Isolation and Identification of Marine Bacterium Cytophaga sp. AYK301 and Optimal Culture Conditions for the Production of Agarase

해양세균 Cytophaga sp. AYK301의 분리.동정 및 한천분해효소 생산을 위한 최적배양조건

  • Published : 1999.10.01

Abstract

A marine bacterium with highly effective agar degrading activity was ioslated from the southern sea of Korea (Chonnam, YoChon) and identified as Cytophaga sp. and named as Cytophaga sp. AYK301. This strain produced an extracellular agarase which had a high activity with agar. The optimum culture conditions for the production of agarase have been determined. For the increase of agarase productivity, 0.2% agar, 0.3% beef extract, and 0.05% NH$_4$NO$_3$ were used as carbon, organic and inorganic nitrogen source, respectively. The optimal initial pH, NaCl, culture time and temperature for the agar degrading activity were 7.5, 7.0%, 36 hr and $25^{\circ}C$, respectively. In the optimal conditions, the agarase production was increased up to more than 4.0 folds as compared to that by the basal medium.

해양으로부터 한천분해능이 뛰어난 균주를 한국의 서해(전남, 여천)으로부터 분리하여 Cytophaga sp.인 것으로 동정되었으며, 이 균주를 Cytophaga sp. AYK301이라 명명하였다. 균체성장과 효소 생산성 향상을 위한 탄소원의 영향을 단당 및 복합당류를 이용하여 살펴본 결과 본 균주는 agar기질을 사용하여 배양하였을 때 효소생산량이 가장 높은 것으로 확인되었으며, agar를 0.0~0.6%(w/v) 농도별로 첨가하여 효소생산능의 최적조건을 살펴보았다. 그 결과 0.2%(w/v) agar에서 최대효소활성을 나타내었다. 질소원의 영향은 유기질소원과 무기질소원으로 나누어 실험을 행하였는데 beef extract 0.3% (w/v), $NH_4NO_3$ 0.05%(w/v)에서 각각 최고의 효소활성을 나타내었다. 또한, 초기 pH의 영향에서 pH 7.5에서 효소활성이 가장 높게 나타났고, NaCl 농도의 영향에서는 7%(w/v)에서 가장 높은 효소활성을 나타내었으며, 각종 금속이온에 대한 효소활성은 거의 영향이 없는 것으로 확인되었다. 따라서, 본 연구에서는 최적배양조건하에서 플라스크로 배양한 결과, 231 units/L를 생한하여 기본배지를 이용하였을 때보다 약 4배 정도 효소활성이 증가하였음을 확인하였다.

Keywords

References

  1. 해양수산통계연보 해양수산부
  2. Kor. J. Biotechnol. Bioeng. v.11 no.1 Purification of Extracellular Agarase from Marine Bacterium (Pseudomonas sp. W7) and Molecular Cloning of the Agarase Gene Kong, J. Y.;S. K. Bae;S. H. Hwang;S. D. Ha;H. T. Kim;S. K. Kim;B. J. Kim
  3. The First Asia-Pacific Marine Biotechnology Conference '95 Purification of Extracellular Agarase of Marine Microorganism (Pseudomonas sp. W7) and Molecular Cloning of the Agarase Gene Kong, J. Y.;S. H. Hwang;B. J. Kim;S. D. Ha;S. K. Kim;J. D. Kim
  4. Agric. Biol. Chem. v.55 no.10 Purification and Some Properties of Agarase from Pseudomonas sp. PT-5 Yamaura, I.;T. Matsumoto;M. Funatsu;H. Shigeiri;T. Shibata
  5. Biochim. Biophys. Acta v.537 Purification and Charaction of Two Different Agarose-Degrading Enzymes Malmqvist, M.
  6. Appl. Environ. Microbiol. v.59 no.5 Purification and Characterization of a Nex Agarase from a Marine Bacterium, Vibrio sp. Strain JT0107 Sugano, Y.;I. Terada;M. Arita;M. Noma;T. Matsumoto
  7. Appl. Environ. Microbiol. v.54 no.1 Cloning and Gene Replacement Mutagenesis of a Pseudomonas atlantica Agarase Gene Belas, R.;D. Bartlett;M. Silverman
  8. Mol. Gen. Genet. v.209 The Agarase Gene(dagA) of Streptomyces coelicolor A3(2) : Nucleotide Sequence and Transcriptional Alalysis Buttner, M. J.;I. M. Feamley;M. J. Bibb
  9. Appl. Environ. Microbiol. v.58 no.12 Purification and Properties of an Extracellular Agarase from Alteromonas sp. Strain C-1 Leon, O.;L. Quintana;G. Peruzzo;J. C. Slebe
  10. J. Gen. Microbiol. v.115 The Isolation and Characterization of Agarolytic Bacterium from a Low Land River Agbo, J. A. C.;M. O. Moss
  11. Biochim. Biophys. Acta v.244 Characterization of the Agarlytic System of Agaro Bacterium Pastinator Sampietro, A. R.;M. A. Vattuone de Sanpietro
  12. J. Gen. Microbiol. v.87 Degradation of Agar by a Gram-Negative Bacterium van Hofsten, B.;M. Malmyvist
  13. The 3rd Academic Plaza in International Food Machinery Exhibition '95 Isolation and Purification of Agarase from Marine Microorgansim(Pseudomonas sp. W7) Hwang, S. H.;S. D. Ha;J. D. Kim;S. K. Kim;J. Y. Kong
  14. The 4rd Academic Plaza in International Food Machinery Exhibition '96 海洋衛生物による 機能性食品素材(寒天オリゴオリゴ糖)の生産 Kong, J. Y.;S. K. Bae
  15. Biotech. Lett. v.19 no.1 Cloning and Expression of an agarase Gene from a Marine Bacterium Pseudomonas sp. W7 Kong, J. Y.;S. H. Hwang;B. J. Kim;S. K. Bae;J. K. Kim
  16. Kor. J. Appl. Microbiol. Biotechnol. v.27 no.3 Immobilization of Agarase for the Agarooli-gosaccharide Production Lim, D. J.;B. J. Kim;S. K. Bae;J. D. Kim;J. Y. Kong
  17. Kor. J. Biotechnol. Bioeng. v.14 no.3 Isolation and Its Optimal Culture Condition for High Agarase-Producing Mutant Hwang, S. H.;S. D. Ha;B. J. Kim;H. J. Kim;J. Y. Kong
  18. Bergey's Manual of Systematic Bacteriology Baumann, P.;A. L. Fumiss;J. V. Lee
  19. J. Biol. Chem. v.195 Notes on Sugar Determination Somogyi, M.