Bioprocess and Biosystems Engineering最新文献

The efficient and eco-friendly removal of lignin is a critical challenge for bioethanol production from lignocellulosic biomass. Herein, we report the integration of laccase with deep eutectic solvents (DESs) for the pretreatment of corn stover to enhance the production of reducing sugars. Three betaine-based DESs were prepared and tested for their effects on the activity and stability of a bacterial laccase from Bacillus amyloliquefaciens LC02. The aqueous solution of DESs showed no adverse influence on laccase activity, and the laccase thermostability was improved in the presence of DESs. More than 95% of the laccase activity was retained in the DESs solution during the first hour of incubation at 70 °C. A red shift in the fluorescence spectra was observed for the laccase in the presence of DESs, indicating conformational changes. The laccase was able to degrade a dimeric lignin model compound by cleaving its β-O-4 bond. The transformation products were identified using LC-MS. The maximal lignin removal from corn stover was achieved by pretreatment using laccase in combination with the betaine-glycerol DES, which also resulted in a yield of fermentable sugar that was 130% higher than the control. This combination strategy provides guidance on the application of laccase and DESs in the pretreatment of lignocellulosic biomass.

The study focused on rhamnolipid production by batch fermentation of Pseudomonas aeruginosa USM-AR2 in a 3-L stirred-tank reactor (STR) using palm sludge oil (PSO) as the sole carbon source. The impact of various agitation rates towards the dispersion of PSO in the medium was evaluated to improve biomass growth and rhamnolipid production. A mechanical foam collection and recycling system was designed and retrofitted to the STR to overcome severe foam formation during fermentation. The maximum biomass produced was 11.29 ± 0.20 g/L obtained at 400 rpm, while the maximum rhamnolipid production was 5.06 ± 1.17 g/L at 600 rpm, giving a rhamnolipid productivity of 0.023 g/L/h. High agitation enhances substrate availability by breaking the hydrophobic semi-solid PSO into smaller substrate particles, increasing surface contact area, thus facilitating the PSO utilisation by P. aeruginosa USM-AR2, thereby inducing rhamnolipid production. This study further demonstrates the ability of rhamnolipid to solubilize and disperse sludge oil, which typically remains a solid at room temperature, in the liquid medium. GCMS analysis showed that five fatty acids, namely palmitic acid, myristic acid, stearic acid, methyl ester and linoleic acid, have been utilised. The rhamnolipid showed an oil spreading test result of 160 mm of waste engine oil displacement compared to control using distilled water that remained non-displaced, and a critical micelle concentration (CMC) of 17 mg/L. In emulsification index (E₂₄) assay, the rhamnolipid was shown to emulsify toluene (66.7% ± 7.2), waste engine oil (58.3% ± 7.2), kerosene (41.8% ± 4.8) and n-hexane (33.1% ± 5.7). UPLC analysis on rhamnolipid revealed a congener mixture of rhamnolipid, namely di-rhamnolipid and mono-rhamnolipid mixture. This is the first report on the employment of an integrated foam control reactor system with PSO as the carbon source for rhamnolipid production by P. aeruginosa USM-AR2 culture.

Due to the prevalence of drug-resistant bacteria and the ongoing shortage of novel antibiotics as well as the challenge of treating breast cancer, the therapeutic and clinical sectors are consistently seeking effective nanomedicines. The incorporation of metal oxide nanoparticles with biological macromolecules and an organic compound emerges as a promising strategy to enhance breast cancer treatment and antibacterial activity against drug-resistant bacteria in various biomedical applications. This study aims to synthesize a unique nanocomposite consisting of CeO₂ embedded with folic acid and carboxymethyl cellulose (CFC NC) via a green precipitation method using Moringa oleifera. Various spectroscopic and microscopic analyses are utilized to decipher the physicochemical characteristics of CFC NC and active phytocompounds of Moringa oleifera. Antibacterial study against MRSA (Methicillin-resistant Staphylococcus aureus) demonstrated a higher activity (95.6%) for CFC NC compared to its counterparts. The impact is attributed to reactive oxygen species (ROS), which induces a strong photo-oxidative stress, leading to the destruction of bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of CFC NC are determined as 600 µg/mL and 1000 µg/mL, respectively. The anticancer activity against breast cancer cell resulted in the IC₅₀ concentration of 10.8 μg/mL and 8.2 μg/mL for CeO₂ and CFC NC respectively.The biocompatibility test was conducted against fibroblast cells and found 85% of the cells viable, with less toxicity. Therefore, the newly synthesized CFC NC has potential applications in healthcare and industry, enhancing human health conditions.

Protein engineering is a powerful tool for designing or modifying therapeutic proteins for enhanced efficacy, increased safety, reduced immunogenicity, and improved delivery. Fusion proteins are an important group of therapeutic compounds that often require an ideal linker to combine diverse domains to fulfill the desired function. GGGGS [(G4S)n] linkers are commonly used during the engineering of proteins because of their flexibility and resistance to proteases. However, unexpected truncation was observed in the linker of a bispecific antibody, which presented challenges in terms of production and quality. In this work, a bispecific antibody containing 5*G4S was investigated, and the truncation position of the linkers was confirmed. Our investigation revealed that codon optimization, which can overcome the negative influence of a high repetition rate and high GC content in the (G4S)n linker, may reduce the truncation rate from 5-10% to 1-5%. Moreover, the probability of truncation when a shortened 3* or 4*G4S linker was used was much lower than that when a 5*G4S linker was used in mammalian cells. In the case of expressing a bispecific antibody, the bioactivity and purity of the product containing a shorter G4S linker were further investigated and are discussed.

The present study aims to analyze the interaction between Rhodotorula toruloides and magnetic nanoparticles and evaluate their effect on carotenoid production. The manganese ferrite nanoparticles were synthesized without chitosan (MnFe₂O₄) and chitosan coating (MnFe₂O₄-CS) by the co-precipitation method assisted by hydrothermal treatment. XRD (X-ray diffraction), Magnetometry, Dynamic Light Scattering (DLS) and FTIR (Fourier-Transform Infrared Spectroscopy), are used to characterize the magnetic nanoparticles. The crystallite size of MnFe₂O₄ was 16 nm for MnFe₂O₄ and 20 nm for MnFe₂O₄-CS. The magnetic saturation of MnFe₂O₄-CS was lower (39.6 ± 0.6 emu/g) than the same MnFe₂O₄ nanoparticles (42.7 ± 0.3 emu/g), which was attributed to the chitosan fraction presence. The MnFe₂O₄-CS FTIR spectra revealed the presence of the characteristic chitosan bands. DLS demonstrated that the average hydrodynamic diameters were 344 nm for MnFe₂O₄ and 167 nm for MnFe₂O₄-CS. A kinetic study of cell immobilization performed with their precipitation with a magnet demonstrated that interaction between magnetic nanoparticles and R. toruloides was characterized by an equilibrium time of 2 h. The adsorption isotherm models (Langmuir and Freundlich) were fitted to the experimental values. The trypan blue assay was used for cell viability assessment. The carotenoid production increased to 256.2 ± 6.1 µg/g dry mass at 2.0 mg/mL MnFe₂O₄-CS. The use of MnFe₂O₄-CS to stimulate carotenoid yeast production and the magnetic separation of biomass are promising nanobiotechnological alternatives. Magnetic cell immobilization is a perspective technique for obtaining cell metabolites.

This research investigated the physicochemical properties and biological activities of green-synthesized copper oxide nanoparticles (CuO NPs) via Moringa peregrina extract, graphene oxide (GO), and their composite (CuO-GO). SEM revealed the morphology and structure, indicating polygonal CuO NPs, thin wrinkled sheets of GO, and a combination of CuO NPs and GO in the nanocomposite. EDS confirmed the elemental composition and distribution. XRD analysis confirmed the crystalline monoclinic structure of CuO NPs and GO, as well as their composite, CuO-GO, with characteristic peaks. DLS analysis exhibited distinct size distributions, with CuO NPs showing the narrowest range. BET surface area analysis revealed mesoporous structures for all materials, with the nanocomposite showing enhanced surface area and pore volume. Anticancer assays on MCF-7 and normal NIH/3T3 cells demonstrated CuO-GO's superior cytotoxicity against cancer cells, with minimal effects on normal cells, suggesting selective cytotoxicity. Moreover, antibacterial assays against Pseudomonas aeruginosa and Staphylococcus aureus indicated CuO-GO's potent inhibitory activity. The composite's synergistic effects were evidenced by its lower minimum inhibitory concentration (MIC) compared to individual components. In conclusion, this study elucidated the promising biomedical applications of CuO NPs, GO, and their nanocomposite, particularly in cancer treatment and antibacterial therapies, showcasing their potential as multifunctional nanomaterials.

The septic tank is the most commonly used decentralized wastewater treatment systems for household wastewater treatment in on-site applications. The removal rate of various pollutants is lower in different septic tank configurations. The integration of a microbial electrolysis cells (MEC) into septic tank or biofilm-based reactors can be a green and sustainable technology for household wastewater treatment and energy production. In this study, a 50-L septic tank was converted into a 50-L MEC coupled with biofilm-based reactor for simultaneous household wastewater treatment and hydrogen production. The biofilm-based reactor was integrated by an anaerobic packed-bed biofilm reactor (APBBR) and an aerobic moving bed biofilm reactor (aeMBBR). The MEC/APBBR/aeMBBR was evaluated at different organic loading rates (OLRs) by applying voltage of 0.7 and 1.0 V. Result showed that the increase of OLRs from 0.2 to 0.44 kg COD/m³ d did not affect organic matter removals. Nutrient and solids removal decreased with increasing OLR up to 0.44 kg COD/m³ d. Global removal of chemical oxygen demand (COD), biochemical oxygen demand (BOD), total nitrogen (TN), ammoniacal nitrogen (NH₄⁺), total phosphorus (TP) and total suspended solids (TSS) removal ranged from 81 to 84%, 84 to 85%, 53 to 68%, 88 to 98%, 11 to 30% and 76 to 88% respectively, was obtained in this study. The current density generated in the MEC from 0 to 0.41 A/m² contributed to an increase in hydrogen production and pollutants removal. The maximum volumetric hydrogen production rate obtained in the MEC was 0.007 L/L^.d (0.072 L/d). The integration of the MEC into biofilm-based reactors applying a voltage of 1.0 V generated different bioelectrochemical nitrogen and phosphorus transformations within the MEC, allowing a simultaneous denitrification-nitrification process with phosphorus removal.

Fruits are a very good source of various nutrients that can boost overall human health. In these days, the recovery of therapeutic compounds from different fruit wastes is trending in research, which might not only minimize the waste problem but also encounter a higher demand for various enzymes that could have antimicrobial properties against infectious diseases. The goal of this review is to focus on the recovery of therapeutic enzymes from fruit wastes and its present-day tendency for utilization. Here we discussed different parts of fruit waste, such as pulp, pomace, seed, kernel, peel, etc., that produce therapeutic enzymes like amylase, cellulose, lipase, laccase, pectinase, etc. These bioactive enzymes are present in different parts of fruit and could be used as therapeutics against various infectious diseases. This article provides a thorough knowledge compilation of therapeutic enzyme isolation from fruit waste on a single platform, distinctly informative, and significant review work on the topic that is envisioned to encourage further research ideas in these areas that are still under-explored. This paper explains the various aspects of enzyme isolation from fruit and vegetable waste and their biotherapeutic potential that could provide new insights into the development of biotherapeutics and attract the attention of researchers to enhance translational research magnitude further.

Immobilized fillers have been increasingly utilized in biotrickling filters (BTFs) due to their positive impact on shock load resistance and recovery performance. However, due to the inherent characteristics of its immobilized carrier, the immobilized filler is prone to swelling during the long-term operation of the system, resulting in increased pressure drop. Polyurethane (PU) sponge was used as the cross-linked skeleton of immobilized filler and compared with direct emulsified cross-linked immobilized filler for treating ethylbenzene gas. In the early stage, both fillers can maintain good performance despite changes in the inlet concentration and short-term stagnation. However, on the 107th day of operation, the immobilized filler experienced swelling, and the pressure drop sharply increased to 137.2 Pa, while the PU immobilized filler was still able to maintain a low-pressure drop level. The results of the microbial diversity analysis revealed that the microbial community structure of PU immobilized fillers remained relatively stable when responding to the fluctuations in operating conditions. PU sponges as the skeleton can effectively prolong the service life of the immobilized filler and improve the performance of the biotrickling filter.

To explore the impact of microbial interactions on outcomes from three prevalent algorithms (Flux Balance Analysis (FBA), community FBA (cFBA), and SteadyCom) analyzing microbial community metabolic networks, five toy community models representing common microbial interactions were designed. These include commensalism, mutualism, competition, mutualism-competition, and commensalism-competition. Various scenarios, considering different biomass yields and substrate constraints, were examined for each type. In commensal communities, all algorithms consistently produced similar results. However, changes in biomass yields and substrate constraints led to variable abundances (0.33-0.8) and community growth rates (2-5 1/h) within a broad range. For competitive communities, all algorithms predicted growth of fastest-growing member. To comply with the natural coexistence of members, suboptimal solutions over optimal point are recommended. FBA faced challenges in modeling mutualism, consistently predicting growth of only one member. Although cFBA and SteadyCom resulted in a lower community growth rate, coexistence of both members were satisfied. In toy models with dual interactions, more realistic outcomes were achieved contrary to purely competitive model as the dependency fosters the coexistence which was missing in the competitive only scenarios. These findings emphasize the importance of algorithm choice based on specific microbial interaction types for reliable community behavior predictions.​.