蚕豆叶片下表皮ABA结合蛋白提取及分离条件的选择

Conditional Selection of the Extraction and Isolation of ABA Binding Protein(s) from Vicia faba Abaxial Epiderm

The abaxial epiderm of Vicia faba L. was chosen to prepare the crude extract of ABA binding protein (ABA-BP). The activity of ABA-BP was dependent on different extraction methods. The specific binding capacity of ABA-BP extracted with 0.5% Triton X-100 (binding activity (B) = 0. 487 nmoL/g protein) was higher than that with extracted cold acetone (0. 325 nmol/g protein) or (NH4)2SO4(0. 223 nmol/ g protein). The activity duration of ABA-BP extracted with Triton X-100 (60% Bmax after 40 h) was longer than that with cold acetone (30% Bmax after 10 h). The ABA-BP activity of binding ABA was sensitive to pH change with an optimum pH of 6.5. The ABA-BP specific binding activity decreased quickly under high concentration of NaC1 ( > 300 mmoL/L), but increased 12% with 5 mmol/L KC1. Some divalent cations like Ca2+ and Mg2 + were required for enhancing the ABA-BP activity. These optimum conditions are primordial for ABA-BP purification.