دورية أكاديمية
Simplification of Corticosteroids Biosynthetic Pathway by Engineering P450BM3
| العنوان: | Simplification of Corticosteroids Biosynthetic Pathway by Engineering P450BM3 |
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| المؤلفون: | Qihang Chen, Zikai Chao, Ke Wang, Xinglong Wang, Hao Meng, Xirong Liu, Xiaoyu Shan, Jingwen Zhou |
| سنة النشر: | 2024 |
| مصطلحات موضوعية: | Biophysics, Biochemistry, Microbiology, Biotechnology, Ecology, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, Information Systems not elsewhere classified, steroidal pharmaceutical production, step process involving, multienzyme cascade process, molecular dynamics simulations, molar conversion rate, cortexolone (< b, developed p450 variants, l ), respectively, cytochrome p450 reductase, hydrocortisone (< b, p450bm3 variants revealed, least two enzymes, engineering p450bm3 synthesis, corticosteroids biosynthetic pathway, p450bm3 variants, hydrocortisone synthesis, sufficient p450bm3, six enzymes, simplified two, cytochrome p450s, biosynthetic steps |
| الوصف: | Synthesis of corticosteroids, particularly hydrocortisone, is challenging owing to the complex network requiring pairing of cytochrome P450s with cytochrome P450 reductase (CPR) for achieving regionally selective hydroxylation modifications at multiple sites. Herein, we engineered a self-sufficient P450BM3 (CYP102A1 from Bacillus megaterium) for effectively reducing the traditionally complex, multienzyme cascade process (three steps and six enzymes) of hydrocortisone synthesis from progesterone (PG) to a simplified two-step process involving at least two enzymes. Driven by computational simulation-guided substrate access channel and heme center pocket engineering, a series of P450BM3 variants were gradually designed with the ability to catalyze C16β, C17α, C21, and C17α/21 oxidation of PG and C11α oxidation of cortexolone ( c ). Subsequently, molecular dynamics simulations with an oxy-ferrous model of P450BM3 variants revealed that the glycine mutations of residues that are repulsive to the substrate allow for more stable exposure of the substrate above FeO. Finally, the developed P450 variants were employed to construct efficient Escherichia coli catalytic systems, which further achieved 11α/β-hydrocortisone ( f / e ) production in one pot from 1 g/L PG at a molar conversion rate of 81 and 84% (912 and 955 mg/L), respectively. Thus, this study provides feasible strategies for simplifying the biosynthetic steps and biocatalysts for steroidal pharmaceutical production. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | unknown |
| العلاقة: | https://figshare.com/articles/journal_contribution/Simplification_of_Corticosteroids_Biosynthetic_Pathway_by_Engineering_P450BM3/25334784Test |
| DOI: | 10.1021/acscatal.3c06137.s001 |
| الإتاحة: | https://doi.org/10.1021/acscatal.3c06137.s001Test https://figshare.com/articles/journal_contribution/Simplification_of_Corticosteroids_Biosynthetic_Pathway_by_Engineering_P450BM3/25334784Test |
| حقوق: | CC BY-NC 4.0 |
| رقم الانضمام: | edsbas.5E5049FD |
| قاعدة البيانات: | BASE |
| DOI: | 10.1021/acscatal.3c06137.s001 |
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