Biodegradation of Lignin Monomers Vanillic, p-Coumaric, and Syringic Acid by the Bacterial Strain, Sphingobacterium sp. HY-H.

Many bacterial strains have been demonstrated to biodegrade lignin for contaminant removal or resource regeneration. The goal of this study was to investigate the biodegradation amount and associated pathways of three lignin monomers, vanillic, p-coumaric, and syringic acid by strain Sphingobacterium sp. HY-H. Vanillic, p-coumaric, and syringic acid degradation with strain HY-H was estimated as 88.71, 76.67, and 72.78%, respectively, after 96 h. Correspondingly, the same three monomers were associated with a COD removal efficiency of 87.30, 55.17, and 67.23%, and a TOC removal efficiency of 82.14, 61.03, and 43.86%. The results of GC-MS, HPLC, FTIR, and enzyme activities show that guaiacol and o-dihydroxybenzene are key intermediate metabolites of the vanillic acid and syringic acid degradation. p-Hydroxybenzoic acid is an important intermediate metabolite for p-coumaric and syringic acid degradation. LiP and MnP play an important role in the degradation of lignin monomers and their intermediate metabolites. One possible pathway is that strain HY-H degrades lignin monomers into guaiacol (through decarboxylic and demethoxy reaction) or p-hydroxybenzoic acid (through side-chain oxidation); then guaiacol demethylates to o-dihydroxybenzene. The p-hydroxybenzoic acid and o-dihydroxybenzene are futher through ring cleavage reaction to form small molecule acids (butyric, valproic, oxalic acid, and propionic acid) and alcohols (ethanol and ethanediol), then these acids and alcohols are finally decomposed into CO2 and H2O through the tricarboxylic acid cycle. If properly optimized and controlled, the strain HY-H may play a role in breaking down lignin-related compounds for biofuel and chemical production.

Gel beads prepared by polyvinyl alcohol cross-linking with phytic acid and Fe as novel microbial carriers for simultaneous partial nitrification, anammox and denitrification.

Enhancing the stability of biomass and ensuring a stable activity of anaerobic ammonia oxidizing bacteria are crucial for successful operation of the simultaneous partial nitrification, Anammox, and denitrification (SNAD) process. In this study, gel beads of polyvinyl alcohol/phytic acid (PVA/PA) and polyvinyl alcohol/phytic acid/Fe (PVA/PA/Fe) were prepared as innovative bio-carriers. Theoretical simulations and analyses revealed that these carriers are predominantly connected via hydrogen and borate bonds, with PVA/PA/Fe also featuring metal coordination bonds. The total nitrogen removal efficiency of reactors with PVA/PA/Fe and PVA/PA increased by 13.5 % and 9.0 %, respectively, compared to reactor without carriers. The iron-enriched PVA/PA/Fe carriers significantly improve SNAD by promoting Anammox, Feammox, and nitrate-dependent Fe2+ oxidation reactions, leading to faster nitrogen conversion and higher nitrogen removal rate than reactor without carriers and with PVA/PA. Using of PVA/PA and PVA/PA/Fe gel beads as bio-carriers offers benefits to the SNAD process, including cost-effective and low carbon requirement.

Insight: High intensity and activity carrier granular sludge cultured using polyvinyl alcohol/chitosan and polyvinyl alcohol/chitosan/iron gel beads.

The newly developed carrier granular sludge (CGS) with polyvinyl alcohol/chitosan (PVA/CS) and PVA/CS/Fe gel beads assistance showed higher intensity and anammox activity than the natural granular sludge (NGS). Through comprehensive investigation, it was found: (1) the gel beads provided a stable framework of cells entangle with extracellular polymeric substances (EPS) to enhance the sludge intensity. In this framework, ß-polysaccharides are distributed at the edge of CGS as a protection layer, α-polysaccharides and proteins are spread in the whole cross-section as backbones, and Fe2+/Fe3+ in CGS-PVA/CS/Fe act as bridges to link with the negatively charged groups on bacterial surfaces and proteins. (2) The porous gel beads satisfied a relatively unimpeded mass transfer. Thus, the sludge activity, microbe's metabolism, membrane transportation and environmental adaption in CGS were apparently improved. The results improved the understanding about the advantages of the CGS and indicated their possible application in full-scale anammox processes.

Enhancing anammox resistance to low operating temperatures with the use of PVA gel beads.

Low temperatures, or a sudden decrease in operating temperature, can seriously inhibit anammox activity, it is, therefore, important to maintain anammox activities at a low temperature. In this study, the use of gel beads to enhance the resistance of anammox biomass to a low temperature was investigated. The performance of three reactors: R1 without gel beads; R2 with polyvinyl alcohol/chitosan (PVA/CS); R3 with PVA/CS/Fe, was studied and compared in a temperature transition from 35 to 8 °C. When the operating temperature was ≥25 °C, there was little difference in nitrogen removal among the three reactors. Decreasing the temperature to < 25 °C created obvious difference between R1 and R2/R3. R1 had a nitrogen removal efficiency (NRE) of 33.1 ± 25.3% at 10 °C, significantly lower than that of R2 (90.5 ± 2.5%) or R3 (87.7 ± 11.1%). Unclassified Candidatus Brocadiaceae was the dominant genus at 10 °C, with an abundance of 44.4, 56.5 and 58.7% in R1, R2 and R3, respectively. These differences were attributed to the use of gel beads, which promoted the granulation of both the non-immobilized sludge and the immobilized biomass, resulting in higher anammox activities in R2/R3. The non-immobilized sludge of R1 was dominated by small particles (<300 µm) at 10 °C, while in R2 and R3 large particles (1000-2000 µm) were the main components. Furthermore, the immobilized biomass on gel beads exhibited much higher anammox activity and maintained a relatively high level of nitrate reductase and nitrite reductase in response to the temperature decrease. The Fe2+/Fe3+ in the PVA/CS/Fe gel beads further promoted microbial aggregation and led to an improved performance in R3 compared to R2. The results of this study demonstrate an effective approach to increase anammox resistance at low operating temperatures.

Achieving reliable partial nitrification and anammox process using polyvinyl alcohol gel beads to treat low-strength ammonia wastewater.

In this study, polyvinyl alcohol (PVA) gel beads were used to aid the partial nitrification and anammox process (PN/A) for treating low-strength ammonia wastewater. When treating synthetic and municipal wastewater, the reactor amended with PVA gel beads achieved a total nitrogen removal efficiency of 75.1 ± 8.4% and 66.6 ± 7.0% respectively, while the control reactor without PVA gel beads achieved 63.2 ± 7.8% and 28.2 ± 11.5% respectively. Dissolved oxygen (<0.5 mg O2 L-1) and substrate diffusion in porous PVA gel beads facilitated the formation of microbial stratification in the gel beads. Unclassified Candidatus Brocadiaceae (major anammox bacteria) and Ignavibacterium (major ammonia oxidizing bacteria) enriched in the inside and outside layers of PVA gel beads, which benefited the synergetic cooperation of these bacteria and protecting them from environmental fluctuations. This study provides a promising solution for achieving a reliable PN/A process in mainstream wastewater treatment.

Granule-based immobilization and activity enhancement of anammox biomass via PVA/CS and PVA/CS/Fe gel beads.

Granule-based immobilization of anammox biomass assisted by polyvinyl alcohol/chitosan (PVA/CS) and PVA/CS/Fe gel beads was studied, via the operation of three identical up-flow reactors (R1 without gel beads, R2 with PVA/CS, R3 with PVA/CS/Fe) for 203 days. In the end, the nitrogen removal rates (NRR) were 5.3 ± 0.4, 10.0 ± 0.3 and 13.9 ± 0.5 kg-N m-3 d-1 for R1, R2 and R3, respectively. The porous PVA/CS and PVA/CS/Fe created a suitable eco-niche for anammox bacteria to grow and attach, thus being retained in the reactor. The EPS entangles newly grown cells within the gel beads, resulting in compact aggregation. The interaction between Fe ions added to PVA/CS/Fe gel beads and negatively charged EPS groups strongly promoted granule strength and compactness. The immobilization method proposed by this study was found to effectively improve biomass retention in the reactors, which is promising for advanced anammox process applications.

PVA/CS and PVA/CS/Fe gel beads' synthesis mechanism and their performance in cultivating anaerobic granular sludge.

Biomass washout from high-speed anaerobic suspended bed bio-reactors is still a challenge to their stable operation. Preserving active biomass to efficiently retain biomass in the reactor is one of the solutions to this problem. Herein, two carriers (polyvinyl alcohol/chitosan (PVA/CS) and PVA/CS/Fe gel beads) were prepared using the cross-linking method. The fourier transform infrared (FTIR) and 13C nuclear magnetic resonance (13C NMR) analyses showed that PVA/CS gel beads formed mainly through hydrogen-bonds (NH2OH-). Furthermore, FTIR, 13C NMR, energy dispersive spectrum (EDS), X-ray diffractometer (XRD) and X-ray photoelectron spectroscopy (XPS) analyses showed that PVA/CS/Fe gel beads formed mainly through chelate bond (NH2-FeM+OH-). The scanning electron microscope (SEM) results affirmed that the gel beads had rough and well-developed porous structure for the attachment of microbes. Furthermore, the abilities of gel beads on the cultivation of granular sludge in an up-flow anaerobic sludge bed (UASB) reactor were effectively demonstrated while treating wastewater polluted with glucose and alkali lignin. The results showed that the gel beads-assisted reactors had a higher performance than those without the gel beads. The cultivation of granules in these reactors was accelerated, while the granules became bigger and exhibited better settling velocities. The reactor with gel beads was easier to withstand a higher organic loading rate due to dense microbial aggregates, which were caused by more humic-like substance. Particularly, the reactor with PVA/CS/Fe gel beads was able to improve the overall robustness of the system due to stronger mechanical properties of gel beads, and also prevented cells detachment.

Kraft Lignin Biodegradation by Dysgonomonas sp. WJDL-Y1, a New Anaerobic Bacterial Strain Isolated from Sludge of a Pulp and Paper Mill.

Wastewater containing kraft lignin (KL) discharged from pulp and paper industries could cause serious environmental contamination. Appropriate effluent treatment is required to reduce the pollution. Investigations on anaerobic bacteria capable of degrading KL are beneficial to both lignin removal and biofuel regeneration from the effluent. In this paper, an anaerobic strain capable of degrading KL was isolated from the sludge of a pulp and paper mill and identified as Dysgonomonas sp. WJDL-Y1 by 16S rRNA analysis. Optimum conditions for KL degradation by strain WJDL-Y1 were obtained at initial pH of 6.8, C:N ratio of 6 and temperature of 33°C, based on statistical analyses by response surface methodology. For a 1.2 g/l KL solution, a COD removal rate of 20.7% concomitant with biomass increase of 17.6% was achieved after 4 days of incubation under the optimum conditions. After the treatment by strain WJDL-Y1, KL was modified and degraded.