A Hitchhiker’s Guide to Supplying Enzymatic Reducing Power into Synthetic Cells

Michele Partipilo, Nico J. Claassens, Dirk Jan Slotboom*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)


The construction from scratch of synthetic cells by assembling molecular building blocks is unquestionably an ambitious goal from a scientific and technological point of view. To realize functional life-like systems, minimal enzymatic modules are required to sustain the processes underlying the out-of-equilibrium thermodynamic status hallmarking life, including the essential supply of energy in the form of electrons. The nicotinamide cofactors NAD(H) and NADP(H) are the main electron carriers fueling reductive redox reactions of the metabolic network of living cells. One way to ensure the continuous availability of reduced nicotinamide cofactors in a synthetic cell is to build a minimal enzymatic module that can oxidize an external electron donor and reduce NAD(P)+. In the diverse world of metabolism there is a plethora of potential electron donors and enzymes known from living organisms to provide reducing power to NAD(P)+ coenzymes. This perspective proposes guidelines to enable the reduction of nicotinamide cofactors enclosed in phospholipid vesicles, while avoiding high burdens of or cross-talk with other encapsulated metabolic modules. By determining key requirements, such as the feasibility of the reaction and transport of the electron donor into the cell-like compartment, we select a shortlist of potentially suitable electron donors. We review the most convenient proteins for the use of these reducing agents, highlighting their main biochemical and structural features. Noting that specificity toward either NAD(H) or NADP(H) imposes a limitation common to most of the analyzed enzymes, we discuss the need for specific enzymes─transhydrogenases─to overcome this potential bottleneck.

Original languageEnglish
Pages (from-to)947–962
Number of pages16
JournalACS synthetic biology
Issue number4
Publication statusPublished - 13 Apr 2023


  • dehydrogenases
  • electron donors
  • minimal metabolism
  • nicotinamide adenine dinucleotides
  • redox reactions
  • synthetic cells
  • vesicles


Dive into the research topics of 'A Hitchhiker’s Guide to Supplying Enzymatic Reducing Power into Synthetic Cells'. Together they form a unique fingerprint.

Cite this