近日，南京大学黄硕团队研究了基于纳米孔传感的所有蛋白质原性氨基酸对映体的手性识别。该研究于2025年10月21日发表在《德国应用化学》杂志上。

D-氨基酸是更为普遍的l型氨基酸的立体异构体，普遍分布于微生物、植物和哺乳动物系统中。最近的进展揭示了它们在生物过程中不可或缺的作用，但D-氨基酸的直接单分子检测长期以来一直没有得到充分的研究。同时辨别所有氨基酸对映体仍然是一个艰巨的分析挑战。

研究组报道了一种镍离子螯合异八聚耻垢分枝杆菌孔蛋白a (MspA) （MspA-NTA-Ni）纳米孔传感器，该传感器可以同时识别所有19种D-氨基酸，这些氨基酸对应于蛋白质生成的l-异构体。通过整合监督机器学习算法，该系统的整体识别准确率达到99.5%。该方法在分析酸水解D肽产品中得到进一步验证，得到了氨基酸对映体的清晰组成。值得注意的是，该技术能够同时检测所有39种氨基酸对映体，总体精度为98.9%。这种方法不仅可以直接表征DL-氨基酸混合物，而且还为基于纳米孔的D肽和D蛋白外显子测序提供了关键的传感器组件。

附：英文原文

Title: Chiral Discrimination of all Proteinogenic Amino Acid Enantiomers by Nanopore Sensing

Author: Hanhan Zhang, Kefan Wang, Xiao Zhou, Yifan Wang, Panke Zhang, Wenfei Li, Shuo Huang

Issue&Volume: 2025-10-21

Abstract: D-amino acids, the stereoisomers of the more prevalent L-forms, are ubiquitously distributed across microorganisms, plants, and mammalian systems. Recent advances have uncovered their indispensable roles in biological processes, yet direct, single-molecule detection of D-amino acids has long been understudied. The simultaneous discrimination of all amino acid enantiomers remains a formidable analytical challenge. Here, we report a nickel-ion chelated hetero-octameric Mycobacterium smegmatis porin A (MspA) (MspA-NTA-Ni) nanopore sensor that achieves simultaneous identification of all 19 D-amino acids corresponding to proteinogenic L-isomers. By integrating supervised machine learning algorithms, this system demonstrated an overall recognition accuracy of 99.5%. The method was further validated in analyzing acid-hydrolyzed D-peptide products, yielding clear compositional profiles of amino acid enantiomers. Notably, the technology enables simultaneous sensing of all 39 amino acid enantiomers with an overall accuracy of 98.9%. This approach not only allows direct characterization of DL-amino acid mixtures but also provides a critical sensor component for nanopore-based exo-sequencing of D-peptides and D-proteins.

DOI: 10.1002/anie.202515531

Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202515531