Particulate photocatalysts developed for the solar energy-driven reduction of the greenhouse gas CO 2 have a small product range and low specificity. Hybrid photosynthesis expands the number of products with photocatalysts harvesting sunlight and transferring charges to microbes harboring versatile metabolisms for bioproduction. Besides CO 2 , abiotic photocatalysts have been employed to increase microbial production yields of reduced compounds from organic carbon substrates. Most single-reactor hybrid photosynthesis systems comprise CdS assembled in situ by microbial activity. This approach limits optimization of the morphology, crystal structure, and crystallinity of CdS for higher performance, which is usually done via synthesis methods incompatible with life. Here, shape and activity optimized CdS nanorods were hydrothermally produced and subsequently applied to Cupriavidus necator for the heterotrophic and autotrophic production of the bioplastic polyhydroxybutyrate (PHB). C. necator with CdS NR under light produced 1.5 times more PHB when compared to the same bacterium with suboptimal commercially-available CdS. Illuminated C. necator with CdS NR synthesized 1.41 g PHB from fructose over 120 h and 28 mg PHB from CO 2 over 48 h. Interestingly, the beneficial effect of CdS NR was specific to C. necator as the metabolism of other microbes often employed for bioproduction including yeast and bacteria was negatively impacted. These results demonstrate that hybrid photosynthesis is more productive when the photocatalyst characteristics are optimized via a separated synthesis process prior to being coupled with microbes. Furthermore, bioproduction improvement by CdS-based photocatalyst requires specific microbial species highlighting the importance of screening efforts for the development of performant hybrid photosynthesis.
Wei Xinlei,Yang Xue,Hu Congcong,Li Qiangzi,Liu Qianqian,Wu Yue,Xie Leipeng,Ning Xiao,Li Fei,Cai Tao,Zhu Zhiguang,Zhang Yi-Heng P. Job,Zhang Yanfei,Chen Xuejun,You Chun
In vitro biotransformation (ivBT) facilitated by in vitro synthetic enzymatic biosystems (ivSEBs) has emerged as a highly promising biosynthetic platform. Several ivSEBs have been constructed to produce poly-3-hydroxybutyrate (PHB) via acetyl-coenzyme A (acetyl-CoA). However, some systems are hindered by their reliance on costly ATP, limiting their practicality. This study presents the design of an ATP-free ivSEB for one-pot PHB biosynthesis via acetyl-CoA utilizing starch-derived maltodextrin as the sole substrate. Stoichiometric analysis indicates this ivSEB can self-maintain NADP + /NADPH balance and achieve a theoretical molar yield of 133.3%. Leveraging simple one-pot reactions, our ivSEBs achieved a near-theoretical molar yield of 125.5%, the highest PHB titer (208.3 mM, approximately 17.9 g/L) and the fastest PHB production rate (9.4 mM/h, approximately 0.8 g/L/h) among all the reported ivSEBs to date, and demonstrated easy scalability. This study unveils the promising potential of ivBT for the industrial-scale production of PHB and other acetyl-CoA-derived chemicals from starch.