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. 1987 Sep;169(9):4171–4176. doi: 10.1128/jb.169.9.4171-4176.1987

Nucleotide sequence of the glucoamylase gene GLU1 in the yeast Saccharomycopsis fibuligera.

T Itoh, I Ohtsuki, I Yamashita, S Fukui
PMCID: PMC213725  PMID: 3114236

Abstract

The complete nucleotide sequence of the glucoamylase gene GLU1 from the yeast Saccharomycopsis fibuligera has been determined. The GLU1 DNA hybridized to a polyadenylated RNA of 2.1 kilobases. A single open reading frame codes for a 519-amino-acid protein which contains four potential N-glycosylation sites. The putative precursor begins with a hydrophobic segment that presumably acts as a signal sequence for secretion. Glucoamylase was purified from a culture fluid of the yeast Saccharomyces cerevisiae which had been transformed with a plasmid carrying GLU1. The molecular weight of the protein was 57,000 by both gel filtration and acrylamide gel electrophoresis. The protein was glycosylated with asparagine-linked glycosides whose molecular weight was 2,000. The amino-terminal sequence of the protein began from the 28th amino acid residue from the first methionine of the putative precursor. The amino acid composition of the purified protein matched the predicted amino acid composition. These results confirmed that GLU1 encodes glucoamylase. A comparison of the amino acid sequence of glucoamylases from several fungi and yeast shows five highly conserved regions. One homology region is absent from the yeast enzyme and so may not be essential to glucoamylase function.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bennetzen J. L., Hall B. D. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase. J Biol Chem. 1982 Mar 25;257(6):3018–3025. [PubMed] [Google Scholar]
  2. Boel E., Hjort I., Svensson B., Norris F., Norris K. E., Fiil N. P. Glucoamylases G1 and G2 from Aspergillus niger are synthesized from two different but closely related mRNAs. EMBO J. 1984 May;3(5):1097–1102. doi: 10.1002/j.1460-2075.1984.tb01935.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  4. De Mot R., Verachtert H. Purification and Characterization of Extracellular Amylolytic Enzymes from the Yeast Filobasidium capsuligenum. Appl Environ Microbiol. 1985 Dec;50(6):1474–1482. doi: 10.1128/aem.50.6.1474-1482.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  6. Jensen R., Sprague G. F., Jr, Herskowitz I. Regulation of yeast mating-type interconversion: feedback control of HO gene expression by the mating-type locus. Proc Natl Acad Sci U S A. 1983 May;80(10):3035–3039. doi: 10.1073/pnas.80.10.3035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Julius D., Brake A., Blair L., Kunisawa R., Thorner J. Isolation of the putative structural gene for the lysine-arginine-cleaving endopeptidase required for processing of yeast prepro-alpha-factor. Cell. 1984 Jul;37(3):1075–1089. doi: 10.1016/0092-8674(84)90442-2. [DOI] [PubMed] [Google Scholar]
  8. Kreil G. Transfer of proteins across membranes. Annu Rev Biochem. 1981;50:317–348. doi: 10.1146/annurev.bi.50.070181.001533. [DOI] [PubMed] [Google Scholar]
  9. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Manjunath P., Shenoy B. C., Raghavendra Rao M. R. Fungal glucoamylases. J Appl Biochem. 1983 Aug-Oct;5(4-5):235–260. [PubMed] [Google Scholar]
  12. Nunberg J. H., Meade J. H., Cole G., Lawyer F. C., McCabe P., Schweickart V., Tal R., Wittman V. P., Flatgaard J. E., Innis M. A. Molecular cloning and characterization of the glucoamylase gene of Aspergillus awamori. Mol Cell Biol. 1984 Nov;4(11):2306–2315. doi: 10.1128/mcb.4.11.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Struck D. K., Lennarz W. J., Brew K. Primary structural requirements for the enzymatic formation of the N-glycosidic bond in glycoproteins. Studies with alpha-lactalbumin. J Biol Chem. 1978 Aug 25;253(16):5786–5794. [PubMed] [Google Scholar]
  15. Sørensen S. H., Norén O., Sjöström H., Danielsen E. M. Amphiphilic pig intestinal microvillus maltase/glucoamylase. Structure and specificity. Eur J Biochem. 1982 Sep 1;126(3):559–568. doi: 10.1111/j.1432-1033.1982.tb06817.x. [DOI] [PubMed] [Google Scholar]
  16. Takahashi T., Tsuchida Y., Irie M. Purification and some properties of three forms of glucoamylase from a Rhizopus species. J Biochem. 1978 Nov;84(5):1183–1194. doi: 10.1093/oxfordjournals.jbchem.a132235. [DOI] [PubMed] [Google Scholar]
  17. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Wilson J. J., Ingledew W. M. Isolation and characterization of Schwanniomyces alluvius amylolytic enzymes. Appl Environ Microbiol. 1982 Aug;44(2):301–307. doi: 10.1128/aem.44.2.301-307.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Yamashita I., Maemura T., Hatano T., Fukui S. Polymorphic extracellular glucoamylase genes and their evolutionary origin in the yeast Saccharomyces diastaticus. J Bacteriol. 1985 Feb;161(2):574–582. doi: 10.1128/jb.161.2.574-582.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yamashita I., Nakamura M., Fukui S. Gene fusion is a possible mechanism underlying the evolution of STA1. J Bacteriol. 1987 May;169(5):2142–2149. doi: 10.1128/jb.169.5.2142-2149.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yamashita I., Suzuki K., Fukui S. Nucleotide sequence of the extracellular glucoamylase gene STA1 in the yeast Saccharomyces diastaticus. J Bacteriol. 1985 Feb;161(2):567–573. doi: 10.1128/jb.161.2.567-573.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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