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Overproduction of Bacterial Trypsin in Streptomyces - Optimization for Streptomyces griseus Trypsin Production by Recombinant Streptomyces (미생물을 이용한 트립신 과대 생산 연구 - Streptomyces용 숙주-벡터계를 이용한 트립신 유전자의 대량발현 최적화 -)

  • Kim, Jong-Hee;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.28-33
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    • 2008
  • The expression vector (pWHM3-TR1R2) for sprT gene encoding Streptomyces griseus trypsin (SGT) followed by two regulatory genes, sgtR1 and sgtR2, was introduced into Streptomyces lividans TK24 and Streptomyces griseus IFO 13350. Various media with different compositions were used to maximize the productivity of SGT in the recombinant trains. he SGT productivity was best when the transformant of S. lividans TK24 was cultivated in R2YE medium (0.74 unit/mL) at 5 days of cultivation. C5/L (0.66 unit/mL) medium also gave a good productivity, but Livid (0.08 unit/mL) and NDSK (0.06 unit/mL) yielded poor productivities. S. griseus IFO 13350/pWHM3-TR1R2 produced SGT by 1.518 unit/mL (C5/L), 1.284unit/mL (R2YE),0.932 unit/mL (NDSK), and 0.295 unit/mL (Livid) at 7 days of cultivation, which was much higher than those from S. lividans TK24/TR1R2. The SGT protein was purified from the culture broth of S. griseus IFO 13350/pWHM3-TR1R2 in C5/L to homogeneity via ammonium sulfate fractionation, and CM-sepharose and SP-sepharose column chromatographies. The specific activity of purified SGT was 69,252 unit/mg, and the final purification fold and recovery yield were 6.5 and 1.4%, respectively.

Kinetics Studies on the Mechanism of Hydrolysis of S-Phenyl-S-vinyl-N-p-tosylsulfilimine Derivatives

  • Pyun, Sang-Yong;Kim, Tae-Rin;Lee, Chong-Ryoul;Kim, Whan-Gi
    • Bulletin of the Korean Chemical Society
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    • v.24 no.3
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    • pp.306-310
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    • 2003
  • Hydrolysis reactions of S-phenyl-S-vinyl-N-p-tosylsulfilimine (VSI) and its derivatives at various pH have been investigated kinetically. The hydrolysis reactions produced phenylvinylsulfoxide and p-toluene sulfonamide as the products. The reactions are first order and Hammett ρ values for pH 1.0, 6.0, and 11.0 are 0.82, 0.45, and 0.57, respectively. This reaction is not catalyzed by general base. The plot of k vs pH shows that there are three different regions of the rate constants $(k_t)$ in the profile.; At pH < 2 and pH > 10, the rate constants are directly proportional to the concentrations of hydronium and hydroxide ion catalyzed reactions, respectively. The rate constant remains nearly the same at 2 < pH < 10. On the bases of these results, the plausible hydrolysis mechanism and a rate equation have been proposed: At pH < 2.0, the reaction proceeds via the addition of water molecule to sulfur after protonation at the nitrogen atom of the sulfilimine, whereas at pH > 10.0, the reaction proceeds by the addition of hydroxide ion to sulfur directly. In the range of pH 2.0-10.0, the addition of water to sulfur of sulfilimine appears to be the rate controlling step.

Crossability, Variation in Chromosome Number, and Agronomic Characters in Hybrid of Hexaploid Triticale with Tetraploid Rye (6배체 트리티케일과 4배체호밀 교잡에서의 교잡율, 염색체수 변이 및 작물학적 특성)

  • 황종진;하용웅;이홍석
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.37 no.4
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    • pp.305-312
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    • 1992
  • This experiment was carried out to investigate seed set, variation in chromosome number, and agronomic characteristics of the progeny in the cross between hexaploid triticale variety, Sinkihomil(P$_1$) and tetraploid rye variety, Dooroohomil(P$_2$). Seed set rate obtained was 30.5% in the cross of Sinkihomil with Dooroohomil, whereas 3.26% in reciprocal cross using Dooroohomil as female. Alsoseed set was 8.75% in F$_1$/P$_1$, 7.20% in F$_1$/P$_2$, and 1.53% in F$_2$(=F$_1$ /F$_1$, respectively. Germination rate of crossed seed was 37% in cross of P$_1$ with P$_2$, 39.0% in F$_1$/P$_1$(BC$_1$), 50% in F$_1$/P$_2$(BC$_2$) and 43.0% in F$_1$/F$_1$(F$_2$), and 1,000 grain wight was 20.7g in the cross of P$_1$ with P$_2$, which have 41.9g and 47.7g, respectively, 24.5g in F$_1$/P$_1$, 23.6g in F$_1$/P$_2$, and 24.5g in F$_1$/F$_1$, respectively. In pollen fertility of F$_1$ plant, 69.8% turned out to be abnormal or sterile pollen grains, whereas 30.2% was fertile or normal. In meiosis of pollen mother cell of F$_1$ plant, 13.5 univalents, 8.89 bivalent and 1.24 trivalent were appeared. Somatic chromosome number of 35 in F$_1$, both 32 to 33 and 35 to 36 in F$_2$, 35 to 39 in BC$_1$ and 28 to 36 in BC$_2$ which mean producing female gamate was 14 to 18 chromosome in PMC of F$_1$ plant. Rate of fertile plant turned out to be 100% in F$_1$, 4.5% in F$_2$, 42.9% in BC$_1$, and 50.0% in BC$_2$, respectively. Number of seed set per spike appeared to be 1.17 in F$_1$ plant, 13.3 in F$_2$, 2.36 in BC$_1$, and 3.75 in BC$_2$, respectively. Days to heading of F$_1$ was intermediate, but F$_2$ was later than both parents. Plant height of F$_1$ , BC$_1$ ,and BC$_2$ was shorter than both parent, but F$_2$, longer than both parents.

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Effects of Water Temperature and Photoperiod on the Oxygen Consumption Rate of Juvenile Dark-banded Rockfish, Sebastes inermis (수온과 광주기에 따른 볼락, Sebastes inermis 치어의 산소 소비율)

  • Oh Sung-Yong;Noh Choong-Hwan
    • Journal of Aquaculture
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    • v.19 no.3
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    • pp.210-215
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    • 2006
  • An experiment was conducted to investigate the effects of four water temperatures (10, 15, 20, and $25^{\circ}C$) in combination with three photoperiods (24L:0D, 12L: 12D, and OL:24D) on the oxygen consumption rate of juvenile dark-banded rockfish, Sebastes inermis (mean body weight $20.5{\pm}0.7g$). The oxygen consumption rates of S. inermis were measured in triplicate for 24 hours using a continuous flow-through respirometer. Different combinations of water temperatures and photoperiods resulted in significant differences in the mean oxygen consumption rate of S. inermis (P<0.001). The oxygen consumption increased with increasing water temperatures for all photoperiod treatments (P<0.01). Mean oxygen consumption rates at 10, 15,20 and $25^{\circ}C$ ranged $178.3\sim283.5,\;386.7\sim530.7,\;529.2\sim754.3$ and $590.0\sim785.5mg\;O_2kg^{-1}h^{-1}$, respectively. $Q_{10}$ values ranged $3.17\sim5.51$ between 10 and $15^{\circ}C,\;1.87\sim2.10$ between 15 and $20^{\circ}C$ and $1.08\siml.24$ between 20 and $25^{\circ}C$, respectively. Fish held in continuous darkness (OL:24D) used consistently less okygen than fish exposed to continuous light (P<0.05). The mean oxygen consumption offish in a 12L:12D photoperiod was higher than that offish in 24L:0D and 0L:24D photoperiods under all temperature treatments except $10^{\circ}C$. The oxygen consumption of fish exposed to the 12L:12D photoperiod was significantly higher during the light phase than during the dark phase under all temperature treatments except $10^{\circ}C\;(P<0.05)$. This study provides empirical data for estimating oxygen consumption of S. inermis under given condition. This result has application for culture management and bioenergetic model for growth of this species.

인두 신경중 환자에서 Esophageal manometry와 24hr double pH-metry 검사 및 위식도 역류와의 관계

  • 김선태;형본승;최창준;차홍억
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1996.04a
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    • pp.84-84
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    • 1996
  • 인후두 이상감각을 주소로 내원한 환자들에서 주요한 원인 중의 하나로 위식도 역류가 관련되어 있다고 보고되고 있다. 이에 저자들은 이학적 검사 및 Esophagogram을 시행하여 기질적 원인을 배제한 후 인두신경증으로 진단받은 50명과 정상 대조군 30명을 이용하여 다음과 같은 결과를 얻었다. 1) 식도내압 검사상 상부식도압과 하부식도압에서 인두신경중 환자군과 대조군에서 유의한 차이는 없었다(UES : P = 0.56, LES P = 0.98). 2) 24시간 보행성 식도 pH검사에서 환자군과 대조군에서 앙와위시와 기립시 각각의 총역류 횟수, pH 4 미만으로 내려가는 역류횟수, 백분율, 5분이상 지속된 역류횟수, 최장역류시간 등을 비교한 바 양군간의 유의한 차이를 발견할 수 있었다(P<0.05). 3) 50명의 인두신경증 환자중 DeMester Scoring에 의해 의식도 역류 질환으로 진단받은 사람은 12명(24%)으로 위식도 역류가 인두신경증의 중요한 원인이 될 수 있음을 보여주었다.

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Acronyculatin P, A New Isoprenylated Acetophenone from the Stem Bark of Acronychia pedunculata

  • Tanjung, Mulyadi;Nurmalasari, Intan;Wilujeng, Aisyah Kanti;Saputri, Ratih Dewi;Rachmadiarti, Fida;Tjahjandarie, Tjitjik Srie
    • Natural Product Sciences
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    • v.24 no.4
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    • pp.284-287
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    • 2018
  • A new isoprenylated acetophenone, acronyculatin P (1) as well as two known compounds, 3',5'-diisoprenyl-2',4'-dihydroxy-6'-methoxyphenylethanone (2) and 3'-isoprenyl-2',4',6'-trihydroxyphenylethanone (3) were isolated from the stem bark of Acronychia pedunculata (L.) Miq. The structures were determined by HRESIMS, 1D and 2D NMR. The inhibitory activity of the isoprenylated acetophenone derivatives against murine leukemia P-388 cells showed compound 1 moderate activity with $IC_{50}$ $15.42{\mu}M$.

Pomegranate extract on eroded dentin: antioxidant action, bond strength and morphology of the adhesive interface after aging

  • Thiago Vinicius Cortez;Nathalia Mancioppi Cerqueira;Julia Adornes Gallas;Wanderley Pereira Oliveira;Silmara Aparecida Milori Corona;Aline Evangelista Souza-Gabriel
    • Restorative Dentistry and Endodontics
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    • v.49 no.1
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    • pp.9.1-9.14
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    • 2024
  • Objectives: This study aimed to evaluate the effect of pomegranate solution (Punica granatum) on eroded dentin through antioxidant action, shear bond strength (SBS) and interface morphology. Materials and Methods: The 10% pomegranate peel extract was prepared by the lyophilization method. Punicalagin polyphenol was confirmed by high-performance liquid chromatography. Antioxidant activity was evaluated by capturing the 2,2-diphenyl1-picrylhydrazyl (DPPH) radical. For the SBS, 48 dentin fragments were divided into sound or eroded, and subdivided according to the pretreatment (n = 12): water or P. granatum. The surfaces were restored with self-etch adhesive and a bulk-fill resin (Ecosite; DMG). The SBS was done immediately (24 hours) and after thermal cycling + water storage (12 months). For scanning electron microscopy, 48 dentin fragments (24 sound and 24 eroded) received the same treatments as for SBS (n = 6), and they were analyzed after 24 hours and 12 months. Results: The P. granatum had antioxidant action similar (p = 0.246) to the phenolic standard antioxidants. After 24 hours, eroded dentin had lower SBS than sound dentin (p < 0.001), regardless of the pretreatment. After 12 months, P. granatum maintained the SBS of sound dentin (13.46 ± 3.42 MPa) and eroded dentin (10.96 ± 1.90 MPa) statistically similar. The lowest values were found on eroded dentin treated with water (5.75 ± 1.65 MPa) (p < 0.001). P. granatum on eroded dentin caused peritubular demineralization and hybrid layer with resin tags. Conclusions: The pomegranate extract had antioxidant action and preserved the adhesive interface of the eroded dentin.

Effect of Defaunation on In Vitro Fermentation Characteristics and Methane Emission When Incubated with Forages

  • Qin, Wei-Ze;Choi, Seong-Ho;Lee, Seung-Uk;Lee, Sang-Suk;Song, Man-Kang
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.3
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    • pp.197-205
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    • 2013
  • An in vitro study was conducted to determine the effects of defaunation (removal of protozoa) and forage sources (rice straw, ryegrass and tall fescue) on ruminal fermentation characteristics, methane ($CH_4$) production and degradation by rumen microbes. Sodium lauryl sulfate, as a defaunation reagent, was added into the mixed culture solution to remove ruminal protozoa at a concentration of 0.375 mg/ml. Pure cellulose (0.64 g, Sigma, C8002) and three forage sources were incubated in the bottle of culture solution of mixed rumen microbes (faunation) or defaunation for up to 24 h. The concentration of ammonia-N was high under condition of defaunation compared to that from faunation in all incubations (p<0.001). Total VFA concentration was increased at 3, 6 and 12 h (p<0.05~p<0.01) but was decreased at 24 h incubation (p<0.001) under condition of defaunation. Defaunation decreased acetate (p<0.001) and butyrate (p<0.001) proportions at 6, 12 and 24 h incubation times, but increased propionate (p<0.001) proportion at all incubation times for forages. Effective degradability of dry matter was decreased by defaunation (p<0.001). Defaunation not only decreased total gas (p<0.001) and $CO_2$ (p<0.01~0.001) production at 12 and 24 h incubations, but reduced $CH_4$ production (p<0.001) at all incubation times for all forages. The $CH_4$ production, regardless of defaunation, in order of forage sources were rice straw > tall fescue > ryegrass > cellulose (p<0.001) up to 24 h incubation.

Optical Properties of InP/InGaP Quantum Structures Grown by a Migration Enhanced Epitaxy with Different Growth Cycles

  • Oh, Jae Won;Cho, Il-Wook;Ryu, Mee-Yi;Song, Jin Dong
    • Applied Science and Convergence Technology
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    • v.24 no.3
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    • pp.67-71
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    • 2015
  • InP/InGaP quantum structures (QSs) were grown on GaAs (001) substrates by a migration-enhanced molecular beam epitaxy method. Temperature-dependent photoluminescence (PL) and emission wavelength-dependent time-resolved PL (TRPL) were performed to investigate the optical properties of InP/InGaP QSs as a function of migration enhanced epitaxy (MEE) growth cycles from 2 to 8. One cycle for the growth of InP QS consists of 2-s In and 2-s P supply with an interruption time of 10 s after each source supply. As the MEE growth cycle increases from 2 to 8, the PL peak is redshifted and exhibited different (larger, comparable, or smaller) bandgap shrinkages with increasing temperature compared to that of bulk InP. The PL decay becomes faster with increasing MEE cycles while the PL decay time increases with increasing emission wavelength. These PL and TRPL results are attributed to the different QS density and size/shape caused by the MEE repetition cycles. Therefore, the size and density of InP QSs can be controlled by changing the MEE growth cycles.

Effect on Nutrients Removal of Algae in Aerobic High Rate Pond by Irradiance Period and pH (호기성 고율 안정조에서 빛의 조사 기간과 pH가 조류의 영양물질 제거에 미치는 영향)

  • 공석기;안승구
    • Journal of Environmental Science International
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    • v.6 no.2
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    • pp.141-152
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    • 1997
  • The pilot plant had been made so as to be an association system from the various items managed to have degrees of efficiency and It have been done to consider the experimental result with irradiance period and pH influence of all major things to treatment function of Waste Stabilization Pond. The results are as following. The attained results for continuous & cyclic irradiance 1. 24L.-reactor was prior to 12L.-12D.-reactor on oxygen generation & algal production ability. 2. 24L.-reactor was prior to 12L.-12D.-reactor on nutrients removal efficiency. 3. In 24L.-reactor it maintained 5mg/L∼6mg/L, DO concent enough to a fish's survival. The attained results for pH condition 1. Oxygen generation ^ algal production in pH 4-reactor were higher than those in pH 10-reactor. 2. The acidic condition at pH 4 and alkalic condition at pH 10 did not so much affect an algal growth and nutrients removal. The attained results for whole 1. In view of the results appeared as [(NH3-N)+(NO3-N)] removal efficiency, 89.1%∼93.9% and PO4-P removal efficiency, 34.3%∼83.7% & COD removal efficiency, 88.5%∼93.9%. It is possible to treat the wastewater with starch and pH which have been known as thedifficult problem. 2. At the point of non using methanol to nitrificate NO3-N, the nutrients removal method by using an algal growth is the most economical method in the whole nutrients removal methods. 3. The nutrients removal method by using an algal growth contributes to natural ecosystem. 4. The nutrients removal method by using an algal growth is excellant in the prevention against the eutrophication.

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