Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Overproduction of Lignin Peroxidase from Phanerochaete chrysosporium PSBL-1

Phanerochaete chrysosporium PSBL-1의 배양조건 최적화를 통한 Lignin Peroxidase의 과량생산

  • 정병철 (명지대학교 생명과학과/환경생명공학원) ;
  • 한윤전 (명지대학교 생명과학과/환경생명공학원) ;
  • 장승욱 (명지대학교 생명과학과/환경생명공학원) ;
  • 정욱진 (화학공학과 /청정기술원) ;
  • 원유정 (화학공학과/인수환경)
  • Published : 2000.09.01
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Abstract

Until now, it was diIliculi to overproduce lignin peroxidase(LiP) fiom Pl~anemchaete ch~ysosporium since the lack of optimized growth conditions. In this paper, we optimized the LIP production conditions and monitored LIP isozyines of fl chqsospoi.ium PSBL-1. The optimized condition includes sponge matrix support, no addition of $MnSO_4$, excess addition of niixogen source(48 inM diarmnonium), and addition of stabilizer(2 mM verakyl alcohol). Finally we obtained Lip activity of 1,800 unitsll. HI isozyne was overproduced when inyceliuin was cultivated in media containing $Mn^{2+}$ (2.73 inM) and excess nitrogen(48 11d4 diannnonium). Three azo dyes(acid yellow 9, congo ued, orange IT; each concenimtion of50 $\mu$M) we1-e rapidly decolorized within 2 inins by 0.4 un~t or Lip.

지금까지의 Phanerochaete chrysosporium을 이용한 Lignin Peroxidase(LiP) 의 생산은 최적화 되지 못한 배양 조건으로 인해 매우 낮은 활성만을 얻을수 있었으며. 그 배양조건 또한 100% 산소의 주입을 요하는 일반적으로는 재현하기 어려운 조건이었다. 본 연구에서는 Phanerochaete chrysosporium PSBL-1 균줄르 이용, 다양한 배양조건의 변화를 통해 LiP의 과량생산과 그에 따른 isozyme 조성의 변화를 확인하고자 하였다. 최종적으로 $Mn^{2+}$를 제거한 배지에 질소 원인 diammonium을 48mM 로 첨가하고, 안정제로서 veratryI alcohol을 2 mM로 첨가하여, sponge에 의한 고착배양을 실시한 결과 1,800 units/l의 LiP가 생산되었다. 이러한 활성의 증가는 기존에 보고된 PSBL-1 균주의 LiP 최대 생산량인 700units/l와 비교 , 약 2.57배의 생산량 증대를 나타낸다. 생산된 LiP isozyme 분석결과 $Mn^{2+}$(2.73 mM)와 diammonium(48 mM)을 첨가하였을 경우 H1 isozyme의 과량 생산이 확인되었다. 생산된 LiP(0.4 units)를 이용하여 각각의 아조계 염료(acid yellow 9, congo red, orange II; each 50$\mu$M)에 대한 탈생능을 확인한 결과, 2분 이내에 이들 염료에 대한 탈색이 이루어지는 것을 확인할 수 있었고 분해 산물이 축적되는 것을 확인할 수 있었다.

Keywords

References

  1. Appl. Envrion. Microbiol. v.58 A new assay for lignin-type peroxidases employing the dye azure B. Archibald,F.S.
  2. J. Biotechnol v.30 Influence of primary and secondary proteases produced by frdd or immobilized cells of the white-rot fungus Phanerochate chryosporium on lignin peroixdase activity Bonnarme,P.
  3. Appl. Environ. Microbiol. v.56 Mn(Ⅱ)regulation of lignin peroxidases and Mn-dependent peroxidases from lignin-degrading white rot fungi Bonnarme,P.;T.W.Jeffries
  4. J, Bacteriol v.172 Manganese regulates expression of manganese peroxidase by Phanerochate chrysosporum Brown,J.A.;J.K.Glenn;M.H.Gold
  5. Appl. Environ Microbiol v.63 Stabilization of lignin peroxidases in white rot fungi by trypophan Collins,P.J.;J.A.Field;A.Teunissen;A.D.W.Dohson
  6. Appl. Environ. Microbiol. v.56 Biodegradation of azo and heterocyclic dyes by Phanerochaete chrysosporium Cripps,C.;D.A.Steven
  7. Gene v.60 Characterization of extracellular peroxidases produced by acetate-buffered cultures of the lignin degrading basidiomycete Phanerochate chrysoporium Dass,S.B.;C,A,Reddy
  8. Arch. Microbiol. v.163 Extracellular proteases produced by the wood-degrading fungus Phanerochaete chrysosporum(BKMF1767)under ligninolytic and non-ligninolytic cinditions Dosoretz,C.G.
  9. Appl. Environ Microbiol v.59 Overproduction of lignin peroxidase by Phanerochaete chrysporitim(BKMF1767)under nonlimiting nutrient condition Dosoretz,C.G.;N.Rothschild;Y.Hadar
  10. Arch. Biochem. Biophys. v.251 oxidation is the principal funtion of the extracellular Mn-peroxidase from Phanerochaete chrysosporium Glenn,J.K.;L.Akileswaran;M.H.Gold
  11. Arch. Biochem. Biopys v.234 Purification and characterization for an extracelluar H₂O₂-requiring diarylpropane oxygenase from the white rot basidiomycete Phanerochaete chrysosporum Glod M.H.;M.Kuwahara;A.A.Chiu;J.K.Glenn
  12. Enzyme Microbiol. Technol v.11 Organopollutant degradation by ligninolytic fungi Hammel,K.E.
  13. Appl. Environ. Microbiol v.58 Determination of the respiration kinetics for mycelial pellets of Phanerochaete chrysosporium Michael.F.C.Jr.;S.B.Dass;E.A.Grulke;C.A.Reddy
  14. Appl. Environ. Microbiol. v.57 Role of manganese peroxidases and lignin peroxidases of Phanerochaete chrysoporium in the decolorization of kraft bleach plant dffluent Michael,F.C.Jr.;S.B.Dass;E.A.grulke;C.A.Reddy
  15. Appl. Environ. Microbiol v.57 Overproduction of lignin degrading enzymes by and isolate of Phanerochaete chrysosporium Orth, A.B.;M.Denny;M.Tien
  16. Biological degradation and bioremediation of toxic chemicals Properties of lignin degrading peroxidases and their use in bioremediation Orth,A.B.;E.A.Pease;M.Tien;Chaudhry,G.R.(ed.)
  17. Arch. Biochem. Biophys. v.372 Extrcelluar mannose-6-phosphatase of Phanenchaete chrysosporium Rothschild,N.;L.Ayala;H.Yitzhak;C.Dosoretz
  18. Arch Biochem. Biophys v.373 Substrate specificity of lignin peroxidase and a SI68W variant of manganese peroxidase Sergei,L.;R.Scott;D.A.Steven
  19. Methods Enzymil v.161 Lignin peroxidase of Phanerochaete chrysosporium Tien,M.;T.K.Kirk
  20. Appl. Environ. Microbiol v.65 Lignin modifying enzymes of the white rot basidiomycete Ganoderma lucidum Trevor,M.D.;S.M.Carlos;C.R.Admarayana
  21. J. Biol. Chem. v.265 Lignin peroxidase compoundⅢ Mechanism of formation and decomposition Wanshi,H.;M.H.Gold
  22. FEMS Microbiol Lett. v.107 Decolorization of industrial effluents containing reactive dyes by acinomucetes Zhou,W.;W.Zimmermann