An acid phosphatase from Manihot glaziovii as an alternative to alkaline Phosphatase for molecular cloning experiments

Biotechnol Lett. 2005 Dec;27(23-24):1865-8. doi: 10.1007/s10529-005-3894-z.

Abstract

An acid phosphatase, free of deoxyribonuclease activity, was isolated from Manihot glaziovii leaves. It had a Mr of 78 kDa and was optimally active at pH 4.3 and 52 degrees C. It was inactivated at 65 degrees C over 15 min. It had a broad substrate specificity with strongest activity towards p-nitrophenyl phosphate. The enzyme dephosphorylated linearized pUC18 DNA and preventing self-ligation under the same conditions used for calf intestine alkaline phosphatase.

Publication types

  • Comparative Study

MeSH terms

  • Acid Phosphatase / chemistry
  • Acid Phosphatase / isolation & purification
  • Acid Phosphatase / metabolism*
  • Alkaline Phosphatase / metabolism*
  • Anions / pharmacology
  • Cations, Divalent / pharmacology
  • Cloning, Molecular / methods*
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Kinetics
  • Manihot / enzymology*
  • Molecular Weight
  • Nitrophenols / metabolism
  • Organophosphorus Compounds / metabolism
  • Phosphorylation / drug effects
  • Plant Leaves / enzymology
  • Plasmids / genetics
  • Plasmids / metabolism
  • Reducing Agents / pharmacology
  • Sodium Dodecyl Sulfate / pharmacology
  • Substrate Specificity
  • Temperature

Substances

  • Anions
  • Cations, Divalent
  • Nitrophenols
  • Organophosphorus Compounds
  • Reducing Agents
  • nitrophenylphosphate
  • Sodium Dodecyl Sulfate
  • Alkaline Phosphatase
  • Acid Phosphatase