Selection of organic Nitrogen Source and Optimization of Culture Conditions for the Production of Arachidonic Acid from Mortierella alpina

Mortierella alpina를 이용한 아라키돈산의 생산에서 유기질소원의 선정과 배양 조건의 최적화

  • 유연우 (아주대학교 분자과학기술학과) ;
  • 하석진 (아주대학교 분자과학기술학과) ;
  • 박장서 (두산바이오텍)
  • Published : 2004.02.01

Abstract

Experiments were carried out to select an organic nitrogen source and optimize the culture conditions for the production of arachidonic acid by Mortierella alpina DSA-12. Corn steep powder(CSP) was selected as an organic nitrogen source based on arachidonic acid production and raw material price. The optimum C/N ratio was in the range of 15 to 17 with the medium containing glucose as carbon source and CSP as nitrogen source. The optimum culture conditions for arachidonic acid production showed 500 rpm agitation and 25$^{\circ}C$ culture temperature at 0.5 vvm aeration. Under the optimum conditions, the concentration of cell, total lipid and arachidonic acid were 21.8 g/L, 10.2 g/L and 3.70 g/L, respectively, from 50 g/L glucose and 18 g/L CSP. In the 500 L fermenter with 0.5 vvm aeration and 200 rpm agitation, the concentration of cell, total lipid and arachidonic acid were 19.8 g/L, 9.1 g/L and 3.67 g/L, respectively, from 50 g/L glucose and 18 g/L CSP. This result showed that an arachidonic acid production could be possible with a bench-scale fermenter using corn steep powder as a nitrogen source.

곰팡이인 Mortierella alpina DSA-12를 이용한 arachidonic acid의 생산을 위한 유기질소원의 선정과 배양 조건의 결정에 연구를 수행하였다. Corn steep powder (CSP)를 원료의 가격과 arachidonic acid의 생산을 기준으로 유기질소원으로 선정하였다. 탄소원으로 glucose와 질소원으로 CSP를 사용한 경우에 최적의 C/N ratio는 15-17 범위이다. Arachidonic acid의 생산을 위한 최적의 배양조건은 $25^{\circ}C$에서 500 rpm의 교반과 0.5 vvm의 통기이며, 이 때 50 g/L의 glucose와 18 g/L의 CSP로부터 21.8 g/L의 균사체량에 10.2 g/L의 총 지질을 얻을 수 있었으며, arachidonic acid의 농도는 3.70 g/L이었다. 500 L의 발효조에서 0.5 vvm과 200 rpm의 교반으로 실험을 수행한 결과 19.8 g/L의 균사체량과 9.1 g/L의 총 지질 및 3.67 g/L의 arachidonic acid를 얻었다. 이러한 결과는 bench-scale의 발효조에서도 질소원으로 CSP를 이용하여 arachidonic acid의 생산이 가능함을 보여주었다.

Keywords

References

  1. Biotechnol. Bioproc. Eng. v.7 no.5 Production of arachidonic acid by Mortierella fungi Higashiyma,K.;s.Fujikawa;E.Y.Park;S.Shimizu https://doi.org/10.1007/BF02932833
  2. Trends in Biotechnol v.15 Polyunsaturated fatty acid, part 1: occurrence, biological activities and applicaions Gill,I.;R.Valivety https://doi.org/10.1016/S0167-7799(97)01076-7
  3. Proc. Natl. Acad. Sci. USA v.90 Arachidonic acid status correlates with the first year growth in preterm infants Calson,S.E.;S.H.Werkman;J.M.Peeples;R.J.Cooke;E.A.Tolley https://doi.org/10.1073/pnas.90.3.1073
  4. Mycotaxon v.94 Chemo-taxonomic significance of fatty acid composition in the genus Mortierella (Zygomycetes, Mortierellaceae) Amano,N.;Y.Shinmen;K.Akimoto;H.Kawashima;T.Amachi;S.Shimizu;H.Yamada
  5. Appl. Microbiol. Biotechnol. v.31 Production of arachidonic acid by Mortierella fungi: selection of a potent producer and optimization of culture conditions for large-acale production Shinmen,Y.;S.Shimizu;K.Akimoto;H.Kawashima;H.Yamada
  6. Biotechnol. Lett. v.10 Arachidoinic acid production by Motierella alpina sp. S-17. Influence of C/N ratio Sajibidor,J.;S.Dobronova;M.Certik
  7. Appl. Environ. Microbial v.57 no.4 Arachidonic acid production by fungi Ward,O.P.;P.K.Bajpai;P.Bajipai
  8. Industrial applications of single cell oils industrial production of arachidonic acid by Mortierella Totani,N.;K.Someya;K.Oba
  9. Appl. Microbiol. Biotechnol. v.28 Effect of culture conditions on mycelial growth and production of γ-lineolenic acid by the fungus Motierella ramanniana Hansson,L.;M.ostalek
  10. Mortierella alliacea strain YN-15, J. Am. Oil Chem. Soc. v.78 Production of arachidonic acid by filamentous fungus Aki,T.;Y.Nagahata;K.Ishihara;Y.Tanaka;T.Morinaga;K.Higashiyama;K.Akimoto;S.Fujikawa;S.Kawamoto;S.Shigeta;K.Ono;O.Suzuki
  11. J. Biosci. Bioeng. v.88 Effect of nitrogen sources on mycelial morphology and arachidomic acid production in cultures of Motierella alpina Park,E.Y.;Y.Koike;K.Higashiyama;S.Fujikawa;M.Okabe https://doi.org/10.1016/S1389-1723(99)80177-7
  12. J. Am. Oil Chem. Sco. v.74 no.5 Optimization of arachidomic acid production by Motierella alpina Wuji-H4 isolate Chen,H.C.;C.C.Chang;C.X.Chen https://doi.org/10.1007/s11746-997-0182-1
  13. J. Ind. Microbiol. Biotechnol. v.30 An inexpensive medium for production of arachidonic acid by Mortierella alpina Zhu,M.;L.J.Yu;Y.X.Wu https://doi.org/10.1007/s10295-002-0013-1
  14. J. Biosci. Bioeng. v.91 no.4 Effect of consumed carbon to nitrogen ratio on mycelial morphology and arachidomic production in cultures of Mortierella alipina Park,E.Y.;Y.Koike;K.Higashiyama;S.Fujikawa https://doi.org/10.1263/jbb.91.382
  15. Appl. Microbiol. Biotechnol. v.39 Effect of temperature and glucose supply on the production of polyunsaturated fatty acids by fungus Mortierella alpina CBS343.66 in fermentor cultures Lindberg,A.M.;G.Molin https://doi.org/10.1007/BF00205031
  16. Appl. Microbiol. Biotechnol. v.37 Lipid formation in oleaginous mold Entomophthora exitalis growth in comtinuous culture: effects of growth rate, temperature and dissolved oxygen tension on polyunsaturated fatty acids Kendrick,A.;C.Ratledge
  17. biotechnol. Boeng. v.63 Effects of dissolved oxygen on the production by Motierella alpina 1S-4 Higashiyama;K.;K.Muarakami;H.Tsujimura;N.Matsumoto;S.Fujikawa https://doi.org/10.1002/(SICI)1097-0290(19990520)63:4<442::AID-BIT7>3.0.CO;2-9
  18. Can. J. Chem. Eng. v.73 Process for production of arachidonic acid concentrate by a strain of Mortierella alpina Li,Z.Y.;Y.Lu;V.B.Yadwad;O.P.Ward https://doi.org/10.1002/cjce.5450730115
  19. J. Biosci. Bioeng. v.87 no.4 Image analysis of morphological change during arachidonic acid production by Mortierella alpina 1S-4 Higashiyama,K.;S.Fujikawa;E.Y.Park;M.Okabe https://doi.org/10.1016/S1389-1723(99)80098-X
  20. J. Fermt. Bioeng. v.84 no.4 Optimization and scale up of γ-linolenic acid production by Mortierella ramanniana MN 15-1, a high γ-linolenic acid producing mutant Hiruta,O.;T.Futamura;H.Takeha;A.Satoh;Y.Kanisaka;T.Yokochi;T.Nakahara;O.Suzuki
  21. Proc. Biochem. v.24 Omega-3 fatty acids: alternative sources of production Yongmanitchai,W.;O.P.Ward