Cloning and Expression as a Tool to Easily Purify a New Highly Thermostable Archaebacterial ß-Galactosidase

  • Marco Moracci
  • Mario De Rosa
  • Mose’ Rossi


Since enzymes from extremophiles are thermostable they could, in theory, be purified very easily after cloning and expression of their genes in mesophiles. This paper demonstrates that the gene of a new ß-galactosidase isolated from the extreme thermophile Sulfolobus solfataricus can be cloned and expressed in E. coli under the control of the archaebacterial promoter. The recombinant enzyme is similar to the native and, as expected, can be easily purified by heat treatment and isoelectric point precipitation. The partial purification and the properties of the recombinant enzyme are described.


Recombinant Enzyme Native Enzyme Lactose Hydrolysis Thermophilic Enzyme Thermophilic Organism 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Mozhaev, V.V., Berezin, I.V. and Martinek, K., Structure-stability relationship in proteins: fundamental tasks and strategy for the development of stabilized enzymes for biotechnology. CRC Critic Rev. Biochem., 1988, 23, 235–281.CrossRefGoogle Scholar
  2. 2.
    Fontana, A., Structure and stability of thermophilic enzymes. Biophys. Chem., 1988, 29, 181–185.PubMedCrossRefGoogle Scholar
  3. 3.
    Brock, T.D. and Zeikus, J.G., Thermophiles: general molecular and applied microbiology. Wiley & Sons, New York, 1986.Google Scholar
  4. 4.
    Rossi, M., Enzymes from extreme thermophilic bacteria as models of special catalists. European Conference on Biotechnology, Scientific Technical and Industrial Challanges, EIT and ZAI Verona Agricultural Italy, Nov. 7-8, 1988, 46–50.Google Scholar
  5. 5.
    Kandler, O., Archaebacteria: biotechnological implications. Proc. 3rd Eur. Congr. on Biotechnology, Munchen 1984, Vol. IV, 551–560.Google Scholar
  6. 6.
    Pisani, F.M., Rella, R., Raia, C.A., Rozzo, C., Nucci, R., Gambacorta, A., De Rosa, M. and Rossi, M., Thermostable ß-galactosidase from the archaebacterium S. solfataricus. Eur. J. Biochem., 1990, 187, 321–328.PubMedCrossRefGoogle Scholar
  7. 7.
    Cubellis, M.V., Rozzo, C., Montecucchi, P. and Rossi, M., Isolation sequencing and cloning in E. coli of a new 3-galactosidase archaebacterial gene. Gene, 1990 (in press).Google Scholar

Copyright information

© SCI 1990

Authors and Affiliations

  • Marco Moracci
    • 1
  • Mario De Rosa
    • 2
  • Mose’ Rossi
    • 1
  1. 1.Dipartimento di Chimica Organica e Biologica eIstituto di Biochimica delle Proteine ed Enzimologia, CNRItaly
  2. 2.Istituto di Biochimica delle MacromolecoleUniversità di NapoliItaly

Personalised recommendations