Insulin Inclusion into a Tragacanth Hydrogel: An Oral Delivery System for Insulin

Nanoparticles or microparticles created by physical complexation between two polyelectrolytes may have a prospective use as an excipient for oral insulin administration. Natural polymers such as tragacanth, alginate, dextran, pullulan, hyaluronic acid, gelatin and chitosan can be potential candidates for this purpose. In this research, insulin particles were prepared by the inclusion of insulin into a tragacanth hydrogel. The effect of the pH and concentration relationship involving polyelectrolytes offering individual particle size and zeta potential was assessed by zetasizer and scanning electron microscopy (SEM). Insulin–tragacanth interactions at varying pH (3.7, 4.3, 4.6, or 6), and concentration (0.1%, 0.5%, or 1% w/w) were evaluated by differential scanning calorimetry (DSC) and ATR Fourier transform infrared (ATR-FTIR) analysis. Individual and smaller particles, approximately 800 nm, were acquired at pH 4.6 with 0.5% of tragacanth. The acid gelation test indicated that insulin could be entrapped in the physical hydrogel of tragacanth. DSC thermograms of insulin–tragacanth showed shifts on the same unloaded tragacanth peaks and suggested polyelectrolyte–protein interactions at a pH close to 4.3–4.6. FTIR spectra of tragacanth–insulin complexes exhibited amide absorption bands featuring in the protein spectra and revealed the creation of a new chemical substance

Application of membrane-based technology for purification of bromelain

About 60% of world’s commercial enzyme products are proteases, giving promising opportunity to derive such enzymes sustainably from waste sources. Bromelain is a crude protease occurring naturally in pineapple, and it possesses properties of benefit for pharmaceutical, medical and food products. The production of bromelain involves a purification stage, normally performed by small-scale conventional operations which lead to high operating cost and low product recovery, while being difficult to scale up and produce polluting by-products. Membrane-based technology offers an alternative to produce high quality purified bromelain in a more efficient and sustainable process. This review identified the current state and future needs for utilising membrane processes for sustainable bromelain production at larger scales. It was found that declining membrane flux due to fouling have been reported, but may be effectively overcome with more appropriate (and advanced) membrane types and/or processing conditions. For example, interactions between macromolecules present in the pineapple derived bromelain mixture (particularly polysaccharides) and the membrane may cause performance limiting fouling, but can be overcome by enzymatic pre-treatment. Membrane fouling can be further reduced by the employment of ceramic membrane filters operating at optimised trans-membrane pressure, cross-flow velocity, feed pH and temperature. Two-stage ultrafiltration together with diafiltration or gas sparging was suggested as a means to reduce fouling and improve enzyme purity. Despite these promising technical findings, the review identified the need for a valid economic assessment to properly guide further work towards purifying bromelain from pineapple waste for sustainable production of commercial proteases

Unravelling conformational aspects of milk protein structure-contributions from nuclear magnetic resonance studies

Changes in the molecular structure and association of milk proteins lead to many desirable (under controlled conditions) or undesirable characteristics of dairy products. Several methods have been used to study the structure of milk proteins and changes therein in different environments. Whey proteins are an excellent model for secondary structure studies using circular dichroism (CD), Fourier-transform infrared spectroscopy (FTIR) and tertiary structure studies using X-ray crystallography and nuclear magnetic resonance (NMR). However, caseins, the most abundant protein class in milk, are far more diffcult to characterize. The tertiary structure of caseins cannot be observed by X-ray crystallography due to the inability to crystallize caseins. However, NMR is an appropriate approach for structural elucidation. Thus far, NMR was applied on specific peptides of individual caseins of the molecules including phosphoserine centers and colloidal calcium phosphate. The literature focuses on these parts of the molecule due to its importance in building the sub-unit particles involving individual caseins and calcium phosphate nanoclusters. This review focuses on present structural studies of milk proteins using NMR and their importance in dairy processing.</p

Milk Protein Hydrolysis by Actinidin—Kinetic and Thermodynamic Characterisation and Comparison to Bromelain and Papain

Plant proteases, including actinidin, papain and bromelain, have been widely used in the food industry but with limited application in dairy systems. This research aimed to establish and compare operational parameters (kinetics, temperature, enzyme type, time and thermodynamics) relevant to the applications of these enzymes in the hydrolysis of whey protein isolates (WPI), whey protein concentrates (WPC) or milk protein concentrates (MPC). The degree of hydrolysis (DH) increased with the rise in temperature, and the maximum DH was achieved at 60 °C for all three dairy systems. The addition of papain resulted in a greater %DH of whey proteins in comparison to bromelain. The cleavage of proteins was clearly time-dependent (p &lt; 0.05), while the pH did not change significantly (p &gt; 0.05) during this time. PAGE analysis revealed that all three enzymes mainly acted on α-lactalbumin and αs-casein in WPI and MPC, respectively. Kinetic parameters from the Lineweaver–Burk plot at 60 °C using WPC and MPC as a substrate varied widely, establishing that WPC hydrolysis was characterised by a lower KM, higher kcat, kcat/KM and Vmax compared to MPC in the case of all three enzymes. The difference in kcat/KM values amongst all enzymes (actinidin &gt; papain &gt; bromelain) indicated the difference in the strength of substrate binding sites. The thermodynamic parameters of these enzymes with MPC and WPC were also determined at a temperature range of 15–60 °C, and the results indicate the potential application of papain and actinidin in the dairy industry