Revealing the mechanism of novel nitrogen-doped biochar supported magnetite (NBM) enhancing anaerobic digestion of waste-activated sludge by sludge characteristics.

Anaerobic digestion (AD) is a promising technology in waste treatment and energy recovery. However, it suffers from long retention time and low biogas yield. In this study, novel nitrogen-doped biochar supported magnetite (NBM) was synthesized and applied to enhance the AD of waste-activated sludge. Results showed that NBM increased cumulative methane production and SCOD removal efficiency by up to 1.75 times and 15% respectively at 5 g/L compared with the blank. NBM enhanced both hydrolysis and methanogenesis process during AD and the activities of α-glucosidase, protease, coenzyme F420 and electron transport system were increased by 19%, 163%, 104% and 160% respectively at 5 g/L NBM compared with the blank. NBM also facilitated the secretion of conductive protein in extracellular polymeric substances as well as the formation of conductive pili, leading to 3.18-7.59 times higher sludge electrical conductivity. Microbial community analysis revealed that bacteria Clostridia and archaea Methanosarcina and Methanosaeta were enriched by the addition of NBM, and direct interspecies electron transfer might be promoted between them. This study provides a practical reference for future material synthesis and its application.

Impact of magnesium ions on lysozyme-triggered disintegration and solubilization of waste activated sludge.

Lysozyme can efficiently accelerate solubilization and hydrolysis of waste activated sludge (WAS) for anerobic digestion. However, the effect of lysozyme was easily to be inhibited by metal ions in WAS. The impact of magnesium ions (Mg2+) on lysozyme catalyze WAS disintegration was investigated in this study. The effect of lysozyme on WAS hydrolysis could be hindered by Mg2+. Relatively high concentrations (>50 mg/L) of Mg2+ in sludge significantly reduced the release of soluble polysaccharides and proteins from WAS, while sulfate ions or chloride ions caused no such effect. Proteins were difficult to be extracted from extracellular polymeric substances (EPS) of WAS in the presence of Mg2+ (>10 mg/L) due to the divalent cation bridging (DCB) behavior, while the extraction of polysaccharides was not significantly affected. The polysaccharides and proteins in the inner EPS layer were transferred to the outer layer during the lysozyme treatment, and total quantities of both components maintained constantly. At least 23.1% lysozymes were trapped in the liquid phase of 100 mg Mg2+/L in the first hour. Mg2+ could significantly affect the transfer of lysozyme from liquid phase to the inner layer of sludge. Mg2+ neutralized the negative surface charge of the sludge particles, which hindered the absorption of positively charged lysozyme molecules by sludge flocs from the liquid phase. The proteins of TB-EPS had higher ratios of α-helixes and tighter structures than those in LB-EPS, which could impede the lysozyme transfer to the cell wall.

New insight into enhanced short-chain fatty acids production from waste activated sludge through pretreatment of cation exchange resin coupled NaCl addition.

In this study, a novel pretreatment of cation exchange resin (CER) coupled NaCl addition was proposed to enhance waste activated sludge (WAS) hydrolysis and promote short-chain fatty acids (SCFAs) production in the anaerobic fermentation process. At the optimal pretreatment condition of 3 g/g SS CER and 15 g/L NaCl, considerable SCOD (i.e. 5107 mg/L, 35.4% of TCOD) was released after 2-day coupled treatment, which provided sufficient organic substance for the subsequent SCFAs production. The sludge hydrolysis mechanism was illustrated, i.e. CER triggered extracellular polymeric substances (EPS) disruption and NaCl induced microbial cells lysis. The synergistic interaction between CER and NaCl pretreatment was investigated and application potential of fermentative liquid was evaluated after the coupled pretreatment-enhanced anaerobic fermentation. In the presence of abundant biodegradable substrates in the fermentative liquid, 4742 mg COD/L (i.e. 388 mg COD/g VSS) of SCFAs production was achieved within 6-day anaerobic fermentation, mainly composed of acetic and propionic acids (70.4% of total SCFAs).

An Inhibitor of GSK3B and HDACs Kills Pancreatic Cancer Cells and Slows Pancreatic Tumor Growth and Metastasis in Mice.

BACKGROUND & AIMS: Growth, progression, and drug resistance of pancreatic ductal adenocarcinomas (PDACs) have been associated with increased levels and activity of glycogen synthase kinase 3 beta (GSK3B) and histone deacetylases (HDACs). We designed and synthesized molecules that simultaneously inhibit the activities of both enzymes. We tested the effects of one of these molecules, Metavert, in pancreatic cancer cells and mice with pancreatic tumors. METHODS: We tested the ability of Metavert to bind GSK3B and HDACs using surface plasmon resonance. MIA PaCa-2, Bx-PC3, HPAF-II, and HPDE6 cell lines were incubated with different concentrations of Metavert, with or without paclitaxel or gemcitabine, or with other inhibitors of GSK3B and HDACs; cells were analyzed for apoptosis and migration and by immunoblotting, immunofluorescence, and real-time polymerase chain reaction. Krasþ/LSLG12D;Trp53þ/LSLR172H;Pdx-1-Cre (KPC) mice (2 months old) were given injections of Metavert (5 mg/kg, 3 times/week) or vehicle (control). B6.129J mice with tumors grown from UN-KPC961-Luc cells were given injections of Metavert or vehicle. Tumors and metastases were counted and pancreata were analyzed by immunohistochemistry. Glucose metabolism was measured using 13C-glucose tracer and mass spectroscopy and flow cytometry. Cytokine levels in blood samples were measured using multiplexing enzyme-linked immunosorbent assay. RESULTS: Metavert significantly reduced survival of PDAC cells but not nontransformed cells; the agent reduced markers of the epithelial-to-mesenchymal transition and stem cells in PDAC cell lines. Cells incubated with Metavert in combination with irradiation and paclitaxel or gemcitabine had reduced survival compared with cells incubated with either agent alone; Metavert increased killing of drug-resistant PDAC cells by paclitaxel and gemcitabine. PDAC cells incubated with Metavert acquired normalized glucose metabolism. Administration of Metavert (alone or in combination with gemcitibine) to KPC mice or mice with syngeneic tumors significantly increased their survival times, slowed tumor growth, prevented tumor metastasis, decreased tumor infiltration by tumor-associated macrophages, and decreased blood levels of cytokines. CONCLUSIONS: In studies of PDAC cells and 2 mouse models of PDAC, we found a dual inhibitor of GSK3B and HDACs (Metavert) to induce cancer cell apoptosis, reduce migration and expression of stem cell markers, and slow growth of tumors and metastases. Metavert had synergistic effects with gemcitabine.

Carotid Intraplaque Hemorrhage Imaging with Quantitative Vessel Wall T1 Mapping: Technical Development and Initial Experience.

Purpose To develop a three-dimensional (3D) high-spatial-resolution time-efficient sequence for use in quantitative vessel wall T1 mapping. Materials and Methods A previously described sequence, simultaneous noncontrast angiography and intraplaque hemorrhage (SNAP) imaging, was extended by introducing 3D golden angle radial k-space sampling (GOAL-SNAP). Sliding window reconstruction was adopted to reconstruct images at different inversion delay times (different T1 contrasts) for voxelwise T1 fitting. Phantom studies were performed to test the accuracy of T1 mapping with GOAL-SNAP against a two-dimensional inversion recovery (IR) spin-echo (SE) sequence. In vivo studies were performed in six healthy volunteers (mean age, 27.8 years ± 3.0 [standard deviation]; age range, 24-32 years; five male) and five patients with atherosclerosis (mean age, 66.4 years ± 5.5; range, 60-73 years; five male) to compare T1 measurements between vessel wall sections (five per artery) with and without intraplaque hemorrhage (IPH). Statistical analyses included Pearson correlation coefficient, Bland-Altman analysis, and Wilcoxon rank-sum test with data permutation by subject. Results Phantom T1 measurements with GOAL-SNAP and IR SE sequences showed excellent correlation (R2 = 0.99), with a mean bias of -25.8 msec ± 43.6 and a mean percentage error of 4.3% ± 2.5. Minimum T1 was significantly different between sections with IPH and those without it (mean, 371 msec ± 93 vs 944 msec ± 120; P = .01). Estimated T1 of normal vessel wall and muscle were 1195 msec ± 136 and 1117 msec ± 153, respectively. Conclusion High-spatial-resolution (0.8 mm isotropic) time-efficient (5 minutes) vessel wall T1 mapping is achieved by using the GOAL-SNAP sequence. This sequence may yield more quantitative reproducible biomarkers with which to characterize IPH and monitor its progression. © RSNA, 2017.

Quantification of liver perfusion using multidelay pseudocontinuous arterial spin labeling.

PURPOSE: To develop a free-breathing multidelay pseudocontinuous arterial spin labeling (pCASL) technique for quantitative measurement of liver perfusion of the hepatic artery and portal vein, respectively. MATERIALS AND METHODS: A navigator-gated pCASL sequence with balanced steady-state free precession (bSSFP) readout was developed and applied on five healthy young volunteers at 3T. Two labeling schemes were performed with the labeling plane applied on the descending aorta above the liver, and perpendicular to the portal vein before its entry to liver to label the hepatic artery and portal vein, respectively. For each labeling scheme, pCASL scans were performed at five or six postlabeling delays between 200 and 2000 msec or 2500 msec with an interval of 400 or 500 msec. Multidelay pCASL images were processed offline with nonrigid motion correction, outlier removal, and fitted for estimation of liver perfusion and transit time. RESULTS: Estimated liver perfusion of the hepatic artery and hepatic portal vein were 21.8 ± 1.9 and 95.1 ± 8.9 mL/100g/min, with the corresponding transit time of 1227.3 ± 355.5 and 667.2 ± 85.0 msec, respectively. The estimated liver perfusion and transit time without motion correction were less reliable with greater residual variance compared to those processed with motion correction (P < 0.05). CONCLUSION: The liver perfusion measurement using multidelay pCASL showed good correspondence with values noted in the literature. The capability to noninvasively and selectively label the hepatic artery and portal vein is a unique strength of pCASL as compared to other liver perfusion imaging techniques, such as computed tomography perfusion and dynamic contrast-enhanced MRI.

Mutant poisoning demonstrates a nonsequential mechanism for digestion of double-stranded DNA by λ exonuclease trimers.

λ Exonuclease (λexo) is a highly processive 5'-3' exonuclease that binds double-stranded DNA (dsDNA) ends and digests the 5'-strand into mononucleotides. The enzyme forms a toroidal homotrimer with a central tapered channel for tracking along the DNA. During catalysis, dsDNA enters the open end of the channel, and the 5'-strand is digested at one of the three active sites. It is currently not known if λexo uses a sequential mechanism, in which the DNA moves from one active site to the next around the trimer for each round of catalysis or a nonsequential mechanism, in which the DNA locks onto a single active site for multiple rounds. To understand how λexo uses its three active sites, we used a mutant poisoning approach, in which a 6xHis-tagged K131A inactive mutant of λexo was mixed with untagged wild type (WT) to form hybrid trimers. Nickel-spin pull-down analysis confirmed complete subunit exchange after 1 h at 37 °C. Exonuclease assays revealed an approximately linear decrease in activity with increasing fraction of mutant, as expected for a nonsequential mechanism. By fitting the observed rates of digestion to a simple mathematical model, the individual rates of the two hybrid species of trimer were determined. This analysis showed that trimers containing only one or two WT subunits contribute significantly to the observed activity, in further agreement with a nonsequential mechanism. Finally, purification of hybrid trimer mixtures by Ni-spin chromatography, to remove the contribution from fully WT trimers, also resulted in significant levels of activity, again consistent with a nonsequential mechanism.

A Structure-Activity Analysis for Probing the Mechanism of Processive Double-Stranded DNA Digestion by λ Exonuclease Trimers.

λ exonuclease (λexo) is an ATP-independent 5'-to-3' exonuclease that binds to double-stranded DNA (dsDNA) ends and processively digests the 5'-strand into mononucleotides. The crystal structure of λexo revealed that the enzyme forms a ring-shaped homotrimer with a central funnel-shaped channel for tracking along the DNA. On the basis of this structure, it was proposed that dsDNA enters the open end of the channel, the 5'-strand is digested at one of the three active sites, and the 3'-strand passes through the narrow end of the channel to emerge out the back. This model was largely confirmed by the structure of the λexo-DNA complex, which further revealed that the enzyme unwinds the DNA by 2 bp prior to cleavage, to thread the 5'-end of the DNA into the active site. On the basis of this structure, an "electrostatic ratchet" model was proposed, in which the enzyme uses a hydrophobic wedge to insert into the base pairs to unwind the DNA, a two-metal mechanism for nucleotide hydrolysis, a positively charged pocket to bind to the terminal 5'-phosphate generated after each round of cleavage, and an arginine residue (Arg-45) to bind to the minor groove of the downstream end of the DNA. To test this model, in this study we have determined the effects of 11 structure-based mutations in λexo on DNA binding and exonuclease activities in vitro, and on DNA recombination in vivo. The results are largely consistent with the model for the mechanism that was proposed on the basis of the structure and provide new insights into the roles of particular residues of the protein in promoting the reaction. In particular, a key role for Arg-45 in DNA binding is revealed.

Crystal structure of λ exonuclease in complex with DNA and Ca(2+).

Bacteriophage λ exonuclease (λexo) is a ring-shaped homotrimer that resects double-stranded DNA ends in the 5'-3' direction to generate a long 3'-overhang that is a substrate for recombination. λexo is a member of the type II restriction endonuclease-like superfamily of proteins that use a Mg(2+)-dependent mechanism for nucleotide cleavage. A previous structure of λexo in complex with DNA and Mg(2+) was determined using a nuclease defective K131A variant to trap a stable complex. This structure revealed the detailed coordination of the two active site Mg(2+) ions but did not show the interactions involving the side chain of the conserved active site Lys-131 residue. Here, we have determined the crystal structure of wild-type (WT) λexo in complex with the same DNA substrate, but in the presence of Ca(2+) instead of Mg(2+). Surprisingly, there is only one Ca(2+) bound in the active site, near the position of Mg(A) in the structure with Mg(2+). The scissile phosphate is displaced by 2.2 Å relative to its position in the structure with Mg(2+), and the network of interactions involving the attacking water molecule is broken. Thus, the structure does not represent a catalytic configuration. However, the crystal structure does show clear electron density for the side chain of Lys-131, which is held in place by interactions with Gln-157 and Glu-129. By combining the K131A-Mg(2+) and WT-Ca(2+) structures, we constructed a composite model to show the likely interactions of Lys-131 during catalysis. The implications with regard to the catalytic mechanism are discussed.

Self-cultivating anammox granules for enhancing wastewater nitrogen removal in nitrification-denitrification flocculent sludge system.

The feasibility of self-cultivating anammox granules for enhancing wastewater nitrogen removal was investigated in a nitrification-denitrification flocculent sludge system. Desirable nitrogen removal efficiency of 84 ± 4 % was obtained for the influent carbon to nitrogen ratio of 1-1.3 (NH4+-N: 150-200 mg N/L) via alternate anaerobic/oxic/anoxic mode. Meanwhile, some red granular sludge was formed in the system. The abundance and activity of anaerobic ammonia oxidation bacteria (AnAOB) increased from 'not detected' in seed sludge to 0.57 % and 29.4 ± 0.7 mg N/(g mixed liquor volatile suspended solids·h) in granules, respectively, suggesting successful cultivation of anammox granules. Furthermore, some denitrifying bacteria with capability of partial denitrification were enriched, such as Candidatus Competibacter (2.45 %) and Thauera (5.75 %), which could cooperate with AnAOB, facilitating AnAOB enrichment. Anammox was dominant in nitrogen removal with the contribution to nitrogen removed above 68.8 ± 0.3 %. The strategy of self-cultivating anammox granules could promote the application of anammox.

Novel nitrogen-doped biochar supported magnetite promotes anaerobic digestion: Material characterization and metagenomic analysis.

Although different conductive materials have been applied to anaerobic digestion, there has not been a material that can really combine their merits and make up their shortcoming from each other. In this study, a novel nitrogen-doped biochar supported magnetite (Fe3O4@N-BC) was synthesized. Various material characterizations confirmed that nitrogen atoms were successful doped into the biochar and magnetite precipitated on its surface. 5 g/L Fe3O4@N-BC achieved the highest promotion of cumulative CH4 production by 1.75 times compared with the blank group. Further metagenomic analysis revealed that Fe3O4@N-BC could increase the gene abundances of pilA, MmcA, Fpo, Rnf and HdrEd in bacteria Clotridium, Pseudomonas and Syntrophomonas and archaea Methanosarcina. Redundancy analysis showed that it was electrical conductivity and electron exchange capacity that were the key physicochemical characteristics for Fe3O4@N-BC to facilitating direct interspecies electron transfer. This study provides a reference for future conductive material synthesis and its application for anaerobic digestion.

Enhanced acidogenic fermentation from Al-rich waste activated sludge by combining lysozyme and sodium citrate pretreatment: Perspectives of Al stabilization and enzyme activity.

The accumulation of poly aluminum chloride (PAC) in dewatered waste activated sludge (WAS) can cause severe Al pollution and significantly reduce the production of volatile fatty acids (VFAs) from anaerobic fermentation. Herein, the combination of lysozyme and sodium citrate pretreatment was applied to stabilize the aluminum and enhance the VFAs production via anaerobic fermentation. The complexation and stabilization of aluminum by the citrate was efficient, which is conducive to relieving the inhibition of aluminum on lysozymes and other extracellular hydrolases. Compared with the control group, the lysozyme, protease and α-glucosidase activities were obtained at 1.86, 1.72, and 1.15 times, respectively, following the pretreatment. 129.71 mg/g volatile suspended solids (VSS) of soluble proteins and 26.3 mg/g VSS of polysaccharides were obtained within 4 h, together with the degradation of 124 % more proteins and 75 % more polysaccharides within three days. This provided a sufficient number of substrates for VFA production. 588.4 mg COD/g VSS of total VFAs were obtained after the six-day anaerobic fermentation from Al-rich WAS following the combination of lysozyme and sodium citrate pretreatment, which was 7.3 times higher than that of the control group. This study presents a novel approach for enhancing VFA production in anaerobic fermentation as well as reducing risk of Al hazards from Al-rich WAS.

Investigating the synergic role of asynchronous dosed protease and lysozyme for facilitating excess sludge solubilization and acidogenic fermentation.

Improving the anaerobic fermentation (AF) efficiency of excess sludge (ES) is essential for attaining biosolid minimization, stabilization, resource recovery, and carbon-emission reduction. Along these lines, here, the synergistic mechanism of protease and lysozyme for enhancing hydrolysis and AF efficiency with better recovery of volatile fatty acids (VFAs) was thoroughly investigated. Single lysozyme was capable of reducing the zeta potential and fractal dimension when dosed into the ES-AF system, which was beneficial for increasing the contact probability between proteases and extracellular proteins. Moreover, the weight-averaged molecular weight of the loosely-bound extracellular polymeric substance (LB-EPS) reduced from 1867 to 1490 in the protease-AF group, which facilitated the penetration of EPS by the lysozyme. The soluble DNA and extracellular DNA (eDNA) of the enzyme cocktail pretreated group increased by 23.24 % and 77.09 %, and the cell viability decreased after 6-hour hydrolysis, demonstrating a better hydrolysis efficiency. Remarkably, the asynchronous dosed enzyme cocktail pretreatment was proven a better strategy to enhance both the solubilization and hydrolysis processes since the synergistic effect of these two enzymes can exclude the mutual interference. As a result, the VFAs were increased by 1.26 times higher than the blank group. The underlying mechanism of an environmental-friendly and effective strategy was examined to promote ES hydrolysis and acidogenic fermentation, which was beneficial for the recovery of VFAs and carbon-emission reduction.

Reconsidering operation pattern for cation-exchange resin assistant anaerobic fermentation of waste activated sludge: Shorting residence period towards dosage-reduction and anti-fouling.

Short-chain fatty acids (SCFAs) generation through anaerobic fermentation has been regarded as a promising pathway to achieve carbon recovery and economic benefits in waste activated sludge management. Despite the cation exchange resin (CER) assistant anaerobic fermentation strategy has been previously reported for enhancing anaerobic fermentation, the overlarge CER usage and serious CER pollution have limited its engineering application. This study provided a reconsideration for the operation pattern modification. Through 4-day anaerobic fermentation with CER residence period shrinking to 1 day, 40.9% sludge VSS solubilization and reduction were achieved, triggering a considerable sludge hydrolysis rate of 28.4%. Thereby, SCFAs production was improved to 264.8 mg COD/g VSS. Such performances were approximately 80.2-87.8% of those with conventional CER residence period (8 days). The organic composition distribution and parallel factor analysis demonstrated that similar biodegradability and utilizability of fermentative liquid were achievable with various operation patterns. Compared with the conventional operation pattern, the modified operation pattern with shortened CER residence period (1 day) also displayed satisfying anaerobic fermentation efficiency and numerous engineering bene fits, e.g. decreased CER usage, reduced engineering footprint, relieved CER fouling, and increased operation convenience. The findings might provide sustainable development for CER assistant anaerobic fermentation strategy and enlighten the direction of anaerobic fermentation process.

Metagenomic analysis reveals the size effect of magnetite on anaerobic digestion of waste activated sludge after thermal hydrolysis pretreatment.

Although magnetite has been widely investigated in anaerobic digestion (AD), its role in the practical AD of waste-activated sludge (WAS) after thermal hydrolysis pretreatment (THP) and its size effect remain unclear. In this study, magnetite with four different particle sizes was added during the AD of WAS after THP. With the reduction of magnetite particle size, cumulative methane production was increased, while the optimal dosage of magnetite decreased, with 0.1 µm magnetite at an optimal dosage of 2 g/L achieving the highest cumulative methane production increase of 111.97 % compared with the blank group (without magnetite). Smaller magnetite particles increased α-glucosidase and protease activities, coenzyme F420 concentration, and electron-transport system activity (20.30 %, 173.02 %, 60.39 % and 158.08 % higher respectively than the blank group). The size of magnetite also influenced the establishment of direct interspecies electron transfer (DIET) during AD. Based on the analysis of the pilA gene abundance, magnetite with a large particle size could promote the formation of e-pili in syntrophic electroactive bacteria (Clostridium, Syntrophomonas, and Pseudomonas) and methanogens (Methanospirillum), thereby completing electron transfer. However, small-sized magnetite particles stimulated DIET by enhancing the secretion of conductive proteins in extracellular polymeric substances and membrane-bound enzymes (Fpo) in Methanosarcina.

Insights into carbon recovery from excess sludge through enzyme-catalyzing hydrolysis strategy: Environmental benefits and carbon-emission reduction.

This study introduced the excellent improvement of enzyme cocktail (lysozyme and protease) on hydrolysis efficiency and the role of reducing carbon emission as an alternative carbon source. The best dosing method after optimization was to add four parts of lysozyme at 0 h and one part of protease at 1 h. The extracellular proteins and polysaccharides increased by 118% and 64% respectively under the optimal dosing mode. Enzyme cocktails reduced more organic matters and extended the distribution of sludge particles in the small particle size part. The enzymatic-treated sludge could reduce 21.09 kg CO2/t VSS if utilized to replace methanol for denitrification carbon source. Enzyme cocktails did better in enhancing both solubilization and hydrolysis than single enzymes under the optimal method. This study will provide a more integrated and comprehensive system for enzymatic pretreatment and new insight into the enzymatic pretreatment enhancing hydrolysis and reducing carbon emission.

Impact of divalent cations on lysozyme-induced solubilisation of waste-activated sludge: Perspectives of extracellular polymeric substances and surface electronegativity.

Lysozyme hydrolysis can accelerate waste-activated sludge (WAS) solubilisation, which can significantly shorten the process and promote the efficiency of anaerobic digestion. This study investigated the impact of divalent cations on lysozyme-induced solubilisation of WAS. The performance of lysozyme pretreatment was dramatically inhibited by Mg2+ and Ca2+. Compared to the control group, the amount of net SCOD, protein, and polysaccharides released to the supernatant were reduced by 36.6%, 44.7%, and 35.8%, respectively, in the presence of divalent cations. The extracellular polymeric substance (EPS) matrix became tightly bound, resulting in fewer proteins and polysaccharides being extracted from loosely-bound EPS (LB-EPS) with divalent cations, which was detrimental to the solubilisation of WAS. Divalent cations decreased the surface electronegativity of sludge particles and prolonged the adsorption of lysozymes by sludge flocs. More than 16.6% of total lysozymes remained in the liquid phase of WAS after 240 min Mg2+ and Ca2+ strengthened the binding among proteins and polysaccharides and promoted the intermolecular cross-linking of polysaccharides. The EPS matrix formed a dense spatial reticular structure that blocked the transfer of lysozymes from the EPS matrix to the pellet. As a result, the lysozymes accumulated in LB-EPS rather than hydrolysing the microorganism's cell wall. This study provides a new perspective on the restriction of WAS pretreatment with lysozymes and optimises the method of lysozyme-induced solubilisation of WAS.

An innovative cation regulation-based anaerobic fermentation strategy for enhancing short-chain fatty acids production from waste activated sludge: Metal ion removal coupled with Na+-regulation.

This study proposed a cation-regulation strategy based on metal ion removal coupled Na+-regulation for enhancing anaerobic fermentation of waste activated sludge. The optimal treatment condition was: cation-exchange resin dosage of 1.75 g/g SS for 1-day treatment, followed by Na+-enhanced anaerobic fermentation at NaCl concentration of 20 g/L. The CER induced sludge solubilization and the Na+-regulation treatment triggered secondary hydrolysis of CER-solubilized sludge, causing remarkable sludge disintegration and extracellular polymeric substance (EPS) disruption. Numerous SCOD of 6588 mg/L (SCOD/TCOD = 40.6%) was released within 2 days, and the short-chain fatty acids (SCFAs) of 439.9 mg COD/g VSS was produced through 4-day anaerobic fermentation. More than 59% of the SCFAs was composed of acetate and propionate. Nitrogen-free organic matters (i.e. SCFAs and carbohydrates) accounted for 77.9% of SCOD, while considerable sludge solid reduction (51.6% of total VSS) was achievable, which was beneficial for fermentative liquid utilization and sludge disposal.

Hydrolase activity and microbial community dynamic shift related to the lack in multivalent cations during cation exchange resin-enhanced anaerobic fermentation of waste activated sludge.

The correlation of the lack in multivalent cations with hydrolase activity and microbial community in anaerobic fermentation of waste activated sludge was investigated in this study. It was demonstrated that considerable solid phase reduction of 41 % (7.87 g/L) was achievable through a cation exchange resin-enhanced anaerobic fermentation of 4 days. The protease and α-glucosidase, especially α-glucosidase, were easily influenced by a lack in multivalent cations. Furthermore, species abundance and diversity of microbial community gradually decreased. Meanwhile, the bacteria community structure presented obvious dynamic shifts. Ruminococcaceae_UCG_009, Bacteroides and Macellibacteroides responsible for organic matter biodegradation and SCFAs production became dominant bacteria in cation exchange resin-enhanced anaerobic fermentation, which was less influenced by the lack in multivalent cations, while the SCFA consumers (e.g. methanogens) were inhibited with reduced abundances due to their susceptibility to the lack in multivalent cations. Redundancy analysis revealed that the lack in multivalent cations were responsible for the microbial community evolution, which was proved by the high Grey relational coefficients (0.747-0.820) and significant negative Spearman coefficients (-0.5798 to -0.9429) between multivalent cation and microbial community. Obviously, the cation exchange resin-induced removal of multivalent cations reduced enzyme activity and modified microbial community structure, which created a beneficial environment for enhancing anaerobic fermentation.

Enhanced anaerobic fermentation of waste activated sludge by NaCl assistant hydrolysis strategy: Improved bio-production of short-chain fatty acids and feasibility of NaCl reuse.

This study developed an economical approach for enhancing short-chain fatty acids (SCFAs) production from waste activated sludge (WAS) by NaCl assistant anaerobic fermentation. With NaCl addition at 20 g/L, sludge disintegration with extracellular polymeric substance (EPS) disruption and cell lysis were induced owing to the attack of osmotic pressure, which facilitated WAS solubilization with release of biodegradable organic matters. The SCOD sharply increased to 4092 mg/L (SCOD/TCOD = 23.9%) after 2-day hydrolysis, against 1462 mg/L in the control. After 4-day anaerobic fermentation, considerable SCFAs production of 288.2 mg COD/g VSS was achievable. More than 60% of the SCFAs was composed of acetic and propionic acids. The feasibility of bio-electrogenesis in microbial fuel cell (MFC) utilizing fermentative liquid was assessed. As such, the produced SCFAs could be consumed with energy recovery, thereby the used NaCl was reusable, which created environmental and economic benefits, e.g. reduced NaCl consumption and cost, negligible residual NaCl.