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

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.

[Experimental study on the environmental fate of nitrogen in snail-macrophyte ecosystem for water purification].

A snail-macrophyte simulation system was built and isotope tracer technique was adopted to study the environmental fate of nitrogen in snail-macrophyte purification system, the results showed that: Vallisneria spiralis increased its wet weight by 580% and its number by 6.6 ramets, moreover, Vallisneria spiralis absorbed 1.07% 15N by the roots and 7.74% by stems and leaves, while Bellamya only absorbed 0.06%. And 5.73% 15N was retained in the sediment. Through analyzing of the results, it indicated that: in such simulation system, sediment was the main nutrition source for the growth of Vallisneria spiralis, which absorbed only few dissolved nitrogen from water; ammonium nitrogen in water was transformed mainly in the sediment-water interface, most of which was absorbed by Vallisneria spiralis, a small amount was removed through nitrification and denitrification, and the rest was kept by sediment; Vallisneria spiralis was final vector for removing nitrogen in the system, and Bellamya could also boost the growth of Vallisneria spiralis and strengthen the processes of nitrification and denitrification, thus promoting the nitrogen removal from the system indirectly. So, during the period of culture, rational allocation of snail-macrophyte structure in different stages plays an important role in controlling water quality in ponds.