Efficient Ammonium Nitrogen Metabolization and γ-PGA Production by Bacillus velezensis GY1 Isolated from Swine Manure Digestate

Efficient microbial assimilation of high-concentration ammonium nitrogen and its conversion into value-added bioproducts represent a pivotal yet underexplored strategy for sustainable nitrogen management. Here, we report a newly isolated Bacillus velezensis strain, GY1, with a robust intrinsic capacity for simultaneous NH4+-N assimilation and γ-polyglutamic acid (γ-PGA) biosynthesis. Under optimized conditions (37 °C, pH 7.0, C/N = 12:1), GY1 achieved 76.5% removal of ammonium nitrogen (400 mg/L) with negligible nitrite accumulation (<0.02 mg/L), indicating assimilation rather than nitrification. Transcriptomic analysis revealed a coordinated metabolic flux wherein the glutamine synthetase - glutamate synthase pathway GS-GOGAT pathway supplies glutamate for γ-PGA synthesis, while polymerization further facilitates ammonium sequestration via electrostatic interactions. GY1 produced up to 612.8 mg/L γ-PGA, and genetic overexpression of capB synchronized these pathways, enhancing both ammonium assimilation (87.4%) and γ-PGA yield (843.9 mg/L). Notably, this metabolic coupling remained resilient in complex substrates, achieving 68.8% ammonium removal and 220.7 mg/L γ-PGA production in untreated biogas slurry. Together, these findings establish GY1 as a metabolically robust platform linking nitrogen assimilation with biopolymer synthesis, offering a mechanistic framework for circular nitrogen economies.