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The journal of microbiology v.35 no.3, 1997년, pp.213 - 221  

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

Shim, Jae-Kyoung    (Dongguk University   ); Kim, Kyoung-Sook    (Korea Research Institute of Bioscience and Biotechnology   ); Kim, Cheorl-Ho    (Dongguk University  );
  • 초록

    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.


  • 주제어

    Alkaliphilic Bacillus sp .   S-1 .   intracellular pullulanse .   secretion .   translocation.  

  • 참고문헌 (30)

    1. Secretion mechanism of Bacillus subtlis levansucrase: characterization of the second step , Chambert, R.;M-F. Petit-Glatron , J. Gen. Microbiol. / v.234,pp.1205-1214,
    2. Specific detection of pullulanase type I on polyacrylamide gels , Kim, C.H. , FEMS Microbiol. Lett. / v.116,pp.327-332,
    3. An inductive effector in production of extracellular pullulanase by Aerobacter aerogenes , Ohba, R.;S. Veda , Agric. Biol. Chem. / v.46,pp.2425-2431,
    4. Disk electrophoresis. II. Methods and application to human serum proteins , Davis, B.J. , Ann. N.Y. Acad. Sci. / v.121,pp.404-427,
    5. Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus , Kuriki, T.;T. Imanaka , J. Gen. Microbiol. / v.135,pp.1521-1528,
    6. Nucleotide sequence of the α-amylase-pullulanase gene from Clostridium thermohydrosulfurium , Melaswien, H.;M. Paloheimo;L. Hemio , J. Gen. Microbiol. / v.136,pp.447-454,
    7. Purification and characterization of a thermostable pullulanase from Thermoactinomyces thalpophilus , Obido, F.J.C.;S.K.C. Obi , J. Industrial Microbiol. / v.3,pp.343-350,
    8. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 , Laemmli, U.K. , Nature / v.227,pp.680-685,
    9. 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 , Ann. N.Y. Acad. Sci. / v.434,pp.271-274,
    10. Immobilization od Streptomyces flavochromogenes Pullulanase on tannic acid and TEAE-cellulose , Ohba, R.;H. Chaen;S. Hayashi;S. Veda , Biotechnol. Bioeng. / v.20,pp.665-676,
    11. Molecular characterization of pulA and its product, pullulanase, a secreted enzyme of Klepsiella pneumoniae UNF5023 , Kornacker, M.G.;A.P. Pugsley , Molecular Microbiology / v.4,pp.73-85,
    12. 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 , Molecular Microbiology / v.6,pp.95-105,
    13. Secretion of Bacillus subtilis levansucrase: a possible two-step mechanism , Petit-Glatron, M.-F.;F. Benyahia;R. Chambert , Eur. J. Biochem. / v.163,pp.379-387,
    14. Characterization of pullulanase and α-amylase activities of a Thermus sp. AMD33 , Nakamura, N.;N. Sashihara;H. Nagayama;K. Horikoshi , Starch/Starke / v.41,pp.112-117,
    15. Kinetics of the unfolding-folding transition of Bacillus subtilis levansucrase precursor , Scotti, P.A.;R. Chambert;M.-F. Petit-Glatron , FEBS Lett. / v.360,pp.307-309,
    16. 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 , J. Industrial Microbiol. / v.12,pp.48-57,
    17. Gooderham, K.;Walkerand, J. M.(ed.);Gaastra, W.(ed.) , Techniques in Molecular Biology / v.,pp.49-87,
    18. Cloning and characterization of a Bacillus subtilis gene homologous to E. coli SecY , Nakamura, K.;A. Nakamura;H. Takamatsu;H. Yoshikawa;Yamane, K. , J. Biochem. / v.107,pp.603-607,
    19. 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 , Molecular Microbiology / v.6,pp.389-394,
    20. Amylases, α and β , Bernfeld, P. , Methods Enzymol. / v.1,pp.149-158,
    21. Intracellular and extracellular forms of alkaline pullulanase of an alkalophilic Bacillus sp. S-1 , Lee, M.J.;Y.C. Lee;C.H. Kim , Arch. Biochem. Biophys. / v.337,pp.308-316,
    22. Five genes at the 3 end of the Klepsiella pneumoniae pulC operon are required for pullulanase secretion , Pugsley, A.P.;I. Reyss , Molecular Microbiology / v.4,pp.365-379,
    23. Purification and biochemical properties of extracellular alkaline pullulanase from an alkalophilic Bacillus sp. S-1 , Kim, C.H.;H.I. Choi;D.S. Lee , Biosci. Biotechnol. Biochem. / v.57,pp.1632-1638,
    24. 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 , Eur. J. Biochem. / v.227,pp.687-693,
    25. Protein measurement with the Folin phenol reagent , Lowry, O.H.;N.H. Rosenbrough;A.L. Farr;A. Randall , J. Biol. Chem. / v.193,pp.265-275,
    26. The isolation of homogeneous leader peptidase from a strain of Escherichia coli which overproduces the enzyme , Wolfe, P.B.;P. Silver;W. Wickner , J. Biol. Chem. / v.257,pp.7898-7902,
    27. Purification and some properties of an alkaline pullulanase from alkalophilic Bacillus sp. KSM-1876 , Ara, K.;K. Igarashi;K. Saeki;S. Kawai;S. Ito , Biosci. Biotech. Biochem. / v.56,pp.62-65,
    28. 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 , J. Biochem. Molec. Biol. / v.30,pp.46-54,
    29. 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 , Appl. Microbiol. Biotechnol. / v.34,pp.707-714,
    30. A highly stable pullulanase from Thermus aquaticus YT-1 , Plant, A.R.;H.W. Morgan;R.M. Daniel , Enz. Microl. Technol. / v.8,pp.668-672,

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