Response of nitrite accumulation to elevated C/NO– 3-N ratio during partial denitrification process: Insights of extracellular polymeric substance, microbial community and metabolic function

This study investigated the response of nitrite accumulation to elevated COD/NO3–-N ratio (C/N) during partial denitrification (PD). Results indicated nitrite was gradually accumulated and remained stable (C/N = 1.5 ∼ 3.0), while that rapidly declined after reaching the peak (C/N = 4.0 ∼ 5.0). The polysaccharide (PS) and protein (PN) content of tightly-bound extracellular polymeric substances (TB-EPS) reached the maximum at C/N of 2.5 ∼ 3.0, which might be stimulated by high level of nitrite. Illumina MiSeq sequencing showed Thauera and OLB8 were dominated denitrifying genera at C/N of 1.5 ∼ 3.0, while Thauera was further enriched with fading OLB8 at C/N of 4.0 ∼ 5.0. Meanwhile, the highly-enriched Thauera might enhance the activity of nitrite reductase (nirK) promoting further nitrite reduction. Redundancy analysis (RDA) showed positive correlations between nitrite production and PN content of TB-EPS, denitrifying bacteria (Thauera and OLB8) and nitrate reductases (narG/H/I) in low C/N. Finally, their synergistic effects for driving nitrite accumulation were comprehensively elucidated.