ATP-free in vitro biotransformation of starch-derived maltodextrin into poly-3-hydroxybutyrate via acetyl-CoA

• Published in: Nature communications Vol. 15; no. 1; p. 3267
• Main Authors: 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
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16-04-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/45/603; 631/92/552; 82/16; 82/29; 82/80; 82/83; Acetyl Coenzyme A - metabolism; ATP; Biosynthesis; Biotransformation; Coenzyme A; Humanities and Social Sciences; Hydroxybutyrates - metabolism; Maltodextrin; multidisciplinary; NADP - metabolism; Poly-3-hydroxybutyrate; Polyesters - metabolism; Polyhydroxybutyrates; Polyhydroxybutyric acid; Polysaccharides; Science; Science (multidisciplinary); Starch; Starch - metabolism; Substrates
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-46871-y

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.
Several in vitro synthetic enzymatic biosystems (ivSEBs) to produce poly-3-hydroxybutyrate (PHB) via acetyl-coenzyme A (acetyl-CoA) have been reported, but suffer from complicated operation procedures, low yields, and/or dependence on costly ATP. Here, the authors report the design of an ATP-free ivSEB for one-pot, high-yield PHB biosynthesis via acetyl-CoA utilizing starch-derived maltodextrin as the sole substrate.