Structural Elucidation and Immunostimulatory Activities of Quinoa Non-starch Polysaccharide Before and After Deproteinization.

Non-starch polysaccharides derived from natural resources play a significant role in the field of food science and human health due to their extensive distribution in nature and less toxicity. In this order, the immunostimulatory activity of a non-starch polysaccharide (CQNP) from Chenopodium quinoa was examined before and after deproteination in murine macrophage RAW 264.7 cells. The chemical composition of CQNP and deproteinated-CQNP (D-CQNP) were spectrometrically analysed that revealed the presence of carbohydrate (22.7 ± 0.8% and 39.5 ± 0.8%), protein (41.4 ± 0.5% and 20.8 ± 0.5%) and uronic acid (8.7 ± 0.3% and 6.7 ± 0.2%). The monosaccharide composition results exposed that CQNP possesses a high amount of arabinose (34.5 ± 0.3) followed by galactose (26.5 ± 0.2), glucose (21.9 ± 0.3), rhamnose (7.0 ± 0.1), mannose (6.0 ± 0.1) and xylose (4.2 ± 0.2). However, after deproteination, a difference was found in the order of the monosaccharide components, with galactose (41.1 ± 0.5) as a major unit followed by arabinose (34.7 ± 0.5), rhamnose (10.9 ± 0.2), glucose (6.6 ± 0.2), mannose (3.4 ± 0.2) and xylose (3.2 ± 0.2). Further, D-CQNP potentially stimulate the RAW 264.7 cells through the production of nitric oxide (NO), upregulating inducible nitric oxide synthase (iNOS) and various pro-inflammatory cytokines including interleukin (IL)-1ß, IL-6, IL-10, and tumor necrosis factor-alpha (TNF-α). Moreover, stimulation of RAW 264.7 cells by D-CQNP takes place along the NF-κB and the MAPKs signaling pathways through the expression of cluster of differentiation 40 (CD40). This results demonstrate that RAW 264.7 cells are effectively stimulated after removal of the protein content in C. quinoa non-starch polysaccharides, which could be useful for develop a new immunostimulant agent.

Eliminating protein interference when quantifying potato reducing sugars with the miniaturized Somogyi-Nelson assay.

Reducing sugar (RS) quantification is essential in the potato industry because RS content plays a vital role in potato quality, acrylamide formation, post-harvest management, and new variety development. A miniaturized Somogyi-Nelson (SN) analysis can effectively and accurately quantify RS. However, soluble proteins in potatoes interfere with SN analysis. Our research goal was to develop an applicable deprotinization procedure without influencing the precision of the SN analysis. Results showed ethanol effectively removed potato proteins and, unlike other chemicals (salts, acids), ethanol did not affect SN accuracy. Protein removal also can be achieved by heating and pH adjustment, but the ethanol-based procedure provides a simpler alternative. RS content measured by the miniaturized SN assay after deproteinization by ethanol was precise and validated by HPAEC-PAD. Data from 118 potao juicies showed that a commonly used biochemical analyzer obtained a lower reducing sugar content than the deprotinization-SN assay because fructose was not identified by the biochemical analyzer. Results demonstrate the reliability of quantifying potato RS with the SN assay following the ethanol-based deproteinization.

Enzymatic approaches in the bioprocessing of shellfish wastes.

Several tonnes of shellfish wastes are generated globally due to the mass consumption of shellfish meat from crustaceans like prawn, shrimp, lobster, crab, Antarctic krill, etc. These shellfish wastes are a reservoir of valuable by-products like chitin, protein, calcium carbonate, and pigments. In the present scenario, these wastes are treated chemically to recover chitin by the chitin and chitosan industries, using hazardous chemicals like HCl and NaOH. Although this process is efficient in removing proteins and minerals, the unscientific dumping of harmful effluents is hazardous to the ecosystem. Stringent environmental laws and regulations on waste disposal have encouraged researchers to look for alternate strategies to produce near-zero wastes on shellfish degradation. The role of enzymes in degrading shellfish wastes is advantageous yet has not been explored much, although it produces bioactive rich protein hydrolysates with good quality chitin. The main objective of the review is to discuss the potential of various enzymes involved in shellfish degradation and their opportunities and challenges over chemical processes in chitin recovery.

Sustainable and eco-friendly strategies for shrimp shell valorization.

Among the seafood used globally, shellfish consumption is in great demand. The utilization of these shellfish such as prawn/shrimp has opened a new market for the utilization of the shellfish wastes. Considering the trends on the production of wealth from wastes, shrimp shell wastes seem an important resource for the generation of high value products when processed on the principles of a biorefinery. In recent years, various chemical strategies have been tried to valorize the shrimp shell wastes, which required harsh chemicals such as HCl and NaOH for demineralization (DM) and deproteination (DP) of the shrimp wastes. Disposal of chemicals by the chitin and chitosan industries into the aquatic bodies pose harm to the aquatic flora and fauna. Thus, there has been intensive efforts to develop safe and sustainable technologies for the management of shrimp shell wastes. This review provides an insight about environmentally-friendly methods along with biological methods to valorize the shrimp waste compared to the strategies employing concentrated chemicals. The main objective of this review article is to explain the utilization shrimp shell wastes in a productive manner such that it would be offer environment and economic sustainability. The application of valorized by-products developed from the shrimp shell wastes and physical methods to improve the pretreatment process of shellfish wastes for valorization are also highlighted in this paper.

Batch anaerobic digestion of deproteinated malt whisky pot ale using different source inocula.

A novel process has been developed for the selective removal of protein from pot ale with recovered protein holding potential as a value-added by-product for the whisky industry. The purpose of this work was to assess the effect of deproteination on pot ale physicochemical characterisation and anaerobic digestion (AD) treatment. Pot ales were taken from five malt whisky distilleries and tested untreated, after centrifugation/filtration and after deproteination at laboratory or pilot scale. At laboratory scale, the deproteination process removed around 20% of total chemical oxygen demand (tCOD) from untreated pot ale and at least 30% dissolved copper from centrifuged pot ale. Biochemical methane potential of untreated, filtered and deproteinated pot ale obtained at pilot scale has been determined using two types of inocula from different source. Average methane yield values of 554±67, 586±24 and 501±23 Nl CH4 kg-1 VS were obtained for untreated, filtered and deproteinated pot ale respectively. A significant difference in methane yield was only observed for untreated pot ale using the two types of inocula. Specifically, when using a non-adapted inoculum untreated pot ale biogas yield was significant lower suggesting inhibition of the AD process. As no significant differences were found for treated pot ale (filtered and deproteinated) with the two inocula it suggests, deproteination may have a positive effect on AD start-up. The results present a clear case for continuation of this work and evaluating the effect on continuous AD.

Study of Endochondral Ossification in Human Fetalcartilage Anlagen of Metacarpals: Comparative Morphology of Mineral Deposition in Cartilage and in the Periosteal Bone Matrix.

The progression of mineral phase deposition in hypertrophic cartilage and periosteal bone matrix was studied in human metacarpals primary ossification centers before vascular invasion began. This study aimed to provide a morphologic/morphometric comparative analysis of the calcification process in cartilage and periosteal osteoid used as models of endochondral ossification. Thin, sequential sections from the same paraffin inclusions of metacarpal anlagen (gestational age between the 20th and 22nd weeks) were examined with light microscopy and scanning electron microscopy, either stained or heat-deproteinated. This process enabled the analysis of corresponding fields using the different methods. From the initial CaPO4 nucleation in cartilage matrix, calcification progressed increasing the size of focal, globular, randomly distributed deposits (size range 0.5-5 µm), followed by aggregation into polycyclic clusters and finally forming a dense, compact mass of calcified cartilage. At the same time, the early osteoid calcification was characterized by a fine granular pattern (size range 0.1-0.5 µm), which was soon compacted in the layer of the first periosteal lamella. Scanning electron microscopy of heat-deproteinated sections revealed a rod-like hydroxyapatite crystallite pattern, with only size differences between the early globular deposits of the two calcifying matrices. The morphology of the early calcium deposits was similar in both cartilage and osteoid, with variations in size and density only. However, integration of the reported data with the actual hypotheses of the mechanisms of Ca concentration suggested that ion transport was linked to the progression of the chondrocyte maturation cycle (with recall of H2 O from the matrix) in cartilage, while ions transport was an active process through the cell membrane in osteoid. Other considered factors were the collagen type specificity and the matrix fibrillar texture. Anat Rec, 301:571-580, 2018. © 2017 Wiley Periodicals, Inc.

Effects of ultrasound irradiation on enzymatic hydrolysis of protein and application for the determination of tetracyclines in complex matrices.

The widespread use of tetracyclines (TCs) in food animals has led to concerns regarding unsafe residue levels in feed, food and manure. The determination of TCs in such matrices suffers from interference by the interactions between proteins and TCs. Three deproteination methods were compared in this study. In contrast with acid deproteination, which caused a large loss of TCs due to the strong adsorption of TCs on protein precipitates, a normal enzymatic hydrolysis was confirmed to have a merit of effectively releasing TCs from protein matrices, but required treatment time as long as 16 h. The adoption of ultrasound irradiation was proposed to shorten the enzymatic hydrolysis time. After investigating the effects of ultrasound power and irradiation time, the conditions of the ultrasound-enhanced enzymatic hydrolysis were optimized as ultrasound power of 100 W and irradiation time of 6 min. The ultrasound-enhanced enzymatic hydrolysis treatment of 6 min yielded recovery of TCs (from protein-containing matrices) as high as that obtained by the normal enzymatic hydrolysis treatment of 16 h. The acceleration effect of ultrasound irradiation was attributed to ultrasound-induced cavitation, which increased exposure of both the functional groups of trypsin and the C-terminal amino acid in the protein that was a cleavage site for trypsin attack. When the ultrasound-enhanced enzymatic hydrolysis method was used to determine TCs in complex matrices, it was found that this new method achieved recoveries of 89.5, 117.7, 110.4 and 100.0% for oxytetracycline, tetracycline, chlortetracycline and doxycycline, being much higher than those (29.6-39.4%) obtained using the common acid deproteination process. Copyright © 2017 John Wiley & Sons, Ltd.

Deproteination of whole blood for LC-MS/MS using paramagnetic micro-particles.

OBJECTIVES: Liquid chromatography tandem mass spectrometry has become increasingly popular in clinical laboratories over the last decade due to the inherent sensitivity and specificity of the technology. Nevertheless, full automation and hence application in routine laboratories is still hampered by laborious and difficult-to-automate sample pre-treatment protocols. Functionalized paramagnetic micro-particles could simplify sample pre-treatment and ease automation. We evaluated the applicability of a pre-commercial, straightforward paramagnetic micro-particle based kit for the lysis and deproteination of whole blood for the LC-MS/MS analysis of everolimus and compared the performance to our routine protein precipitation method. DESIGN AND METHODS: Samples were prepared for LC-MS/MS everolimus analysis on an Acquity UPLC chromatographic system coupled to a TQD mass spectrometer (both Waters Ltd.) using a pre-commercial MagSi-TDMprep kit and a routine protein precipitation respectively. Both pre-treatment methods were validated for imprecision, accuracy, linearity, limit of quantification, matrix effect, recovery and process efficiency. A method comparison (n=63) between both pre-treatment methods was performed. RESULTS: Imprecision and bias were within pre-defined criteria (15%) for both pre-treatment methods. Both methods were linear from 1.2 to 14.8µg/L with a limit of quantification of 0.6µg/L. Process efficiency was on average 65% for protein precipitation pre-treatment and was substantially higher for the MagSi-TDMprep method (85%). A Passing-Bablok regression showed no significant difference between the two pre-treatment methods. CONCLUSION: For everolimus in whole blood, the MagSi-TDMprep sample pre-treatment was applicable and comparable to protein precipitation for LC-MS/MS with the possible advantage of easier automation.