BackgroundSolid-binding peptides (SBPs) bind strongly to a diverse range of solid materials without the need for any chemical reactions. They have been used mainly for the functionalisation of nanomaterials but little is known about their use for the immobilisation of thermostable enzymes and their feasibility in industrial-scale biocatalysis.ResultsA silica-binding SBP sequence was fused genetically to three thermostable hemicellulases. The resulting enzymes were active after fusion and exhibited identical pH and temperature optima but differing thermostabilities when compared to their corresponding unmodified enzymes. The silica-binding peptide mediated the efficient immobilisation of each enzyme onto zeolite, demonstrating the construction of single enzyme biocatalytic modules. Cross-linked enzyme aggregates (CLEAs) of enzyme preparations either with or without zeolite immobilisation displayed greater activity retention during enzyme recycling than those of free enzymes (without silica-binding peptide) or zeolite-bound enzymes without any crosslinking. CLEA preparations comprising all three enzymes simultaneously immobilised onto zeolite enabled the formation of multiple enzyme biocatalytic modules which were shown to degrade several hemicellulosic substrates.ConclusionsThe current work introduced the construction of functional biocatalytic modules for the hydrolysis of simple and complex polysaccharides. This technology exploited a silica-binding SBP to mediate effectively the rapid and simple immobilisation of thermostable enzymes onto readily-available and inexpensive silica-based matrices. A conceptual application of biocatalytic modules consisting of single or multiple enzymes was validated by hydrolysing various hemicellulosic polysaccharides.
The use of fluoride is effective in preventing dental caries. However, an excessive intake of fluoride leads to dental fluorosis, making it necessary to regularly monitor the fluoride intake especially for infants. There is hitherto a lack of information on fluoride content in infant foods from an Australian perspective. Therefore, this study aims to estimate the amount of fluoride content from a range of commercially available ready-to-eat (RTE) infant foods and drinks available in Australia. Based on an external calibration method, potentiometry involving a fluoride ion selective electrode and a silver|silver chloride reference electrode was conducted to analyse the fluoride content of a total of 326 solid food samples and 49 liquid food samples in this work. Our results showed an overall median (range) fluoride content of 0.16 (0.001–2.8) µg F/g of solid food samples, and 0.020 (0.002–1.2) µg F/mL of liquid food samples. In addition, ~77.5% of the liquid samples revealed a fluoride content < 0.05% µg F/mL. The highest variation of fluoride concentration (0.014–0.92 µg F/g) was found in formulas for ≥6 month-old infants. We have attributed the wide fluoride content variations in ready-to-eat infant foods and drinks to the processing steps, different ingredients and their origins, including water. In general, we found the fluoride content in most of the collected samples from Australian markets to be high and may therefore carry a risk of dental fluorosis. These results highlight the need for parents to receive appropriate information on the fluoride content of ready-to-eat infant food and drinks.
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