Strategies for regeneration of nicotinamide coenzymes emphasizing self-sufficient closed-loop recycling systems

• Different strategies to regenerate the reduced coenzymes NAD(P)H are presented.
• Besides two standard methods this contribution focusses on a further method used rarely so far.
• This method referred to as “closed-loop” or “self-sufficient” method is illustrated by several examples.
• Neither an additional substrate nor a further regenerating enzyme are required for this regenerating method.
• The current status as well as advantages and disadvantages of this method are discussed.

Biocatalytic reduction reactions depending on nicotinamide coenzymes require an additional reaction to regenerate the consumed cofactor. For preparative application the preferred method is the simultaneous coupling of an in situ regeneration reaction. There are different strategically advantageous routes to achieve this goal. The standard method uses a second enzyme and a second co-substrate, for example formate and formate dehydrogenase or glucose and glucose dehydrogenase. Alternatively, a second substrate is employed which is converted by the same enzyme used for the primary reaction. For example, alcohol dehydrogenase catalyzed reactions are often coupled with excess 2-propanol which is oxidized to acetone during the regeneration of NAD(P)H. A third method utilizes a reaction-internal sequence by the direct coupling of an oxidizing and a reducing enzyme reaction. Neither an additional substrate nor a further regenerating enzyme are required for the recycling reaction. This kind of “closed-loop” or “self-sufficient” redox process for cofactor regeneration has been used rarely so far. Its most intriguing advantage is that even redox reactions with unstable precursors can be realized provided that this compound is produced in situ by an opposite redox reaction. This elegant method is applicable in special cases only but increasing numbers of examples have been published during the last years.