A facile analytical method for reliable selectivity examination in cofactor NADH regeneration

Area of Science:

• Biochemistry
• Analytical Chemistry
• Catalysis

Background:

• NADH regeneration is crucial for enzymatic catalysis and biocatalysis.
• Accurate quantification of selective (1,4-NADH) and unselective (1,2- and 1,6-NADH) products is essential for process optimization.
• Current methods may lack the precision to differentiate between NADH isomers.

Purpose of the Study:

• To develop and validate a novel, combined spectroscopic and biological assay method for quantifying selective and unselective NADH products.
• To assess the efficiency and selectivity of NADH regeneration under specific catalytic conditions.

Main Methods:

• Utilized combined UV-Vis spectroscopy and biological assays for product quantification.
• Employed platinum on carbon (Pt/C) as a catalyst for NAD+ hydrogenation.
• Used hydrogen gas as the reducing agent in the reaction system.

Main Results:

• Successfully demonstrated a novel method for quantifying 1,4-NADH, 1,2-NADH, and 1,6-NADH.
• Validated the method's accuracy in the context of Pt/C catalyzed NAD+ hydrogenation.
• The method provides a reliable tool for assessing NADH regeneration processes.

Conclusions:

• The developed method offers a robust approach for distinguishing and quantifying different NADH isomers.
• This advancement facilitates better understanding and optimization of NADH regeneration systems.
• The findings are applicable to various biocatalytic and chemical processes relying on NADH regeneration.