Biochemical Engineering Journal最新文献

Denitrification of wastewater with a low organic carbon to NO₃^--N ratio (C/N ratio) faces challenges due to slow rates and low efficiency. This study reported that carbon-based conductive carriers are able to enhance the removal of nitrogen from wastewater with low C/N ratio by coupling Fe(II)-driven autotrophic and heterotrophic bioelectrochemical denitrification. When Fe(II) was the sole electron donor, the bioreactor using conductive carrier achieved a denitrification rate constant (k_DN) of 0.016 h⁻¹, 1.7 times of that with non-conductive materials. This enhancement was due to the conductive carrier boosting direct electron transfer and supporting the growth of electroactive microorganisms. For wastewater with a low C/N ratio of 0.76, the bioreactor featuring both Fe(II) and the conductive carrier reached a k_DN of 0.095 h⁻¹, five times higher than without Fe(II). The presence of Fe(II) promoted denitrification by enhancing electron transfer and serving as a mediator. Microbial analysis showed that adding Fe(II) enriched electroactive bacteria like Comamonas and denitrifiers such as Chryseobacterium. Our findings suggest a promising strategy to enhance denitrification in wastewater treatment systems with low C/N ratios.

Mathematical models are indispensable for designing, optimizing, and controlling large-scale bioprocesses. In the present work, the production of pectinases was studied experimentally using an internal loop airlift bioreactor, pectin as a substrate, and Aspergillus flavipes FP-500 as a biocatalyst. The N-tanks-in-series (NTIS) model was implemented to predict the behavior of fungal growth, pectin and dissolved oxygen consumption, pectinases production, and the oxygen mass transfer rate from gas phase to liquid culture medium. A double Monod-Logistic kinetic model was used to describe the biomass growth rate as a function of biomass, pectin, and dissolved oxygen concentrations. In contrast, a Luedeking-Piret kinetic model was used to describe the production rate of endo and exo pectinases. A hydrodynamic model was utilized to estimate gas hold-ups, volumetric mass transfer coefficients, and air inflow velocities. Good agreement was observed between the experimental data and the theoretical results, demonstrating the predictive capacity of the NTIS model to describe pectinase production, the oxygen consumption rate, and the oxygen evolution in the gas phase. The model highlighted its robust capability to capture the critical parameters of aerobic fermentation processes. Therefore, it could be used as a tool for the scalability of the airlift bioreactors.

Cordyceps sinensis is widely known for its therapeutic properties. Enhancing the yield of exopolysaccharides (EPS), which is crucial for its medicinal efficacy, is a major challenge. In this study, we applied high initial glucose concentrations with talc particles to enhance EPS production and assessed the cell morphology, intracellular biochemical reactants, and bioactivity contribution of glycoproteins. The use of 150 g/L glucose and 10 g/L 2000 mesh talc increased the EPS yield by 1.8-fold to 4.21 g/L. The addition of talc regulated cell morphology, facilitated the entry of oxygen molecules into the cells to produce a large amount of ATP for polysaccharide synthesis, and altered the cell wall structure to facilitate the secretion of EPS. Moreover, environmental stress resulted in a notable increase in intracellular reactive oxygen species levels, which can potentially enhance cell membrane permeability and promote EPS synthesis. Furthermore, the highest protein content in crude EPS corresponded to the maximum activation of alcohol dehydrogenase (ADH) of 44.2 %, suggesting a mechanistic relationship between the proteins and polysaccharides in the glycoproteins that influence the activation of ADH. These findings elucidate the intricate interplay between fermentation conditions and EPS production and provide new avenues for optimizing the fermentation process of CS-HKI to enhance its therapeutic applications.

In this study, the effluent from recycled paper mill was treated using a combined ozone (O₃) and biological aerated filter (BAF) process. Key operational parameters such as ozone dosage, pH, hydraulic retention time (HRT), volume load and gas-to-water ratio were optimized. Under optimal conditions, with a total ozone dosage of 100 g/m, a gas-to-water ratio of 4:1, and an HRT of 3.0 hours in the BAF, the chemical oxygen demand (COD) and chroma of the treated wastewater were reduced to 44–55 mg/L and 2–4 PCU, achieving removal efficiencies of 70 % and 95 %, respectively. The discharge effluent not only satisfy the new discharge standard of China (GB3544–2008), but also can be used as recycling water. Additionally, the treatment cost of wastewater was ca. 1.3 ¥/m³ in pilot-scale test, significantly decreasing the cost. Ozone pretreatment has a significant effect on wastewater decolorization by disrupting the molecular chemical structure of pollutants, which increase the biochemical properties of biofilm and is beneficial to the sequential BAF treatment. The sludge in the O₃/BAF system exhibited increased biomass with minimal filamentous bacteria and higher dehydrogenase activity, confirming stable and robust bacterial growth. GC-MS analysis revealed substantial reduction in pollutant content and diversity post-treatment, although the recalcitrant compound (Z)-13-docosenamide remained relatively high, decreasing from 27.37 % to 21.14 %. The mechanism of the O₃/BAF process for the pollutant degradation were also proposed. This study demonstrated that a combination of ozone and fixed biofilm treatment is an efficient and cost-effective treatment, providing the theory and practical applicability for the industrial wastewater.

Despite extensive efforts in biotherapeutics production, there is a pressing need to enhance the biomanufacturing process to meet the growing demand for recombinant products. Ubiquitous Chromatin Opening Element (UCOE), chromatin-modifying elements derived from CpG islands, are crucial in regulating gene expression. Therefore, to explore the UCOE effect on gene expression, we cloned codon-optimized Darbepoetin alfa (DPO) sequence into the pOptiVEC then the Darbepoetin alfa-IRES-DHFR fragment was digested, and cloned into the UCOE vector. Both pOptiVEC-DPO and UCOE-DPO-IRES-DHFR cassettes were linearized and transfected into the two cell pools of CHO DG44 cells. The transfected cells were adapted with CD OptiCHO™ medium after 20 days. Recombinant DPO expression was evaluated by qRT-PCR technique. Qualitative and quantitative protein expression analysis was also carried out by western blotting and ELISA assays. Our findings showed a boosted DPO expression rate at both mRNA and protein levels in the UCOE-DPO-IRES-DHFR pool compared to pOptiVEC-DPO. These data suggest that UCOE and codon optimization result in high and stable gene expression in the CHO DG44 cell line.

The lack of denitrifying bacteria and organic carbon sources, and the inhibition of dissolved oxygen (DO) result in nitrate accumulation in aquaculture wastewater. In order to solve this problem, an encapsulation method was introduced to prepare a novel bioactive capsule, which can provide organic carbon source, denitrifying bacteria, and anoxic microenvironment for aquaculture wastewater denitrification. And can reduce the recovery time of the enclosed denitrifying bacteria. The morphology of the capsule, its nitrate removal rate, and nitrogen conversion pathway in synthetic aquaculture wastewater were investigated. The capsule had a porous surface and the pore diameter ranged from 150.0 nm to 300.0 nm. The enclosed denitrifying bacteria had a reduced recovery time and excellent denitrification performance. The nitrate removal rate reached 86.2 % on the first day and was maintained at 99.7 %. Nitrogen conversion pathways in the capsule include denitrification, assimilatory/dissimilatory nitrate reduction, and nitrogen fixation. The denitrifying capsule has short recovery time and good denitrification performance, which would help to achieve denitrification in aquaculture wastewater or other low C/N wastewater.

This study examined the impact of different biochar (BC) as an anaerobic digestion (AD) additive on antibiotic-resistant bacteria (ARB) survival and AD performance using dairy cow manure. Bamboo BC and Olive BC with different particle sizes were added into the mesophilic AD at 15 g/L and 30 g/L dosages (Bamboo-15, Bamboo-30, Olive-15, and Olive-30). The study provides a detailed analysis of biogas production, organic metabolism, and ARB and microbial dynamics, elucidating the mechanisms by which BC influences AD. Findings reveal significant reductions in CEZ-resistant bacteria (CEZ-r) across all reactors, ranging from 12.88 % to 76.47 %. Both Bamboo and Olive BC increased CEZ-r removal by 3.08–5.94 times compared to the control. Additionally, BC supplementation prevented the rise in CEZ-r percentage within the total bacteria count observed in the control reactor. Bamboo BC outperformed Olive BC in enhancing biogas yield, with Bamboo-15 and Bamboo-30 showing significant increases of 43.2 % and 48.0 %, respectively, compared to the control. Adding BC in AD regulates ARB by decreasing potential ARG hosts and impeding the transmission of resistance. It also enhances biogas production by improving the efficiency of methanogenic bacteria and optimizing the methanogenic pathway. This research provides insights into how BC can be used to enhance AD performance and mitigate ARB proliferation, offering a sustainable approach to waste management and energy production.

Longan (Dimocarpus longan Lour.) is a nutrient-rich fruit, widely cultivated in China. However, its seeds and pericarp, often discarded as waste, are rich in bioactive components, particularly polyphenolics. In this research, we develop microbiota-assisted extraction technique, which is efficient and environmentally friendly for extracting these valuable components. These results showcased superior efficiency in the diversity and richness of extracted bioactive components, although yielding slightly lower TP content than ethanol extraction. The active components extracted by microbiota-assisted extraction technique increased to 2.45 mg/g, which was higher 2.13 times and 3.10 times compared to optimized ethanol and water extraction, respectively. The numbers of active components extracted by these three methods were 39, 24 and 30, separately, and Quercetin 3-D-xyloside and Kaempferol 3-O-alpha-L-rhamnopyranosyl were identified in longan pericarp extracts by microbiota-assisted extraction. Metagenomics analysis revealed stable and diverse microbiota with functional capabilities for enhancing extraction efficiency. Functional annotations indicated significant roles in biosynthesis, carbohydrate degradation, and fatty acid metabolism. Microbiota-assisted extracts exhibited a broad spectrum of bioactive compounds with potential health benefits, including antioxidant and anti-inflammatory properties. These results provide a theoretical foundation for developing sustainable extraction technologies.

Significant progress has been achieved in large-scale mammalian cell culture technology for biotherapeutics manufacturing over the past decades, necessitating the Process Analytical Technology (PAT) for the real-time measurement of critical quality attributes and the guidance for precise process control to ensure productivity, quality, and consistency. The Oxygen Uptake Rate ($\mathrm{OUR}$) serves as a crucial indicator for characterizing the energy metabolism of mammalian cells, offering insights into cellular state and metabolism dynamics. However, current cellular $\mathrm{OUR}$ monitoring in antibody production depends mainly on costly gas analyzers or periodic manual sampling. Here, we introduce a novel method for in-line monitoring of cellular $\mathrm{OUR}$ in bioreactors based on the stationary liquid phase balance (SLPB) theory, which extends its applicability to diverse aeration and foam conditions without additional equipment or labor expenditures. We modeled the $k_{L}a$ of the aerated stirred bioreactor, assessed the influence of foam on liquid surfaces induced by gas sparging on oxygen transfer, and processed raw $\mathrm{OUR}$ data using a sliding filter. The established method was applied to monitoring the real-time $\mathrm{OUR}$ of Chinese Hamster Ovary (CHO) cell cultures for antibody production, demonstrating its excellent accuracy, sensitivity and readability. Aligned with the Quality by Design (QbD) concept, this real-time $\mathrm{OUR}$ estimation enables rapid detection of metabolic changes, revealing cellular physiology and facilitating precise feedback control in biotherapeutics manufacturing.

Complex bispecific antibody formats tend to form more product-related impurities than monoclonal antibodies. The primary constraints are yield and purity in the polishing stage. The purpose of this study was to enhance the understanding of the optimal working window for four mixed-mode resins and to reduce the burden of resin screening and parameter optimization during process development. This study optimized the loading and elution conditions of four different mixed-mode cationic resins to enhance the yield and purity by integrating Design of Experiments with High-Throughput Screening. It was observed that despite being weakly acidic mixed-mode cationic resins, these four resins exhibited significant differences in their adsorption and elution performances and varied tolerances to salt concentrations. Capto MMC demonstrated strong hydrophobicity, while the performance profiles of MX-Trp-650 M and Nuvia cPrime were similar, with Nuvia cPrime showing superior purification effects. Eshmuno CMX, by extending its side chain ligand, achieved higher binding efficiency and capacity. Through the optimization of salt concentrations or the application of dual-gradient elution strategies, the target protein with high yield (77 %) and purity over 99 % was successfully obtained. This research not only provides in-depth insights into the application of mixed-mode chromatography in the biopharmaceutical field but also offers practical optimization strategies for the industrial-scale purification of bispecific antibodies.