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Studies on the Cellulase produced by Myriococcum of albomyces (Myriococcum albomyces가 생산하는 Cellulase에 관한 연구)

  • Chung, Dong-Hyo
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.59-97
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    • 1971
  • As a study on the cellulase of Myriococcum albomyces the culture media for enzyme formation and properties of its crude preparation were investigated and the crude enzyme preparation was further fractionated. The results are summarized as follows: 1. Wheat bran solid culture produced stronger activities of cellulase than rice bran or defatted soy bean meal solid culture. 2. Shaking culture with wheat bran, rice bran or defatted soy bean meal produced higher cellulase activities than solid culture with the corresponding media. 3. The enzyme formation was higher at $45^{\circ}C$ than at $37^{\circ}C$ or $50^{\circ}C$ regardless of the kind of culture medium. 4. The formation of CMCase activity was more promoted by organic nitrogen source than inorganic nitrogen source. 5. The formation of cellulase activities were increased 1.5 to 3.0-fold by adding CMC, Avicel or cellulose powder as an inducer into 5% wheat bran basal medium. 6. Cellulase production using a tank culture procedure with addition of CMC or Avicel as an inducer was the highest at fifth day and thereafter decreased slightly. 7. The crude enzyme preparation showed pH optimum in 4.0 to 4.5, and pH stability in the range of 3.5 to 8.0. Optimum temperature for the activity was $65^{\circ}C$ which was higher than among other cellulases and it was stable at $60^{\circ}C$ for 120 minutes. 8. Dialyzed crude enzyme was activated by $Ca^{++}$ and $Mg^{++}$, but inhibited by $Hg^{++}$, $Cu^{++}$ and $Ag^{+}$. 9. Four different types of cellulase, i. e., fraction I, fraction II-a, fraction II-b, and fraction III were purified from the culture filtrate of Myriococcum albomyces through a sequence of ammonium sulfate fractionation, and elution chromatography on DEAE-Sephadex A-25, Amberlite CG-25 type 2 and hydroxyapatite columns. 10. These four cellulase fractions were showed to be homogenous by electrophoresis and ultracentrifugation and also gave a typical ultraviolet absorption spectrum of protein. 11. Four purified fraction showed different specificity toward substrates, fraction I has a stronger activity toward Avicel, cellulose powder, and gauze than that of other cellulase fractions. Fraction II-a had a powerful activity toward cellobiose but it was almost inactive agaisnt fibrous cellulose contrary to fraction I. On the contrary, the main component fraction II-b had a fairly higher activity on CMC and Avicel. Activity of fraction II-b toward cellobiose was about one-third of that of fraction II-a and activity on Avicel was lower than that of fraction I. Fraction III had a more powerful activity in decreasing viscosity of CMC. 12. Final hydrolysis products of fibrous cellulose by each fraction were cellobiose and glucose. Whereas oligosaccharides were predominant in the early stage of hydrolysis, prolonged reaction produced more glucose than cellobiose. Fraction I and fraction II-a acted synergically on Avicel. 13. Optimum pH for the activities of cellulase fraction I, fraction II-a, fraction II-b and fraction III were found to be 5.5, 5.0, 4.0 and $4.0{\sim}4.5$, respectively. These fractions were found to be stable in the range of pH $3.0{\sim}7.5$. 14. Optimum temperature for the activities of fraction I, fraction II-a, fraction II-b, and fraction III were $50^{\circ}C$, $55^{\circ}C$, $60^{\circ}C$ and $55^{\circ}C$, respectively. No less of activity was found by heating 120 minutes at $55^{\circ}C$ and fraction II-a was more stable than the others at $60^{\circ}C$. 15. Fraction I and fraction II-b were activated by $Ca^{++}$ and $Mg^{++}$ but inhibited by $Hg^{++}$ and $Ag^{+}$.

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An Anatomical Study of the Posterior Tympanum (한국인 중이강후벽에 관한 형태해부학적 고찰)

  • 양오규;윤강묵;심상열;김영명
    • Proceedings of the KOR-BRONCHOESO Conference
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    • 1982.05a
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    • pp.17.2-19
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    • 1982
  • The sinus tympani is subject to great variability in the size, shape and posterior extent. A heavy compact bony zone, especially in the posterior portion and the narrow space between the facial nerve and posterior semicircular canal are the limitation of surgical approach. The facial recess should be opened, creating a wide connection between the mesotympanum and mastoid in the Intact canal wall tympanoplasty with mastoidectomy. The surgically created limits of the facial recess are the facial nerve medially, the chorda tympani laterally and the bone adjacent to the incus superiorly. Using adult Korean's thirty-five temporal bones, the authors measured the osteologic reslationship in the posterior tympanum, especially sinus tympani and facial recess. The result was as followed. 1. The average distance from the anterior end of the pyramidal eminence. 1) to the edge of the sinus tympani directly posterior was 2.54(1.05-5.40)mm. 2) to the maximum posterior extent was 3.22(1.25-7.45)mm. 3) to the maximum cephaled extent was 0.67 (0.40-1.75)mm. 2. The boundary of the sinus tympani was 82.9% from the lower margin oval window to the upper margin round window niche. 3. The deepest part of the sinus tympani was 62.9% in the mid portion, between the ponticulus and subiculum. 4. The oblique dimension from the fossa incudis above to the hypotympanum below was 8.13(7.90-9.55)mm. 5. The transverse dimensions midway between the oval window above and round window below was 3.00(2.85-3.45)mm. 6. The transverse dimension at the level of the fossa incudis was 1.81(1.40-2.15)mm. 7. The facial nerve dehiscence was 14.3%. 8. Anterior-posterior diameter of the footplate was 2.98(2.85-3.05) mm. 9. The average distance from the footplate. 1) to the cochleariform process was 1.42(1.35-1.55) mm. 2) to the round window niche was 1.85(1.45-2.10) mm.

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