摘要: n-Caproate,which is produced via chain elongation(CE)using waste biomass,can supply various fossilderived products,thus advancing the realization of carbon neutrality.Ammonia released from the degradation of nitrogen-rich waste biomass can act as a nutrient or an inhibitor in anaerobic bioprocesses,including CE,with the distinction being primarily dependent on its concentration.Currently,the optimal concentration of ammonia and the threshold of toxicity for open-culture n-caproate production using ethanol as an electron donor,along with the underlying mechanisms,remain unclear.This study revealed that the optimal concentration of ammonia for n-caproate production was 2.0 g∙L^(-1),whereas concentrations exceeding this threshold markedly suppressed the CE performance.Exploration of the mechanism revealed the involvement of two forms of ammonia(i.e.,ammonium ions and free ammonia)in this inhibitory behavior.High ammonia levels(5.0 g∙L^(-1))induced excessive ethanol oxidation and suppressed the reverse β-oxidation(RBO)process,directly leading to the enhanced activities of enzymes(phosphotransacetylase and acetate kinase)responsible for acetate formation and diminished activities of butyryl-coenzyme A(CoA):acetyl-CoA transferase,caproyl-CoA:butyryl-CoA transferase,and caproyl-CoA:acetyl-CoA transferase that are involved in the syntheses of n-butyrate and n-caproate.Furthermore,the composition of the microbial community shifted from Paraclostridium dominance(at 0.1 g∙L^(-1)ammonia)to a co-dominance of Fermentimonas,Clostridium sensu stricto 12,and Clostridium sensu stricto 15 at 2.0 g∙L^(-1)ammonia.However,these CE-functional bacteria were mostly absent in the presence of excessive ammonia(5.0 g∙L^(-1)ammonia).Metagenomic analysis revealed the upregulation of functions such as RBO,fatty acid synthesis,K^(+)efflux,adenosine triphosphatase(ATPase)metabolism,and metal cation export in the presence of 2.0 g∙L^(-1)ammonia,collectively contributing to enhanced n-caproate production.Conversely,the aforementioned functions(excluding metal cation export)and K^(+)influx were suppressed by excessive ammonia,undermining both ammonia detoxification and n-caproate biosynthesis.The comprehensive elucidation of ammonia-driven mechanisms influencing n-caproate production,as provided in this study,is expected to inspire researchers to devise effective strategies to alleviate ammonia-induced inhibition. 摘要译文