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β-Cyclodextrin-Functionalized Mycobacterium smegmatis Porin A for Steroid Discrimination and Gut Metabolite
Ruidong Li1,2, Zixuan Wang1,2, Jialu Chen1,2
1State Key Laboratory of Analytical Chemistry for Life Sciences, School of Chemistry, Nanjing University, 210023Nanjing, China.
Nano Letters
|August 12, 2026
Summary
This study presents a new MspA-βCD nanopore sensor for distinguishing structurally similar steroid metabolites like estradiol and bile acids. The sensor achieves high accuracy, enabling potential point-of-care diagnosis of metabolic disorders.
Area of Science:
- Biomolecular Engineering
- Nanotechnology
- Analytical Chemistry
Background:
- Steroid metabolites, including estradiol (E2) and bile acids (BAs), are crucial for human metabolism.
- Distinguishing these structurally similar molecules presents a significant analytical challenge.
Purpose of the Study:
- To develop a high-resolution nanopore sensor for differentiating analogous steroid metabolites.
- To investigate the structural factors influencing bile acid-β-cyclodextrin complex interactions.
Main Methods:
- Integration of β-cyclodextrin (βCD) into the MspA nanopore via hetero-octameric assembly.
- Utilizing Michael addition for sensor construction, termed MspA-βCD.
- Employing automated machine learning for classification of single-molecule events.
Main Results:
- The MspA-βCD sensor achieved high-resolution discrimination of 11 steroid metabolites.
- Machine learning classification yielded 99.7% validation accuracy.
- Identified key structural determinants (chirality, steric hindrance, side-chain conjugation) affecting BA-βCD complex dissociation rates.
Conclusions:
- The MspA-βCD platform offers precise control over βCD conformation for enhanced sensing.
- Demonstrated successful bile acid profiling in fecal samples, indicating diagnostic potential.
- Highlights the potential for point-of-care diagnosis of metabolic disorders using nanopore technology.
