Global expansion of mariculture generates large volumes of saline wastewater that require sustainable and decentralized treatment to protect coastal ecosystems. Gravity-driven membrane bioreactors (GMBRs) are an attractive low-energy option for this purpose; however, severe membrane fouling in saline environments usually limits water flux. In this study, a microalgal-bacterial consortium (MBC) was introduced to reshape biofilm architecture and improve the long-term performance of GMBRs. Four ceramic membrane-integrated GMBRs were operated for 160 days to treat synthetic mariculture wastewater, including two MBC systems with bacteria-to-microalgae inoculation ratios of 3:2 and 5:2 and two bacteria-only controls. The optimized MBC system (5:2) achieved a stable flux of 14.9 LMH, which was 2.4 times that of the conventional GMBRs (∼6 LMH), while also showing superior nutrient removal (TN removal: 67.9-73.3%, effluent TN: 4.65-5.73 mg/L; TP removal: 92.0-96.3%, effluent TP: 0.1-0.3 mg/L). The improved performance was associated with the formation of larger, more porous aggregates (>20 μm), the development of a synergistic MBC architecture, and enhanced degradation of extracellular polymeric substances. Confocal laser scanning microscopy and scanning electron microscopy revealed a dynamic "loose-dense-loose" structural evolution of the biofilm in the MBC systems, which prevented the dense and irreversible layering observed in the conventional systems. Microbial community analysis further showed that the optimized system enriched functional bacteria (e.g., Nitrosomonas and Nitrospira) while maintaining high microalgal viability. These findings demonstrate that MBC is an effective strategy for improving GMBR performance and offers a sustainable approach for mariculture wastewater treatment.
Xing Du, Zihong Liao, Wei Xie et al.· Water Research· 0 citations
Cyanidin-3-O-glucoside (C3G) is a water-soluble, value-added natural flavonoid with extensive applications in the nutraceutical and cosmetic industries. However, its efficient bioproduction is hampered by intermediate instability, metabolic imbalance and enzyme kinetic constraints. Here, we successfully constructed an efficient biosynthesis pathway from the dihydroquercetin (DHQ) to C3G in Escherichia coli through a multilevel engineering strategy. Initially, the integration of glutathione S-transferase (GST) redirected the metabolic flux towards target cyanidin formation. To minimize the dissipation of labile intermediates, pathway enzymes were spatially organized within a protein cage to enhance cascade efficiency. Furthermore, genomic integration of Glycine max sucrose synthase established an in-situ UDP-glucose regeneration module to ensure a continuous precursor supply for the final glycosylation step. After these pathway-level optimizations, the key enzyme dihydroflavonol 4-reductase (FaDFR) emerged as a new rate-limiting bottleneck due to substrate inhibition under increased DHQ loading. Structure-guided and evolution-informed engineering generated FaDFR variants with improved high-substrate tolerance, as supported by in vitro activity profiling and molecular dynamics simulations. Through combined pathway and enzyme engineering, the G130C-containing strain achieved a C3G titer of 1.34 g/L, representing a 23-fold improvement over the GST-assisted baseline strain. Our platform enables efficient, value-added C3G production and provides a promising framework for constructing downstream pathways toward structurally diverse anthocyanin derivatives.