Structural extracellular polymeric substances (SEPS) as valuable biopolymers, can be extracted from waste activated sludge (WAS). However, the extraction yield is typically low, and detailed information on SEPS characterizations, as well as proper treatment of the sludge after SEPS extraction, remains limited. This study aimed to optimize the conditions of heating-Na₂CO₃ extraction process to increase the yield of SEPS extracted from WAS. Subsequently, SEPS were characterized, and, for the first time, insights into their protein composition were uncovered by using proteomics. A maximum SEPS yield of 209 mg g^-1 volatile solid (VS) was obtained under optimal conditions: temperature of 90 °C, heating time of 60 min, Na⁺ dosage of 8.0 mmol/g VS, and pH required to precipitation of 4.0, which was comparable to that from the aerobic granular sludge reported in literature. Proteomics analysis unveiled that the proteins in SEPS primarily originated from microorganisms involved in nitrogen fixation and organic matter degradation, including their intracellular and membrane-associated regions. These proteins exhibited various catalytic activities and played crucial roles in aggregation processes. Besides, the process of SEPS extraction significantly enhanced volatile fatty acid (VFA) production during the anaerobic fermentation of residual WAS after SEPS extraction. A maximum VFA yield of 420 ± 14 mg COD/g VS_added was observed in anaerobic fermentation of 10 d, which was 77.2 ± 0.1 % higher than that from raw sludge. Mechanism analysis revealed that SEPS extraction not only improved WAS disintegration and solubilization but also reduced the relative activity of methanogens during anaerobic fermentation. Moreover, SEPS extraction shifted the microbial population during anaerobic fermentation in the direction towards hydrolysis and acidification such as Fermentimonas sp. and Soehngenia sp. This study proposed a novel strategy based on SEPS extraction and VFA production for sludge treatment, offering potential benefits for resource recovery and improved process efficiency.

In recent years, volatile fatty acid (VFA) production through anaerobic fermentation of sewage sludge, instead of methane production, has been regarded as a high-value and promising roadmap for sludge stabilization and resource recovery. This review first presents the effects of some essential factors that influence VFA production and composition. In the second part, we present an extensive analysis of conventional pretreatment and co-fermentation strategies ultimately addressed to improving VFA production and composition. Also, the effectiveness of these approaches is summarized in terms of sludge degradation, hydrolysis rate, and VFA production and composition. According to published studies, it is concluded that some pretreatments such as alkaline and thermal pretreatment are the most effective ways to enhance VFA production from sewage sludge. The possible reasons for the improvement of VFA production by different methods are also discussed. Finally, this review also highlights several current technical challenges and opportunities in VFA production with spectrum control, and further related research is proposed.

White rot fungi using P. sajor-caju and T. versicolor was examined to pretreat raw champost (lignin-rich) and oyster champost (cellulose-rich) for enhancement of fermentative volatile fatty acid (VFA) production. Results showed that the efficiency of pretreatment and VFA production was influenced by the fungal strains and substrates. P. sajor-caju pretreatment showed preferential lignin degradation on raw champost and obtained the maximum VFA yield (203 ± 9 mg COD/g VS_added), which increased by 60% and 74% compared to that of control and unpretreated champost, respectively. For cellulose-rich oyster champost, however, fungal pretreatment decreased VFA yield compared to unpretreated champost. Further mechanisms analysis demonstrated the two strains grow and secreted ligninolytic enzymes, which substantially influenced the characteristics of two champosts such as cellulose/lignin ratio and morphology in different extents. P. sajor-caju was highly efficient to lignin-rich champost on selectively degrading lignin and further enhancing digestibility such as VFA production.

Anaerobic digestion has been widely applied throughout the world for lignocellulosic biomass treatment and energy recovery. However, the solid digestate from anaerobic digestion still contains a rather large fraction of poorly anaerobic degradable lignocellulosic fibers due to inhibition of lignin, which deeply limits the bioenergy production from lignocelullosic biomass. Therefore, a novel fungal pretreatment method using P. sajor-caju and T. versicolor was investigated to advance the solid-state fermentation of solid digestate and improve the production of fermentative volatile fatty acids (VFAs). The results showed that a maximum VFA yield of 240 mg COD/g VS was obtained from solid digestate pretreated by P. sajor-caju in 6 weeks, which was 1.17-fold and 1.24-fold higher than that of the autoclaved group and raw substrate, respectively. The mechanisms indicated that these fungal strains could grow on the solid digestate and secrete ligninolytic enzymes such as laccase and manganese peroxidase to degrade lignin in different extents. Besides, fungal pretreatment substantially changed the solid digestate characteristics such as cellulose/lignin ratio and the presence of specific functional groups. Moreover, fungal pretreatment using P. sajor-caju effectively damaged the structure and increased surface area and pore size of the solid digestate, which is beneficial to further VFA production.

In order to improve the efficiency of anaerobic sludge digestion, alkaline and high pressure homogenization (HPH) were combined to pre-treat the excess activated sludge. The effect of HPH operating parameters, including homogenization pressure and cycle number, on the performances of anaerobic sludge digestion was studied. The results demonstrated that the performances of sludge disintegration and anaerobic digestion were markedly enhanced by increasing the homogenization pressure. After pretreatment at a homogenization pressure of 60 MPa with one homogenization cycle combined with an alkaline dosage of 0.04 mol/L, the sludge TCOD, VS removal and cumulative biogas production in a mesophilic anaerobic digestion system increased by 24.68%, 18.95% and 95.81%, respectively, in comparison with that with the alkaline pretreatment alone. But the sludge disintegration and biogas production only slightly increased with the increase of homogenization cycle. Considering biogas production and energy-saving, the suitable homogenization operation was selected as homogenization pressure of 60 MPa with once cycle. Relationships between methane yield and sludge disintegration showed that the improved methane production was mainly attributed to the sludge disintegration resulted from combined sludge pretreatment.