Effects of nitrite on anammox activities have been of widespread concern. However, the molecular mechanisms of specific microorganisms in anammox systems responding to nitrite remain unexplored. This study investigated the effects of excess nitrite on core metabolism of AnAOB and symbiotic bacteria, and further elucidated the response mechanism of these effects on microbial growth and nitrogen removal performance. Specifically, nitrogen removal process in a continuous-flow anaerobic ammonia oxidation membrane bioreactor completely collapsed when the nitrite concentration reached 243 mg N/L. Integrated meta-omics analyses demonstrated that excess nitrite disrupted the energy metabolism of Ca. Brocadia sapporoensis (AMXB1), reducing the energy available for establishing tolerance. It disrupted cell replication by impairing biosynthesis process of AMXB1, especially DNA replication and the formation of vital cell structures, e.g., cell membrane and cell wall, as well as the cellular protection system, leading to the collapse of the anammox system. In addition, the cross-feeding of glycogen, lipopolysaccharide and amino acid between AMXB1 and symbiotic bacteria was hindered by excess nitrite, which also contributed to the anomalous cell proliferation and metabolism of AMXB1. These findings contribute to our understanding of the ability of anammox consortia to respond to nitrite stress and process stability in engineered ecosystems. HighlightsO_LINO --N concentration of 243 mg N/L caused the performance collapse of a continuous-flow anammox MBR. C_LIO_LIExcess nitrite likely disrupted the energy metabolism of AMXB1, reducing the energy availability for mitigating nitrite toxicity. C_LIO_LIThe cross-feeding between AMXB1 and symbiotic bacteria was hindered by excess nitrite. C_LIO_LIThe hindrance of cross-feeding was reversed as the concentration of nitrite decreased. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=132 SRC="FIGDIR/small/568540v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@6b199forg.highwire.dtl.DTLVardef@15f2a74org.highwire.dtl.DTLVardef@dc2d8eorg.highwire.dtl.DTLVardef@132deb1_HPS_FORMAT_FIGEXP M_FIG C_FIG
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