초록
<P><B>ABSTRACT</B></P><P>The photosynthetic bacterium <I>Rhodobacter capsulatus</I> normally photoproduces H<SUB>2</SUB> as a by‐product of its nitrogenase‐catalyzed nitrogen‐fixing activity. Such H<SUB>2</SUB> production, however, is expensive from a metabolic perspective, requiring nearly four times as many photons as the equivalent algal hydrogenase‐based system (Ghirardi et al., 2009 Photobiological hydrogen‐producing systems. Chem Soc Rev 38(1):52–61). Here, we report the insertion of a <I>Clostridium acetobutylicum</I> [FeFe]‐hydrogenase and its three attendant hydrogenase assembly proteins into an <I>R. capsulatus</I> strain lacking its native uptake hydrogenase. Further, this strain is modified to fluoresce upon sensing H<SUB>2</SUB>. The resulting strain photoproduces H<SUB>2</SUB> and self‐reports its own H<SUB>2</SUB> production through fluorescence. This model system represents a unique method of developing hydrogenase‐based H<SUB>2</SUB> production in <I>R. capsulatus</I>, may serve as a powerful system for in vivo directed evolution of hydrogenases and hydrogenase‐associated genes, and provides a means of screening for increased metabolic production of H<SUB>2</SUB>. Biotechnol. Bioeng. 2017;114: 291–297. © 2016 Wiley Periodicals, Inc.</P>