Characterization of the Bacterial Cell Wall Lytic Enzyme Produced by Aspergillus sp. HCLF-4

Aspergillus sp. HCLF-4에 의해 생성되는 세균세포벽 분해효소의 특성

  • Published : 2001.03.01

Abstract

In this study, we have isolated bacterial cell wall lytic enzyme in the culture supernatant of Aspergillus sp. HCLF-4. This hydrolase showed cell wall lytic activity against Anabaena cylindrica. The extracellular enzyme was produced by Aspergillus sp. HCLF-4 when it was grown in a PDB media containing 0.05% heat killed Micrococcus luteus cells. The molecular weight of lytic enzyme was about 14.3 kDa. The optimal pH and temperature for the activity of this enzyme were 3.0~4.0 and $30^{\circ}C$, respectively. This hydrolase activity was reduced by $Na^{+}$, $Li^{+}$, $Ca^{2+}$, $Cu^{2+}$, $Fe^{3+}$, EDTA, and PMSF, whereas it was increased by $Mg^{2+}$, $Mn^{2+}$>. The enzyme has N-acetylmuramyl-L-amidase or endopeptidase activity.

Aspergillus sp. HCLF-4가 생성하는 세균 세포벽 분해효소의 특성을 규명하였다. 본 세포벽 분해 효소는 Anabaena cylindrica 세포벽 분해능을 보였다. 이 세포벽 분해 효소는 Aspergillus sp. HCLF-4를 기질성분으로 0.05% heat killed Micrococcus luteus가 포함된 PDB 배지에 키웠을 때 생성되는 inducible enzyme으로 분자량은 약 14.3 kDa 이었다. 본 세포벽 분해효소는 pH 3.0-4.0, 온도 $30^{\circ}C$ 조건에서 최고의 활성을 보였고 $Mg^{2+}$와, $Mn^{2+}$의 2가 이온에서 분해 효소의 활성이 촉진되었다. 반면, 1가 양이온 $Na^{+}$$Li^{+}$, 2강 양이온 $Ca^{2+}$$Cu^{2+}$, 3가 양이온 $Fe^{3+}$에서는 활성이 억제되었으며 EDTA와 PMSF 또한 분해 효소의 활성을 억제 시켰다. 이 효소는 N-acetylmuramyl-L-amidase 또는 endopeptidase와 같은 활성을 보였다.

Keywords

References

  1. 효소단백질 정제법 안용근
  2. 한국미생물학회지 v.33 Moraxella sp. CK-1의 세포외 autolysin의 특성 안준익;김철호;최영길
  3. 한국미생물학회지 v.35 Penicillium oxalicium(HCLF-34)으로부터 분비되는 Anabaena cylindrica 세포벽 분해효소의 특성 현성희;최영길
  4. 한국환경생물학회지 v.5 시안세균 생장 억제 곰팡이의 분리 및 동정 현성희;성주연;최영길
  5. Biotechnol. Appl. Biochem. v.8 Natural antimicrobial systems and their potential in food preservation in the future Banks, J.G.;R.G. Board;N.H. Sparks
  6. J. Bacteriol v.177 Molecular cloning and nucleotide sequence of the gene encoding the major peptidoglycan hydrolase of Latococcus lactis, a mulamidase needed for cell separation Buist, G.;J. Kok.;K.J. Leenhonuts;M.D. Abrowska;G. Venema;A.J. Haandrikman
  7. Eur. J. Biochem v.274 Purification and properties of a basic endo-α-1,6-glucanase(BGN 16.1) from the antagonistuc fungus Trichoderma harzianum de la Cruz J.;A. Llobell
  8. Arch. Microbiol v.159 Carbon source control on α-glucanases, chiotobiase and chitinase from Trichoderma harzianum de la Cruz J.;M. Rey;J.M. Lora;A. Hidalgo-Gallego;F. Dominguez;J.A. Pintor-Toro;A. Llobell;T. Benitez
  9. J. Bacteriol. v.171 The second peptidoglycan hydrolase of Streptococcus faecium ATCC 9790 covalently binds penicillin Dolinger, D.L.;L. Daneo-moore;G.D. Shockman
  10. Polymer Degradation and Stablity v.57 Influence of 1-sorbose and the cell-wall-lytic Micrococcus sp. on the major polymers of Aspergillus fumigatus El-Shafei H.A.
  11. J. Bacteriol v.174 Analysis of the autolysis of Bacillus subtilis 168 during vegetative growth and differentiation by using renaturing polyacylamide gel electrophoresis Foster, S.J.
  12. Bacteriol. Rev. v.32 Use of bacteriolytic enzyme in determination of wall structure and their role in cell metabolism Ghuysen, J.M.
  13. Method. in Enzymol v.8 Enzymes that degrade bacterial cell wall Ghuysen, J.M.;D.J. Tipper;J.L. Strominger
  14. J. Bacteriol v.176 Analysis of the sodium dodecyl sulfate-srable peptidoglycan autolysins of select gram-negative pathogens by using renaturing polyacry1-amide gel electrophoresis Gilad Bernadsky;T.J. Beveridge;A.J. Clarke
  15. App. Environ. Microbiol. v.53 Antimicrobial activity of lysozyme against bacteri involved in food spoilage and food-borne disease Hughey, V.L.;E.A. Johnson
  16. Arch. Oral. Biol. v.24 Enzymiclysis and structure of the cell walls of the oral Streptococcus mutanms BH1 Inone, M.;S. Hamada;S. Kotani;K. Kato
  17. J. Microbiol. Biotechnol. v.1 Purification and characterization of a bacteriolytic enzyme from Alkalophilic Bacillus sp. Jung, M.H.;S. Kong;D.H. Bai;J.H. Yu
  18. Kor. J. Environ. Biol. v.15 Lysis of Anabaena cylindrica(Cyanobacterium) cell wall by extracelluar enzyme of Moraxella sp. CK-1 Kim, C.H.;Y.K. Choi;B.R. Min
  19. Nature(London) v.227 Cleavage of structural proteins during the assembly of the head of bacteriaophage T4 Laemmli, U.K.
  20. J. Biol. Chem. v.193 Protein measurement with the folin phenol regent Lowry, O.H.;N.J. Rosenbrough;A.L. Farr;R.J. Randall
  21. Biotechnol. Appl. Biochem. v.28 α-Galactosidases of Penicillium simplicissimum: production, purification characterization of the gene encoding AGLI Luonteri E.;E. Alatalo;M. Siika-aho;M. Penttila;M. Trnkanen
  22. Mole. and Cell. v.7 Cloning and expression of a gene encoding Moraxella sp. CK-1 autolysin Ohn, T.B.;C.H. Kim;B.R. Min;Y.K. Choi
  23. J. Gen. Microbiol v.12 Isolation of Streptomyces spp. capable of decomposing preparations of cell walls from various microorganisms and a comparison of their lytic activities with those of certain actinomyctes and myxobacteria Salton, M.R.J.
  24. Ann. Inst. Pasteur. Microbiol. v.136 The autolytic peptidoglycan hydrolases Streptococcus faecium Shockman, G.D.;T. Kawamura;J.F. Barrett;D.L. Dolinger
  25. J. Biol. Chem. v.266 The primary structure of Aspergillus niger acid proteinase A. Takahashi, K.;J. Inoue;K. Sakai;T. Kohama;S. Kitahara;K. Takishima;M. Tanji;S.B.P. Athauda;T. Takahashi;H. Akanuma;G. Mamiya;M. Yamasaki
  26. Anal. biochem. v.22 A modification of the Park and Johnson reducing sugar determination suitable for the assay of insoluble materials: Its application to bacterial cell walls Thompson J.S.;G.D. Shockman
  27. J. Bacteriol v.176 Initial characterization of two extracellular autolysins from Pseudomonas aeruginosa PAO1 Watt S.R.;A.J. Clarke