Optimal Culture Condition and Extracellular Enzyme Activity of Metarhizium anisopliae

녹강균(Metarhizium anisopliae)의 최적 배양조건 및 효소활성

  • 박영일 (동국대학교 자연과학대학 생물학과) ;
  • 한영환 (동국대학교 자연과학대학 생물학과)
  • Published : 2000.06.01

Abstract

In order to use Metarhizizmn~ anisopliae as a biological pesticide, effect of envrionmental factors on nlycelial growth, spore formation, and extracellular enzyme activity in culture broth of M. anisopliae DGUM 35001 was determined. Optimal temperature was 26^{\circ}C.$ and optimal pH ranged from 5 to 9. Among the complex media tested, MCM and SDPY media were the most favorable for mycelial growth. When Czapek-Dox agar was used as a mnimal medium, glucose and sucrose among the saccharides were very excellent source of carbohydrate. Among the biopolyners tested. chitin was the most favorable source for mycelial growth and produced high aerial inycelia. Urea and ammonium phosphate as an inorganic nitrogen source and bacto-peptone and soytone as an organic nitrogen source enhanced the mycelial growth When serine as a source of amino acid was supplemented, excellent mycelial growth was shown. Large amount of spores could be obtained from the aerial mycelia of starch medium. When the culture broth was filtrated and then the concentrate with ammonium sulfate was used as a crnde enzyme solution, high enzyme activities of amylase and protease were shown. However, lipase and chitinase activities were comparatively low.

녹장균(Metarhizium ansiopliae)을 생물살충제로서 활용할 목적으로 녹강균의 균사생육 및 포자형성에 미치는 환경요인과 균사외 분비 효소 활성에 대하여 조사였다. M. anisopliae DGUM35001의 균사 생장을 보여주었으며, 공중균사는 SDPY 사용시 가장 우수하엿다. 최소배지로 Czapek-Dox 한천배지를 사용하여 탄소원 이용성 실험의 결과, 단당류에서 glucoserk 이당류에서 sucrose가 가장 우수한 탄소원이었다. 고분자 물질로 chitin 사용시 우수한 균사 생육을 보여주었다. 유기질소원으로 bacto-peptone과 soytone아 무기질소원으로 urea와 ammonium phosphate가 우수한 균사생육을 나타내었으며, 아미노산으로 serine 첨가시 가장 우수한 균사 생육을 나타내었다. Starch배지를 사용하였을 때 가장 우수한 포자형성의 결과를 얻었다. 액체배양후 군사를 제거한 상등액을 조효소액으로 사용시 analyase와 prorease 효소 활성이 우수하였다. 그러나,;ipase와 chitinase 효소활동은 상대적으로 낮았다.

Keywords

References

  1. 한국의 동충하초 성재모
  2. Anal. Biochem. v.72 A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding Bradford, M.M.
  3. Arch. Microbiol. v.124 Excretion of protease of Serratia marcescens Brown, V.; Schmitz, G.
  4. J. Invertebr. Pathol v.31 Growth and sporulation of Beauveria bassiana and Metarrizuim anisopliae on media containing various amino acids Campbell, R.K.; Perring, T.M.;Barnes, G.L.;Eikenbary, R.D.;Gentry, C.R.
  5. hemibapha DGUM 28001. Dongguk J. v.38 Optimization of culture condition for enhanced mycelial growth of Amanita hemibapha subsp Han, Y.H.;Ham, J.H.
  6. Appl. Microbiol. v.29 Powdered chitin agar as a selective medium for enumeration of Actinomycetes in water soil Hsu, S.C.;Lockwood, L.
  7. Kor. J. Appl. Micro-biol. Biotechnol. v.23 Isolation of microorganism producing chitinase for chitooligosaccharides production purfication of chitinase and its enzymatic characteristics Jeong, E.U.; Lee, Y.H.
  8. J. Ferment Technol. v.60 Purification and properties of an endocellulase of avicelase type from Irpex Iacteus(Polyporus tuliferae) Kanda, T.;Wakabayashi, K.; Nisizawa, K.
  9. Kor. J. Appl. Microbiol. Biotechnol. v.23 Isolation of a thermophilic Bacillus sp. producing the thermostable cellualse-free xylanase and properties of the enzyme Kim, D.J.;Shin, H.J.;Min, B.H.;Yoon, K.H.
  10. Anal. Chem. v.31 Use of dinirtrosalicylic acid reagent for determination of reducing sugar Miller, G. L.
  11. J. Invertebr. Pathol. v.8 Toxins from the entomogenous fungus in Metarrhisium anisopliae. I Production in submerged and surface cultures and in inorganic and organic nitrogen media Roberts, D.W.
  12. Enzyme. Microb. Tech-nol v.14 Formation of extracellular lipases by filamentous fungi, yeasts, and bacteria Rapp,P.;Backhaus, S.
  13. v.3 Bacterial and mold amylases Takaki, T.;Toda, H.;Isemura, T.
  14. J. Agri. Sci. Camb. v.93 Cellulase production by Penicillium purpurogenum Tokao, S.;Kamagata, Y.; Sasaki, S.
  15. Kor. J. Microbiol. v.33 Development of lipase hyper-producing strain from hybrids between Aspergillus niger and Penicillium chrysogenum by nuclear transfer Yang, Y.K.;Moon, M.N.;Lee, Y.H.;Lim, C.Y.