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Bacillus coagulans DL-1이 생산하는 고분자 중합체의 특성과 생산조건의 최적화

Characterization of biopolymer produced by Bacillus coagulans DL-1 and optimization of its production

  • 김지모 (동아대학교 생명자원과학대학 생물공학전공) ;
  • 손창우 (동아대학교 생명자원과학대학 생물공학전공) ;
  • 서형필 (동아대학교 생명자원과학대학 생물공학전공) ;
  • 문병주 (동아대학교 생명자원과학대학 응용생물학전공) ;
  • 도성국 (동아대학교 생활과학대학 섬유공학전공) ;
  • 이진우 (동아대학교 생명자원과학대학 생물공학전공)
  • 발행 : 2003.04.01

초록

고분자 중합체를 생산 능력이 있는 균주를 토양에서부터 분리하여 형태학적 및 생화학적인 특성을 확인하고 Bacillus 속 균주를 동정하는 API-50CH/B kit를 사용하여 동정한 결과, Bacillus coagulans로 확인되어 B. coagulans DL-1으로 명명하였며 고분자 중합체를 생산하여 체외로 분비하는 균주임을 확인하였다. 플라스크 실험을 통하여 고분자 중합체 생산을 위한 B. coagulans DL-1의 최적 탄소원과 질소원 및 각각의 최적 농도는 2% (w/v) 포도당과 0.25%(w/v) yeast extract임을 확인하였다. 플라스크 실험에서 확립한 최적 배지조건과 1.0 vvm의 통기량 및 500 rpm의 교반속도로 $7\ell$ 생물배양기를 사용하여 수행한 회분식 배양에서의 최대 고분자 중합체 생산량은 5.00$\pm$0.15 g/$\ell$이었다. 가스 크로마토그라피를 사용하여 B. coagulans DL-1이 생산한 고분자 중합체의 구성 성분을 분석한 결과, 포도당과 람노오스로 구성되어 있으며 포도당과 람노오스의 구성 비율은 약 9 : 1 이었다. 또한 평균 분자량을 gel permeation chromatography로 측정한 결과, 2.80$\times$$10^5$이었다.

The microorganism isolated from soil was identified as Bacillus coagulans by morphological and biochemical analyses and API-50CH/B kit, which was an identification kit for Bacillus species, and named as B. coagulans DL-1. It produced an extracellular biopolymer. Maximum production of biopolymer was 5.00 $\pm$0.15 g/$\ell$ in a $7\ell$bioreactor with an aeration rate of 1.0 vvm and an agitation speed of 500 rpm when concentrations of glucose and yeast as the optimal carbon and nitrogen sources were 2.0% (w/v) and 0.25% (w/v), which were optimized with a flask scale. Gas chromatographic analysis showed that the biopolymer producded by B. coagulans DL-1 consisted of glucose and rhanmose and their molar ratios was about 9 : 1. Its average molecular weight was 2.80$\times$$10^5$ with gel permeation chromatographic (GPC) analysis.

키워드

참고문헌

  1. J. Microbiol. Biotechnol. v.8 Isolation and characterization of a novel polysaccharide producing Bacillus polymyxa A49 KCTC 4648P Ahn,S.G.;H.H.Suh;C.H.Lee;S.H.Moon;H.S.Kim;K.H.Ahn;G.S.Kwon;H.M.Oh;B.D.Yoon
  2. Appl. Microbiol. Biotechnol. v.45 Isolation of mutants of Penicillium purpurogen resistant to catabolite repression Anwar,M.N.;M.Suto;F.Tomita https://doi.org/10.1007/s002530050748
  3. Anal. Biochem. v.123 A rapid and sensitive method for the analysis of carbohydrate components in glycoproteins using gas-liqid chromatography Chaplin,M. https://doi.org/10.1016/0003-2697(82)90455-9
  4. Biotechnol. Bioprocess Eng. v.6 Improved production of curdlan with concentrated cells of Agrobacterium sp Chung,D.Y.;Y.S.Cho;C.H.Chung;D.I.Chung;K.Kim;J.W.Lee https://doi.org/10.1007/BF02931955
  5. Fungal Gen. Biol. v.25 Carbon catabolite repression of gene expression and condition in Neurospora crassa Ebbole,D.J. https://doi.org/10.1006/fgbi.1998.1088
  6. Carbohydr. Polym. v.34 Exopolymer production by Bacillus species Gandhi,H.P.;R.M.Ray;R.M.Patel https://doi.org/10.1016/S0144-8617(97)00132-X
  7. Enz. Microbiol. Technol. v.30 Production of heteropolysaccharide-7 by Beijernckia indica from agro-industrial byproducts Jin,H.;H.S.Kim;S.K.Kim;M.K.Shin;J.H.Kim;J.W.Lee https://doi.org/10.1016/S0141-0229(02)00064-9
  8. J. Life Science v.9 Microbial modification of extracellular polysaccharides Lee,J.W.
  9. Appl. Environ. Microbiol. v.65 Biosynthesis of novel exopolymers by Aureobasidium pullulans Lee,J.W.;W.G.Yeomans;A.L.Allen;F.Deng;R.A.Gross;D.L.Kaplan
  10. Can. J. Microbiol. v.43 Exopolymers from curdlan production: incorporation of glucose-related sugars by Agrobacterium sp. strain ATCC 31749 Lee,J.W.;W.G.Yeomans;A.L.Allen;D.L.Kapaln;D.Frank;R.A.Gross https://doi.org/10.1139/m97-020
  11. J. Gen. Microbiol. v.130 Identification of Bacillus strain using the API system Logan,N.A.;R.C.Berkeley
  12. Biotechnol. Adv. v.4 Microbial polysaccharides with actual potential industrial applications Paul,F.;M.Andre;P.Monsan https://doi.org/10.1016/0734-9750(86)90311-3
  13. Virology v.226 Role of the V2,V3, and CD4-binding domains of GP120 in curdlan sulfate neutralization sensitivity of HIV-1 during infection of T lymphocytes Paul,P.J.;W.Jason;K.Dariusz;J.W.Thomas;F.Andrzej;L.S.Aleksander;T.Mitsuo;T.Takashi;T.Mimura;S.F.Michael https://doi.org/10.1006/viro.1996.0649
  14. J. Microbiol. Meth. v.4 A simple plate assay for detection of group A streptococcus proteinase Hynes,W.L.;J.R.Tagg https://doi.org/10.1016/0167-7012(85)90004-1
  15. Immunopharmacol. v.7 A soluble β-1, 3-D-glucan derivative potentiates the cytostatic and cytolytic capacity of mouse peritoneal macrophages in vitro Seljelid,R.;J.Boegwald;J.Hoffman;O.Larm https://doi.org/10.1016/0162-3109(84)90009-2
  16. Korean J. Biotechnol. Bioeng. v.17 Seo,H.P.;J.M.Kim;H.D.Shin;T.K.Kim;H.J.Chang;B.R.Park;J.W.Lee
  17. Can. J. Microbiol. v.35 Effect of yeast extract on the degradation of organophosphorous insecticides by soil enrichment and bacterial cultures Sharmila,M.;K.Ramanans;N.Sethunathan https://doi.org/10.1139/m89-185
  18. Appl. Microbiol. Biotechnol. v.39 Properties of anaerobic sludge as affected by yeast extract, cobalt and iron supplements Shen,C.F.;N.Kosaric;R.Blaszezyk https://doi.org/10.1007/BF00166862
  19. Adv. Appl. Microbiol. v.23 Production of microbial polysaccharide Slodki,M.E.;M.C.Cadmus https://doi.org/10.1016/S0065-2164(08)70064-9
  20. Carbohydr. Res. v.276 Synthesis of curdlan sulfates having inhibitory effects in vitro against AIDS viruses HIV-1 and HIV-2 Soung,C.Y.;Y.Takashi;M.Toru;Y.Kaneko;N.Hideki;Y.Naoki;U.Toshiyuki https://doi.org/10.1016/0008-6215(95)00186-W
  21. Microbial, Technology, Current State, Future Prospects Microbial exopolysaccharides in polymers of current and future potential Sutherland,I.W.;D.C.Ellwood;Bull,A.T.(ed.);D.C.Ellwood(ed.);C.Ratledge(ed.)
  22. Appl. Microbiol. v.11 Polysaccharide produced by the genus pullularia Ueda,S.;K.Fusita;K.Komatsu;Z.Nakashima
  23. Food Microbiol. v.19 Characterization of Bacillus cereus isolates from fresh vegetables and refrigerated minimally processed foods by biochemical and physiological tests Valero,M.;L.A.Hemandez-Herrero;P.S.Femandez;M.C.Salmeron https://doi.org/10.1006/fmic.2002.0507
  24. Methods of soil analysis, Part 2: Chemical and microbiological properties(2nd eds.) Wollum,A.G.