An acid phosphatase from Manihot glaziovii as an alternative to alkaline Phosphatase for molecular cloning experiments.
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.
Tham
S C
SC
Faculty of Science, Department of Biological Sciences, National University of Singapore, Kent Ridge, 119260, Singapore, Singapore.
Lim
S H
SH
Yeoh
H H
HH
D000135:Acid Phosphatase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000378:metabolism*; D000838:Anions / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods*; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D002365:Manihot / Q000201:enzymology*; D008970:Molecular Weight; D009596:Nitrophenols / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D018515:Plant Leaves / Q000201:enzymology; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D019163:Reducing Agents / Q000494:pharmacology; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature
D000838:Anions; D002413:Cations, Divalent; D009596:Nitrophenols; D009943:Organophosphorus Compounds; D019163:Reducing Agents; C008644:nitrophenylphosphate; D012967:Sodium Dodecyl Sulfate; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase