期刊
BIOTECHNOLOGY LETTERS
卷 43, 期 12, 页码 2233-2241出版社
SPRINGER
DOI: 10.1007/s10529-021-03188-w
关键词
BCMO7211; beta-carotene; beta-carotene 15; 15 '-monooxygenase; Retinal
资金
- Program for Science & Technology Innovation Talents in the University of Henan Province [18HASTIT040]
- Science and Technology Project of China Guangxi Tobacco Industry Co. Ltd. [2019450000340011]
- Science and Technology Project of Overseas Chinese in Henan Province [2019-03]
A novel thermophilic beta-carotene 15,15'-monooxygenase BCMO7211 was characterized from marine bacterium Candidatus Pelagibacter sp. HTCC7211. BCMO7211 showed broad carotenoid substrate specificity and potential for enzymatic synthesis of retinal in biotechnological applications, producing 888.3 mg/L retinal with a conversion rate of 89.0% (w/w) under optimal conditions.
To characterize a novel thermophilic beta-carotene 15,15 '-monooxygenase BCMO7211 isolated from the marine bacterium Candidatus Pelagibacter sp. HTCC7211. BCMO7211 was functionally overexpressed in Escherichia coli and purified to homogeneity by Ni-NTA affinity chromatography and Superdex-200 gel filtration chromatography. Labeling experiments with (H2O)-O-18 demonstrated that the oxygen atom in the terminal aldehyde group of the produced retinal molecules was provided from both molecular oxygen and water, indicating that BCMO7211 is the first characterized bacterial beta-carotene 15,15 '-monooxygenase. BCMO7211 exhibited broad carotenoid substrate specificity toward alpha-carotene, beta-cryptoxanthin, beta-carotene, zeaxanthin, and lutein. The optimum temperature, pH, and concentrations of the substrate and enzyme for retinal production were 60 degrees C, 9.0, 500 mg beta-carotene/L, and 2.5 U/ml, respectively. Under optimum conditions, 888.3 mg/L retinal was produced in 60 min with a conversion rate of 89.0% (w/w). BCMO7211 is a potential candidate for the enzymatic synthesis of retinal in biotechnological applications.
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