Abstract
A soluble enzyme system from suspension cultures of Acer pseudoplatanus L. converts d-glucose 6-phosphate to myoinositol. A Mg(2+)-dependent phosphatase, present in the crude extract, hydrolyzes the product of the cyclization, myoinositol monophosphate, to free myoinositol. Further purification of the enzyme system by precipitation with (NH(4))(2)SO(4) followed by diethylaminoethyl cellulose chromatography eliminates the phosphatase and makes it necessary to add alkaline phosphatase to the reaction mixture in order to assay for free myoinositol. Gel filtration on Sephadex G-200 increases the specific activity of the cycloaldolase to 8.8 x 10(-4) units per milligram protein (1 unit = 1 micromole of myoinositol formed per minute). The cycloaldolase has an absolute requirement for nicotinamide adenine dinucleotide and a maximum activity at pH 8 with 0.1 mm nicotinamide adenine dinucleotide. The reaction rate is linear for 2.5 hours when d-glucose 6-phosphate is below 4 mm and has a K(m) of 1.77 mm. The diethylaminoethyl cellulose-purified enzyme is stable for 6 to 8 weeks in the frozen state.