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Molecular Engineering of Nicotinamide Riboside Kinase and Process Optimization for Efficient Nicotinamide

Dai Ma1, Rui Liu1, Tong Bao1

  • 1School of Food and Biological Engineering, Hefei University of Technology, Hefei 230000, China.

Foods (Basel, Switzerland)
|June 12, 2026
PubMed
Summary

Researchers enhanced nicotinamide riboside kinase (NRK) enzyme activity and stability through semi-rational design. This breakthrough enables more efficient and greener industrial-scale production of nicotinamide mononucleotide (NMN).

Keywords:
ATP regeneration systemdual-enzyme couplingmolecular engineeringnicotinamide mononucleotide synthesisnicotinamide riboside kinase

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Area of Science:

  • Biotechnology
  • Enzyme Engineering
  • Biochemical Synthesis

Background:

  • Nicotinamide mononucleotide (NMN) is a vital vitamin B derivative.
  • Nicotinamide riboside kinase (NRK) is crucial for efficient NMN synthesis.
  • Current NMN production methods require optimization for industrial scale.

Purpose of the Study:

  • To engineer the Hi-NRK enzyme for improved activity and stability.
  • To establish an efficient and environmentally friendly NMN production process.
  • To achieve high yields of NMN for potential industrial applications.

Main Methods:

  • Semi-rational design was used to modify the Hi-NRK enzyme.
  • A critical mutant, Hi-NRKG8S, was identified and characterized.
  • Reaction conditions were optimized, including substrate concentration and enzyme co-factor addition.

Main Results:

  • The Hi-NRKG8S mutant showed a twofold increase in enzymatic activity.
  • Thermal stability was significantly enhanced, with half-life at 40 °C extending from 4 to 8 hours.
  • Optimized conditions and ADK addition achieved NMN yields of 94.17% and 97.24% respectively.

Conclusions:

  • Engineered Hi-NRKG8S provides a robust platform for NMN synthesis.
  • The optimized process demonstrates high efficiency and yield for NMN production.
  • This study lays the groundwork for scalable, green industrial NMN manufacturing.