• Title/Summary/Keyword: HI분해

Search Result 89, Processing Time 0.027 seconds

Isolation and Characterization of Recombinant Vibrio parahaemolyticus Collagenase (재조합 Vibrio parahaemolyticus 콜라겐분해효소의 분리 및 특성 분석)

  • 차재호;김수광;전인준;이재원
    • Journal of Life Science
    • /
    • v.13 no.3
    • /
    • pp.308-313
    • /
    • 2003
  • The collagenase gene from Vibrio parahaemolyticus 04 was subcloned into an expression vector pET-29b. The recombinant collagenase was expressed in Escherichia coli BL2l(DE3) and partially purified by Hi-Trap affinity and Sephacryl S-100 size exclusion chromatographies. The recombinant enzyme was purified by 43.7-fold and the yield was 73%. SDS-PAGE revealed that the molecular weight of the enzyme was approximately 35 kDa. Substrate specificity study of the enzyme displayed that the enzyme showed the highest activity with the type I collagen and the synthetic peptide, Z-GPGGPA, indicating that the enzyme was indeed a collagenase. The enzyme showed broad pH optimum around pH 6-12 and was stable between pH 5.5 and 11.5. The optimum temperature for the type I collagen degradation was $35^{\circ}C$. The thermostability measurement of the enzyme indicated that the enzyme was stable up to $55^{\circ}C$, but the activity was diminished quickly above $60^{\circ}C.$

Isolation of dhlA Gene Responsible for Degradation of 1, 2-dichloroethane from Metagenomic Library Derived from Daecheong Reservoir (대청호로부터 제작한 메타지놈 라이브러리에서 1, 2-dichloroethane의 분해에 관여하는 dhlA 유전자의 분리)

  • Kang, Cheol-Hee;Moon, Mi-Sook;Song, Ji-Sook;Lee, Sang-Mhan;Kim, Chi-Kyung
    • Korean Journal of Ecology and Environment
    • /
    • v.38 no.2 s.112
    • /
    • pp.137-145
    • /
    • 2005
  • Traditional screening techniques have missed up to 99% of microbial resources existing in the nature. Strategies of direct cloning of environmental DNAs comprising tine genetic blueprints of entire microbial metagenomes provide vastly more genetic information than is contained in the culturable. Therefore, one way to screening the useful gene in a variety of environments is the construction of metagenomic DNA library. In this study, the water samples were collected from Daecheong Reservoir in the mid Korea, and analyzed by T-RFLP to examine the diversity of the microbial communities. The crude DNAs were extracted by SDS-based freezing-thawing method and then further purified using an $UltraClean^{TM}kit$ (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoRI, BamHI, and SacII in Escherichia coli DH10B using the pBACe3.6 vector. About 14.0 Mb of metagenomic libraries were obtained with average inserts 13 ${\sim}$ 15 kb in size. The genes responsible for degradation of 1, 2-dichloroethane (1, 2-DCE) via hydrolytic dehalogenation were identified from the metagenomic libraries by colony hybridization. The 1, 2-dichloroethane dehalogenase gene (dhlA) was cloned and its nucleotide sequence was analyzed. The activity of the 1, 2-DCE dehalogenase was highly expressed to the substrate. These results indicated that the dhlA gene identified from the metagenomes derived from Deacheong Reservoir might be useful to develop a potent strain for degradation of 1, 2-DCE.

Hydrogen production by catalytic decomposition of propane over carbon black catalyst in a fluidized bed (유동층 반응기에서 카본블랙 촉매를 이용한 프로판의 촉매 분해에 의한 수소생산 연구)

  • Nam, Woo-Seok;Jung, Jae-Uk;Yoon, Ki-June;Lee, Dong-Hyun;Han, Gui-Young
    • 한국신재생에너지학회:학술대회논문집
    • /
    • 2006.11a
    • /
    • pp.388-391
    • /
    • 2006
  • A fluidized bed reactor made of quartz with 0.055m I.D. and 1.0m in height was employed for the thermocatalytic decomposition of propane to produce $CO_2-free$ hydrogen. The fluidized bed was proposed for the continuous withdraw of product carbons from the reactor The propane decomposition rate used carbon black DCC-N330, Hi-900L as a catalyst. The propane decomposition reaction was carried out at the temperature range of $600-800^{\circ}C$, propane gas velocity of $1.0U_{mf}$ and the operating pressure of 1.0 atm. Effect of operating parameters such as reaction temperature on the reaction rates was investigated. Resulting production in our experiment were not only hydrogen but also several by products such as methane, ethylene, ethane, and propylene.

  • PDF

Formation of Fungal Protoplast by the Reaction of Cell Wall Lytic Enzymes (섬유소 가수분해효소와 Chitinase 처리에 의한 균류 원형질체 생성에 관한 연구)

  • Oh, Sung-Hi;Chang, Myung-Ho
    • Korean Journal of Food Science and Technology
    • /
    • v.13 no.2
    • /
    • pp.101-106
    • /
    • 1981
  • Osmotically sensitive fungal protoplasts were liberated from the mycelium of several kinds of molds by mixed enzyme system of cellulase from Trichoderma viride $TO_4$ and chitinase from the culture filtrate of Streptomyces sp. 115-5. Relatively higher number of protoplast were released from young mycelium of Zygomycetes strains than Ascomycetes strains by using 10 mM phosphate buffer (pH 6.0) and 0.6 M NaCl as osmotic stabilizer. Protoplasts were released through ruptures in the wall, initially at the apices, but later also from old party of the hyphae.

  • PDF

Isolation of whey protein and hydrolysis pattern of whey protein by proteolytic enzyme (유청단백질의 분리 및 단백질 분해 효소에 의한 유청단백질의 가수분해 양상)

  • Renchinkhand, Renchinkhand;Bae, Hyoung Churl;Jeong, Seok Geun;Nam, Myoung Soo
    • Korean Journal of Agricultural Science
    • /
    • v.39 no.4
    • /
    • pp.561-568
    • /
    • 2012
  • The aim of this study was to introduce a simple method for isolation of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin from cow's milk, and peptides produced by enzymatic hydrolysis of ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Whey protein were precipitated from whey by ammonium sulfate and, ${\alpha}$-lactalbumin and ${\beta}$-lactoglobulin were isolated using Hi Prep 26/60 Sephacryl S-100 column gel filtration chromatography. Bovine serum albumin and ${\beta}$-lactoglobulin were isolated by Mono-Q 5/50 GL column anion exchange chromatography of the 50% Ammonium Sulfate-supernatant. Isolated whey proteins were hydrolyzed by proteolytic alcalase. Tricine SDS-PAGE and reverse-phase HPLC analyses revealed that almost hydrolyzed all the ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin and bovine serum albumin with alcalase. Molecular weight of various peptides derived from alcalase hydrolysate were small molecular weight than 3.5 kDa.

Design and Optimization of Pilot-Scale Bunsen Process in Sulfur-Iodine (SI) Cycle for Hydrogen Production (수소 생산을 위한 Sulfur-Iodine Cycle 분젠반응의 Pilot-Scale 공정 모델 개발 및 공정 최적화)

  • Park, Junkyu;Nam, KiJeon;Heo, SungKu;Lee, Jonggyu;Lee, In-Beum;Yoo, ChangKyoo
    • Korean Chemical Engineering Research
    • /
    • v.58 no.2
    • /
    • pp.235-247
    • /
    • 2020
  • Simulation study and validation on 50 L/hr pilot-scale Bunsen process was carried out in order to investigate thermodynamics parameters, suitable reactor type, separator configuration, and the optimal conditions of reactors and separation. Sulfur-Iodine is thermochemical process using iodine and sulfur compounds for producing hydrogen from decomposition of water as net reaction. Understanding in phase separation and reaction of Bunsen Process is crucial since Bunsen Process acts as an intermediate process among three reactions. Electrolyte Non-Random Two-Liquid model is implemented in simulation as thermodynamic model. The simulation results are validated with the thermodynamic parameters and the 50 L/hr pilot-scale experimental data. The SO2 conversions of PFR and CSTR were compared as varying the temperature and reactor volume in order to investigate suitable type of reactor. Impurities in H2SO4 phase and HIX phase were investigated for 3-phase separator (vapor-liquid-liquid) and two 2-phase separators (vapor-liquid & liquid-liquid) in order to select separation configuration with better performance. The process optimization on reactor and phase separator is carried out to find the operating conditions and feed conditions that can reach the maximum SO2 conversion and the minimum H2SO4 impurities in HIX phase. For reactor optimization, the maximum 98% SO2 conversion was obtained with fixed iodine and water inlet flow rate when the diameter and length of PFR reactor are 0.20 m and 7.6m. Inlet water and iodine flow rate is reduced by 17% and 22% to reach the maximum 10% SO2 conversion with fixed temperature and PFR size (diameter: 3/8", length:3 m). When temperature (121℃) and PFR size (diameter: 0.2, length:7.6 m) are applied to the feed composition optimization, inlet water and iodine flow rate is reduced by 17% and 22% to reach the maximum 10% SO2 conversion.

Acid-Catalyzed Hydrolysis Reaction of Poly(vinyl acetate) (폴리(비닐 아세테이트)의 산촉매 가수분해 반응)

  • Park, Sang-Soo;Yoon, Hi-Sook
    • Polymer(Korea)
    • /
    • v.29 no.3
    • /
    • pp.304-307
    • /
    • 2005
  • The acid-catalyzed hydrolysis reaction of poly(vinyl acetate) (PVAc) in water/acetic acid solution at $35^{circ}C$ was studied at two different solvent compositions. The mole fractions of vinyl acetate (Vac) and vinyl alcohol (VA) during the course of the reaction were determined by NMR, and the equilibrium constant $K_{eq}$ of the reaction was determined using the molar ratio of VAc to VA at the chemical equilibrium. $K_{eq}$ was 0.75 (${\pm}0.01$) when the VAc mole faction at the equilibrium was 0.78 (${\pm}0.01$) and it was 0.69 (${\pm}0.01$) when the VAc mole fraction was 0.57 (${\pm}$0.02). The reaction was found to be a pseudo 1-st order reaction with the rate coefficient at $3.4{\times}10^{-6}/sec$.

Cyanide Degradation by Two Recombinant Cyanide Hydratases (Recombinant Cyanide Hydratases에 의한 시안화물 분해)

  • Kwon, Sung-Hyun;Cho, Dae-Chul
    • Journal of the Korea Academia-Industrial cooperation Society
    • /
    • v.10 no.6
    • /
    • pp.1287-1291
    • /
    • 2009
  • The genes of cyanide hydratase(CHT), a kind of nitrilases whichhydrolyze cyanide to formamide were extracted from N. crassa and A. nidulans, the two fungal strains. The recombinant forms of the CHT originated from N. crassa and A. nidulans were prepared with N-terminal hexahistidine purificationtags or no tags, and expressed in E. coli. The enzymes were purified using immobilized metal affinity chromatography. They were compared according to their pH activity profiles, and kinetic parameters. The N. crassa CHT has the wider pH range of activity above 50% and three-fold higher turnover rate (6.6 ${\times}$ $10^8$ $min^{-1}$) than the A. nidulans, meanwhile the CHT of A. nidulans has the higher $K_m$ value. Expression of CHT in both N. crassa and A. nidulans were induced by the presence of KCN, regardless of any presence of nitrogen sources. Max. 82% of KCN was degraded in 60 min for biological degradation tests.

Molecular Weight and Structural Changes of Enzymatic Hydrolysate from Bombyx mori Fibroin (효소처리에 의한 Bombyx mori fibroin 가수분해물의 분자량 및 구조 변화)

  • Yeo, Joo-Hong;Park, Kyung-Ho;Lee, Kwang-Gill;Woo, Soon-Ok;Kweon, Hae-Yong;Han, Sang-Mi;Lee, Heui-Sam;Lee, Jin-Ah;Lee, In-Seok;Cho, Yun-Hi
    • Korean Journal of Food Science and Technology
    • /
    • v.40 no.4
    • /
    • pp.470-473
    • /
    • 2008
  • This study examined the enzymatic digestion, average molecular weight distribution and structural changes of Bombyx mori silk gland fibroin using gel penneation chromatography and nuclear magnetic resonance methods. The pure-separation of calcium chloride-treated fibroin hydrolysates was carried out by gel filtration chromatography. Also, the effects of fibroin's enzymatic hydrolysis were investigated using an edible enzyme. The average molecular weight of three hydrolysate samples (silk gland fibroin (SF), SF-calcium chloride (SFC), and SFC-enzyme) were measured to compare their characteristics. The molecular weights of SF, SFC, and SFC-enzyme were approximately 108,000, 65,000, and 1,000 Da, respectively. Finally, we determined the structural characteristic changes of the different enzymatically digested samples by $^{13}C$ nuclear magnetic resonance methods. For the enzymatically digested fibroin, the glycine $^{13}C^{\alpha}$ resonance indicated that the amino acid was dramatically changed and/or separated out; however, this was not shown for the normal Bombyx mori silk gland fibroin.

Crystal Sinking Modeling for Designing Iodine Crystallizer in Thermochemical Sulfur-Iodine Hydrogen Production Process (열화학 황-요오드 수소 생산 공정의 요오드 결정화기 설계를 위한 결정 침강 모델링)

  • Park, Byung Heung;Jeong, Seong-Uk;Kang, Jeong Won
    • Korean Chemical Engineering Research
    • /
    • v.52 no.6
    • /
    • pp.768-774
    • /
    • 2014
  • SI process is a thermochemical process producing hydrogen by decomposing water while recycling sulfur and iodine. Various technologies have been developed to improve the efficiency on Section III of SI process, where iodine is separated and recycled. EED(electro-electrodialysis) could increase the efficiency of Section III without additional chemical compounds but a substantial amount of $I_2$ from a process stream is loaded on EED. In order to reduce the load, a crystallization technology prior to EED is considered as an $I_2$ removal process. In this work, $I_2$ particle sinking behavior was modeled to secure basic data for designing an $I_2$ crystallizer applied to $I_2$-saturated $HI_x$ solutions. The composition of $HI_x$ solution was determined by thermodynamic UVa model and correlation equations and pure properties were used to evaluate the solution properties. A multiphysics computational tool was utilized to calculate particle sinking velocity changes with respect to $I_2$ particle radius and temperature. The terminal velocity of an $I_2$ particle was estimated around 0.5 m/s under considered radius (1.0 to 2.5 mm) and temperature (10 to $50^{\circ}C$) ranges and it was analyzed that the velocity is more dependent on the solution density than the solution viscosity.