Skip to main content
SpringerLink
Log in

Purification and characterisation of an Aspergillus niger invertase and its DNA sequence

  • Original Articles
  • Published:
Current Genetics Aims and scope Submit manuscript

Abstract

A secreted invertase was purified 23-fold by ultrafiltration, ion-exchange, and gel filtration chromatography from the culture supernatant of 18h sucrose-grown cultures of Aspergillus niger. The purified enzyme hydrolysed sucrose and raffinose but there was no detectable hydrolysis of inulin, melezitose or PNPG. Invertase activity was optimal at pH 5.5 and 50°C. The molecular mass of reduced invertase was 115 kDa, as determined by SDS gel electrophoresis. The native molecular weight of between 225 kDa and 250 kDa, estimated by electrophoresis under non-denaturing conditions, suggests that the protein is a dimer of identical subunits. The suc1 gene encoding this protein was completely sequenced. The translated sequence yields a protein of 566 amino acids with a calculated molecular mass of 61 kDa, suggesting that carbohydrates represent about 50% of the mass of the protein.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ballance DJ (1986) Yeast 2:229–236

    Google Scholar 

  • Ballance DJ, Buxton FP, Turner G (1983) Biochem Biophys Res Comm 112:284–289

    Google Scholar 

  • Bégueret J, Razanamparany V, Perrot M, Barreau C (1984) Gene 32:487–492

    Google Scholar 

  • Benoist C, O'Hare K, Breathnach R, Chambon P (1980) Nucleic Acids Res 8:127–142

    Google Scholar 

  • Bergès T, Barreau C, Peberdy JF, Boddy LM (1993) Curr Genet (in press)

  • Buxton FP, Gwynne DI, Garven S, Sibley S, Davies RW (1987) Gene 60:255–266

    Google Scholar 

  • Carlson C, Botstein D (1982) Cell 28:145–154

    Google Scholar 

  • Edmann P, Begg G (1967) Eur J Biochem 1:80–91

    Google Scholar 

  • Ettalibi M, Baratti JC (1987) Appl Microbiol Biotechnol 26:13–20

    Google Scholar 

  • Fouet A, Klier A, Rapoport G (1986) Gene 45:221–225

    Google Scholar 

  • Frederick KR, Tung J, Emerick RS, Masiarz FR, Chamberlain SH, Vasavada A, Rosenberg S, Chakraborty S, Schopter LM, Massey V (1990) J Biol Chem 265:3793–3802

    Google Scholar 

  • Frischauf A-M, Lehrach H, Poustka A, Murray N (1983) J Mol Biol 170:827–842

    Google Scholar 

  • Gannon F, O'Hare K, Perrin F, Le Pennec JP, Benoist C, Cochet M, Breathnach R, Royal A, Garaspin A, Cami B, Chambon P (1979) Nature 278:428–434

    Google Scholar 

  • Gascón S, Lampen JO (1968) J Biol Chem 243:1567–1572

    Google Scholar 

  • Gascón S, Neumann NP, Lampen JO (1968) J Biol Chem 243:1573–1577

    Google Scholar 

  • Gavel Y, Heijne von G (1990) Protein Eng 3:433–442

    Google Scholar 

  • Gunasekaran P, Karunakaran T, Cami B, Mukundan AG, Preziosi L, Baratti J (1990) J Bacteriol 172:6727–6735

    Google Scholar 

  • Gupta AK, Bhatia IS (1982) Biochemistry 21:1249–1253

    Google Scholar 

  • Gurr SJ, Unkles SE, Kinghorn JR (1987) In: Kinghorn JR (ed) Gene structure in eukaryotic microbes, vol 2. Special publication of the Society for General Microbiology, IRL Press, Oxford, pp 91–139

    Google Scholar 

  • Iizuka M, Tsuji Y, Yamamoto T (1974) Agric Biol Chem 38:213–215

    Google Scholar 

  • Iizuka M, Yamamoto T (1979) Agric Biol Chem 43:217–222

    Google Scholar 

  • Klein RD, Deibel MR, Jr., Sarcich JL, Zurcher-Neely HA, Reardon IM, Heinrikson RL (1989a) Prep Biochem 19:293–319

    Google Scholar 

  • Klein RD, Poorman RA, Favreau MA, Shea MH, Hatzenbuhler NT, Nulf SC (1989b) Curr Genet 16:145–152

    Google Scholar 

  • Kos A, Kujivenhoven J, Hessing HGM, Pouwels PH, van den Hondel CAMJJ (1988) Curr Genet 13:137–144

    Google Scholar 

  • Kubicek CP, Röhr M (1978) Eur J Appl Microbiol Biotechnol 5:263–271

    Google Scholar 

  • Laemmli UK (1970) Nature 227:680–683

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Laboratory, Cold Spring Harbour, New York

    Google Scholar 

  • Martin I, Dabarbouille M, Ferrari E, Papopart G (1987) Mol Gen Genet 208:177–184

    Google Scholar 

  • McKnight GL, Kato H, Upshall A, Parker ML, Saan G, O'Hara PJ (1985) EMBO J 4:2093–2099

    Google Scholar 

  • Metzenberg RL (1963) Biochim Biophys Acta 77:455–465

    Google Scholar 

  • Moreno S, Sanchez Y, Rodriguez L (1990) Biochem J 267:697–702

    Google Scholar 

  • Neumann NP, Lampen JO (1967) Biochemistry 6:468–475

    Google Scholar 

  • Onodera K, Maruyama Y (1979) J Gen Appl Microbiol 25:335–337

    Google Scholar 

  • Perlman D, Halvorson HO (1981) Cell 25:525–536

    Google Scholar 

  • Pontecorvo G, Roper JA, Hemmons LM, MacDonald KD, Bufton AWJ (1953) Adv Genet 5:141–238

    Google Scholar 

  • Proudfoot NJ, Brownlee GG (1976) Nature 263:211–214

    Google Scholar 

  • Raeder U, Broda P (1985) Lett Appl Microbiol 1:17–20

    Google Scholar 

  • Rambosek J, Leach J (1987) CRC Crit Rev Biotechnol 6:357–374

    Google Scholar 

  • Razanamparany V, Bégueret J (1986) Curr Genet 10:811–817

    Google Scholar 

  • Sanger F, Milken S, Coulson AR (1977) Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Sato Y, Kuramitsu H (1988) Infect Immunol 56:1956–1960

    Google Scholar 

  • Scholle R, Robb S, Robb F, Wood D (1989) Gene 80:49–56

    Google Scholar 

  • Soberon X, Covarrubias L, Bolivar F (1980) Gene 9:287–305

    Google Scholar 

  • Sutton DD, Lampen JO (1962) Biochim Biophys Acta 56:303–312

    Google Scholar 

  • Tarentino AL, Plummer TH Jr, Maley F (1974) J Biol Chem 249:818–824

    Google Scholar 

  • Taussig R, Carlson M (1983) Nucleic Acids Res 11:1943–1954

    Google Scholar 

  • Trimble RB, Maley F (1977) J Biol Chem 252:4409–4412

    Google Scholar 

  • Vainstein MH (1990) PhD thesis, University of Nottingham

  • Vainstein MH, Peberdy JF (1990) FEMS Microbiology Lett 71:265–270

    Google Scholar 

  • Ward M, Wilson LJ, Carmona CL, Turner G (1988) Curr Genet 14:37–42

    Google Scholar 

  • Whittington H, Kerry-Williams S, Bidgood K, Dodsworth N, Peberdy J, Dobson M, Hinchliffe E, Ballance DJ (1990) Curr Genet 18:531–536

    Google Scholar 

  • Yannisch-Perron C, Vieira J, Messing J (1985) Gene 33:103–119

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire de Génétique, Université de Bordeaux II UA CNRS 542, Avenue des Facultés, F-33405, Talence Cedex, France

    L. M. Boddy, T. Bergès & C. Barreau

  2. Microbial Biochemistry and Genetics Group, Department of Life Science, University of Nottingham, University Park, NG7 2RD, Nottingham, UK

    L. M. Boddy, M. H. Vainstein, M. J. Dobson & J. F. Peberdy

  3. Castle Court, Delta Biotechnology Ltd., Casle Boulevard, NG7 1FD, Nottingham, UK

    D. J. Ballance

Authors
  1. L. M. Boddy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. Bergès
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Barreau
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. H. Vainstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. M. J. Dobson
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D. J. Ballance
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J. F. Peberdy
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by P.P. Slonimski

Rights and permissions

Reprints and permissions

About this article

Cite this article

Boddy, L.M., Bergès, T., Barreau, C. et al. Purification and characterisation of an Aspergillus niger invertase and its DNA sequence. Curr Genet 24, 60–66 (1993). https://doi.org/10.1007/BF00324666

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00324666

Key words

Search

Navigation