Biomaterial Database

Directed Evolution of Escherichia coli Nissle 1917 to Utilize Allulose as Sole Carbon Source. 2024

Bo Xu, and Li-Hua Liu, and Shijing Lai, and Jingjing Chen, and Song Wu, and Wei Lei, and Houliang Lin, and Yu Zhang, and Yucheng Hu, and Jingtao He, and Xipeng Chen, and Qian He, and Min Yang, and Haimei Wang, and Xuemei Zhao, and Man Wang, and Haodong Luo, and Qijun Ge, and Huamei Gao, and Jiaqi Xia, and Zhen Cao, and Baoxun Zhang, and Ao Jiang, and Yi-Rui Wu

School of Basic Medical Sciences, Hubei University of Science and Technology, Xianning, 437100, P. R. China.

Sugar substitutes are popular due to their akin taste and low calories. However, excessive use of aspartame and erythritol can have varying effects. While D-allulose is presently deemed a secure alternative to sugar, its excessive consumption is not devoid of cellular stress implications. In this study, the evolution of Escherichia coli Nissle 1917 (EcN) is directed to utilize allulose as sole carbon source through a combination of adaptive laboratory evolution (ALE) and fluorescence-activated droplet sorting (FADS) techniques. Employing whole genome sequencing (WGS) and clustered regularly interspaced short palindromic repeats interference (CRISPRi) in conjunction with compensatory expression displayed those genetic mutations in sugar and amino acid metabolic pathways, including glnP, glpF, gmpA, nagE, pgmB, ybaN, etc., increased allulose assimilation. Enzyme-substrate dynamics simulations and deep learning predict enhanced substrate specificity and catalytic efficiency in nagE A247E and pgmB G12R mutants. The findings evince that these mutations hold considerable promise in enhancing allulose uptake and facilitating its conversion into glycolysis, thus signifying the emergence of a novel metabolic pathway for allulose utilization. These revelations bear immense potential for the sustainable utilization of D-allulose in promoting health and well-being.