Production of β-apo-10′-carotenal from β-carotene by human β-carotene-9′,10′-oxygenase expressed in E. coli

Kim, Yeong-Su; Yeom, Soo‐Jin; Oh, Deok‐Kun

doi:10.1007/s10529-011-0556-1

Cited by 13 publications

(10 citation statements)

References 18 publications

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“…Of the various detergents, Tween 40 at 2% (w/v) afforded the highest EhLCD enzymatic activity and was selected for use in forming detergent micelles. These results were similar to those obtained with human BCO2 [38]. Incubation with Triton X-100, Span 20, and Span 40 at 1% (w/v) reduced the enzymatic activity by less than 20% (Table 4).…”

Section: Effects Of Organic Solvent and Detergent On Enzymatic Activitysupporting

confidence: 87%

Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from Enterobacter hormaechei YT-3 for the Enzymatic Synthesis of 3-Hydroxy-β-ionone from Lutein

et al. 2021

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3-Hydroxy-β-ionone, a flavor and fragrance compound with fruity violet-like characteristics, is widely applied in foodstuff and beverages, and is currently produced using synthetic chemistry. In this study, a novel lutein cleavage enzyme (EhLCD) was purified and characterized from Enterobacter hormaechei YT-3 to convert lutein to 3-hydroxy-β-ionone. Enzyme EhLCD was purified to homogeneity by ammonium sulfate precipitation, Q-Sepharose, phenyl-Sepharose, and Superdex 200 chromatography. The molecular mass of purified EhLCD, obtained by SDS-PAGE, was approximately 50 kDa. The enzyme exhibited the highest activity toward lutein, followed by zeaxanthin, β-cryptoxanthin, and β-carotene, suggesting that EhLCD exhibited higher catalytic efficiency for carotenoid substrates bearing 3-hydroxy-ionone rings. Isotope-labeling experiments showed that EhLCD incorporated oxygen from O2 into 3-hydroxy-β-ionone and followed a dioxygenase reaction mechanism for different carotenoid substrates. These results indicated that EhLCD is the first characterized bacterial lutein cleavage dioxygenase. Active EhLCD was also confirmed to be a Fe2+-dependent protein with 1 molar equivalent of non-haem Fe2+. The purified enzyme displayed optimal activity at 45 °C and pH 8.0. The optimum concentrations of the substrate, enzyme, and Tween 40 for 3-hydroxy-β-ionone production were 60 mM lutein/L, 1.5 U/mL, and 2% (w/v), respectively. Under optimum conditions, EhLCD produced 3-hydroxy-β-ionone (637.2 mg/L) in 60 min with a conversion of 87.0% (w/w), indicating that this enzyme is a potential candidate for the enzymatic synthesis of 3-hydroxy-β-ionone in biotechnological applications.

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Section: Effects Of Organic Solvent and Detergent On Enzymatic Activitysupporting

confidence: 87%

Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from Enterobacter hormaechei YT-3 for the Enzymatic Synthesis of 3-Hydroxy-β-ionone from Lutein

et al. 2021

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“…Consistent with the in vitro chicken BCO2 results (36), Bco2 knockout mice accumulate xanthophylls (49), and Bco2 expression was inversely associated with lutein and zeaxanthin concentrations in nonhuman primate brain (52). Alternatively, 1 group found that, in vivo, a lack of macular BCO2 activity in the primate eye underlies macular accumulation of lutein and zeaxanthin (53), but the exact structural causes of this inactivation are unclear (53)(54)(55).…”

Section: Overview Of Carotenoid Bioavailability Serum or Plasma Halfmentioning

confidence: 52%

Intrinsic and Extrinsic Factors Impacting Absorption, Metabolism, and Health Effects of Dietary Carotenoids

Moran

Mohn

Hason

et al. 2018

Advances in Nutrition

153

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Carotenoids are orange, yellow, and red lipophilic pigments present in many fruit and vegetables, as well as other food groups. Some carotenoids contribute to vitamin A requirements. The consumption and blood concentrations of specific carotenoids have been associated with reduced risks of a number of chronic conditions. However, the interpretation of large, population-based observational and prospective clinical trials is often complicated by the many extrinsic and intrinsic factors that affect the physiologic response to carotenoids. Extrinsic factors affecting carotenoid bioavailability include food-based factors, such as co-consumed lipid, food processing, and molecular structure, as well as environmental factors, such as interactions with prescription drugs, smoking, or alcohol consumption. Intrinsic, physiologic factors associated with blood and tissue carotenoid concentrations include age, body composition, hormonal fluctuations, and variation in genes associated with carotenoid absorption and metabolism. To most effectively investigate carotenoid bioactivity and to utilize blood or tissue carotenoid concentrations as biomarkers of intake, investigators should either experimentally or statistically control for confounding variables affecting the bioavailability, tissue distribution, and metabolism of carotene and xanthophyll species. Although much remains to be investigated, recent advances have highlighted that lipid co-consumption, baseline vitamin A status, smoking, body mass and body fat distribution, and genetics are relevant covariates for interpreting blood serum or plasma carotenoid responses. These and other intrinsic and extrinsic factors are discussed, highlighting remaining gaps in knowledge and opportunities for future research. To provide context, we review the state of knowledge with regard to the prominent health effects of carotenoids.

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“…The 18 kDa Trx-His-S Tag protein of the vector pET-32a promoted highlevel expression of the target proteins while having little effect on the expressed proteins (Yang et al 2003;Gasparian et al 2007). At present, many measures have been reported for the purification of the recombinant proteins expressed in Escherichia coli (Li 2011;Kim et al 2011;Tzou et al 2011). In our study, high-purity recombinant pAPN-C proteins were obtained from inclusion bodies by the gel-cutting purification method combined with dialysis refolding, which had been verified as a simple, time-saving, and inexpensive method (Liu et al 2009;Sun et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Virus-binding activity of the truncated C subunit of porcine aminopeptidase N expressed in Escherichia coli

et al. 2011

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Seven overlapping truncated forms of the C subunit of porcine aminopeptidase N (pAPN-C) were expressed in Escherichia coli. By western blotting and ELISA test, all recombinant proteins were recognized by the antibody against native porcine aminopeptidase N. Recombinant proteins, rpAPN-C2 (aa 623-722) and rpAPN-C3 (aa 673-772), had the highest binding activity with swine transmissible gastroenteritis virus among the truncated pAPN-C recombinant proteins. The overlapping region (aa 673-722) between rpAPN-C2 and rpAPN-C3 is indicated to play a key role in viral binding.

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Production of β-apo-10′-carotenal from β-carotene by human β-carotene-9′,10′-oxygenase expressed in E. coli

Cited by 13 publications

References 18 publications

Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from Enterobacter hormaechei YT-3 for the Enzymatic Synthesis of 3-Hydroxy-β-ionone from Lutein

Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from Enterobacter hormaechei YT-3 for the Enzymatic Synthesis of 3-Hydroxy-β-ionone from Lutein

Intrinsic and Extrinsic Factors Impacting Absorption, Metabolism, and Health Effects of Dietary Carotenoids

Virus-binding activity of the truncated C subunit of porcine aminopeptidase N expressed in Escherichia coli

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