Purification and Characterization of an Alkaline Phosphatase Induced by Phosphorus Starvation in Common Bean (Phaseolus vulgaris L.) Roots

Authors

  • Lorena Morales Universidad Nacional Autónoma de México
  • Natalia Gutiérrez Universidad Nacional Autónoma de México
  • Vanessa Maya Universidad Nacional Autónoma de México
  • Carmen Parra Universidad Nacional Autónoma de México
  • Eleazar Martínez-Barajas Universidad Nacional Autónoma de México
  • Patricia Coello Universidad Nacional Autónoma de México

DOI:

https://doi.org/10.29356/jmcs.v56i1.279

Keywords:

Alkaline phosphatase, Phaseolus vulgaris, phosphate deficiency, pyrophosphate, purification

Abstract

Two phosphatase isoforms from roots of the common bean (Phaseolus vulgaris L.) showed an increase in activity in response to phosphate deficiency. One of them (APIII) was chosen for further purification through ionic exchange chromatography and preparative electrophoresis. The estimated molecular mass of APIII was 35 kDa by both SDS-PAGE and gel filtration analyses, suggesting a monomeric form of the active enzyme. The phosphatase was classified as an alkaline phosphatase based on the requirement of pH 8 for optimum catalysis. It not only exhibited broad substrate specificity, with the most activity against pyrophosphate, but also effectively catalyzed the hydrolysis of polyphosphate, glucose-1-phosphate and phosphoenolpyruvate. Activity was completely inhibited by molybdate, vanadate and phosphate but was only partially inhibited by fluoride. Although divalent cations were not essential for the pyrophosphatase activity of this enzyme, the hydrolysis of pyrophosphate increased substantially in the presence of Mg2+.

Downloads

Download data is not yet available.

Author Biographies

Lorena Morales, Universidad Nacional Autónoma de México

Departamento de Bioquímica, Facultad de Química.

Natalia Gutiérrez, Universidad Nacional Autónoma de México

Departamento de Bioquímica, Facultad de Química.

Vanessa Maya, Universidad Nacional Autónoma de México

Departamento de Bioquímica, Facultad de Química.

Carmen Parra, Universidad Nacional Autónoma de México

Departamento de Bioquímica, Facultad de Química.

Eleazar Martínez-Barajas, Universidad Nacional Autónoma de México

Departamento de Bioquímica, Facultad de Química.

Patricia Coello, Universidad Nacional Autónoma de México

Departamento de Bioquímica, Facultad de Química.

Downloads

Published

2017-10-12