Mechanisms of metabolic regulation and enhanced methane production by hydrolytic nanozyme in sludge anaerobic digestion

Artificial nanozymes have demonstrated potential as natural enzyme substitutes due to their efficient biomacromolecule hydrolysis. However, the metabolic and catalytic mechanisms of nanozyme during the biodegradation of complex organics remain inadequately understood. This study evaluated the effect of the hydrolytic mechanism of cerium-based metal-organic frameworks (Ce-MOFs) hydrolytic nanozyme on shaping microbiome and metabolic functions. Biochemical methane potential test revealed that Ce-MOF-based hydrolytic nanozyme increased methane production by 7.8 - 22.2% as the hydrolytic nanozyme dosage increased from 50 to 150 mg/ g VS. The highest methane production observed in 150 mg/g VS dosage of Ce-MOF (Ce-MOF-150) could be attributed to the enhanced sludge hydrolysis, acidogenesis, and methanogenesis processes, proven by the increased relative abundance of dominant acidogens and methanogens. Furthermore, 150 mg/g VS hydrolytic nanozyme bolstered the relative abundance of genes encoding acetyl-CoA carboxylase (accABCD) and involving in glycolysis (hxk and pfkA). These quantities were higher (8.31 - 34.14%) than that without nanozyme addition. The enhanced relative abundance of ackA, pta, cdhCDE, mvhAD, fpoABCDHIJKLMNO, vhoC, rnfABCDEG, mtrABCDEFGH, and hdrABC in the Ce-MOF-150 test indicated that the high proton conductivity of nanozyme facilitated acetoclastic methanogenesis and membrane-bound electron transport system, thereby enhancing methane production. These findings contribute to the expanding understanding of the application of artificial nanozymes as biocatalysts for solid waste resource recovery, providing a promising approach for methane production from sewage sludge.