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Purification of β‐carotene 15,15′‐monooxygenase from pig intestine and its enzymatic hydrolysis of pigment in soybean oil
Author(s) -
Yu Dianyu,
Zhang Ruchun,
Wang Yuqi,
Zou Dezhi,
Li Tingting,
Tang Honglin,
Jiang Lianzhou,
Wang Liqi
Publication year - 2019
Publication title -
international journal of food science and technology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.831
H-Index - 96
eISSN - 1365-2621
pISSN - 0950-5423
DOI - 10.1111/ijfs.13960
Subject(s) - soybean oil , carotene , chemistry , chromatography , vitamin , enzyme , pigment , hydrolysis , size exclusion chromatography , molecular mass , enzymatic hydrolysis , sodium , monooxygenase , biochemistry , food science , organic chemistry , cytochrome p450
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.