Purification of β-carotene 15,15′-monooxygenase from pig intestine and its enzymatic hydrolysis of pigment in soybean oil

Summary β-Carotene 15,15′-monooxygenase was isolated and purified from the intestinal mucosa of pigs, and then, it was applied to hydrolyse the pigment in soybean oil, and thus, vitamin A fortified soybean oil was obtained. The pig intestinal mucosal protein solution was purified to a specific activity of 2.487 × 10 −4 U mg −1 , maximum reaction velocity ( V max ) of 8.42 × 10 −9 mol/h and Michaelis constant (K m ) of 2.03 × 10 −5 m . The protein solution had a molecular mass of 156 kDa by gel filtration. The sodium deoxycholate concentrations, optimum pH, Tween 40 amount and enzyme amount for vitamin A production in soybean oil were determined to be 6.0 m m , 8.0, 3.0% (w/v) and 0.2 U/mL enzyme, respectively. Under these conditions, β-carotene 15,15′-monooxygenase produced 14.65 mg/L vitamin A after 20 h, with a conversion yield of β-carotene of 33.29% (w/w). Therefore, the nutrients in soybean oil were improved.