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Volume 139, Issue 10

Cloning and sequencing of a gene encoding acidophilic amylase from Bacillus acidocaldarius Free

Abstract

SUMMARY: Two starch-degrading enzymes produced by (renamed as ) were identified. According to SDS-PAGE, the apparent molecular masses of the enzymes were 90 and 160 kDa. Eight peptide fragments and the N-terminal end of the 90 kDa polypeptide were sequenced. An oligonucleotide, based on the amino acid sequence of a peptide fragment of the 90 kDa protein, was used to screen a lDgt10 bank of and the region encoding the 90 kDa protein was cloned. Unexpectedly, the ORF continued upstream of the N terminus of the 90 kDa protein. The entire ORF was 1301 amino acids (aa) long (calculated molecular mass 140 kDa) and it was preceded by a putative ribosomal binding site and a promoter. Computer analysis showed that the 1301 aa protein was closely related to an a-amylase-pullulanase of We suggest that the starch-degrading 160 kDa protein of is an a-amylase-pullulanase, and the 90 kDa protein is a cleavage product of the 160 kDa protein. Another ORF, apparently in the same transcription unit, was found downstream from the amylase gene. It encoded a protein that was closely related to the maltose-binding protein of

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Society for General Microbiology, 1993
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1993-10-01
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References

  1. Binder F., Huber O., BÖck A. 1986; Cyclodextrin- glucosyltransferase from Klebsiella pneumoniae M5al: cloning, nucleotide sequence and expression.. Gene 47:269–277
     [Google Scholar]
  2. Boyer E.W., Ingle M.B., Mercer G.D. 1979; Isolation and characterization of unusual bacterial amylases.. Starch Stärke 31:166–171
     [Google Scholar]
  3. Buonocore V., Caporale C., De ROSA M., Gambacorta A. 1976; Stable, inducible thermoacidophilic α-amylase from Bacillus acidocaldarius. . Journal of Bacteriology 128:515–521
     [Google Scholar]
  4. Darland G., Brock T.D. 1971; Bacillus acidocaldarius sp. nov., an acidophilic thermophilic spore-forming bacterium.. Journal of General Microbiology 67:9–15
     [Google Scholar]
  5. Devereux J., Haeberli P., Smithies O. 1984; A comprehensive set of sequence analysis programs for the VAX.. Nucleic Acids Research 12:387–395
     [Google Scholar]
  6. Duplay P., Bedouelle H., Fowler A., Zabin I., Saurin W., Hofnung M. 1984; Sequences of the malE gene and its product, the maltose-binding protein of Escherichia coli K12.. Journal of Biological Chemistry 259:10606–10613
     [Google Scholar]
  7. Feng D.F., Johnson M.S., Doolittle R.F. 1985; Aligning amino acid sequences: comparison of commonly used methods.. Journal of Molecular Evolution 21:112–125
     [Google Scholar]
  8. Haima P., Bron S., Venema G. 1987; The effect of restriction on shotgun cloning and plasmid stability in Bacillus subtilis Marburg.. Molecular and General Genetics 209:335–342
     [Google Scholar]
  9. Hengge R., Boos W. 1983; Maltose and lactose transport in Escherichia coli. Examples of two different types of concentrative transport systems.. Biochimica et Biophysica Acta 737:443–478
     [Google Scholar]
  10. Horinouchi S., Fukusumi S., Ohshima T., Beppu T. 1988; Cloning and expression in Escherichia coli of two additional amylase genes of a strictly anaerobic thermophile, Dictyoglomus thermo- philum. . European Journal of Biochemistry 176:243–253
     [Google Scholar]
  11. Kalkkinen N. 1986; Radio-sequence analysis: an ultra-sensitive method to align protein and nucleotide sequences.. InAdvanced Methods in Protein Microsequence Analysis, pp. 194–206 Edited by Wittmann-Liebold B., Salnikow J., Erdman V. A. Berlin, Heidelberg: Springer-Verlag;
     [Google Scholar]
  12. Kalkkinen N., Tilgmann C. 1988; A gas-pulsed-liquid-phase sequencer constructed from a Beckman 890D instrument by using Applied Biosystems delivery and cartridge blocks.. Journal of Protein Chemistry 7:242–243
     [Google Scholar]
  13. Kanno M. 1986; A Bacillus acidocaldarius a-amylase that is highly stable to heat under acidic conditions.. Agricultural and Biological Chemistry 50:23–31
     [Google Scholar]
  14. Koivula T., Sibakov M., Palva I. 1991; Isolation and characterization of Lactococcus lactis subsp. lactis promoters.. Applied and Environmental Microbiology 57:333–340
     [Google Scholar]
  15. Kuriki T., Imanaka T. 1989; Nucleotide sequence of the neopullulanase gene from Bacillus stearothermophilus. . Journal of General Microbiology 135:1521–1528
     [Google Scholar]
  16. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature, London 227:680–685
     [Google Scholar]
  17. Marmur J. 1961; A procedure for the isolation of deoxyribonucleic acid from microorganisms.. Journal of Molecular Biology 3:208–218
     [Google Scholar]
  18. Melasniemi H. 1987; Effect of carbon source on production of thermostable α-amylase, pullulanase and α-glucosidase by Clost-ridium thermohydrosulfuricum. . Journal of General Microbiology 133:883–890
     [Google Scholar]
  19. Melasniemi H. 1988; Purification and some properties of the extracellular a-amylase-pullulanase produced by Clostridium thermo- hydrosulfuricum. . Biochemical Journal 250:813–818
     [Google Scholar]
  20. Melasniemi H., Paloheimo M., Hemio L. 1990; Nucleotide sequence of the a-amylase-pullulanase gene from Clostridium thermohydrosulfuricum. . Journal of General Microbiology 136:447–454
     [Google Scholar]
  21. Mozdzanowski J., Speicher D.W. 1990; Quantitative electrotransfer of proteins from polyacrylamide gels onto PVDF membranes.. In Current Research in Protein Chemistry, pp. 87–93 Edited by Villafranca J. New York: Academic Press;
     [Google Scholar]
  22. Nelson N. 1944; A photometric adaptation of the Somogyi method for the determination of glucose.. Journal of Biological Chemistry 153:375–380
     [Google Scholar]
  23. Pearson W.R., Lipman D.J. 1988; Improved tools for biological sequence comparison.. Proceedings of the National Academy of Sciences of the United States of America 852444–2448
     [Google Scholar]
  24. Sakano Y., Sano M., Kobayashi T. 1985; Hydrolysis of a-1,6- glucosidic linkages by a-amylases.. Agricultural and Biological Chemistry 49:3041–3043
     [Google Scholar]
  25. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A.R. 1977; DNA sequencing with chain-terminating inhibitors.. Proceedings of the National Academy of Sciences of the United States of America 745463–5467
     [Google Scholar]
  27. Sibakov M., Palva I. 1984; Isolation and the 5’-end nucleotide sequence of Bacillus licheniformis α-amylase gene. . European Journal of Biochemistry 145:567–572
     [Google Scholar]
  28. Siezen R.J., De Vos W.M., Leunissen J.A.M., Dijkstra B.W. 1991; Homology modelling and protein engineering strategy of subtilases, the family of subtilisin-like serine proteases.. Protein Engineering 4:719–737
     [Google Scholar]
  29. Somogyi M. 1952; Notes on sugar determination.. Journal of Biological Chemistry 195:19–23
     [Google Scholar]
  30. Spurlino J.C., Lu G.Y., Quiocho F.A. 1991; The 2-3-Å resolution structure of the maltose- or maltodextrin-binding protein.. Journal of Biological Chemistry 266:5202–5219
     [Google Scholar]
  31. Tapio S., Yeh F., Shuman H.A., Boos W. 1991; The malZ gene of Escherichia coli, a member of the maltose regulon, encodes a maltodextrin glucosidase.. Journal of Biological Chemistry 266:19450–19458
     [Google Scholar]
  32. Uchino F. 1982; A thermophilic and unusually acidophilic amylase produced by a thermophilic acidophilic Bacillus sp.. Agricultural and Biological Chemistry 46:7–13
     [Google Scholar]
  33. Vihinen M., Mäntsälä P. 1989; Microbial amylolytic enzymes.. Critical Reviews in Biochemistry and Molecular Biology 24:329–418
     [Google Scholar]
  34. Wells J.A., Estell D.A. 1988; Subtilisin - an enzyme designed to be engineered.. Trends in Biochemical Sciences 13:291–297
     [Google Scholar]
  35. Wisotzkey J.D., Jurtshuk P, Deinhard G., Poralla K. 1992; Comparative sequence analysis on the 16S rRNA (rDNA) of Bacillus acidocaldarius, Bacillus acidoterrestris,and Bacillus cycloheptanicus and proposal for creation of a new genus, Alicyclobacillus gen. nov.. International Journal of Systematic Bacteriology 42:263–269
     [Google Scholar]
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Cloning and sequencing of a gene encoding acidophilic amylase from Bacillus acidocaldarius
Microbiology 139, 2399 (1993); https://doi.org/10.1099/00221287-139-10-2399
/content/journal/micro/10.1099/00221287-139-10-2399
/content/journal/micro/10.1099/00221287-139-10-2399
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