Investigating enhancement of protease and lysozyme combination pretreatment on hydrolysis of sludge organics under humic acid inhibition

This study investigated the impact of humic acid (HA) on enzymatic pretreatment efficiency, focusing on sludge properties and HA molecular structure. The results showed that enzymatic pretreatment alleviates HA inhibition, improving hydrolysis efficiency. In the presence of HA, soluble proteins and polysaccharides in the enzyme- cocktail group reached 27.7 mg/L and 23.9 mg/L, 1.4 and 1.3 times higher than the blank group, respectively. The enzyme-cocktail group also had the highest soluble DNA concentration (19.4 mg/L) and the lowest viable cell proportion (69.3 %), indicating effective cell lysis. Enzyme-cocktail pretreatment reduced electrostatic repulsion, enhancing the mobility of extracellular organics. Enzyme interactions with HA released internal hydrolases and decreased amide groups on the HA surface, increasing the availability of biodegradable substrates. Overall, enzymatic pretreatment proves effective in mitigating HA-induced inhibition, thereby improving sludge biodegradation and enhancing carbon recovery in anaerobic fermentation.