Characterization of Two NMN Deamidase Mutants as Possible Probes for an NMN Biosensor

Characterization of Two NMN Deamidase Mutants as Possible Probes for an NMN Biosensor

Article Summary:

Title: Identification of Inactive Mutants of NMN Deamidase as Molecular Recognition Elements for an NMN-Specific Biosensor


  • Nicotinamide mononucleotide (NMN) is an important intermediate in the production of nicotinamide adenine dinucleotide (NAD+), a key molecule involved in cellular metabolism.
  • Supplementation of NMN has shown positive effects on various diseases, such as metabolic, vascular, and neurodegenerative disorders.

Study Objective:

  • This study aimed to identify inactive mutants of NMN deamidase (PncC) as molecular recognition elements (MREs) for an NMN-specific biosensor.


  • Thermal stability assays and fluorescence spectroscopy were used to analyze the binding of NMN and related metabolites to mutated PncC variants.
  • The S29A and K61Q PncC variants were chosen for further analysis.


  • Both S29A and K61Q PncC variants can bind to NMN, but only S29A also binds to NaMN, a product of the PncC reaction.
  • K61Q PncC shows potential as a specific probe for an NMN biosensor.


  • A biosensor for NMN could provide insights into its role in cellular metabolism and disease.
  • The study identified mutants of NMN deamidase that still bind NMN, making them potential candidates for an NMN biosensor.


  • S29A and K61Q PncC mutants have the ability to bind NMN and may be used as molecular recognition elements in an NMN-specific biosensor.

Further Considerations:

  • Future studies should investigate ligand-induced conformational changes, protein stability under different conditions, and factors like pH, temperature, ionic strength, and solvent polarity for biosensor design.
  • Additional data and insights related to this study can be found in the supplementary materials provided online.

Author Information:

  • Alessandra Camarca, S.D., and N.R. were involved in project conceptualization, formal analysis, and funding acquisition.
  • G.M., A.P., Alessandro Capo (A.C.), and A.A. contributed to the methodology.
  • S.D. and N.R. supervised the project.
  • A.C., A.P., and N.R. drafted the original manuscript, which was reviewed and edited by A.C., S.D., and N.R.
  • All authors have approved the manuscript for publication.


  • The funding for this work was provided by Ministero Dell’Università e della Ricerca for the Prin 2017 CBNCYT project.

Ethical Considerations:

  • The document states that the Institutional Review Board (IRB) statement is not applicable, suggesting that the research does not involve human subjects or falls outside the IRB's jurisdiction.
  • Researchers should ensure that their work complies with ethical considerations, even when an IRB statement is not applicable.

Data Availability:

  • The datasets used in the study can be accessed by contacting the corresponding author.
  • Data availability is subject to the permission and discretion of the corresponding author.
  • Interested individuals should reach out directly to the corresponding author to request access to the datasets.

Conflicts of Interest:

  • The authors have declared that they have no conflicts of interest.
  • This increases the credibility and objectivity of their work.

Publisher Note:

MDPI, the publisher, remains neutral regarding jurisdictional claims and institutional affiliations.

Title of paper: Characterization of Two NMN Deamidase Mutants as Possible Probes for an NMN Biosensor

Author(s): Camarca A, Minazzato G, Pennacchio A, Capo A, Amici A, D'Auria S, Raffaelli N.

Year published: 2021

Published in: Int J Mol Sci

Original article can be found here.

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