This work demonstrated the optimization and scale up of microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) of bioactive compounds from Orthosiphon stamineus using energy-based parameters such as absorbed power density and absorbed energy density (APD-AED) and response surface methodology (RSM). The intensive optimum conditions of MAE obtained at 80% EtOH, 50mL/g, APD of 0.35W/mL, AED of 250J/mL can be used to determine the optimum conditions of the scale-dependent parameters i.e. microwave power and treatment time at various extraction scales (100-300mL solvent loading). The yields of the up scaled conditions were consistent with less than 8% discrepancy and they were about 91-98% of the Soxhlet extraction yield. By adapting APD-AED method in the case of UAE, the intensive optimum conditions of the extraction, i.e. 70% EtOH, 30mL/g, APD of 0.22W/mL, AED of 450J/mL are able to achieve similar scale up results.
The present study was conducted to optimize the extraction yield of starch from sago (Metroxylon sagu) pith waste (SPW) with the assistance of ultrasound ensued by the transformation of extracted starch into a higher value-added bioplastic film. Sago starch with extraction yield of 71.4% was successfully obtained using the ultrasound-assisted extraction, with the following conditions: particle size <250 µm, solid loading of 10 wt.%, ultrasonic amplitude of 70% and duty cycle of 83% in 5 min. The rapid ultrasound approach was proven to be more effective than the conventional extraction with 60.9% extraction yield in 30 min. Ultrasound-extracted starch was found to exhibit higher starch purity than the control starch as indicated by the presence of lower protein and ash contents. The starch granules were found to have irregular and disrupted surfaces after ultrasonication. The disrupted starch granules reduced the particle size and increased the swelling power of starch which was beneficial in producing a film-forming solution. The ultrasound-extracted sago starch was subsequently used to prepare a bioplastic film via solution casting method. A brownish bioplastic film with tensile strength of 0.9 ± 0.1 MPa, Young's modulus of 22 ± 0.8 MPa, elongation at break of 13.6 ± 2.0% and water vapour permeability (WVP) of 1.11 ± 0.1 × 10-8 g m-1 s-1 Pa-1 was obtained, suggesting its feasibility as bioplastic material. These findings provide a means of utilization for SPW which is in line with the contemporary trend towards greener and sustainable products and processes.
Bioplastic has been perceived as a promising candidate to replace petroleum-based plastics due to its environment-friendly and biodegradable characteristics. This study presents the chitosan reinforced starch-based bioplastic film prepared by the solution casting and evaporation method. The effects of processing parameters, i.e., starch concentration, glycerol loading, process temperature and chitosan loading on mechanical properties were examined. Optimum tensile strength of 5.19 MPa and elongation at break of 44.6% were obtained under the combined reaction conditions of 5 wt.% starch concentration, 40 wt.% glycerol loading, 20 wt.% chitosan loading and at a process temperature of 70 °C. From the artificial neural network (ANN) modeling, the coefficient of determination (R2) for tensile strength and elongation at break were found to be 0.9955 and 0.9859, respectively, which proved the model had good fit with the experimental data. Interaction and miscibility between starch and chitosan were proven through the peaks shifting to a lower wavenumber in FTIR and a reduction of crystallinity in XRD. TGA results suggested the chitosan-reinforced starch-based bioplastic possessed reasonable thermal stability under 290 °C. Enhancement in water resistance of chitosan-incorporated starch-based bioplastic film was evidenced with a water uptake of 251% as compared to a 302% registered by the pure starch-based bioplastic film. In addition, the fact that the chitosan-reinforced starch-based bioplastic film degraded to 52.1% of its initial weight after 28 days suggests it is a more sustainable alternative than the petroleum-based plastics.
A sequencing batch reactor (SBR) seeded with flocculated sludge and fed with synthetic wastewater was operated for an enhanced biological phosphorus removal (EBPR) process. Eight weeks after reactor startup, sludge granules were observed. The granules had a diameter of 0.5 to 3.0 mm and were brownish in color and spherical or ellipsoidal in shape. No significant change was observed in sludge granule size when operational pH was changed from 7 to 8. The 208-day continuous operation of the SBR showed that sludge granules were stably maintained with a sludge volume index (SVI) between 30 to 55 mL/g while securing a removal efficiency of 83% for carbon and 97% for phosphorus. Fluorescent in situ hybridization (FISH) confirmed the enrichment of polyphosphate accumulating organisms (PAOs) in the SBR. The observations of sludge granulation in this study encourage further studies in the development of granules-based EBPR process.
The applicability of the enhanced biological phosphorus removal (EBPR) process for the removal of phosphorus in warm climates is uncertain due to frequent reports of EBPR deterioration at temperature higher than 25 °C. Nevertheless, a recent report on a stable and efficient EBPR process at 28 °C has inspired the present study to examine the performance of EBPR at 24 °C-32 °C, as well as the PAOs and GAOs involved, in greater detail. Two sequencing batch reactors (SBRs) were operated for EBPR in parallel at different temperatures, i.e., SBR-1 at 28 °C and SBR-2 first at 24 °C and subsequently at 32 °C. Both SBRs exhibited high phosphorus removal efficiencies at all three temperatures and produced effluents with phosphorus concentrations less than 1.0 mg/L during the steady state of reactor operation. Real-time quantitative polymerase chain reaction (qPCR) revealed Accumulibacter-PAOs comprised 64% of the total bacterial population at 24 °C, 43% at 28 °C and 19% at 32 °C. Based on fluorescent in situ hybridisation (FISH), the abundance of Competibacter-GAOs at both 24 °C and 28 °C was rather low (<10%), while it accounted for 40% of the total bacterial population at 32 °C. However, the smaller Accumulibacter population and larger population of Competibacter at 32 °C did not deteriorate the phosphorus removal performance. A polyphosphate kinase 1 (ppk1)-based qPCR analysis on all studied EBPR processes detected only Accumulibacter clade IIF. The Accumulibacter population shown by 16S rRNA and ppk1 was not significantly different. This finding confirmed the existence of single clade IIF in the processes and the specificity of the clade IIF primer sets designed in this study. Habitat filtering related to temperature could have contributed to the presence of a unique clade. The clade IIF was hypothesised to be able to perform the EBPR activity at high temperatures. The clade's robustness most likely helps it to fit the high-temperature EBPR sludge best and allows it not only to outcompete other Accumulibacter clades but coexist with GAOs without compromising EBPR activity.
Development of new materials for different applications especially as bio-composites has received great attention. This study concentrates on development of a biopolymer based on chitosan (CT) and halloysite nanotubes (HNT) and evaluates the copper removal intake as a potential application of this bio-composite. In this study, CT/HNT beads were prepared by ultrasonic-assisted extrusion-dripping method for the first time. Two sources of HNTs (i.e. Dragonite and Matauri Bay) were added into a chitosan solution (2wt.%) at various loading fractions (25, 50, 75wt.%). The effect of ultrasound as a mixing device was also studied by varying the amplitude at constant frequency of 25%, 50% and 75%. Characteristics and physical properties of the prepared CT/HNT beads were also analyzed by SEM, FTIR, TGA and BET the results show that introducing HNT to chitosan increases the adsorption capacity toward copper ions; however HNT loading fraction above 50wt.% resulted in a decrease in adsorption capacity attributed to limited accessibility of the amino groups. The adsorption capacity of the CT/HNT beads prepared from Dragonite source had a larger adsorption capacity of 14.2mg/g as compared to that of Matauri Bay, 10.55mg/g. It was observed that the adsorption capacity of the beads toward copper ions decreased when the loading fraction of HNT is increased at constant ultrasound amplitude. The result of this study helps to understand the links between the characteristics and adsorption abilities of CT/HNT beads.
Many developing countries, mostly situated in the tropical region, have incorporated a biological nitrogen removal process into their wastewater treatment plants (WWTPs). Existing wastewater characteristic data suggested that the soluble chemical oxygen demand (COD) in tropical wastewater is not sufficient for denitrification. Warm wastewater temperature (30 °C) in the tropical region may accelerate the hydrolysis of particulate settleable solids (PSS) to provide slowly-biodegradable COD (sbCOD) for denitrification. This study aimed to characterize the different fractions of COD in several sources of low COD-to-nitrogen (COD/N) tropical wastewater. We characterized the wastewater samples from six WWTPs in Malaysia for 22 months. We determined the fractions of COD in the wastewater by nitrate uptake rate experiments. The PSS hydrolysis kinetic coefficients were determined at tropical temperature using an oxygen uptake rate experiment. The wastewater samples were low in readily-biodegradable COD (rbCOD), which made up 3-40% of total COD (TCOD). Most of the biodegradable organics were in the form of sbCOD (15-60% of TCOD), which was sufficient for complete denitrification. The PSS hydrolysis rate was two times higher than that at 20 °C. The high PSS hydrolysis rate may provide sufficient sbCOD to achieve effective biological nitrogen removal at WWTPs in the tropical region.
This study investigates the use of nanodiamonds (ND) as a promising carrier for enzyme immobilization and compares the effectiveness of immobilized and native enzymes. Three different enzyme types were tested, of which Rhizopus niveus lipase (RNL) exhibited the highest relative activity, up to 350 %. Under optimized conditions (1 h, pH 7.0, 40 °C), the immobilized ND-RNL showed a maximum specific activity of 0.765 U mg-1, significantly higher than native RNL (0.505 U mg-1). This study highlights a notable enhancement in immobilized lipase; furthermore, the enzyme can be recycled in the presence of a natural deep eutectic solvent (NADES), retaining 76 % of its initial activity. This aids in preserving the native conformation of the protein throughout the reusability process. A test on brine shrimp revealed that even at low concentrations, ND-RNL had minimal toxicity, indicating its low cytotoxicity. The in silico molecular dynamics simulations performed in this study offer valuable insights into the mechanism of interactions between RNL and ND, demonstrating that RNL immobilization onto NDs enhances its efficiency and stability. All told, these findings highlight the immense potential of ND-immobilized RNL as an excellent candidate for biological applications and showcase the promise of further research in this field.