Chemical and Biomolecular Engineering - Research Publications

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    The effect of pH on the fat and protein within cream cheese and their influence on textural and rheological properties
    Ong, L ; Pax, AP ; Ong, A ; Vongsvivut, J ; Tobin, MJ ; Kentish, SE ; Gras, SL (Elsevier BV, 2020-12-01)
    The effect of variation in acid gel pH during cream cheese production was investigated. The gel microstructure was denser and cheese texture firmer, as the pH decreased from pH 5.0 to pH 4.3, despite the viscoelasticity of these gels remaining similar during heating. Protein hydration and secondary structure appeared to be key factors affecting both cheese microstructure and properties. Proteins within the matrix appeared to swell at pH 5.0, leading to a larger corpuscular structure; greater β-turn structure was also observed by synchrotron-Fourier transform infrared (S-FTIR) microspectroscopy and the cheese was softer. A decrease in pH led to a denser microstructure with increased aggregated β-sheet structure and a firmer cheese. The higher whey protein loss at low pH likely contributed to increased cheese hardness. In summary, controlling the pH of acid gel is important, as this parameter affects proteins in the cheese, their secondary structure and the resulting cream cheese.
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    Effects of shredding on the functionality, microstructure and proteolysis of low-moisture mozzarella cheese
    Pax, AP ; Ong, L ; Kentish, SE ; Gras, SL (ELSEVIER SCI LTD, 2021-06)
    Low-moisture mozzarella cheese (LMMC) is commonly shredded before packaging, however, the effects of shredding are not fully understood. Industrially-produced block and shredded LMMC were studied during 8 weeks of storage at 4 °C. Cheese shredded on 15 d and at 8 weeks of age, coated with microcrystalline cellulose and stored in a modified atmosphere (70% N₂ and 30% CO₂), had an altered microstructure after 8 weeks compared with vacuum-packed block cheese. In the latter case the fat formed a more dispersed phase. Proteolysis was higher in shredded samples and a higher level of two bacterial proteases was detected. Despite these differences, the meltability and stretchability of the block and shredded LMMC were similar. The microstructure and functionality of cheese shredded at 15 d and stored for a further 6 weeks was similar to cheese shredded at 8 weeks, suggesting there is a flexible period for performing cheese shredding processes.
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    Structure and functionality of almond proteins as a function of pH
    Devnani, B ; Ong, L ; Kentish, S ; Gras, SL (ELSEVIER, 2021-10)
    Almond proteins have potential utility in a range of food and beverages but it is not clear how pH affects protein structure and function. The behaviour of almond protein isolate was examined under conditions of neutral and acidic pH (pH 7 and 4). The isolate was highly soluble (70–80%) at either pH. An increase in acidity lead to protein unfolding, an increase in random coil structure and the appearance of lower molecular weight proteins due to acidic hydrolysis. These structural changes at pH 4 increased the capacity for foam formation and foam stability, increased viscosity and led to concentration and age dependent thickening. Gels, similar in strength but with distinct microstructures and properties were obtained following heating. At pH 7, a particulate type gel with an interconnected protein network was formed, while the gel at pH 4 had a dense continuous protein matrix. The gels differed in their susceptibility to chemical disruption, suggesting different underlying molecular interactions. The ability to alter protein structure and properties as a function of pH and heating could be used to broaden the application of almond proteins and develop a variety of food products, such as protein supplements and vegan alternatives to traditional products.
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    Pilot scale concentration of cheese whey by forward osmosis: A short-cut method for evaluating the effective pressure driving force
    Artemi, A ; Chen, GQ ; Kentish, SE ; Lee, J (Elsevier, 2020-11-01)
    Cheese whey was concentrated to a concentration factor of 2.7 in a pilot scale forward osmosis filtration system, using a commercial cellulose triacetate membrane in a spiral-wound configuration. The whey was concentrated in a batch mode, using sodium chloride as the draw solution at initial osmotic pressures of 53–75 bar. During the process, flux was shown to reduce due to the simultaneous decrease in the bulk osmotic pressure of the draw solution, increase in the bulk osmotic pressure of the whey and the effect of concentration polarisation on both sides of the membrane. The flux is known to be driven by the effective osmotic pressures of whey and the draw solution on the surface of the membrane active layer. A short-cut approach that requires minimal information in advance about the osmotic pressure of whey and the geometry of the filtration system was implemented, enabling the determination of these effective osmotic pressures. The results obtained were shown to be in agreement with the fundamental forward osmosis flux model. The short-cut approach can be utilised for estimating effective osmotic pressures of other liquid food streams to be concentrated by forward osmosis, without the need of external measurements.
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    The relevance of critical flux concept in the concentration of skim milk using forward osmosis and reverse osmosis
    Artemi, A ; Chen, GQ ; Kentish, SE ; Lee, J (Elsevier BV, 2020-10-01)
    Skim milk was concentrated at 10 °C using forward osmosis (FO), reverse osmosis (RO) and pressure-assisted forward osmosis (PAFO). A pressure of 40 bar, in the form of draw solution osmotic pressure (FO and PAFO modes) or transmembrane hydraulic pressure (RO mode) was applied; an additional hydraulic pressure of 2 bar was applied in the PAFO mode. More severe protein fouling was observed in RO, followed by PAFO and then FO. This was credited to the difference in the initial permeate flux, induced by the different effective driving pressures, with RO having a greater deviation of the initial flux from the critical flux value. The critical flux was determined for the FO and RO modes using a step-wise increase of draw solution osmotic pressure or hydraulic pressure, at a constant milk solids content. The critical flux was between 5.4 L/m2h (1.5 × 10−6 m3/m2s) and 7.2 L/m2h (2 × 10−6 m3/m2s) for both the FO and RO modes at a cross flow velocity of 0.2 m/s. The similarities in the critical flux for FO and RO suggests that the critical flux does not depend on the nature of pressure applied on the system (hydraulic or osmotic). Therefore, when operated at the same flux and crossflow velocity, FO would not fundamentally provide a lower fouling environment compared to RO. An increase of the solids content from 8.7% to 17.3% caused a reduction in the critical flux from 5.4 L/m2h to 3.1 L/m2h (8.5 × 10−7 m3/m2s).
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    Synthesis of N-Acetyllactosamine and N-Acetyllactosamine-Based Bioactives
    Alavijeh, MK ; Meyer, AS ; Gras, SL ; Kentish, SE (AMER CHEMICAL SOC, 2021-07-14)
    N-Acetyllactosamine (LacNAc) or more specifically β-d-galactopyranosyl-1,4-N-acetyl-d-glucosamine is a unique acyl-amino sugar and a key structural unit in human milk oligosaccharides, an antigen component of many glycoproteins, and an antiviral active component for the development of effective drugs against viruses. LacNAc is useful itself and as a basic building block for producing various bioactive oligosaccharides, notably because this synthesis may be used to add value to dairy lactose. Despite a significant amount of information in the literature on the benefits, structures, and types of different LacNAc-derived oligosaccharides, knowledge about their effective synthesis for large-scale production is still in its infancy. This work provides a comprehensive analysis of existing production strategies for LacNAc and important LacNAc-based structures, including sialylated LacNAc as well as poly- and oligo-LacNAc. We conclude that direct extraction from milk is too complex, while chemical synthesis is also impractical at an industrial scale. Microbial routes have application when multiple step reactions are needed, but the major route to large-scale biochemical production will likely lie with enzymatic routes, particularly those using β-galactosidases (for LacNAc synthesis), sialidases (for sialylated LacNAc synthesis), and β-N-acetylhexosaminidases (for oligo-LacNAc synthesis). Glycosyltransferases, especially for the biosynthesis of extended complex LacNAc structures, could also play a major role in the future. In these cases, immobilization of the enzyme can increase stability and reduce cost. Processing parameters, such as substrate concentration and purity, acceptor/donor ratio, water activity, and temperature, can affect product selectivity and yield. More work is needed to optimize these reaction parameters and in the development of robust, thermally stable enzymes to facilitate commercial production of these important bioactive substances.
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    Purification of organic acids using electrodialysis with bipolar membranes (EDBM) combined with monovalent anion selective membranes
    Wang, Q ; Chen, GQ ; Lin, L ; Li, X ; Kentish, SE (ELSEVIER, 2021-12-15)
    The feasibility of using electrodialysis with bipolar membranes (EDBM), combined with monovalent selective anion exchange membranes was investigated to purify organic acids from fermentation broths or wastewater streams. A simulated beet molasses feed containing a mixture of monobasic lactic acid and polybasic citric acid was used for this purpose. The impact of the feed pH, the configuration of the membrane stack and the voltage applied on the selectivity of the process was investigated. At a pH of 9–10 and an electric field intensity of 9 V/cm, a lactic acid product with a purity of at least 97% can be obtained by using both two chamber (BP-A) and three chamber (BP-A-C) configurations with the monovalent selective membrane. When using the BP-A configuration, the hydroxide ions generated by the bipolar membrane compete with the lactate anions to move into the acid solution and so the energy efficiency is lower than with the BP-A-C configuration. To mimic a multiple pass process and increase the lactic acid concentration of the final product, experiments were performed across a range of volume ratios between the feed and acid solutions. As this ratio is varied, the purity of lactic acid produced remains higher than 95% while the energy consumption is essentially unaffected. Due to the osmotic flow of water during experiments, the highest lactic acid concentration that can be achieved is limited to 153 g/L at a volume ratio (VA: VF) of 1:10.
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    Direct Air Capture of CO2 by Microalgae with Buoyant Beads Encapsulating Carbonic Anhydrase
    Xu, X ; Kentish, SE ; Martin, GJO (AMER CHEMICAL SOC, 2021-07-26)
    Microalgae cultures have promise as a CO2 sink for atmospheric carbon and as a sustainable source of food and chemical feedstocks. However, large-scale microalgae cultivation is currently limited by the need to provide carbon dioxide from point sources, as the diffusion of atmospheric CO2 is too slow. Carbonic anhydrase (CA) is an effective enzyme to facilitate the dissolution of atmospheric CO2 that could be used to enhance the photosynthetic uptake of this greenhouse gas. Here we investigate a means of retaining CA at the surface of algae ponds to facilitate direct air capture by cross-linking CA with glutaraldehyde (GA) before encapsulation into buoyant calcium alginate beads. Coomassie Blue dyeing and Wilbur-Anderson assays confirmed the successful bonding of CA to the beads. Microscopic images showed the paraffin-embedded alginate framework. The CA-GA beads retain virtually all hydrase activity throughout 10 assay cycles. Compared with a natural growth rate of 22.7 ± 0.5 mg L-1 day-1, free CA and CA-GA beads increased the productivity of Nannochloropsis salina to 37 ± 3 mg L-1 day-1 and 40 ± 1 mg L-1 day-1, respectively. The CA-GA beads further provided a stable growth enhancement for three rounds of microalgae cultivation, confirming that these buoyant beads can be readily recovered and re-used, which is promising for industrial biomass production.
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    The influence of propane and n-butane on the structure and separation performance of cellulose acetate membranes
    Liu, L ; Doherty, CM ; Ricci, E ; Chen, GQ ; De Angelis, MG ; Kentish, SE (Elsevier BV, 2021-11-15)
    This work presents the impact of propane and n-butane on the CO2/CH4 separation performance of both cellulose diacetate (CDA) and cellulose triacetate (CTA) membranes by exposing both pristine membranes to either propane (400 kPa) or n-butane (200 kPa) at room temperature (22 ± 2 °C) for 4 weeks. The propane and n-butane sorption isotherms in both membranes were anomalous at 35 °C. X-ray diffraction (XRD) results indicated that the crystalline nature of both polymers was altered by this exposure, although dynamic scanning calorimetry (DSC) did not detect a significant change in the overall crystallinity. Positron Annihilation Lifetime Spectroscopy (PALS) revealed that the average pore size of the CTA polymer and the number of free volume elements of both membranes also increased, even though the sorption uptake was less than 2 wt%. CO2 and CH4 permeabilities at 35 °C were essentially unaffected by the propane or n-butane exposure, indicating that while the crystalline regions of the polymer were affected, plasticization of the glassy amorphous region did not occur. There was a slight decrease in CH4 permeability for the CDA membrane after n-butane exposure, consistent with a slight decline in the CH4 solubility at this feed pressure. The propane and n-butane permeabilities were 0.029 Barrer at 300 kPa and 0.019 Barrer at 125 kPa for the fresh CTA membrane, but these fell significantly after long term exposure to these gases, possibly due to penetrant clustering.
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    Improving β-Galactosidase-Catalyzed Transglycosylation Yields by Cross-Linked Layer-by-Layer Enzyme Immobilization
    Alavijeh, MK ; Meyer, AS ; Gras, SL ; Kentish, SE (AMER CHEMICAL SOC, 2020-11-02)
    The biotransformation of lactose into gut-bioactive glycans catalyzed by β-galactosidase can give economic value to lactose-rich side streams generated in the food or the dairy industry. Herein, we study the immobilization of the commercially used β-galactosidase from Bacillus circulans onto silica particles using an enzyme immobilization technology involving a cross-linked layer-by-layer encapsulation method. The immobilized β-galactosidase was used for the synthesis of N-acetyllactosamine (LacNAc) as an important precursor for numerous bioactive compounds and a prebiotic in itself. Techniques including molecular analysis, enzyme activity determination, secondary structure analysis, thermodynamic characterization, and the determination of thermal and operational stability were conducted to characterize the immobilized enzyme. Changes in the activity of the enzyme after immobilization were attributed to possible changes in electrostatic, covalent, and protein-protein interactions. Immobilization significantly improved the enzymatic LacNAc yield compared to the free enzyme. In turn, this improved the economics and the sustainability of the process. The immobilized enzyme encapsulated in multilayer films was significantly more stable in the presence of divalent cations and its thermostability also substantially increased with the thermal denaturation activation energy increasing from 53 to 294 kJ mol-1. The immobilized enzyme was successfully reused in eight consecutive reaction cycles with no significant reduction in the LacNAc yield. The improved transgalactosylation yield and productivity, higher stability, and reusability obtained with this immobilization method provide new opportunities for industrial applications.