Preparation of homogeneous crystals of myo-inositol 1-phosphate synthase from rat testicles — further data on the chemical and catalytic properties of the enzyme (studies on the biosynthesis cyclitols XXXIX1)

✍ Scribed by Fritz Pittner; Otto Hoffmann-Ostenhof

• Publisher: Springer
• Year: 1979
• Tongue: English
• Weight: 316 KB
• Volume: 28
• Category: Article
• ISSN: 0300-8177
• DOI: 10.1007/bf00223357

✦ Synopsis

Pre-purified preparations of myoinositol-1-phosphate synthase (E.C. 5.5.1.4) from rat testes can be purified to homogeneity by first crystallizing the enzyme according to JAKOBY and then recrystallizing it at a pH value close to the isoelectric point while slowly increasing the temperature from 0 to 15 degrees C. This method gives a much yield of homogeneous enzyme than the previously used purification by affinity chromatography. It was further found that the pure enzyme contains close to 2 mol NAD+ per mol enzyme; it does not contain any metal. At substrate saturation the enzyme binds close to 1 mol substrate per mol enzyme, as determined by using radioactively labelled substrate and binding it to the enzyme by reduction with NaBH4. The reaction catalyzed by the enzyme is irreversible.