The Effect of Oxygen Transfer Rate on the Nebramycin Factor 5' Activity and Component Ratio in Streptoalloteichus hindustanus Fermentation.

Streptoalloteichus hindustanus 발효시 Nebramycin Factor 5' 역가 및 구성비율에 대한 산소전달속도의 영향

  • 김정근 (한국산업기술대학교 생명화학공학과) ;
  • 이병규 (유한양행 중앙연구소) ;
  • 노용택 (영동대학교 생명공학부)
  • Published : 2003.12.01

Abstract

Nebramycin is a complex of aminocyclitol compounds that is produced by aerobic culture in fermentation process. The major antibiotic factors produced by Streptoalloteichus hindustanus are nebramycin factor 2, 4, 5'and kanamycin A. A mutant was selected, producing nebramycin factor 5' activity 16.4 times higher than parent strain by microbiological assay using Pseudomonas aeruginosa CH-U34AF. The component ratio of nebramycin factor 5' was dramatically increased from 34% to 70% by the optimization of fermentation condition. It was found that the component ratio of nebramycin factor 5' in fermentation was especially affected by the oxygen transfer rate. Optimum oxygen transfer rate for maximal nebramycin factor 5' productivity and ratio during S. hindustanus fermentation was elucidated to $0.50 mMO_2$/min.

본 연구에서는 S. hindustanus ATCC 31218로부터 선별된 변이주를 사용하여 배양조건이 nebramycin 주요 생산물인 factor 2, factor 4, factor 5', kanamycin A 등의 구성비율과 역가에 미치는 영향을 조사하였다. 실험결과, nebramycin의 factor들이 구성비율은 온도 및 배지보다는 배양액량, 교반속도 등에 의해 크게 영향을 받는 것으로 나타났다. 따라서 플라스크와 발효조의 산소전달속도를 각각 측정하여 factor 5' 생산의 최적 산소전달조건을 검토한 결과, $0.50 mMO_2$/min으로 배양시 factor 5'의 역가가 가장 높았으며 구성비율도 70% 이상을 나타내었다. 한편 $0.9 mMO_2$/min 이상으로 배양시는 factor 2의 역가와 구성비율이 현저히 감소한 반면 factor 2의 역가와 구성비율이 급격히 증가하였다. 한편 본 연구에 사용된 변이주 S. hindustanus YHT-0001는 factor 5'의 역가는 16.4배, factor 5'구성비율은 약 11% 향상된 균주로 확인되었다. S. hindustanus 발효시 공급되는 산소량은 단지 hydroxyl기와 아미노기의 존재유무에 따라 구조가 결정되는 factor 5', factor 4, kanamycin A의 hydroxyl기 생성을 위한 산소공여자로 역할뿐 만 아니라 배당체 구조에 차이를 보이는 apramycin 생합성과정에 영향을 크게 준다고 사료되었다.

Keywords

References

  1. Ind. Eng. Chem. v.36 Performance of gas-liquid contactors Cooper,F.M.;G.A.Fernstrom;S.A.Miller https://doi.org/10.1021/ie50414a005
  2. Drug Action Drug Resistant Bact. v.2 Chemistry and conformation of aminoglycoside antibiotic: Tobramycin Koch,K.F.
  3. J. Antibiot v.26 Nebramycin:Separation of the complex and identification of factors 4,5 and 5' Koch,K.F.;F.A.Davis;J.A.Rhoades https://doi.org/10.7164/antibiotics.26.745
  4. Antimicrob. Agents Chemother Structure of nebramycin factor 6. a new aminoglycosidic antibiotic Koch,K.F.;J.A.Rhoades
  5. J. Org. Chem v.43 Structures of some of the minor aminoglycoside factors of the nebramycin fermentation Koch,K.F.;K.E.Merkel;S.C.O'Connor;J.L.Occolowitz;J.W.Paschal;D.E.Dorman https://doi.org/10.1021/jo00401a030
  6. Yakhak Hoeji v.37 Determination of nebramycin factor 2,4,5,5',6 and kanamycin A in fermentation broth of Strptoalloteichus hindustranus ATCC 31218 mutant using 2,4-dinitrofluorobenzene (DNFB) as a derivatizing agent by High performance liquid chromatography Park,Y.K.;M.Y.Park;S.C.Kim;H.G.Yang
  7. J. Org. Chem. v.41 Apramycin, a unique aminocyclitol antibiotic Sean O'Connor;L.K.T.Lam;Noel,D.Jones;M.O.Chaney https://doi.org/10.1021/jo00874a003
  8. Antimicrob. Agents Chemother Nebramycin, a new broad-spectrum antibiotic complex.Ⅰ.Detection and biosynthesis Stark,W.M.;M.M.Honexl;K.G.Knox
  9. Folia Microbiol. v.16 Strains of Streptomyces tenebrarius and biosynthesis of nebramycin Stark,W.M.;N.G.Knox;R.M.Wilgus https://doi.org/10.1007/BF02884209
  10. Develop. Indust. Microbiol. v.17 The nebramycin fermentation: Culture and fermentation development Stark,W.M.;N.G.Knox;R.M.Wilgus;R.Dubus
  11. Develop. indust. Microbiol v.21 Fermentation studies with aminoglycoside-producing microorganisms Stark,W.M.;R.M.Wilgus;R.Dubus
  12. Antimicrob. Agents Chemother Nebramycin, a new broad-spectrum antibiotic complex.Ⅲ. Isolation and chemical-physical properties Thompson,R.Q.;E.A.Presti
  13. U.S.Patent 4,032,404 Fermentation process for producing apramycin and nebramycin factor 5'