Current Genetics

, Volume 18, Issue 3, pp 217–222

Cloning and sequence analysis of a cDNA for cellulase (FI-CMCase) from Aspergillus aculeatus

  • Toshihiko Ooi
  • Atsuhiko Shinmyo
  • Hirosuke Okada
  • Saburo Hara
  • Tokuji Ikenaka
  • Sawao Murao
  • Motoo Arai
Original Articles

Summary

Wa have cloned and characterized the cDNA coding for a major component of cellulase, endoglucanase (FI-CMCase), produced by Aspergillus aculeatus. The cDNA was isolated from a A. aculeatus cDNA library using synthetic oligonuceotide mixtures that correspond to the internal amino acid sequence of the mature FI-CMCase protein. Nucleotide sequence analysis of the cloned cDNA insert revealed a 711 bp open reading frame that encoded a protein of 237 amino acid residues. The primary structure of FI-CMCase deduced from the nucleotide sequence of cDNA agreed with that found by amino acid sequencing of peptide fragments obtained by digestion with several proteinases and cyanogen bromide cleavage. There may be a signal peptide sequence of 16 amino acid residues at the N-terminus. The molecular mass of the mature protein calculated from the cDNA is 24002 daltons, which compares favorably with molecular mass estimates of purified FI-CMCase obtained from SDS-PAGE (25000 Da). No distinct homology was found between the amino acid sequence of FI-CMCase and known cellulase sequences of other microorganisms. This study is the first example of cDNA cloning of an endoglucanase from the genus Aspergillus.

Key words

Cellulase Filamentous fungi Aspergillus aculeatus cDNA cloning 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Berget SM (1984) Nature 309:179–182Google Scholar
  2. Bhikhabhai R, Pettersson LG (1984) FEBS Lett 167:301–308Google Scholar
  3. Boel E, Hansen MT, Hjort I, Hoegh I, Fiil NP (1984) EMBO J 3:1581–1585Google Scholar
  4. Chen CM, Gritzali M, Stafford DW (1987) Bio/Technology 5:274–278Google Scholar
  5. Chirgwin JW, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Biochemistry 18:5294–5299Google Scholar
  6. Cigan AM, Donahue TF (1987) Gene 59:1–18Google Scholar
  7. Duff SJB (1985) Biotechnol Lett 7:185–190Google Scholar
  8. Fagerstam LG, Pettersson LG, Engström JÅ (1984) FEBS Lett 167:309–315Google Scholar
  9. Feinberg AP, Vogelstein B (1983) Anal Biochem 132:6–13Google Scholar
  10. Fitzgerald M, Shenk T (1981) Cell 24:251–260Google Scholar
  11. Geoffrey MW, Stern M, Stark GR (1979) Proc Natl Acad Sci USA 76:3683–3687Google Scholar
  12. Glišin V, Crkvenjakov R, Byus C (1974) Biochemistry 18:2633–2637Google Scholar
  13. Gross E, Witokop B (1962) J Biol Chem 237:1856–1860Google Scholar
  14. Grunstein M, Hogness DS (1975) Proc Natl Acad Sci USA 72:3961–3965Google Scholar
  15. Gubler U, Hoffman BJ (1983) Gene 25:263–269Google Scholar
  16. Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA cloning: A practical approach. IRL Press, Oxford, England, pp 109–135Google Scholar
  17. Kamei K, Yamamura Y, Hara S, Ikenaka T (1989) J Biochem 105:979–985Google Scholar
  18. Knowles J, Lehtovaara P, Teeri T (1987) Trends Biotechnol 5:255–261Google Scholar
  19. Kozak M (1984) Nucleic Acids Res 12:857–872Google Scholar
  20. Kozak M (1986) Cell 44:283–292Google Scholar
  21. MacRae WD, Buxton FP, Sibley S, Garven S, Gwynne DI, Davies RW, Arst Jr HN (1988) Gene 71:339–348Google Scholar
  22. McKnight GL, Kato H, Upshall A, Parker MD, Saari G, O'Hara PJ (1985) EMBO J 4:2093–2099Google Scholar
  23. McMaster GK, Carmichael GG (1977) Proc Natl Acad Sci USA 74:4835–4838Google Scholar
  24. Mega T, Hamazume Y, Hong YM, Ikenaka T (1986) J Biochem 100:1109–1116Google Scholar
  25. Minter SJ and Sealey PG (1982) Nucleic acid molecular weight markers. In: Rickwood D and Hames BD (eds) Gel electrophoresis of nucleic acids. IRL Press, Oxford, England, pp 227–235Google Scholar
  26. Nunberg JH, Maede JH, Cole G, Lawyer FC, McCabe P, Schweickart V, Tal R, Wittman VP, Flatgaard JE, Innis MA (1984) Mol Cell Biol 4:2306–2315Google Scholar
  27. Münger K, Germann UA, Lerch K (1985) EMBO J 4:2665–2668Google Scholar
  28. Murao S, Kanamoto J, Arai M (1979) J Ferment Technol 57:151–156Google Scholar
  29. Murao S, Sakamoto R, Arai M (1988) Methods Enzymol 160:274–299Google Scholar
  30. Okada G (1985) Agric Biol Chem 49:1257–1265Google Scholar
  31. Penttilä M, Lehtovaara P, Nevalainen H, Bhikhabhai R, Knowles J (1986) Gene 45:253–263Google Scholar
  32. Penttilä ME, André L, Lehtovaara P, Bailey M, Teeri TT, Knowles JKC (1988) Gene 63:103–112Google Scholar
  33. Proudfoot NJ (1976) Nature 263:211–214Google Scholar
  34. Saloheimo M, Lehtovaara P, Penttilä M, Teeri TT, Stahlberg J, Johansson G, Pettersson G, Claeyssens M, Tomme P, Knowes JKC (1988) Gene 63:11–21Google Scholar
  35. Sanger F, Nicklen S, Coulson AR (1977) Proc Natl Acad Sci USA 74:5463–5467Google Scholar
  36. Schmuck M, Pilz I, Hayn M, Esterbauer H (1986) Biotechnol Lett 8:397–402Google Scholar
  37. Shoemaker S, Schweickart V, Ladner M, Gelfand D, Kwok S, Myambo K, Innis M (1983) Bio/Technology 1:691–696Google Scholar
  38. Stewart JC, Parry JB (1981) J Gen Microbiol 125:33–39Google Scholar
  39. Teeri T, Salovuori I, Knowles J (1983) Bio/Technology 1:696–699Google Scholar
  40. Teeri TT, Lehtovaara P, Kauppinen S, Salovuori I, Knowles J (1987) Gene 51:43–52Google Scholar
  41. Thomas PS (1980) Proc Natl Acad Sci USA 77:5201–5205Google Scholar
  42. Van Arsdell JN, Kwok S, Schweickart VL, Ladner MB, Gelfand DH, Innis MA (1987) Bio/Technology 5:60–64Google Scholar
  43. von Heijne G (1983) Eur J Biochem 133:17–21Google Scholar
  44. Yanisch-Perron C, Vieira J, Messing J (1985) Gene 33:103–119Google Scholar
  45. Zaret KS, Sherman F (1982) Cell 28:563–573Google Scholar
  46. Zaret KS, Choen EH (1984) Mol Cell Biol 4:1515–1520Google Scholar

Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Toshihiko Ooi
    • 1
  • Atsuhiko Shinmyo
    • 2
  • Hirosuke Okada
    • 2
  • Saburo Hara
    • 3
  • Tokuji Ikenaka
    • 3
  • Sawao Murao
    • 1
  • Motoo Arai
    • 1
  1. 1.Department of Agricultural ChemistryUniversity of Osaka PrefectureSakai-shi, OsakaJapan
  2. 2.Department of Fermentation TechnologyOsaka UniversityOsakaJapan
  3. 3.Department of ChemistryOsaka UniversityOsakaJapan
  4. 4.Department of Agricultural ChemistryUniversity of UtsunomiyaUtsunomiyaJapan

Personalised recommendations