PDF(Pubmed)
Abstract：
Enzymatic peptide synthesis is a powerful alternative to solid-phase methods, as enzymes can have high regio- and stereoselectivity and high yield and require mild reaction conditions. This is beneficial in formulation research due to the rise of nucleic acid therapies. Peptide nucleic acids (PNAs) have a high affinity toward DNA and RNA, and their solubility and cellular delivery can be improved via conjugation to peptides. Here, we designed and assessed the viability of the papain enzyme to conjugate four PNA-peptide models in water and an organic solvent using QM/MM metadynamics. We found that the reactions in water yield better results, where three conjugates could potentially be synthesized by the enzyme, with the first transition state as the rate-limiting step, with an associated energy of 14.53 kcal mol-1, although with a slight endergonic profile. The results highlight the importance of considering the enzyme pockets and different substrate acceptivities and contribute to developing greener, direct, and precise synthetic routes for nucleic acid-based therapies. By exploring the enzyme\'s potential in conjunction with chemical synthesis, current protocols can be simplified for the synthesis of longer nucleic acids and peptide sequences (and, by extension, proteins) from smaller oligo or peptide blocks.
摘要：
酶肽合成是固相方法的强大替代方法，酶可以具有高的区域和立体选择性和高产率，并且需要温和的反应条件。由于核酸疗法的兴起,这在制剂研究中是有益的。肽核酸（PNA）对DNA和RNA具有很高的亲和力，并且它们的溶解度和细胞递送可以通过与肽缀合来改善。这里,我们使用QM/MM元动力学设计并评估了木瓜蛋白酶在水和有机溶剂中缀合四种PNA-肽模型的活力。我们发现在水中的反应产生更好的结果，酶可能会合成三种结合物，以第一过渡状态作为限速步骤，具有14.53kcalmol-1的相关能量，尽管具有轻微的内皮特征。结果突出了考虑酶袋和不同底物可接受性的重要性，并有助于开发更环保的产品，直接，以及基于核酸的治疗的精确合成路线。通过探索酶与化学合成结合的潜力，目前的方案可以简化，用于合成更长的核酸和肽序列(和，通过延伸,蛋白质)来自较小的寡核苷酸或肽块。
