기질의 담체화를 이용한 키토산올리고당의 생산

The Production of Chitosanoligosaccharides Using Chitosan Bead

  • 김승모 (여수대학교 생물공학과)
  • 발행 : 2000.08.01

초록

키토산의 가수분해 산물인 키토산 올리고당의 효율적이고 단순화된 분리를 위하여 기질인 키토산을 담체화 하여 효소를 이용한 올리고당의 생산에 있어서 product의 새로운 분리법에 대한 가능성을 연구하였다. 기질로 사용한 chitosan bead 를 W/O emulsion 상분리법을 이용한 유기상의 방법과 키토산 용액을 알칼리 용액에 적하하는 수상의 방법으로 제조하였는데 bead가 효소와 접촉하는 표면적을 최대로 하기 위하여 구형으로 제조하였다. 이때 유기상 bead의 지름은 $200{\mu}m$였고, 수상 bead의 경우 $4000{\mu}m$, $100{\mu}m$, $30{\mu}m$였다. pH 와 온도, 효소의 양을 변화시켜 가면서 반응 시스템에서 최 적의 유기상 bead의 가수분해 조건을 조사한 결과 pH 6.0, 온도는 $50^{\circ}C$, 그리고 효소의 양은 40U이 최적임을 알 수 있 었다. 유기상 bead의 최적 가수분해 조건을 수상 bead에 적 용시켜 반응성을 살펴보았는데 반응에 사용된 수상 bead의 표면적이 유기상 bead에 비하여 낮음에도 불구하고 거의 동 일한 분해도를 나타내었다. 수상 bead의 경우 크기가 작을수록 높은 분해도를 나타냈으며, 각 bead를 사용하였을 때 생산된 최종 산물의 올리고당의 조성을 HPLC와 GPC로 분석하였는데, 유기상 bead로부터 생산된 올리고당의 조성은 주 로 2-4당이었으며, 수상 bead로부터 생산된 올리고당의 조성은 2-5당이었고, 평균 분자량은 M.W. 540과 M.W. 380이었다.

Preparation for the simplified separation of chitosandoligosaccharides from enzymatic hydrolysate was investigated. Two different types of chitosan beads as substrate were prepared as organic-based bead by W/O emulsion method and water-based bead by alkaline treatement. The average size of organic-based bead was $200{\mu}m$, and that of water based beads were $4000{\mu}m$, $100{\mu}m$, $30{\mu}m$, in diameter respectively. Enzyme stability was maintained over 80% at PH 6 after 24 hours. The optimal condition for the production of chitosanoligosaccharides was at pH 6.0, $50^{\circ}C$ and 40U (200U/g-chitosan) According to final oligosaccharide concentration water-based bed showed the similar result with that of organic-based bead even through it had smaller surface area attacked by chitosanse than that of organic-based bead. It is probable that the structure of water-based chitosan bead was looser than that of organic-based bead so enzyme penetrated easily into the bead structure. For the oligosaccharide production versus surface area the different size of water-based beads was investigated, Maxiaml production yield was observed in the $30{\mu}m$ beads. Consequently the water-based chitosan bead was better than the organic-based bead in this reaction system.

키워드

참고문헌

  1. Chitin and Chitosan Production and application of chitin and chitosan in Japan Hirano, S.
  2. Science v.212 Chitin: New facets of research Austin, P. R.;C. J. Brine;J. E. Castle;J. P. Zikakis
  3. Chitin, Chitosan, and related enzymes Zikakis. J. P.
  4. Chitian and Chitosan, The Japanese Society of Chitin and Chitosan Suzuki, S.;Y. Okawa;Y. Okura;K. Hashmoto;M. Suzuki;Hirano, S.(ed.);Tokukara, S.(ed.)
  5. Chem. Pharm. Bull. v.36 Enhancing effect of N-acetyl-chitooligosaccharides on the active oxygen-generating and microbicidal activities of perritoneal exudate cells in mice Sujuki. K.;A. Tokoro;Y. Okawa;S. Suzuki;M. Suzuki
  6. Carbohydr. Res. v.151 Antitumor effect of hexa-N-acetylchitohexaose and chitohexaose Suzuki, K.;Mikami;Y. Okawa;S. Suzuki;M. Susuki
  7. Chem. Pharm. Bull v.36 Growth-inhibitory effect of hexa-N-acetylchitohexaose against Meth-A solid tumor Tokoro, A.;M. Tatewaki;K. Suzuki;T. Mikami;S. Suzuki;M. Suzuki
  8. Microbiol. Immunol. v.28 Protecting effect of chitin and chitosan on experimentally induced murine candidasis Suzuki, K.;Y. Okawa;K. Hashimoto;S. Suzuki;M. Suzuki
  9. Microbiol. Immunol. v.34 Effect of N-acetylchitohexaose against Candida albicans infection of tumor-bearing mice Kobayashi, M.;T. Watanabe;S. Suzuki;M. Suzuki
  10. Microbiol. Immunol. v.33 Protective effect of N-acetyl chitoexaose on Listeria monocytogenes infection in mice Tokoro, A.;M. Kobayashi;N. Tatewaki;K. Suzuki;Y. Okawa;T. Mikami;S. Suzuki;M. Suzuki
  11. Biochim. Biophys. Acta. v.83 The hydrolysis of chitin by concentrated hydrochloric acid, and the preparation of low-molecular-weight substrates for lysozyme Ruply, J. A.
  12. Korean J. of chitin and chitosan v.2 no.3 Development of chitosanase for the Production of Chitosanoligosaccharides Choi, Y. J.;E. J. Kim;Y. S. Kim;Y. C. Shin
  13. Korean J. of chitin and chitosan v.3 no.1 Preparation of chitosan beads, microcapsules, microspheres and vesicles and their biochemical application Ha, B. J.;O. S. Lee;S. K. Park;Y. S. Lee
  14. Korean J. of chitin and chitosan v.4 no.1 Selective adsorption abilities of metal ions by cross-linked N-Methylthiobenzyl-chitosan bead and chitosan beads Han, S. M.;Y. B. Kim
  15. J. of Korean Ind. & Eng. Chemistry v.7 no.1 Swelling properties and releasing charateristics of chitosan beads containing bisamino-PEG Ha, B. J.;O. S. Lee;Y. S. Lee
  16. Anal. Chem. v.31 Use of dinitrosalisylic acid reagent for determination of reducing sugar Miller, G. L.
  17. Korean J. of chitin and chitosan v.3 no.3 Commercial production and applications of chitosanse Shin, Y. C.;E. J. Kim;E. J. Choi;J. J. Kim;Y. C. Yoon;C. M. Choi;Y. S. Kim