Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

A possible mechanism responsible for translocation and secretion an alkaliphilic bacillus sp. S-1 pullulanase

  • Published : 1997.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

The secretion of the alkaliphilic Bacillus sp. S-1 extracellular pullulanase involves translocation across the cytoplasmic membrane of the Gram-positive bacterial cell envelope. Translocation of the intracellular pullulanase PUL-I, was traced to elucidate the mechanism and pathway of protein secretion from an alkaliphilic Bacillus sp. S-1. Pullulanase could be slowly bue quantitatively released into the medium during growth of the cells in medium contianing proteinase K. The released pullulanase lacked the N-terminal domain. The N-terminus is the sole membrane anchor in the pullulanase protein and was not affected by proteases, confirming that it is not exposed on the cell surface. Processing of a 180,000M$\_$r/ pullulanase to a 140,000M$\_$r/ polypeptide has been demonstrated in cell extracts using antibodies raised against 140,000M$\_$r/ extracellular form. Processing of the 180,000 M$\_$r/ protein occured during the preparation of extracts in an alkaline pH condition. A modified rapid extraction procedure suggested that the processing event also occured in vivo. Processing apparently increased the activity of pullulanase. The western blotting analysis with mouse anti-serum against 140-kDa extracellular pullulanase PUL-E showed that PUL-I is processed into PUL-X via intermediate form of PUL-E. Possible explanationa for the translocation are discussed.

Keywords

References

  1. Biosci. Biotech. Biochem. v.56 Purification and some properties of an alkaline pullulanase from alkalophilic Bacillus sp. KSM-1876 Ara, K.;K. Igarashi;K. Saeki;S. Kawai;S. Ito
  2. Methods Enzymol. v.1 Amylases, α and β Bernfeld, P.
  3. J. Gen. Microbiol. v.234 Secretion mechanism of Bacillus subtlis levansucrase: characterization of the second step Chambert, R.;M-F. Petit-Glatron
  4. Ann. N.Y. Acad. Sci. v.121 Disk electrophoresis. II. Methods and application to human serum proteins Davis, B.J.
  5. Techniques in Molecular Biology Gooderham, K.;Walkerand, J. M.(ed.);Gaastra, W.(ed.)
  6. FEMS Microbiol. Lett. v.116 Specific detection of pullulanase type I on polyacrylamide gels Kim, C.H.
  7. J. Industrial Microbiol. v.12 Pullulanases of alkaline and broad pH range from a newly isolated alkalophilic Bacillus sp. S-1 and a Micrococcus sp. Y-1 Kim, C.H.;H.I. Choi;D.S. Lee
  8. Biosci. Biotechnol. Biochem. v.57 Purification and biochemical properties of extracellular alkaline pullulanase from an alkalophilic Bacillus sp. S-1 Kim, C.H.;H.I. Choi;D.S. Lee
  9. Eur. J. Biochem. v.227 Purification and detailed characterization of a bifunctional amylase-pullulanase enzyme from Bacillus circulans having two different active sites on one polypeptide Kim, Y.S.;C.H. Kim
  10. Molecular Microbiology v.4 Molecular characterization of pulA and its product, pullulanase, a secreted enzyme of Klepsiella pneumoniae UNF5023 Kornacker, M.G.;A.P. Pugsley
  11. J. Gen. Microbiol. v.135 Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus Kuriki, T.;T. Imanaka
  12. Nature v.227 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Laemmli, U.K.
  13. J. Biochem. Molec. Biol. v.30 Processing of an intracellular immature pullulanase to the mature form involves enzymatic activation and stabilization in alkalophilic Bacillus sp. S-1 Lee, M.J.;B.S. Kang;D.S. Kim;Y.T. Kim;S.K. Kim;K.H. Chung;J.K. Kim;K.S. Nam;Y.C. Lee;C.H. Kim
  14. Arch. Biochem. Biophys. v.337 Intracellular and extracellular forms of alkaline pullulanase of an alkalophilic Bacillus sp. S-1 Lee, M.J.;Y.C. Lee;C.H. Kim
  15. J. Biol. Chem. v.193 Protein measurement with the Folin phenol reagent Lowry, O.H.;N.H. Rosenbrough;A.L. Farr;A. Randall
  16. J. Gen. Microbiol. v.136 Nucleotide sequence of the α-amylase-pullulanase gene from Clostridium thermohydrosulfurium Melaswien, H.;M. Paloheimo;L. Hemio
  17. J. Biochem. v.107 Cloning and characterization of a Bacillus subtilis gene homologous to E. coli SecY Nakamura, K.;A. Nakamura;H. Takamatsu;H. Yoshikawa;Yamane, K.
  18. Starch/Starke v.41 Characterization of pullulanase and α-amylase activities of a Thermus sp. AMD33 Nakamura, N.;N. Sashihara;H. Nagayama;K. Horikoshi
  19. J. Industrial Microbiol. v.3 Purification and characterization of a thermostable pullulanase from Thermoactinomyces thalpophilus Obido, F.J.C.;S.K.C. Obi
  20. Agric. Biol. Chem. v.46 An inductive effector in production of extracellular pullulanase by Aerobacter aerogenes Ohba, R.;S. Veda
  21. Biotechnol. Bioeng. v.20 Immobilization od Streptomyces flavochromogenes Pullulanase on tannic acid and TEAE-cellulose Ohba, R.;H. Chaen;S. Hayashi;S. Veda
  22. Eur. J. Biochem. v.163 Secretion of Bacillus subtilis levansucrase: a possible two-step mechanism Petit-Glatron, M.-F.;F. Benyahia;R. Chambert
  23. Enz. Microl. Technol. v.8 A highly stable pullulanase from Thermus aquaticus YT-1 Plant, A.R.;H.W. Morgan;R.M. Daniel
  24. Molecular Microbiology v.6 Pullulanase secretion in Escherichia coli K-12 requires a cytoplasmic protein and a putative polytopic cytoplasmic membrane protein Possot, O.;d'Enfert, I.;Reyss;A.P. Pugsley
  25. Molecular Microbiology v.4 Five genes at the 3 end of the Klepsiella pneumoniae pulC operon are required for pullulanase secretion Pugsley, A.P.;I. Reyss
  26. Ann. N.Y. Acad. Sci. v.434 Characterization of a new class of thermophilic pullulanases from Bacillus acidopullulyticus: comparison with Aerobacter aerogense pullulanase; compatibility with Aspergillus niger glucoamylase Schulein, M.;B. Hojer-De
  27. FEBS Lett. v.360 Kinetics of the unfolding-folding transition of Bacillus subtilis levansucrase precursor Scotti, P.A.;R. Chambert;M.-F. Petit-Glatron
  28. Appl. Microbiol. Biotechnol. v.34 A hyperthermostable pullulanase produced by an extreme thermophile, Bacillus flavocaldarius KP1228, and evidence for the proline theory of increasing protein thermostability Suzuki, Y.;K. Hatagaki;H. Oda
  29. Molecular Microbiology v.6 Role of lipid modification on a starchdebranching enzyme, Klepsiella pullulanase: comparison of properties of lipid-modified and unmodified pullulanases Yamashita, M.;A. Nakagawa;N. Katsuragi;Y. Murooka
  30. J. Biol. Chem. v.257 The isolation of homogeneous leader peptidase from a strain of Escherichia coli which overproduces the enzyme Wolfe, P.B.;P. Silver;W. Wickner