Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase
Rhododendrol (RD) is a naturally occurring phenolic compound found in many plants. Tyrosinase (Ty) converts RD to RD-catechol and subsequently RD-quinone via two-step oxidation reactions, after which RD-melanin forms spontaneously from RD-quinone. RD is cytotoxic in melanocytes and lung cancer cells...
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MDPI AG
2020-09-01
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| Series: | Catalysts |
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| Online Access: | https://www.mdpi.com/2073-4344/10/10/1114 |
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| author | Chan Mi Park Hyun Seo Park Gun Su Cha Ki Deok Park Chul-Ho Yun |
| author_facet | Chan Mi Park Hyun Seo Park Gun Su Cha Ki Deok Park Chul-Ho Yun |
| author_sort | Chan Mi Park |
| collection | DOAJ |
| description | Rhododendrol (RD) is a naturally occurring phenolic compound found in many plants. Tyrosinase (Ty) converts RD to RD-catechol and subsequently RD-quinone via two-step oxidation reactions, after which RD-melanin forms spontaneously from RD-quinone. RD is cytotoxic in melanocytes and lung cancer cells, but not in keratinocytes and fibroblasts. However, the function of RD metabolites has not been possible to investigate due to the lack of available high purity metabolites. In this study, an enzymatic strategy for RD-catechol production was devised using engineered cytochrome P450 102A1 (CYP102A1) and Ty, and the product was analyzed using high-performance liquid chromatography (HPLC), LC-MS, and NMR spectroscopy. Engineered CYP102A1 regioselectively produced RD-catechol via hydroxylation at the ortho position of RD. Although RD-quinone was subsequently formed by two step oxidation in Ty catalyzed reactions, L-ascorbic acid (LAA) inhibited RD-quinone formation and contributed to regioselective production of RD-catechol. When LAA was present, the productivity of RD-catechol by Ty was 5.3-fold higher than that by engineered CYP102A1. These results indicate that engineered CYP102A1 and Ty can be used as effective biocatalysts to produce hydroxylated products, and Ty is a more cost-effective biocatalyst for industrial applications than engineered CYP102A1. |
| first_indexed | 2024-03-10T16:03:05Z |
| format | Article |
| id | doaj.art-c97a82958924439f859787d78a2474b3 |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-10T16:03:05Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Catalysts |
| spelling | doaj.art-c97a82958924439f859787d78a2474b32023-11-20T15:06:31ZengMDPI AGCatalysts2073-43442020-09-011010111410.3390/catal10101114Regioselective Hydroxylation of Rhododendrol by CYP102A1 and TyrosinaseChan Mi Park0Hyun Seo Park1Gun Su Cha2Ki Deok Park3Chul-Ho Yun4School of Biological Sciences and Technology, Chonnam National University, 77 Yongbongro, Gwangju 61186, KoreaSchool of Biological Sciences and Technology, Chonnam National University, 77 Yongbongro, Gwangju 61186, KoreaNamhae Garlic Research Institute, 2465-8 Namhaedaero, Namhae, Gyeongsangnamdo 52430, KoreaGwangju Center, Korea Basic Science Center, 77 Yongbongro, Gwangju 61186, KoreaSchool of Biological Sciences and Technology, Chonnam National University, 77 Yongbongro, Gwangju 61186, KoreaRhododendrol (RD) is a naturally occurring phenolic compound found in many plants. Tyrosinase (Ty) converts RD to RD-catechol and subsequently RD-quinone via two-step oxidation reactions, after which RD-melanin forms spontaneously from RD-quinone. RD is cytotoxic in melanocytes and lung cancer cells, but not in keratinocytes and fibroblasts. However, the function of RD metabolites has not been possible to investigate due to the lack of available high purity metabolites. In this study, an enzymatic strategy for RD-catechol production was devised using engineered cytochrome P450 102A1 (CYP102A1) and Ty, and the product was analyzed using high-performance liquid chromatography (HPLC), LC-MS, and NMR spectroscopy. Engineered CYP102A1 regioselectively produced RD-catechol via hydroxylation at the ortho position of RD. Although RD-quinone was subsequently formed by two step oxidation in Ty catalyzed reactions, L-ascorbic acid (LAA) inhibited RD-quinone formation and contributed to regioselective production of RD-catechol. When LAA was present, the productivity of RD-catechol by Ty was 5.3-fold higher than that by engineered CYP102A1. These results indicate that engineered CYP102A1 and Ty can be used as effective biocatalysts to produce hydroxylated products, and Ty is a more cost-effective biocatalyst for industrial applications than engineered CYP102A1.https://www.mdpi.com/2073-4344/10/10/1114rhododendrolcytochrome P450tyrosinasebiocatalysthydroxylationbioconversion |
| spellingShingle | Chan Mi Park Hyun Seo Park Gun Su Cha Ki Deok Park Chul-Ho Yun Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase Catalysts rhododendrol cytochrome P450 tyrosinase biocatalyst hydroxylation bioconversion |
| title | Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase |
| title_full | Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase |
| title_fullStr | Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase |
| title_full_unstemmed | Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase |
| title_short | Regioselective Hydroxylation of Rhododendrol by CYP102A1 and Tyrosinase |
| title_sort | regioselective hydroxylation of rhododendrol by cyp102a1 and tyrosinase |
| topic | rhododendrol cytochrome P450 tyrosinase biocatalyst hydroxylation bioconversion |
| url | https://www.mdpi.com/2073-4344/10/10/1114 |
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