Chemical and Biomolecular Engineering - Research Publications
Permanent URI for this collection
Search Results
1 - 10 of 51
-
ItemThe effect of pH on the fat and protein within cream cheese and their influence on textural and rheological properties(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.
-
ItemEffects of Freeze-Thaw Phenomena on Controlled Nutrient Release: Application to Bioremediation(WILEY-BLACKWELL, 2016-12)Cold region soils are often devoid of sufficient nutrients essential for timely bioremediation of petroleum hydrocarbon contaminants. While materials that release nutrients in a controlled manner have been shown to stimulate biodegradation over extended time periods, the physical and chemical response of these materials to repetitive freeze–thaw stresses typical of cold regions remains poorly understood. This study reports the performance of four controlled release materials in water (Osmocote™, Nutricote™, Polyon™, Zeopro™) exposed to freeze–thaw cycling or control temperature of 4°C. Additionally Zeopro™‐activated carbon mixtures are investigated for application to permeable reactive barriers. Osmocote™ experienced higher nutrient release under control conditions compared with samples exposed to freeze–thaw after 20 days. Nutrient release from Nutricote™ and Polyon™ was similar under freeze–thaw and control conditions. Zeopro™ delivered low nutrient concentrations into solution in both freeze–thaw and control samples with calcium phosphate dissolution accelerated in the presence of activated carbon. Osmocote™, Nutricote™ and Polyon™ revealed strong resistance to breakdown under freeze–thaw. Zeopro™ experienced only partial disintegration, while significant break‐up of activated carbon occurred under freeze–thaw. A physical and chemical understanding of the response of multiple fertilisers can guide the selection of materials for biostimulation and biodegradation of petroleum hydrocarbons in environments exposed to freeze–thaw cycling.
-
ItemExtrusion of a Curcuminoid-Enriched Oat Fiber-Corn-Based Snack Product(WILEY, 2019-02)Extruded snack products were made from an oat fiber-corn flour matrix fortified with 1.5% (w/w) curcuminoids (750 mg curcuminoids/100 g) to improve the solubility and stability of curcuminoids. The effects of extruder feed moisture content (21%, 28%, and 35%) and screw speed (200 and 300 rpm) on the extrusion parameters and physical properties of final snacks were investigated. Curcuminoids lost during extrusion and curcuminoids loss during subsequent drying of extrudates were analyzed, to separate the losses occurring in each unit process. Drying post extrusion (at 50 °C for 4 hr) was essential to obtain a crunchy shelf stable product (5% moisture). Curcuminoids loss during extrusion was from 17% to 84%, with high loss for the extrusion with low feed moisture content (21%). A further curcuminoids loss of 4% to 44% occurred during drying, with much higher loss for the extrudate with high moisture content. Total curcuminoids retained after extrusion and drying was 12% to 41% (59% to 88% loss), equivalent to 180 to 616 mg curcuminoids retained per 100 g snack, levels within recommended daily dose. Curcuminoids retained after drying was stable during 80 days of storage at 25 °C. The results highlighted the importance of understanding the impact of each unit process separately (for example, extrusion and drying) on the stability of curcuminoids for the development of healthier extruded snacks. PRACTICAL APPLICATION: Extruded snacks products were developed by fortifying the snacks with oat fiber and curcuminoids in order to address the need for a healthy ready to eat food products. Some extrusion characteristics were selected to produce snack products which have favorable properties in terms of consumer acceptance.
-
Item
-
Item
-
ItemImproving β-Galactosidase-Catalyzed Transglycosylation Yields by Cross-Linked Layer-by-Layer Enzyme Immobilization(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.
-
ItemCytochrome P450-mediated N-demethylation of noscapine by whole-cell biotransformation: process limitations and strategies for optimisation(Springer Verlag, 2020-07-01)Cytochrome P450 enzymes catalyse reactions of significant industrial interest but are underutilised in large-scale bioprocesses due to enzyme stability, cofactor requirements and the poor aqueous solubility and microbial toxicity of typical substrates and products. In this work, we investigate the potential for preparative-scale N-demethylation of the opium poppy alkaloid noscapine by a P450BM3 (CYP102A1) mutant enzyme in a whole-cell biotransformation system. We identify and address several common limitations of whole-cell P450 biotransformations using this model N-demethylation process. Mass transfer into Escherichia coli cells was found to be a major limitation of biotransformation rate and an alternative Gram-positive expression host Bacillus megaterium provided a 25-fold improvement in specific initial rate. Two methods were investigated to address poor substrate solubility. First, a biphasic biotransformation system was developed by systematic selection of potentially biocompatible solvents and in silico solubility modelling using Hansen solubility parameters. The best-performing biphasic system gave a 2.3-fold improvement in final product titre compared to a single-phase system but had slower initial rates of biotransformation due to low substrate concentration in the aqueous phase. The second strategy aimed to improve aqueous substrate solubility using cyclodextrin and hydrophilic polymers. This approach provided a fivefold improvement in initial biotransformation rate and allowed a sixfold increase in final product concentration. Enzyme stability and cell viability were identified as the next parameters requiring optimisation to improve productivity. The approaches used are also applicable to the development of other pharmaceutical P450-mediated biotransformations.
-
ItemStructure-Dependent Interfacial Properties of Chaplin F from Streptomyces coelicolor(MDPI, 2017-09)Chaplin F (Chp F) is a secreted surface-active peptide involved in the aerial growth of Streptomyces. While Chp E demonstrates a pH-responsive surface activity, the relationship between Chp F structure, function and the effect of solution pH is unknown. Chp F peptides were found to self-assemble into amyloid fibrils at acidic pH (3.0 or the isoelectric point (pI) of 4.2), with ~99% of peptides converted into insoluble fibrils. In contrast, Chp F formed short assemblies containing a mixture of random coil and β-sheet structure at a basic pH of 10.0, where only 40% of the peptides converted to fibrils. The cysteine residues in Chp F did not appear to play a role in fibril assembly. The interfacial properties of Chp F at the air/water interface were altered by the structures adopted at different pH, with Chp F molecules forming a higher surface-active film at pH 10.0 with a lower area per molecule compared to Chp F fibrils at pH 3.0. These data show that the pH responsiveness of Chp F surface activity is the reverse of that observed for Chp E, which could prove useful in potential applications where surface activity is desired over a wide range of solution pH.
-
ItemThe Assembly of Individual Chaplin Peptides from Streptomyces coelicolor into Functional Amyloid Fibrils(PUBLIC LIBRARY SCIENCE, 2011-04-19)The self-association of proteins into amyloid fibrils offers an alternative to the natively folded state of many polypeptides. Although commonly associated with disease, amyloid fibrils represent the natural functional state of some proteins, such as the chaplins from the soil-dwelling bacterium Streptomyces coelicolor, which coat the aerial mycelium and spores rendering them hydrophobic. We have undertaken a biophysical characterisation of the five short chaplin peptides ChpD-H to probe the mechanism by which these peptides self-assemble in solution to form fibrils. Each of the five chaplin peptides produced synthetically or isolated from the cell wall is individually surface-active and capable of forming fibrils under a range of solution conditions in vitro. These fibrils contain a highly similar cross-β core structure and a secondary structure that resembles fibrils formed in vivo on the spore and mycelium surface. They can also restore the growth of aerial hyphae to a chaplin mutant strain. We show that cysteine residues are not required for fibril formation in vitro and propose a role for the cysteine residues conserved in four of the five short chaplin peptides.
-
ItemThe propensity of the bacterial rodlin protein RdlB to form amyloid fibrils determines its function in Streptomyces coelicolor(NATURE PORTFOLIO, 2017-02-17)Streptomyces bacteria form reproductive aerial hyphae that are covered with a pattern of pairwise aligned fibrils called rodlets. The presence of the rodlet layer requires two homologous rodlin proteins, RdlA and RdlB, and the functional amyloid chaplin proteins, ChpA-H. In contrast to the redundancy shared among the eight chaplins, both RdlA and RdlB are indispensable for the establishment of this rodlet structure. By using a comprehensive biophysical approach combined with in vivo characterization we found that RdlB, but not RdlA, readily assembles into amyloid fibrils. The marked difference in amyloid propensity between these highly similar proteins could be largely attributed to a difference in amino acid sequence at just three sites. Further, an engineered RdlA protein in which these three key amino acids were replaced with the corresponding residues from RdlB could compensate for loss of RdlB and restore formation of the surface-exposed amyloid layer in bacteria. Our data reveal that RdlB is a new functional amyloid and provide a biophysical basis for the functional differences between the two rodlin proteins. This study enhances our understanding of how rodlin proteins contribute to formation of an outer fibrillar layer during spore morphogenesis in streptomycetes.