity condition by introducing a jet flow velocity of air. The jet flow velocity of air is estimated using the bubble size where the liquid is quiescent. In a medium without flow, the size of the bubble is estimated by an empirical method where bubble formation regime is divided into a laminar-flow range, a transition range, and a turbulent-flow range based on the Reynolds number of the injected air. The proposed bubble size model is confirmed to be in good agreement with the Computational Fluid Dynamics (CFD) analysis result and the experimental results of the existing literature. Using the acoustic inversion method, the air bubble population is estimated from the insertion loss measured during the air injection experiment of the air- masker model in a large cavitation tunnel. The results of the experiments and the bubble size model are compared in the paper.

Neuroprotective effects of phenolic compounds isolated from Spiraea prunifolia var. simpliciflora (조팝나무(Spiraea prunifolia var. simpliciflora)로부터 분리한 페놀 화합물의 신경세포 보호효과)

  • Oh, Seon Min;Choi, Doo Jin;Kim, Hyoung-Geun;Lee, Jae Won;Lee, Young-Seob;Lee, Jeong-Hoon;Lee, Seung-Eun;Kim, Geum-Soog;Baek, Nam-In;Lee, Dae Young
    • Journal of Applied Biological Chemistry
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    • v.61 no.4
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    • pp.397-403
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    • 2018
  • The leaves of Spiraea prunifolia were extracted with 80% aqueous MeOH and the concentrates were partitioned into EtOAc, n-BuOH, and $H_2O$ fractions. The repeated $SiO_2$ or ODS column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of two phenolic glucosides. The chemical structures of these compounds were determined as isosalicin (1) and crenatin (2) based on spectroscopic analyses including Nuclear magnetic resonance and MS. Extracts were analyzed using UPLC-MS/MS providing a short analysis time within 5 min using MRM technique. The concentration of crenatin was higher as 9.53 mg/g and isosalicin was lower as 0.65 mg/g. Neuroprotective effects of these compounds against hydrogen peroxide ($H_2O_2$)-induced neurotoxicity were evaluated. The results showed that exposure to $H_2O_2$ induced morphological changes, cell death and neurotoxicity in SK-N-MC cells. However, pretreatment with crenatin resulted in inhibition of morphological change, reduction of loss of cell viability and attenuation of neuronal damage. These results suggested that neuroprotective effect of crenatin isolated from S. prunifolia can be a good candidate for the development of health beneficial foods which can ameliorate the degenerative neuronal disease caused by oxidative stress.

Purification and Biochemical Characterization of β-agarase Produced by Marine Microorganism Cellulophga sp. J9-3 (해양미생물 Cellulophga sp. J9-3이 생산하는 베타-아가레이즈의 분리 및 생화학적 특성)

  • Kim, Da Som;Kim, Jong-Hee;Chi, Won-Jae
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.329-336
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    • 2021
  • Cellulophga sp. J9-3, is a gram-negative, aerobic marine bacterium belonging to the family Flavobacteriaceae. In addition to cellulose degradability, the J9-3 strain is also capable of hydrolyzing agar in the solid and liquid medium, and the production of agarase in the presence of agarose can be remarkably induced by the bacterium. From the cell culture broth of Cellulophga sp. J9-3, ammonium sulfate precipitation and three kinds of column chromatography were successively performed to purify a specific agarase protein, the AgaJ93. Purified AgaJ93 showed the strongest hydrolyzing activity towards agarose (approximately 22%), and even displayed activity towards starch. AgaJ93 hydrolyzed agarose into neoagarotetraose and neoagarohexaose via various oligosaccharide intermediates, indicating that AgaJ93 is an endo-type β-agarase. AgaJ93 showed maximum activity at a pH of 7.0 and temperature of 35 ℃. Its activity increased by more than six times in the presence of Co2+ ions. The N-terminal sequence of AgaJ93 showed 82% homology with the heat-resistant endo-type β-agarase Aga2 of Cellulophaga sp. W5C. However, the biochemical properties of the two enzymes were different. Therefore, AgaJ93 is expected to be a novel agarose, different from the previously reported β-agarases.

Effect of Anti-inflammation on Oryeong-san Formulation for Mix Extract Tablet (오령산 정제 개발 및 항염증 효과)

  • Kim, Se Jin;Leem, Hyun Hee;Nam, Won Hee;Son, Su Mi;Choi, Hye Min;Kim, Myung Jin;Kim, Jung Ok;Lee, Hwa Dong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.34 no.6
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    • pp.348-354
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    • 2020
  • Oryeong-san (ORS) is a traditional Korean herbal medicine widely used for renal associated diseases, composed of five medicine herbs; Atractylodes japonica Koidzumi, Cinnamomum cassia Presl, Polyporus umbellatus Fries, Poria cocos Wolf and Alisma orientale Juzepzuk. We studied to improve the convenience of intake and portability by developing modernized dosage forms, and examined the effect on anti-inflammation of ORS. In order to develop the tablet formulation of ORS (ORS-F), the tablets were evaluated on the basis of physical characteristics include diameter, thickness, weight variation, hardness, friability and disintegration. To analyze the marker components of ORS-F, eight index markers from five herbal medicines were chosen. And the method using high performance liquid chromatography (HPLC) with diode-array detector method was established for the simultaneous analysis. The biological activities were examined the effect of ORS-F on pro-inflammation mediated by LPS-stimulation. The production of nitric oxide (NO) and cytokines were determined by reacting cultured medium with griess reagent and enzyme-linked immunosorbent assay (ELISA). The expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) were investigated by Western blot and RT-PCR. The anti-oxidant activities of OJS-F increased markedly, in a dose-dependent manner. and, The total phenolic compound and flavonoids contents of OJS-F were 10.20±0.09 ㎍/㎎ and 12.86±0.86 ㎍/㎎. OJS-F which is LPS has diminished in the LPS-induced release of inflammatory mediators (NO, iNOS, COX2 and PGE2) and pro-inflammatory cytokines (TNF-α, IL-6 and IL-1β) from the RAW264.7 macrophages. Therefore, the developed formulation for tablet of ORS-F provide efficiency and usability, and indicated effect of anti-inflammation.

A simple mid-term preservation method (SMPM) of plant callus under low temperature conditions (저온 보존을 이용한 간편 중기 식물캘러스 저장법)

  • Park, Sung-Chul;Park, Su Hyun;Kim, Soyoung;Jeong, Yu Jeong;Kim, Cha Young;Jeong, Jae Cheol
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.187-192
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    • 2022
  • The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

Expression and Localization of Heat Shock Protein 70 in Frozen-Thawed IVF and Nuclear Transfrred Bovine Embryos

  • Park, Y.J;S.J Song;J.T Do;B.S Yoon;Kim, A.J;K.S Chung;Lee, H.T
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.78-78
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    • 2002
  • The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

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Artificial Cultivation and Antioxidant Activity of Korean Native Mushroom Pycnoporus coccineus (한국산 간버섯의 인공재배 및 항산화 활성 평가)

  • Lee, Y.K.;Kim, M.K.;Yun, B.S.;Seo, G.S.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.21 no.2
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    • pp.35-47
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    • 2019
  • The mycelial growth of P. coccineus strain was good in PDA and YMA, but mycelial growth was low in MEA. Light irradiation during the incubation period affected the pigment formation and density of mycelia. Mushroom of P. coccineus strain was able to produce fruiting bodies in both bottle and bag cultivation, and oak sawdust was found to be the most suitable substrate for spawn culture and cultivation. In artificial cultivation using sawdust medium, fruiting body was grown to the extent that visual observation was possible from the 15th day, and it formed about 5 days fast in the treatment group with low relative humidity. From 40 to 45 days of mushroom development, mature fruiting bodies could be harvested, and the lower relative humidity of the growing room favored mushroom development and growth. Antioxidant activity of fruiting bodies harvested from artificial cultivation showed that ABTS radical scavenging activity of bottle-cultivated and wild fruit bodies were shown at 505㎍/㎖ and 515㎍/㎖, respectively. However, fruiting bodies harvested in bag cultivation were high at 910㎍/㎖. As a result of DPPH radical scavenging activity, all extracts were found to be inactive, exhibiting IC50 value of more than 2,000㎍/㎖ concentration. The ethyl acetate extract of mushrooms obtained from bottle cultivation showed the highest activity with 1,550㎍/㎖ IC50 value. Methanol extract of wild fruit bodies had the highest ABTS radical scavenging activity at the same concentration (10mg/㎖).