I am trying to find out how NADH or NADPH are produced in bulk. These molecules are very important biologically, however, they are also sold as chemical reagents and produced in large quantities. How are these chemically produced industrially? Is that through an organic synthesis (which synthesis?) or do they use engineered bacteria or enzymes and then harvest the NADH produced? I tried to find this information online, but could not find it anywhere.
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2$\begingroup$ search the patent literature. A quick search suggests that state of the art is enzymatic synthesis starting from nicotinamide riboside which is chemically synthesized, but that might not be the dominant method. $\endgroup$– AndrewCommented Feb 6, 2023 at 12:44
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1$\begingroup$ Andrew might be on the right track today, and maybe for the past few decades. However I'd bet that at one time (maybe as recently as 30 or even 20 years ago), the preferred method would have been purifying NAD+ or NADP+ from yeast extract, and if desired, converting to NADH or NADPH enzymatically. $\endgroup$– Curt F.Commented Feb 6, 2023 at 16:51
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2 Answers
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The nicotinamide cofactors, $\ce{NAD+, NADP+, NADH,}$ and $\ce{NADPH,}$ are useful in a variety of enzyme-catalyzed oxidation and reduction reactions in organic synthesis. As Curt F. suggested in the comment, $\ce{NAD+}$ as known as Coenzyme 1 (which is the longest-known coenzyme), is manufactured by a yeast based fermentation process (at least until 1999; Ref.1).
An early chemical synthesis of $\ce{NAD+}$ was reported in 1957 (Ref.2). In that approach, dicyclohexylcarbodiimide (DCC) was used for the coupling of adenosine monophosphate (AMP) and $\beta$-nicotinamide mononucleotide ($\beta$-NMN). This semi-synthesis was envisioned in 1950 (Ref.3) and achieved by 1957 (Ref.2). Finally, in 1999, Jaemoon Lee & coworkers have achieved the complete chemical synthesis of $\ce{NAD+}$ (Ref.4):
Where conditions are: (i) $\ce{HBr(g)/CH2Cl2}$; (ii) nicotinamide/$\ce{SO2},$ $\pu{210 ^\circ C},$ 90% from $\bf{7}$; (iii) $\ce{NH3/MeOH},$ $\pu{25 ^\circ C},$ $\pu{20 h},$ 80%; (iv) $\ce{POCl3}$ (4 equiv.), $\ce{PO(OMe)3},$ $25$ to $\pu{0 ^\circ C}, \pu{7 h},$ 80%; and (v) $\ce{MnCl2–MgSO4, HCONH2,}$ room temp., $\pu{16 h},$ 78% (58% isolated yield, > 99% purity).
Note: There is a peer-reviewed wore of enzymatic procedure capable of atomic labelling (Ref.5). The most recent published procedure is also semi-enzymatic while using $\ce{H2}$ gas to reduce and $\ce{D2O}$ to lable (Ref. 6).
References:
- S. Budavari, editor, The Merck Index, 11th edition; Merck & Co., Inc.: Rahway, NJ, 1989, p. 6259 (ISBN 10: 091191028X).
- N. H. Hughes, G. W. Kenner, and Sir. Alexander Todd, "735. Codehydrogenases. Part 111. A Synthesis of Diphosphopyridine Nucleotide (Coxymase), and Some Observations on the Synthesis of Triphosphopyridine Nucleotide," J. Chem. Soc. 1957, 3733-3738 (DOI: https://doi.org/10.1039/JR9570003733).
- L. J. Haynes and A. R. Todd, "66. Codehydrogenases. Part 1. The synthesis of dihydronicotinamide-D-ribofuranoside [N-D-ribofuranosidyl-1 : 2(or 6)-dihydronicotinamide]," J. Chem. Soc. 1950, 303-308 (DOI: https://doi.org/10.1039/JR9500000303).
- Jaemoon Lee, Hywyn Churchil, Woo-Baeg Choi, Joseph E. Lynch, F. E. Roberts, R. P. Volante, and Paul J. Reider, "A chemical synthesis of nicotinamide adenine dinucleotide ($\ce{NAD+}$)," Chem. Commun. 1999, (8), 729-730 (DOI: https://doi.org/10.1039/A809930H).
- Kathleen A. Rising and Vern L. Schramm, "Enzymic synthesis of $\ce{NAD+}$ with the specific incorporation of atomic labels," J. Am. Chem. Soc. 1994, 116(15), 6531-6536 (DOI: https://doi.org/10.1021/ja00094a006).
- Jack S. Rowbotham, Adam P. Hardy, Holly A. Reeve, and Kylie A. Vincent, "Synthesis of $[4S-\ce{^2H]NADH,}$ $[4R-\ce{^2H]NADH,}$ $[4-\ce{^2H2]NADH}$ and $[4-\ce{^2H]NAD+}$ cofactors through heterogeneous biocatalysis in heavy water," J. Labelled Compounds & Radiopharmaceuticals 2021, 64(15), 181–186 (DOI: https://doi.org/10.1002/jlcr.3899).
answered Feb 6, 2023 at 18:46
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$\begingroup$ I have included 2021 paper as well. $\endgroup$ Commented Feb 6, 2023 at 20:02
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According to a recent patent[1], the chemical synthesis approach is being replaced by an chemoenzymatic synthesis:
$$\text{nicotinamide riboside}\ce{->[NRK; ATP]}\text{nicotinamide mononucleotide}\ce{->[NMNAT; ATP]}\text{NAD}$$
where NRK = nicotinamide ribokinase and NMNAT = nicotinamide mononucleotide adenyl transferase.
The starting material nicotinamide riboside can be chemically synthesized, as described in another patent[2] (though I'm not sure what synthetic approach is used most for current commercial production).
[1] CN112437813A, Method for industrially producing NAD (nicotinamide adenine dinucleotide) by enzyme method
[2] US11242364 Efficient and Scalable Syntheses of Nicotinoyl Ribosides and Reduced Nicotinoyl Ribosides, Modified Derivatives Thereof, Phosphorylated Analogs Thereof, Adenylyl Dinucleotide Conjugates Thereof, and Novel Crystalline Forms Thereof
