Construction of Recombinant Escherichia coli Expressing Ammonia Assimilation Genes and Evaluation of Its Effect on Removing Ammonium Nitrogen (NH4+-N)

作者：Pan Pan, Y Yang, Runxuan Shi, Yulin Kang, Hanli Xu, Xiyu Cheng, Qiong Yan, Honggang Hu · 发表于：Microorganisms · 年份：2025 · DOI：10.3390/microorganisms13122646 · 被引用次数：4 · 研究领域：Wastewater Treatment and Nitrogen Removal、Plant nutrient uptake and metabolism、Microbial Metabolic Engineering and Bioproduction

Treating wastewater with high ammonium nitrogen (NH4+-N) is a public and environmental priority. Unlike nitrification–denitrification, ammonia assimilation channels NH4+-N into the glutamate biosynthetic pathway, avoiding gaseous nitrogen species (NOx, N2). Here we engineered Escherichia coli to enhance ammonia assimilation by co-expressing three key genes, gdhA, glnA, and guaA. The genes were synthesized, cloned into expression plasmids, and introduced into E. coli BL21 for IPTG-inducible expression. Expression of the target proteins at expected sizes was observed, and NH4+-N removal was assessed in flask fermentations. Recombinant strains exhibited significantly higher NH4+-N reduction than the empty vector control; among them, BL21(pET-gdhA-glnA-guaA) performed best, achieving a maximum removal efficiency of 90.09% under the tested conditions. These results indicate that reinforcing the glutamate pathway through multi-gene co-expression can effectively lower NH4+-N in culture and provide a basis for developing recombinant bacteria for practical sewage treatment.