Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein
3-Hydroxy-<i>β</i>-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...
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2021
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oai:doaj.org-article:d8edddf174ce4afab1258f0f07765ff72021-11-25T17:04:59ZCharacterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein10.3390/catal111112572073-4344https://doaj.org/article/d8edddf174ce4afab1258f0f07765ff72021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4344/11/11/1257https://doaj.org/toc/2073-43443-Hydroxy-<i>β</i>-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 <i>Enterobacter hormaechei</i> YT-3 to convert lutein to 3-hydroxy-<i>β</i>-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, <i>β</i>-cryptoxanthin, and <i>β</i>-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 O<sub>2</sub> into 3-hydroxy-<i>β</i>-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 Fe<sup>2+</sup>-dependent protein with 1 molar equivalent of non-haem Fe<sup>2+</sup>. 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-<i>β</i>-ionone production were 60 μM lutein/L, 1.5 U/mL, and 2% (<i>w</i>/<i>v</i>), respectively. Under optimum conditions, EhLCD produced 3-hydroxy-<i>β</i>-ionone (637.2 mg/L) in 60 min with a conversion of 87.0% (<i>w</i>/<i>w</i>), indicating that this enzyme is a potential candidate for the enzymatic synthesis of 3-hydroxy-<i>β</i>-ionone in biotechnological applications.Zhangde LongNaixin DuanYun XueMin WangJigang LiZan SuQibin LiuDuobin MaoTao WeiMDPI AGarticleEhLCD3-hydroxy-<i>β</i>-iononecarotenoid substratesluteindioxygenaseChemical technologyTP1-1185ChemistryQD1-999ENCatalysts, Vol 11, Iss 1257, p 1257 (2021) |
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DOAJ |
| collection |
DOAJ |
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EN |
| topic |
EhLCD 3-hydroxy-<i>β</i>-ionone carotenoid substrates lutein dioxygenase Chemical technology TP1-1185 Chemistry QD1-999 |
| spellingShingle |
EhLCD 3-hydroxy-<i>β</i>-ionone carotenoid substrates lutein dioxygenase Chemical technology TP1-1185 Chemistry QD1-999 Zhangde Long Naixin Duan Yun Xue Min Wang Jigang Li Zan Su Qibin Liu Duobin Mao Tao Wei Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein |
| description |
3-Hydroxy-<i>β</i>-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 <i>Enterobacter hormaechei</i> YT-3 to convert lutein to 3-hydroxy-<i>β</i>-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, <i>β</i>-cryptoxanthin, and <i>β</i>-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 O<sub>2</sub> into 3-hydroxy-<i>β</i>-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 Fe<sup>2+</sup>-dependent protein with 1 molar equivalent of non-haem Fe<sup>2+</sup>. 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-<i>β</i>-ionone production were 60 μM lutein/L, 1.5 U/mL, and 2% (<i>w</i>/<i>v</i>), respectively. Under optimum conditions, EhLCD produced 3-hydroxy-<i>β</i>-ionone (637.2 mg/L) in 60 min with a conversion of 87.0% (<i>w</i>/<i>w</i>), indicating that this enzyme is a potential candidate for the enzymatic synthesis of 3-hydroxy-<i>β</i>-ionone in biotechnological applications. |
| format |
article |
| author |
Zhangde Long Naixin Duan Yun Xue Min Wang Jigang Li Zan Su Qibin Liu Duobin Mao Tao Wei |
| author_facet |
Zhangde Long Naixin Duan Yun Xue Min Wang Jigang Li Zan Su Qibin Liu Duobin Mao Tao Wei |
| author_sort |
Zhangde Long |
| title |
Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein |
| title_short |
Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein |
| title_full |
Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein |
| title_fullStr |
Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein |
| title_full_unstemmed |
Characterization of a Novel Lutein Cleavage Dioxygenase, EhLCD, from <i>Enterobacter hormaechei</i> YT-3 for the Enzymatic Synthesis of 3-Hydroxy-<i>β</i>-ionone from Lutein |
| title_sort |
characterization of a novel lutein cleavage dioxygenase, ehlcd, from <i>enterobacter hormaechei</i> yt-3 for the enzymatic synthesis of 3-hydroxy-<i>β</i>-ionone from lutein |
| publisher |
MDPI AG |
| publishDate |
2021 |
| url |
https://doaj.org/article/d8edddf174ce4afab1258f0f07765ff7 |
| work_keys_str_mv |
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