Microalgae are rich in valuable biomolecules and grow on non-arable land with rapid growth rate, which has a host of new possibility as alternative protein sources. In the present study, extraction of proteins from Chlorella vulgaris via an efficient technique, Liquid Triphasic Flotation (LTF) system, was studied. The optimized conditions in LTF system were 70% v/v of t-butanol, 40% w/v of salt solution, 0.5% w/v of biomass, pH 5.54, 1:1 of salt to t-butanol solution, and 10 min of air flotation time to attain 87.23% of protein recovery and 56.72% of separation efficiency. Besides, the study on recycling t-butanol has demonstrated that only one run was sufficient to maintain the performance of system. The efficiency of LTF in extracting protein has performed better than just Three Phase Partitioning (TPP) system. LTF system is hence an effective protein extraction and purification method with minimum operation unit and processing time.
In this study, the bacterial lipoxygenase (LOX) gene from Pseudomonas aeruginosa ATCC27853 (pse-LOX) was cloned, sequenced and heterologous expressed in Escherichia coli by auto-induction expression strategy. Production of the recombinant pse-LOX (pse-rLOX) gene up to 23,850 U/mL (264 mg pure protein/L bacterial culture fluid) was observed in the end of this process. To the best of our knowledge, this is the first attempt to manipulate LOX heterologous expression process using auto-induction expression approach, and it is the highest production of recombinant LOX compared with other reports. Subsequently, the resulted pse-rLOX was proved to efficiently degrade triphenylmethane dyes such as malachite green, brilliant green and aniline blue. Generally, an overproduction of the LOX from P. aeruginosa was observed in E. coli, and this recombinant gene is a potential candidate as biocatalyst for triphenylmethane dyes decolorization.
An ideal scaffold should be biocompatible, having appropriate microstructure, excellent mechanical strength yet degrades. Chitosan exhibits most of these exceptional properties, but it is always associated with sub-optimal cytocompatibility. This study aimed to incorporate graphene oxide at wt % of 0, 2, 4, and 6 into chitosan matrix via direct blending of chitosan solution and graphene oxide, freezing, and freeze drying. Cell fixation, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, alkaline phosphatase colorimetric assays were conducted to assess cell adhesion, proliferation, and early differentiation of MG63 on chitosan-graphene oxide scaffolds respectively. The presence of alkaline phosphatase, an early osteoblast differentiation marker, was further detected in chitosan-graphene oxide scaffolds using western blot. These results strongly supported that chitosan scaffolds loaded with graphene oxide at 2 wt % mediated cell adhesion, proliferation, and early differentiation due to the presence of oxygen-containing functional groups of graphene oxide. Therefore, chitosan scaffolds loaded with graphene oxide at 2 wt % showed the potential to be developed into functional bone scaffolds.
In this research, a protein nanofiber membrane (P-COOH-CEW) was developed to treat the dye waste. Initially, polyacrylonitrile nanofiber membrane (PAN) was prepared by electrospinning, followed by heat treatment, alkaline treatment, and neutralization to obtain weak cation exchange nanofiber membrane (P-COOH). The P-COOH membrane was chemically coated with chicken egg white (CEW) proteins to obtain a 3D structure of complex protein nanofiber membrane (P-COOH-CEW). The composite prepared was characterized with Fourier Transform Infrared analysis (FTIR), Scanning Electron Microscopy (SEM), and thermogravimetric analysis (TGA). Further, the composite was evaluated by investigating the removal of Toluidine Blue O (TBO) from aqueous solutions in batch conditions. Different operating parameters - coupling of CEW, shaking rate, initial pH, contact time, temperature, and dye concentration were studied. From the results, maximum removal capacity and equilibrium association constant was determined to be 546.24 mg/g and 10.18 mg/mg, respectively at pH 10 and 298 K. The experimental data were well fitted to pseudo-second order model. Furthermore, desorption studies revealed that the adsorbed TBO can be completely eluted by using 50% ethanol or 50% glycerol in 1 M NaCl solution. Additionally, the reuse of P-COOH-CEW membrane reported to have 97.32% of removal efficiency after five consecutive adsorption/desorption cycles.
Recently, supercritical fluid CO2 extraction (SFE) has emerged as a promising and pervasive technology over conventional extraction techniques for various applications, especially for bioactive compounds extraction and environmental pollutants removal. In this context, temperature and pressure regulate the solvent density and thereby effects the yield, selectivity, and biological/therapeutic properties of the extracted components. However, the nature of plant matrices primarily determines the extraction mechanism based on either density or vapor pressure. The present review aims to cover the recent research and developments of SFE technique in the extraction of bioactive plant phytochemicals with high antioxidant, antibacterial, antimalarial, and anti-inflammatory activities, influencing parameters, process conditions, the investigations for improving the yield and selectivity. In another portion of this review focuses on the ecotoxicology and toxic metal recovery applications. Nonpolar properties of Sc-CO2 create strong solvent strength via distinct intermolecular interaction forces with micro-pollutants and toxic metal complexes. This results in efficient removal of these contaminants and makes SFE technology as a superior alternative for conventional solvent-based treatment methods. Moreover, a compelling assessment on the therapeutic, functional, and solvent properties of SFE is rarely focused, and hence this review would add significant value to the SFE based research studies. Furthermore, we mention the limitations and potential of future perspectives related to SFE applications.
In this study, four different selected wall materials (namely gelatin, soy protein isolate, maltodextrin and Arabic gum) were applied for blueberry anthocyanin extract encapsulation. The effect of these wall material types on the release and degradation of anthocyanin and the modulation of gut microbiota during in vitro simulated gastrointestinal digestion and colonic fermentation were investigated. It was found that the encapsulation of anthocyanin extract using appropriate wall material could significantly enhance the colonic accessibility of anthocyanins. Soy protein isolate and gelatin delayed the release of anthocyanins, whereas the other two wall materials displayed no significant effect on the release time of anthocyanins. Gut microbiota mainly metabolized some phenolic compounds such as 4-hydroxycinnamic acid and chlorogenic acid. Meanwhile, different fermented anthocyanin extract microcapsule broth could significantly decrease the composition and abundance of Firmicutes and increase that of Bacteroidetes. Furthermore, the presence of anthocyanin extract microcapsules, especially those encapsulated with soy protein isolate, promoted the biosynthesis of short-chain fatty acids by gut microbiota. It is concluded that, amongst the wall materials studied, soy protein isolate appeared to be a functional and suitable candidate to delay anthocyanin release and prevent disease through the promotion of gut health.
Adsorption of lysozyme on the dye-affinity nanofiber membranes was investigated in batch and dynamic modes. The membrane matrix was made of electrospun polyacrylonitrile nanofibers that were grafted with ethylene diamine (EDA) and/or chitosan (CS) for the coupling of Reactive Blue 49 dye. The physicochemical properties of these dye-immobilized nanofiber membranes (P-EDA-Dye and P-CS-Dye) were characterized microscopically, spectroscopically and thermogravimetrically. The capacities of lysozyme adsorption by the dye-affinity nanofiber membranes were evaluated under various conditions, namely pH, dye immobilized density, and loading flow rate. The adsorption of lysozyme to the dye-affinity nanofiber membranes was well fitted by Langmuir isotherm and pseudo-second kinetic models. P-CS-Dye nanofiber membrane had a better performance in the dynamic adsorption of lysozyme from complex chicken egg white solution. It was observed that after five cycles of adsorption-desorption, the dye-affinity nanofiber membrane did not show a significant loss in its capacity for lysozyme adsorption. The robustness as well as high dynamic adsorption capability of P-CS-Dye nanofiber membrane are promising for the efficient recovery of lysozyme from complex feedstock via nanofiber membrane chromatography.
In this study, three potential probiotic strains were selected to ferment blueberry and blackberry juices. The viable cell counts of selected strains were increased by 0.4-0.7 log CFU/mL in berry juices environments after 48-h fermentation. Meanwhile, the contents of cyanindin-3-glucoside and peonidin-3-glucoside decreased over 30%. Heatmap presented an upgrade trend of syringic acid, ferulic acid, gallic acid and lactic acid during fermentation. However, the contents of p-coumaric acid, protocatechuic acid, chlorogenic acid, critic acid and malic acid showed downgrade trend. The metabolism of phenolics probably contributed to the enhancement of the ABTS radical scavenging activity (40%-60%) in fermented berry juices. Moreover, the three strains presented different capacities on changing the quality of berry juices according to the PCA and LDA analysis. The contents of individual organic acids had positive correlations with sensory quality, especially for sourness. Overall, probiotic fermentation could improve the sensory quality of berry juices.
Electrospinning technology was applied for the preparation of polyacrylonitrile (PAN) nanofiber membrane in this work. After hot pressing, alkaline hydrolysis and neutralization treatment, a weak acid cation exchange membrane (P-COOH) was prepared. By the covalent coupling reaction between the acidic membrane and aminomethane sulfonic acid (AMSA), a strong acidic nanofiber membrane (P-SO3H) was obtained. The surface morphology, chemical structure, and thermal stability of the prepared ion exchange membranes were analyzed via SEM, FTIR and TGA. Analytical results showed that the membranes were prepared successfully and thermally stable. The ion exchange membrane (IEX) was conducted with the newly designed membrane reactor, and different operating conditions affecting the adsorption efficiency of Toluidine Blue dye (TBO) were investigated by dynamic flow process. The results showed that dynamic binding capacity (DBC) of weak and strong IEX membranes for TBO dye was ~170 mg/g in a dynamic flow process. Simultaneously, the ion exchange membranes were also used for purifying lysozyme from chicken egg white (CEW). Results illustrated that the recovery yield and purification factor of lysozyme were 93.43% and 29.23 times (P-COOH); 90.72% and 36.22 times (P-SO3H), respectively. It was revealed that two type ion exchange membranes were very suitable as an adsorber for use in dye waste treatment and lysozyme purification process. P-SO3H strong ion-exchange membrane was more effective either removal of TBO dye or purification of lysozyme. The ion exchange membranes not only effectively purified lysozyme from CEW solution, but also effectively removed dye from wastewater.
Carbon dioxide (CO2) emission will increase due to the increasing global plastic demand. Statistical data shows that plastic production alone will contribute to at least 20% of the annual global carbon budget in the near future. Hence, several alternative methods are recommended to overcome this problem, such as bio-product synthesis. Algae consist of diverse species and have huge potential to be a promising biomass feedstock for a range of purposes, including bio-oil production. The convenient cultivation method of algae could be one of the main support for algal biomass utilization. The aim of this study is to forecast and outline the strategies in order to meet the future demand (year 2050) of plastic production and, at the same time, reduce CO2 emission by replacing the conventional plastic with bio-based plastic. In this paper, the analysis for 25%, 50% and 75% CO2 reduction has been done by using carbon emission pinch analysis. The strategies of biomass utilization in Malaysia are also enumerated in this study. This study suggested that the algal biomass found in Malaysia coastal areas should be utilized and cultivated on a larger scale in order to meet the increasing plastic demand and, at the same time, reduce carbon footprint. Some of the potential areas for macroalgae sea-farming cultivation in Sabah coastline (Malaysia), comprised of about 3885 km2 (388,500 ha) in total, have been highlighted. These potential areas have the potential to produce up to 14.5 million tonnes (Mt)/y of macroalgae in total, which can contribute 370 Mt of phenol for bioplastic production.
The richness of high-value bio-compounds derived from microalgae has made microalgae a promising and sustainable source of useful product. The present work starts with a review on the usage of open pond and photobioreactor in culturing various microalgae strains, followed by an in-depth evaluation on the common harvesting techniques used to collect microalgae from culture medium. The harvesting methods discussed include filtration, centrifugation, flocculation, and flotation. Additionally, the advanced extraction technologies using ionic liquids as extractive solvents applied to extract high-value bio-compounds such as lipids, carbohydrates, proteins, and other bioactive compounds from microalgae biomass are summarized and discussed. However, more work needs to be done to fully utilize the potential of microalgae biomass for the application in large-scale production of biofuels, food additives, and nutritive supplements.
Microalgae have become imperative for biological wastewater treatment. Its capability in biological purification of wastewaters from different origins while utilizing wastewater as the substrate for growth has manifest great potentials as a sustainable and economical wastewater treatment method. The wastewater grown microalgae have also been remarked in research to be a significant source of value-added bioproducts and biomaterial. This paper highlights the multifaceted roles of microalgae in wastewater treatment from the extent of microalgal bioremediation function to environmental amelioration with the involvement of microalgal biomass productivity and carbon dioxide fixation. Besides, the uptake mechanism of microalgae in wastewater treatment was discussed in detail with illustrations for a comprehensive understanding of the removal process of undesirable substances. The performance of different microalgae species in the uptake of various substances was studied and summarized in this review. The correlation of microalgal treatment efficacy with various algal strain types and the bioreactors harnessed for cultivation systems was also discussed. Studies on the alternatives to conventional wastewater treatment processes and the integration of microalgae with accordant wastewater treatment methods are presented. Current research on the biological and technical approaches for the modification of algae-based wastewater system and the maximization of biomass production is also reviewed and discussed. The last portion of the review is dedicated to the assertion of challenges and future perspectives on the development of microalgae-based wastewater treatment technology. This review serves as a useful and informative reference for readers regarding the multifaceted roles of microalgae in the application of wastewater biotreatment with detailed discussion on the uptake mechanism.
Ionic liquids (ILs) have emerged as an alternative solvent used in the bioprocessing of microalgae for recovery of valuable biomolecules. The aim of this work is to extract fucoxanthin from Chaetoceros calcitrants (C. calcitrans) by using the readily distillable CO2-based alkyl carbamate ILs. The degree of cell permeabilization was analysed by the quantification of extracted fucoxanthin and the analyses of cell surface morphology. Among the tested CO2-based alkyl carbamate ILs, diallylammonium diallylcarbamate (DACARB) extraction system gave the maximal yield of fucoxanthin at 17.51 mg/g under the optimal extraction conditions [90% (v/v), 3 min and 25 °C]. Moreover, the extracted fucoxanthin fraction exhibited the satisfactory antioxidant activities. The recyclability of DACARB was demonstrated in the multiple batches of fucoxanthin extraction. Hence, CO2-based alkyl carbamate ILs can prospectively substitute conventional organic solvents in the downstream processing of bioactive compounds from microalgae.
At present, polyhydroxyalkanoates (PHAs) have been considered as a promising alternative to conventional plastics due to their diverse variability in structure and rapid biodegradation. To ensure cost competitiveness in the market, thermoseparating aqueous two-phase extraction (ATPE) with the advantages of being mild and environmental-friendly was suggested as the primary isolation and purification tool for PHAs. Utilizing two-level full factorial design, this work studied the influence and interaction between four independent variables on the partitioning behavior of PHAs. Based on the experimental results, feed forward neural network (FFNN) was used to develop an empirical model of PHAs based on the ATPE thermoseparating input-output parameter. In this case, bootstrap resampling technique was used to generate more data. At the conditions of 15 wt % phosphate salt, 18 wt % ethylene oxide⁻propylene oxide (EOPO), and pH 10 without the addition of NaCl, the purification and recovery of PHAs achieved a highest yield of 93.9%. Overall, the statistical analysis demonstrated that the phosphate concentration and thermoseparating polymer concentration were the most significant parameters due to their individual influence and synergistic interaction between them on all the response variables. The final results of the FFNN model showed the ability of the model to seamlessly generalize the relationship between the input⁻output of the process.
Nowadays, downstream bioprocessing industries inclines towards the development of a green and high efficient bioseparation technology. Betacyanins are presently gaining higher interest in the food science as driven by their high tinctorial strength and health promoting functional properties. In this study, a novel green integration process of liquid biphasic electric partitioning system (LBEPS) was proposed for betacyanins extraction from peel and flesh of red-purple pitaya. Initially, the betacyanins extraction using LBEPS with initial settings was compared with that of liquid biphasic partitioning system (LBPS), and the results revealed that both systems demonstrated a comparable betacyanins extraction. This was followed by further optimizing the LBEPS for better betacyanins extraction. Several operating parameters including operation time, voltage applied, and position of graphitic electrodes in the system were investigated. Moreover, comparison between optimized LBEPS and LBPS with optimized conditions of electric system (as post-treatment) as well as color characterization and antioxidant properties assessment were conducted. Overall, the betacyanins extraction employing the optimized LBEPS showed the significant highest values of betacyanins concentration in alcohol-rich top phase (C t ) and partition coefficient (K) of betacyanins from peel (99.256 ± 0.014% and 133.433 ± 2.566) and flesh (97.189 ± 0.172% and 34.665 ± 2.253) of red-purple pitaya. These results inferred that an optimal betacyanins extraction was successfully achieved by this approach. Also, the LBEPS with the peel and flesh showed phase volume ratio (V r ) values of 1.667 and 2.167, respectively, and this indicated that they have a clear biphasic separation. In addition, the peel and flesh extract obtained from the optimized LBEPS demonstrated different variations of red color as well as their antioxidant properties were well-retained. This article introduces a new, reliable, and effective bioseparation approach for the extraction of biomolecules, which is definitely worth to explore further as a bioseparation tool in the downstream bioprocessing.
Third generation biofuels, also known as microalgal biofuels, are promising alternatives to fossil fuels. One attractive option is microalgal biodiesel as a replacement for diesel fuel. Chlamydomonas sp. Tai-03 was previously optimized for maximal lipid production for biodiesel generation, achieving biomass growth and productivity of 3.48 ± 0.04 g/L and 0.43 ± 0.01 g/L/d, with lipid content and productivity of 28.6 ± 1.41% and 124.1 ± 7.57 mg/L/d. In this study, further optimization using 5% CO2 concentration and semi-batch operation with 25% medium replacement ratio, enhanced the biomass growth and productivity to 4.15 ± 0.12 g/L and 1.23 ± 0.02 g/L/d, with lipid content and productivity of 19.4 ± 2.0% and 239.6 ± 24.8 mg/L/d. The major fatty acid methyl esters (FAMEs) were palmitic acid (C16:0), oleic acid (C18:1), and linoleic acid (C18:2). These short-chain FAMEs combined with high growth make Chlamydomonas sp. Tai-03 a suitable candidate for biodiesel synthesis.
The coexistence of algae and bacteria in nature dates back to the very early stages when life came into existence. The interaction between algae and bacteria plays an important role in the planet ecology, cycling nutrients, and feeding higher trophic levels, and have been evolving ever since. The emerging concept of algal-bacterial consortia is gaining attention, much towards environmental management and protection. Studies have shown that algal-bacterial synergy does not only promote carbon capture in wastewater bioremediation but also consequently produces biofuels from algal-bacterial biomass. This review has evaluated the optimistic prospects of algal-bacterial consortia in environmental remediation, biorefinery, carbon sequestration as well as its contribution to the production of high-value compounds. In addition, algal-bacterial consortia offer great potential in bloom control, dye removal, agricultural biofertilizers, and bioplastics production. This work also emphasizes the advancement of algal-bacterial biotechnology in environmental management through the incorporation of Industry Revolution 4.0 technologies. The challenges include its pathway to greener industry, competition with other food additive sources, societal acceptance, cost feasibility, environmental trade-off, safety and compatibility. Thus, there is a need for further in-depth research to ensure the environmental sustainability and feasibility of algal-bacterial consortia to meet numerous current and future needs of society in the long run.
In this study, a sustainable NH2-MIL-101(Al) is synthesized and subjected to characterization for cryogenic CO2 adsorption, isotherms, and thermodynamic study. The morphology revealed a highly porous surface. The XRD showed that NH2-MIL-101(Al) was crystalline. The NH2-MIL-101(Al) decomposes at a temperature (>500 °C) indicating excellent thermal stability. The BET investigation revealed the specific surface area of 2530 m2/g and the pore volume of 1.32 cm3/g. The CO2 adsorption capacity was found to be 9.55 wt% to 2.31 wt% within the investigated temperature range. The isotherms revealed the availability of adsorption sites with favorable adsorption at lower temperatures indicating the thermodynamically controlled process. The thermodynamics showed that the process is non-spontaneous, endothermic, with fewer disorders, chemisorption. Finally, the breakthrough time of NH2-MIL-101(Al) is 31.25% more than spherical glass beads. The CO2 captured by the particles was 2.29 kg m-3. The CO2 capture using glass packing was 121% less than NH2-MIL-101(Al) under similar conditions of temperature and pressure.
With the growing interest in food safety and in environmental protection, it is more attractive to develop novel biodegradable packaging films. In this regard, one new blending film was prepared with curdlan (CD)/polyvinyl alcohol (PVA)/thyme essential oil. Our results demonstrated that the mechanical properties of the blending film were the best when the ratio of the CD and PVA was 4:1. Further, the barrier properties of the film were optimized by incorporating with thyme essential oil. It was proved that not only water vapor permeability was lower, but also the elongation at break was improved, when 2% (w/w) thyme essential oil used. The potential interactions of the film matrix were analyzed by FTIR, XRD and Cryo-scanning electron microscopy. Importantly, both the antioxidant activity and antibacterial activity were improved. Finally, the blending film was employed for the preservation of chilled meat, while the shelf life was extended up to 10 days.
Despite of ongoing research programs and numerous clinical trials, seasonal influenza epidemics remain a major concern globally. Vaccination remains the most effective method to prevent influenza infection. However, current flu vaccines have several limitations, including limited vaccine capacity, long production times, inconsistence efficacy in certain populations, and lack of a "universal" solution. Different next-generation approaches such as cell line-based culture, reverse genetics, and virus expression technology are currently under development to address the aforementioned challenges in conventional vaccine manufacture pipeline. Such approaches hope for safe and scalable production, induce broad-spectrum immunity, create premade libraries of vaccine strains, and target nonvariable regions of antigenic proteins for "universal" vaccination. Here, we discuss the process and challenges of the current influenza vaccine platform as well as new approaches that are being investigated. These developments indicate that an exciting future lies ahead in the influenza vaccine field.