Production of protein extracts from Swedish red, green, and brown seaweeds, Porphyra umbilicalis Kützing, Ulva lactuca Linnaeus, and Saccharina latissima (Linnaeus) J. V. Lamouroux using three different methods

peer-reviewedThe demand for vegetable proteins increases globally and seaweeds are considered novel and promising protein sources. However, the tough polysaccharide-rich cell walls and the abundance of polyphenols reduce the extractability and digestibility of seaweed proteins. Therefore, food grade, scalable, and environmentally friendly protein extraction techniques are required. To date, little work has been carried out on developing such methods taking into consideration the structural differences between seaweed species. In this work, three different protein extraction methods were applied to three Swedish seaweeds (Porphyra umbilicalis, Ulva lactuca, and Saccharina latissima). These methods included (I) a traditional method using sonication in water and subsequent ammonium sulfate-induced protein precipitation, (II) the pH-shift protein extraction method using alkaline protein solubilization followed by isoelectric precipitation, and (III) the accelerated solvent extraction (ASE®) method where proteins are extracted after pre-removal of lipids and phlorotannins. The highest protein yields were achieved using the pH-shift method applied to P. umbilicalis (22.6 ± 7.3%) and S. latissima (25.1 ± 0.9%). The traditional method resulted in the greatest protein yield when applied to U. lactuca (19.6 ± 0.8%). However, the protein concentration in the produced extracts was highest for all three species using the pH-shift method (71.0 ± 3.7%, 51.2 ± 2.1%, and 40.7 ± 0.5% for P. umbilicalis, U. lactuca, and S. latissima, respectively). In addition, the pH-shift method was found to concentrate the fatty acids in U. lactuca and S. latissima by 2.2 and 1.6 times, respectively. The pH-shift method can therefore be considered a promising strategy for producing seaweed protein ingredients for use in food and feed

Switchable surfactant-assisted carbon nanotube coatings: innovation through pH shift

The inner surface of fused-silica capillaries has been coated with a dense/homogeneous coating of commercial multi-wall carbon nanotubes (MWCNTs) using a stable ink as deposit precursor. Solubilization of the MWCNTs was achieved in water/ethanol/dimethylformamide by the action of a surfactant, which can switch between a neutral or an ionic form depending on the pH of the medium, which thus becomes the driving force for the entire deposition process. Careful control of the experimental conditions has allowed us to selectively deposit CNTs on the inner surface of insulating silica capillaries by a simple, reproducible, and easily adaptable method.The authors gratefully acknowledge the funding from the Ministerio de Economía y Competitividad, Generalitat Valenciana, and FEDER (Projects CTQ2012-31762, MAT2013-42007-P, PrometeoII/2014/10, JCI-2012-12664, and RyC 2009-03913)

In vitro bioaccessibility of proteins and lipids of pH-shift processed Nannochloropsis oculata microalga

The pH-shift process fractionates biomass into soluble proteins and insoluble fractions, followed by precipitation and recovery of the solubilized proteins. Nannochloropsis oculata in seawater was subjected to the pH-shift process, followed by digestion of various intermediates and product fractions of the process, using the Infogest in vitro digestion model (Minekus et al., 2014) with added gastric lipase. As measures for protein and lipid accessibility, degrees of protein hydrolysis and fatty acid liberation were assessed post-digestion and compared to the amounts of peptide bonds and total fatty acids present in the raw materials. Results showed that neither proteins nor lipids of intact Nannochloropsis cells were accessible to the mammalian digestive enzymes used in the digestion model. Cell disruption, and to a lesser extent, further pH-shift processing with protein solubilisation at pH 7 or pH 10, increased the accessibility of lipids. For proteins, differences amongst the pH-shift processed materials were non-significant, though pre-freezing the product prior to digestion increased the accessibility from 32% to 47%. For fatty acids, pH-shift process-products gave rise to 43% to 52% lipolysis, with higher lipolysis for products solubilised at pH 10 as opposed to pH 7. Our results indicate the importance of processing to produce an algal product that has beneficial nutritional properties when applied as food or feed

Application of Filtration to Recover Solubilized Proteins During pH-Shift Processing of Blue Whiting (Micromesistius poutassou); Effects on Protein Yield and Qualities of Protein Isolates

Previous studies of the pH-shift protein isolation process have shown that substantial amounts of solubilized proteins can be trapped in the sediments formed in the first centrifugation step of this process. As a strategy to improve the protein yield during pH-shift processing, the aim of this study was to evaluate how filtration as an alternative to centrifugation in the first separation step of pH-shift processing of blue whiting affected proteins yield and protein isolate characteristics (basic composition, polypeptide profiles, surimi gel quality and color attributes). The study comprised both the acid and alkaline versions of the method, and also fresh as well as frozen fish raw material. Results showed that the replacement of centrifugation with filtration substantially improved the protein yield by from about 38% to 62%, but also reduced the removal of lipid. There were no significant effects on gel quality. Protein isolates from fresh raw material were about 5 % whiter and frozen raw materials about 3% whiter with centrifugation as compared to filtration in the pHshift process. For surimi from fresh raw material centrifugation gave about 2 % whiter gels, while the gels from frozen raw material were about 3% whiter for filtered compared to centrifuged material. The whitest isolates and gels were obtained with acid processing of fresh blue whiting. Slight proteolytic breakdown resulting in fragments of 83 and 152 kDa was however noted with the acid process, especially when centrifugation was used

<i>Clitoria ternatea</i> L. flower extract inhibits α-amylase during <i>in vitro </i>starch digestion

This study aimed to investigate the inhibitory effect of Clitoria ternatea flower against α-amylase during simulated in vitro wheat starch digestion. The dark-blue tropical flower is used as a food colorant but its ability to modulate starch digestion has not been tested before. The aqueous extract of the flower containing anthocyanins was a competitive inhibitor against α-amylase with an IC50 value (concentration of inhibitor required to reduce the enzyme activity by half) and inhibition constant, Ki, of 0.91 mg/mL and 0.75 mg/mL,respectively. Subjecting the extract to pasteurisation (72oC for 15 s) and boiling (for 30 min) it significantly (P&lt;0.05) decreased the anthocyanin content as determined by a pH-shift method, although the light absorbance profile of the extract remained virtually unchanged, suggesting that the equilibrium mixture of anthocyanin species was unaffected. The thermal degradation of the anthocyanins explained the partial loss of inhibition activity of the extract, as indicated by the decrease in Michaelis-Menten constant, Km, from 14.8 mg/mL in thesystems with unheated extract to 11.3 and 6.1 mg/mL in pasteurised and boiled extracts, respectively. The thermal treatments, however, did not change the type (competitive) of inhibition. The results from this work demonstrated the potential of C. ternatea flower extract in inhibiting α-amylase during starch digestion, which might lead to development of functional food/drink for controlling postprandial blood glucose level

Comparative study of the potential of poly(2-ethyl-2-oxazoline) as carrier in the formulation of amorphous solid dispersions of poorly soluble drugs

Despite the fact that solid dispersions are gaining momentum, the number of polymers that have been used as a carrier during the past 50 years is rather limited. Recently, the poly(2-alkyl-2-oxazoline) (PAOx) polymer class profiled itself as a versatile platform for a wide variety of applications in drug delivery, including their use as amorphous solid dispersion (ASD) carrier. The aim of this study was to investigate the potential of poly(2-ethyl-2-oxazoline) (PEtOx) by applying a benchmark approach with well-known, commercially available carriers (ie. polyvinylpyrrolidone (PVP) K30, poly(vinylpyrrolidone-co-vinyl acetate) (PVP-VA) 64 and hydro-xypropylmethylcellulose (HPMC)). For this purpose, itraconazole (ITC) and fenofibrate (FFB) were selected as poorly water-soluble model drugs. The four polymers were compared by establishing their supersaturation maintaining potential and by investigating their capability as carrier for ASDs with high drug loadings. Spray drying, as well as hot melt extrusion and cryo-milling were implemented as ASD manufacturing technologies for comparative evaluation. For each manufacturing technique, the formulations with the highest possible drug loadings were tested with respect to in vitro drug release kinetics. This study indicates that PEtOx is able to maintain supersaturation of the drugs to a similar extent as the commercially available polymers and that ASDs with comparable drug loadings can be manufactured. The results of the in vitro dissolution tests reveal that high drug release can be obtained for PEtOx formulations. Overall, proof-of-concept is provided for the potential of PEtOx for drug formulation purposes

Possibilities and limitations of direct detection of protein charges by means of an immunological field-effect transistor

An outline of the requirements for the construction of an immunological field-effect transistor (ImmunoFET) which should operate on the direct potentiometric sensing of protein charges is given. Selectivity of the ImmunoFET can be obtained by immobilizing antibodies on the gate area of the ISFET, enhancing the surface affinity to the corresponding antigens over other molecules in the solution. A theoretical approach is given based on the Donnan equilibrium description, which provides an insight into the potential and ion distribution in the protein layer on the ImmunoFET. It is shown that the Donnan potential and the internal pH shift, induced by the protein charges, compensate each other to a great extent. If the ISFET shows Nernstian behaviour, it is concluded that a direct detection of protein charge is impossible. In order to construct an ImmunoFET, a reference FET (REFET) or ISFET with low sensitivity would satisfy the detection of the partially compensated Donnan potential in the presence of an adsorbed protein layer. However, the application of such an ImmunoFET is limited to samples with low ionic strength