Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, P. R. China.
Key Laboratory of Plant Resources, Institute of Botany, Chinese Academy of Sciences, 20 Nanxincun, Xiangshan, Beijing, 100093, P. R. China.
Angew Chem Int Ed Engl. 2022 Jan 3;61(1):e202111054. doi: 10.1002/anie.202111054. Epub 2021 Nov 22.
Many existing in vitro biosystems harness power from the chemical energy contained in substrates and co-substrates, and light or electric energy provided from abiotic parts, leading to a compromise in atom economy, incompatibility between biological and abiotic parts, and most importantly, incapability to spatiotemporally co-regenerate ATP and NADPH. In this study, we developed a light-powered in vitro biosystem for poly(3-hydroxybutyrate) (PHB) synthesis using natural thylakoid membranes (TMs) to regenerate ATP and NADPH for a five-enzyme cascade. Through effective coupling of cofactor regeneration and mass conversion, 20 mM PHB was yielded from 50 mM sodium acetate with a molar conversion efficiency of carbon of 80.0 % and a light-energy conversion efficiency of 3.04 %, which are much higher than the efficiencies of similar in vitro PHB synthesis biosystems. This suggests the promise of installing TMs as a green engine to drive more enzyme cascades.
许多现有的体外生物系统利用基质和共基质中所含的化学能量以及来自非生物部分的光或电能来获取能量，这导致原子经济性受到妥协、生物和非生物部分之间不兼容，最重要的是，无法时空协同再生 ATP 和 NADPH。在这项研究中，我们开发了一种使用天然类囊体膜（TMs）为五酶级联反应再生 ATP 和 NADPH 的光驱动体外生物系统，用于合成聚（3-羟基丁酸酯）（PHB）。通过有效偶联辅助因子再生和质量转化，从 50 mM 醋酸钠中得到 20 mM PHB，碳摩尔转化率为 80.0%，光能量转化率为 3.04%，均高于类似的体外 PHB 合成生物系统的效率。这表明将 TMs 作为绿色引擎来驱动更多的酶级联反应具有广阔的应用前景。
