A novel thermophilic β-carotene 15,15'-monooxygenase with broad substrate specificity from the marine bacterium Candidatus Pelagibacter sp. HTCC7211

Abstract

To characterize a novel thermophilic β-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 H₂¹⁸O 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 β-carotene 15,15'-monooxygenase. BCMO₇₂₁₁ exhibited broad carotenoid substrate specificity toward α-carotene, β-cryptoxanthin, β-carotene, zeaxanthin, and lutein. The optimum temperature, pH, and concentrations of the substrate and enzyme for retinal production were 60 °C, 9.0, 500 mg β-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.

Keywords: BCMO7211; Retinal; β-carotene; β-carotene 15,15′-monooxygenase.