1887

Abstract

A gene encoding the periplasmic α-amylase of K-11151 was cloned into using pUC19 as a vector. An ORF of 1578 bp was deduced to be the amylase structural gene. The primary structure of the enzyme had little identity with other α-amylases, except with the enzyme from . The enzyme was expressed in from the promoter of pUC19 and was found to be transported to the periplasmic space. The expressed enzyme showed the same thermal stability, optimum temperature and substrate specificity as the enzyme from . The enzyme formed maltotetraose, but not 6- nor 6-maltosyl-maltose, from maltose by the reverse reaction, and the tetraose was then hydrolysed to maltotriose and glucose. The addition of maltotriose enhanced the production of glucose from maltose. In addition, maltose was formed by the condensation of glucose by the enzyme. Thus, the periplasmic α-amylase of was shown to produce glucose from maltose by hydrolysing maltotetraose and possibly higher maltooligosaccharides, which were the products of a condensation reaction, as a major pathway, and by direct hydrolysis of maltose as a minor pathway.

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1996-06-01
2021-10-17
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References

  1. Abe J., Hizukuri S., Koizumi K., Kubota Y., Utamura T. Enzymic syntheses of doubly-branched cyclomaltoheptaose through the reverse action of Pseudomonas isoamylase. Carbohydr Res 1988; 176:87–95
    [Google Scholar]
  2. Abe J., Onitsuka N., Nakano T., Shibata Y., Hizukuri S., Entani E. Purification and characterization of periplasmic a-amylase from Xanthomonas campestris K-11151. J Bacteriol 1994; 176:3584–3588
    [Google Scholar]
  3. Brumm P.J., Hebeda R.E., Teague W.M. Purification and characterization of the commercialized, cloned Bacillus mega-terium a-amylase. Part I. Purification and hydrolytic properties. Starch Staerke 1991a; 43:315–319
    [Google Scholar]
  4. Brumm P.J., Hebeda R.E., Teague W.M. Purification and characterization of the commercialized, cloned bacillus mega-terium a-amylase Part II. Transferase properties. Starch Staerke 1991b; 43:319–323
    [Google Scholar]
  5. Bush K., Sykes R.B. p- Lactamase (penicillinase, cephalo-sporinase). In Methods of Enzymatic Analysis 1984 Edited by Bergmeyer H.U., Bergemeyer H.J., Grabl M. Weinheim: Verlag Chemie; 5 pp 280–285
    [Google Scholar]
  6. Davis B.J. Disc electrophoresis. II. Method and application to human serum proteins. Ann NY Acad Sci 1964; 121:404–427
    [Google Scholar]
  7. Gough C.L., Dow J.M., Keen J., Henrissat B., Daniels M.J. Nucleotide sequence of the engXCA gene encoding the major endoglucanase of Xanthomonas campestris pv campestris. Gene 1990; 89:53–59
    [Google Scholar]
  8. Hizukuri S., Takeda Y., Yasuda M., Suzuki A. Multibranched nature of amylose and the action of debranching enzyme. Carbohydr Res 1981; 94:205–213
    [Google Scholar]
  9. Hu N.T., Hung M.N., Huang A.M., Tsai H.F., Yang B.Y., Chow T.Y., Tseng Y.H. Molecular cloning, characterization and nucleotide sequence of the gene for secreted a-amylase from Xanthomonas campestris pv campestris. J Gen Microbiol 1992; 138:1647–1655
    [Google Scholar]
  10. Jaspersen H.M., MacGregor E.A., Sierks M.R., Svensson B. Comparison of the domain-level organization of starch hydrolases and related enzymes. Biochem J 1991; 280:51–55
    [Google Scholar]
  11. Kuriki T., Imanaka T. Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus. J Gen Microbiol 1989; 135:1521–1528
    [Google Scholar]
  12. Lansky S., Kooi M., Shhoch T.J. Properties of the fractions and linear subfractions from various starches. J Am Chem Soc 1949; 71:4066–4075
    [Google Scholar]
  13. Matsuura Y., Kusunoki M., Harada W., Kakudo M. Structure and possible catalytic residues of Taka-amylase A. J Biochem 1984; 95:697–702
    [Google Scholar]
  14. Metz R.J., Allen L.N., Cao T.M., Zeman N.W. Nucleotide sequence of an amylase gene from Bacillus megaterium. Nucleic Acids Res 1988; 16:5203–1
    [Google Scholar]
  15. Nakajima R., Imanaka T., Aiba S. Comparison of amino acid sequences of eleven different a-amylases. Appl Microbiol Biotechnol 1986; 23:355–360
    [Google Scholar]
  16. Parker J.E., Barber C.E., Mi-Jiao F., Daniels M.J. Interaction of Xanthomonas campestris with Arabidopsis thaliana: characterization of a gene from Xanthomonas campestris pv raphani that confers avirulence to most A. thaliana accessions. Mol Plant-Microbe Interact 1992; 6:216–224
    [Google Scholar]
  17. Pasero L., Mazzei-Pierron Y., Abadie B., Chicheportiche Y., Marchis-Mouren G. Complete amino acid sequence and location of the five disulfide bridges in porcine pancreatic a-amylase. Biochim Biophys Acta 1986; 869:147–157
    [Google Scholar]
  18. Podkovyrov S.M., Zeikus I.G. Structure of the gene encoding cyclomaltodextrinase from Clostridium thermobydrosul-furicum 39E and characterization of the enzyme purified from Escherichia coli. J Bacteriol 1992; 174:5400–5405
    [Google Scholar]
  19. Pugsley A.P. The complete general secretory pathway in Gram-negative bacteria. Microbiol Rev 1993; 57:50–108
    [Google Scholar]
  20. Sambrook J., Fritsch E.F., Maniatis T. Molecular Cloning: a Laboratory Manual 1989 2nd edn Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  21. Smith A.F. Malate to oxaloacetate reaction. In Methods of Enzymatic Analysis 1983 Edited by Bergmeyer H.U., Bergemeyer H.J., Grabl M. Weinheim: Verlag Chemie; 3 pp 166–171
    [Google Scholar]
  22. Toda H., Kondo K., Narita K. The complete amino acid sequence of Taka-amylase A. Proc Jpn Acad 1982; 58:208–212
    [Google Scholar]
  23. Yamazaki H., Ohmura K., Nakayama A., Takeichi Y., Otozai K., Yamasaki M., Tamura G., Yamane K. a-Amylase genes (amyR2 and amyE(+)) from an a-amylase-hyperproducing Bacillus subtilis strain: molecular cloning and nucleotide sequences. J Bacteriol 1983; 156:327–337
    [Google Scholar]
  24. Weber K., Osborn M. The reliability of molecular weight determination by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem 1969; 244:4406–4412
    [Google Scholar]
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