Purification and characterization of ceql restriction endonuclease

Zsuzsa Izsvák, Zsolt Jobbágy, E. Duda

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Ceql, a type II restriction endonuclease, an isoschizomer of EcoRV was purified to apparent homogeneity by a combination of salt precipitation, ion exchange, dye affinity and hydrophobic interaction chromatographies. The crude enzyme was present in the form of large aggregates that could be pelleted by high speed centrifugation. The enzyme was not associated with cellular membranes, though non-ionic detergents lowered the apparent size of the aggregates. The purified enzyme also showed a tendency to form large molecular mass (66-600 kDa) complexes under physiological conditions, in the absence of cleavable DNA. The enzyme formed smaller complexes in the presence of DNA and non-ionic detergents and dissociated into subunits (and undergoes reversible loss of activity) in the presence of high concentrations of salts. According to SDS gel electrophoresis and sedimentation analysis the molecular mass of the monomer 32 ± 2 kDa. The enzyme had a rather broad pH optimum, extending into the alkaline range and lost specificity and activity in buffers below pH 6.

Original languageEnglish
Pages (from-to)830-834
Number of pages5
JournalZeitschrift fur Naturforschung - Section C Journal of Biosciences
Volume47
Issue number11-12
DOIs
Publication statusPublished - Dec 1 1992

Fingerprint

DNA Restriction Enzymes
Purification
Enzymes
Molecular mass
Detergents
Type II Site Specific Deoxyribonucleases
Salts
Centrifugation
Ion Exchange
DNA
Chromatography
Electrophoresis
Hydrophobic and Hydrophilic Interactions
Sedimentation
Ion exchange
Buffers
Coloring Agents
Monomers
Gels
Membranes

Keywords

  • Corynebacterium
  • Endonuclease
  • Multimeric Enzyme
  • Purification
  • Restriction Enzyme

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Purification and characterization of ceql restriction endonuclease. / Izsvák, Zsuzsa; Jobbágy, Zsolt; Duda, E.

In: Zeitschrift fur Naturforschung - Section C Journal of Biosciences, Vol. 47, No. 11-12, 01.12.1992, p. 830-834.

Research output: Contribution to journalArticle

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