Purification and properties of a phosphatase in French bean (Phaseolus vulgaris L.) leaves that hydrolyses 2'-carboxy-D-arabinitol 1-phosphate

Kingston-Smith, A. H. and Major, I. and Parry, M. A J and Keys, A. J. (1992) Purification and properties of a phosphatase in French bean (Phaseolus vulgaris L.) leaves that hydrolyses 2'-carboxy-D-arabinitol 1-phosphate. Biochemical Journal, 287 (3). pp. 821-825. ISSN 0264-6021

Abstract

An enzyme that releases P. from 2-carboxy-D-arabinitol l-phosphate, a naturally occurring tightly binding inhibitor of ribulose 1,5-bisphosphate carboxylase/oxygenase (EC 4. 1.1. 39), was purified from leaves of French bean seedlings. It was a monomeric protein of M(r) about 56000. Catalytic activity was stimulated by increased concentrations of inorganic salts to a maximum at an ionic strength above 0.2. NADPH and D-fructose 1,6-bisphosphate increased the activity of the enzyme in both the presence and absence of 0.2 M-KCl. The pure enzyme did not require dithiothreitol for activity. The pH optimum was 7, the K(m) for 2-carboxy-D-arabinitol l-phosphate was 0.43 mM and the specific activity 6.8 μmol/min per mg of protein. The enzyme had little or no activity against phosphate ester intermediates of photosynthetic metabolism and glycolysis but hydrolysed the 1,5-bisphosphates of 2'-carboxy-D-ribitol and 2'-carboxy-D-arabinitol more rapidly than 2'-carboxy-D-arabinitol 1-phosphate.