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引用次数: 0
摘要
分解代谢β-氧化酶级联的招募及其天然产物生物合成的反应逻辑仍未得到充分探索,这代表了合成生物学为结构独特的代谢物设计新途径的重要机会。在这项研究中,首次报道了真菌β-氧化级联的功能重构,该级联负责组装免疫抑制剂霉酚酸(MPA)。通过体外酶分析,确定了5种过氧化物酶体酶协同介导生物合成前体MFDHMP-3C的两轮反复侧链切割,揭示了MPA药效团形成的关键氧化策略。这些酶催化了顺序氧化、脱氢、水合、还原、异构化和反向克拉森缩合反应,反映了典型的β-氧化,同时使其适应于生物合成目的。此外,短压缩青霉菌NRRL864中限制性过氧化物酶体酰基辅酶a氧化酶PbACOX323、过氧化物酶体生物生成因子PbPex337和内质网定位加氧酶MpaB’的综合过表达使MPA产量增加了50%(从0.77 g L-1增加到1.15 g L-1),证明了途径优化的生物技术有效性。这项工作建立了真菌天然产物生物合成中完整的β-氧化样酶级联的第一个例子,为分解代谢模块的进化再利用提供了一个范例,以推动合成创新。
Enhancing the β-Oxidation-Like Pathway for the Optimal Production of the Immunosuppressant Mycophenolic Acid.
The recruitment of catabolic β-oxidation enzyme cascades and their reaction logic for natural product biosynthesis remains underexplored, representing a significant opportunity for synthetic biology to engineer novel pathways for structurally unique metabolites. In this study, the first functional reconstitution of the fungal β-oxidative cascade responsible for assembling the immunosuppressant mycophenolic acid (MPA) is reported. Through in vitro enzyme assays, five peroxisomal enzymes are identified that cooperatively mediate two iterative rounds of side-chain cleavage of the biosynthetic precursor MFDHMP-3C and revealed a key oxidative strategy for pharmacophore formation of MPA. These enzymes catalyzed sequential oxidation, dehydrogenation, hydration, reduction, isomerization, and reverse Claisen condensation reactions, mirroring canonical β-oxidation while adapting it for biosynthetic purposes. Furthermore, integrated overexpression of the rate-limiting peroxisomal acyl-CoA oxidase PbACOX323, peroxisomal biogenesis factor PbPex337, and endoplasmic reticulum (ER)-localized oxygenase MpaB' in Penicillium brevicompactum NRRL864 increased MPA production by 50% (from 0.77 to 1.15 g L-1), demonstrating the biotechnological efficacy of pathway optimization. This work establishes the first example of a full β-oxidation-like enzyme cascade in fungal natural product biosynthesis, providing a paradigm for the evolutionary repurposing of catabolic modules to drive synthetic innovation.
期刊介绍:
Advanced Science is a prestigious open access journal that focuses on interdisciplinary research in materials science, physics, chemistry, medical and life sciences, and engineering. The journal aims to promote cutting-edge research by employing a rigorous and impartial review process. It is committed to presenting research articles with the highest quality production standards, ensuring maximum accessibility of top scientific findings. With its vibrant and innovative publication platform, Advanced Science seeks to revolutionize the dissemination and organization of scientific knowledge.