• Title/Summary/Keyword: OOO

Search Result 34, Processing Time 0.017 seconds

Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
    • /
    • v.20 no.5
    • /
    • pp.519-526
    • /
    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

  • PDF

An Experimental Study on Friction Welding and Heat Treatment of Engine Exhaust Valve Steels ( SCr4-21-4 N , SUH3-21-4-N (기관배기 밸브용 강 ( SCr4-21-4N , SUH3-21-4N ) 의 마찰압접과 열처리에 관한 실험적 연구)

  • 오세규
    • Journal of the Korean Society of Fisheries and Ocean Technology
    • /
    • v.14 no.2
    • /
    • pp.79-87
    • /
    • 1978
  • This is an experimental study on friction welding and heat treatment of engine exhaust valve materials whose welding combination is SCr4 as stem to 21-4N as head and SUH3 to 21-4N. In this study, not only the experiments of friction welding under the selected optimum welding condition and the examination of the mechanical properties were carried out, but also the heat treatment of friction welded specimens under the two selected conditions was taken to obtain the better welding character, eliminating the latent stress and the hardness peak which appeared at the welded zones of heat resisting steel(21-4N, SUH3) and low alloyed steel ($SCr_4$) friction weldments. The results obtained by the experiments and consideration in this study are as follows: I) It was experimentally proved quite reasonable that 'speed=3,OOO rpm, heating pressure Pl=8 kg/ mm2, upsetting pressure p, = 20 kg/mm', heating time $t_1$ = 3 see, upsetting time TEX>$t_2$ = 2.5 sec' was selected as the optimum welding condition for friction-welding the engine exhaust valve materials $SCr_4$ to 21-4 Nand SUH 3 to 21-4 N. 2) The results of the previous study and this one on friction welding of such dissimilar materials as SUH 3-SUH 31, SCr 4-SUH 31, SCr 4-SUH 3, SUH 3-CRK 22, SCr4-21-4 Nand SUH3-21-4 N agreed with each other substantially in the friction welding characteristics at welded interface zones. 3) It was also certified quite satisfactory that '600\ulcornerCX30 min. Xroom air cooling' as an optimum heat treatment condition of the friction welded materials SCr 4-21-4 Nand SUH 3-21-4 N was experimentally determined to eliminate the latent stress and the hardness peak at welded zones. 4) About 20% of the tensile strength before heat treatment of friction welded specimens was decreased after heat treatment 600\ulcornerCX30 min. Xair cooling, but the location of fracture was moved from heat affected zone to parent $SCr_4$ & SUH3. 5) Microscopic examination of the weld joints friction-welded and heat-treated under the above mentioned conditions revealed that the weld zone is very narrow and has a fine grained intermixed structure without any welding defects. 6) The above mentioned conditions can be also utilized as friction welding parameters of the other dissimiar materials for engine valve production.

  • PDF

Immobilization of Burkholderia cepacia Lipase on Weak Base Styrene Resin Using Polyethyleneimine with Cross-linking (PEI(Polyethyleneimine)를 이용하여 음이온계 레진에 고정화된 Lipase AH 제조 및 효소적 Interesterification을 통한 반응 특성 연구)

  • Lee, Chi Woo;Lee, Ki Teak
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.43 no.7
    • /
    • pp.1025-1035
    • /
    • 2014
  • This study assessed the effect of immobilized lipase on weak base styrene resin using polyethyleneimine (PEI) with cross-linking. Two procedures were used in this study. The first one, "mono-layer" lipase immobilization, involves washing PEI after adsorption. The second procedure, "multi-layer" lipase immobilization, has no washing before the cross-linking step. Treverlite XS-100200 (weak base styrene resin) was immersed with PEI solution (2.2 mg/mL). Lipase AH (from Burkholderia cepacia) was adsorbed onto the support coated with PEI before cross-linking with glutaraldehyde. Structured lipid was synthesized by immobilized lipase-catalyzed interesterification using canola oil, palmitic ethyl ester (PEE), and stearic ethyl ester (StEE). Total fatty acid contents of triacylglycerol (TAG) in structured lipids were analyzed to investigate activity, properties, and reusability of immobilized lipases. Activities of immobilized lipases on the multi-layer and mono-layer increased at a high concentration (8 mg/mL) of lipase solution used for immobilization. The results show that immobilized lipase with the mono-layer method at pH 8.0 on resin had the highest total saturated fatty acid content (26.17 area%). Activity of immobilized lipase with the multi-layer method at pH 7.5 on support was lower than that of the mono-layer, but total saturated fatty acid content was 16.79 area% higher than that of lipase AH (15.01 area%).

Characterization of Scaled-up Low-Trans Shortening from Rice Bran Oil and High Oleic Sunflower Seed Oil with Batch Type Reactor (회분식반응기를 이용한 미강유, 팜스테아린과 고올레인산 해바라기씨유 유래 대량 제조된 저트랜스 쇼트닝의 특성 연구)

  • Kim, Ji-Young;Lee, Ki-Teak
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.38 no.3
    • /
    • pp.338-345
    • /
    • 2009
  • Scaled-up low-trans shortening (LTS) was produced by lipase-catalyzed interesterification. Blend of rice bran oil (RBO), palm stearin (PS) and high oleic sunflower seed oil (HO) with 1:2:0.9 (w/w/w) ratio was interesterified using immobilized lipase from Thermomyces lanuginosus (TLIM) in the batch type reactor at $65^{\circ}C$ for 24 hr, and physicochemical melting properties of LTS were compared with commercial shortening. Solid fat content (SFC) of commercial shortening (used as control) and LTS was similar at 9.56 and 8.77%, respectively, at $35^{\circ}C$. Major fatty acids in LTS were C16:1 (33.7 wt%), C18:1 (45.7 wt%) and C18:2 (13.4 wt%). Trans fatty acid content in the commercial shortening (4.8 wt%) was higher than that of LTS (0.5 wt%). After reverse-phase HPLC analysis, major triacylglycerol (TAG) species in LTS were POO, POP and PLO. Total tocopherol, ${\gamma}$-oryzanol and phytosterol contents in the LTS were 12.37, 0.43 and 251.38 mg/100 g, respectively. Hardness of LTS was similar to that of commercial shortening. Also, x-ray diffraction analysis showed coexistence of ${\beta}'$ and ${\beta}$ form in the LTS.