Protein Removal Rate Research Articles

Optimization of welan gum extraction and purification using lysozyme and alkaline protease.

Welan gum, a natural polysaccharide produced by Sphingomonas sp. ATCC 31555, has attracted considerable attention in the scientific community due to its desirable properties. However, challenges, such as high viscosity, residual bacterial cells, carotenoids, and protein complexation, hinder the widespread application of welan gum. In this study, we established a method for the extraction and purification of welan gum using a synergistic approach with lysozyme and alkaline protease. Lysozyme hydrolysis conditions were optimized by applying response surface methodology, and the best results for bacterial cell removal were achieved at 11 000 U/g, 44°C, and pH 9 after 3h of treatment. Subsequently, we evaluated protein hydrolysis through computer simulation and identified alkaline protease as the most suitable enzyme. Through experimental investigations, we found that the optimal conditions for alkaline protease hydrolysis were 7500 U/g, 50°C, pH 10, and 600rpm. These conditions resulted in a sugar recovery rate of 76.1%, carotenoid removal rate of 89.5%, bacterial removal rate of 95.2%, and protein removal rate of 87.3% after 3h of hydrolysis. The purified welan gum exhibited high transparency and purity. Structural characterization and antioxidant activity evaluation revealed that enzymatically purified welan gum has potential application prospects. Our study provides valuable insights into the optimal method for the enzymatic extraction and purification of welan gum. Such a method is conducive to the development of the multiple potential applications of welan gum. KEY POINTS: • A novel process for the synergistic purification of welan gum using lysozyme and alkaline protease was established. • In silico virtual digestion was employed to select the purification enzyme. • Welan gum with high transparency and purity was obtained.

Preparation and analysis of polysaccharide from Solanum tuberdsm

The crude Solanum tuberdsm polysaccharides (STP) were extracted with hot water. In the process of extraction, proteins, pigments, small molecules and salts in the mixture were removed by Sevage reagent, diatomite and distilled water dialysis, respectively. In addition, the process conditions of protein removal by response surface methodology (RSM) were optimized, and the optimum process conditions of Sevage method were established as follows: ultrasound power 350 W, ultrasound time 20 min, deproteinization twice, volume ratio of polysaccharide solution to Sevage reagent 1:1 (mL/mL). Under these conditions, the protein removal rate was 93.14%.

A green and efficient technology for sequential extraction of lipid and paramylon from Euglena gracilis

Euglena gracilis is rich in nutrients and high-value metabolites. However, at present, most studies only extract the paramylon of E. gracilis, which is a waste of resources. In this study, a method of sequential extraction of lipid and paramylon from E. gracilis was studied, and the extraction conditions were optimized. Firstly, by optimizing the solid-liquid ratio, extraction time, extraction temperature and volume ratio of n-hexane to ethanol, the lipid extraction efficacy was increased from 73.0 % to 96.0 %. Next, paramylon was extracted from the E. gracilis residue. After optimizing the enzyme type, enzyme concentration, ratio of material to liquid and enzymolysis time, the protein removal rate was 91.3 %, the yield of paramylon was 58.3 % and the purity reached 89.3 %. Compared with the traditional method, this sequential extraction method was greener and more effective for the extraction of paramylon and lipids from E. gracilis, which can promote the comprehensive utilization of algae resources.

The prebiotic properties of polysaccharides obtained by differentiated deproteinization methods from Flos Sophorae Immaturus on Lactobacillus fermentum.

The polysaccharides derived from various deproteinization methods were prepared from Flos Sophorae Immaturus (FSI) to investigate the prebiotic efficacy of Lactobacillus fermentum (L.f ). The implications of polysaccharides from FSI (PFSI) gained after purification performed by non-deproteinization and different deproteinization processes (Savage method, papain method, and TCA method) via one-factor optimization were firstly investigated for the influences on the growth of L.f. The utilization of carbohydrate sources and the synthesis of protein and lactate during its growth were analyzed, as well as the variations of LDH, SOD, and GSH- Px enzyme dynamics. The results showed that the one-factor optimization of the deproteinization process with the protein removal rate and polysaccharide retention rate as the indexes led to the optimal methods of the Sevage method with 5 elution times, papain method with 80 U/mL concentration, and TCA method with 2.5 ratio, respectively. In addition, the PFSI obtained with or without deproteinization purification had a certain effect on promoting L.f proliferation. Moreover, the PFSI gained by the third deproteinization purification, at a concentration of 10 g/L, significantly elevated L.f biomass and growth rate compared with the blank control, and the utilization of reducing sugars and the synthesis of protein and lactic acid were higher than the control (P < 0.05); improved LDH, SOD, and GSH-Px activity in L.f (P < 0.05), and the TCA method could be effectively applied to eliminate the proteins affecting FSI in probiotics, and PFSI may be a potentially beneficial prebiotic and intestinal reinforcer.

Automated preparation of washed platelet concentrates through spinning-membrane filtration.

Washed platelet concentrates (WPC), prepared with an automated system cell processor (ACP), have recently been approved to be manufactured and marketed in Japan. From the perspective of risk management, it is preferable to secure alternative technologies for ACP. Here, we conducted a study to evaluate the quality of WPC prepared using an automated membrane filtration-based system, Lovo. Replaced PCs prepared from apheresis PCs were equally divided into control and test units, and subsequently washed using ACP and Lovo respectively. Work and operational efficiencies were evaluated by in vitro analyses, including total handling time, platelet recovery, and plasma protein removal rate. Product quality, including a set of biochemical and physiological indicators of platelets and supernatants, were assessed before and 3 days after washing. In vitro platelet recovery rates and plasma protein removal rates were >85% and >95%, respectively, in both groups. The pH values on day 0 were significantly high (6.97 vs. 6.86) due to low pCO2 in the test group, while no significant differences in glucose consumption and lactate production were observed between the two groups. The levels of hypotonic shock responses, aggregation response, platelet shape, CD62P expression, and sCD62P concentration were similar in both groups during the 3-day storage period. Platelet washing with Lovo provides platelet quality equivalent to, or better than, conventional washing with ACP. Thus, the new automated system, Lovo, can be considered as an alternative to ACP for WPC preparation.

Small molecule carbon source promoting dairy wastewater treatment of Rhodospirillum rubrum by co-metabolism and the establishment of multivariate nonlinear equation.

Rhodospirillum rubrum water treatment technology could recycle bio-resource. However, the inability to degrade macromolecular organics limited its wide application. This paper discussed the feasibility of small molecular carbon source promoting R. rubrum directly treating dairy machining wastewater (DMW) and accumulations for single cell protein and pigment, and establishment of a mathematical model. Six small molecules promoted the degradation of macromolecules (proteins) in DMW. They promoted protease secretion and non-growth matrix (protein) decomposition in DMW through co-metabolism. Among the molecules, 550 mg/L potassium sodium tartrate was the best, protease activity and protein removal rate were increased by 100% compared with control. Then chemical oxygen demand (COD) and protein removal rates reached 80%, the single cell protein, carotenoid and bacterial chlorophyll yields were increased 2 times. Meanwhile, carbon nitrogen ratio (C/N) and food microbial ratio (F/M) were identified as the most important factors by principal component analysis. A multivariate nonlinear equation model between COD removal rate and C/N, F/M, time was established.

Characterization of Different Bentonites and Their Properties as a Protein-Fining Agent in Wine

Bentonite is a natural clay that is used in different industries as a desiccant, ion-exchange material, or additive to remove impurities. For example, marketed as healing clay and as protein-fining agent in wine, bentonite is expected to adsorb specific compounds while having no negative effects on the quality of the product in which it is used. In this study, 34 commercially available bentonites for different applications were selected and analyzed for their elemental composition, extraction of heavy metals, swelling behavior, and protein removal rate under conditions relevant to wine. The results indicate that bentonites can have a very variable composition that does not correlate with the intended use. The extraction of heavy metals is not directly related to the raw material and depends on swelling behavior and surface area of the clay. Interestingly, there is a similar degree of variability in swelling behavior among wine bentonites as there is for healing clays. This correlates with the protein removal rate but also with the extraction of iron, a transition metal that is known for its catalytic activity for oxidation reactions. Even though the protein removal rate is much higher than for other clays, bentonites that are marketed for wine show an extraction behavior that can have a negative effect on the final product.

Separation and purification of enveloped and non-enveloped viruses from water samples using an aqueous two-phase system

In this study, a new strategy for enriching viruses in water was developed to improve the detection of viruses in environmental samples. The virus was enriched from water by aqueous two-phase system (ATPS), and the recovery and purification were characterized by tissue culture median infection dose (TCID50) and total protein removal rate. The isolation and purification methods of envelope virus (H1N1) and non-envelope virus (human adenovirus type 5, HAdV-5E) were established and optimized. The results of ATPS were compared with those of membrane filtration, polyethylene glycol precipitation and ultrafiltration. Using ATPS, both viruses were enriched in interphase due to the combined action of volume exclusion and bottom stage salting out. Under the optimum conditions, the recoveries of H1N1 and HAdV-5E were 94.87 ± 2.37% and 96.22 ± 4.71% respectively, and the total protein removal rates were 43.02 ± 5.21% and 71.01 ± 6.34% respectively. Compared with conventional methods, ATPS showed superior performance in recovering virus content and water volume, and the average recoveries were 94.06% (H1N1) and 90.07% (HAdV-5E). Therefore, ATPS had superiorities of high recovery, simple operation and maintaining virus particle activity(Fig. 1).

Evaluation of the time-dependent efficacy of commercial denture or orthodontic appliance cleansers: An in vitro study.

The aim of this study was to evaluate the antimicrobial, stain and protein removal efficacy of denture or orthodontic appliance cleansers using in vitro test methods. Experimental cleansers were applied for experimental time in each evaluation method. To evaluate the microorganism removal efficacy, C. albicans and S. mutans removal rate was calculated from the specimen surface. Stain and protein removal rate was calculated using the spectrophotometer. Experimental cleansers significantly affected the microorganism removal rate for both C. albicans and S. mutans, as well as the stain and protein removal rates, at each experimental time (p<0.05). As the application time increased, the stain and protein removal rates of all experimental cleansers significantly increased (p<0.05). The present study provided in vitro evaluation methods to assess the efficacy of denture or orthodontic appliance cleansers. Also, manufacturers and researchers can predict clinical outcomes and ensure proper hygiene management of dentures or orthodontic appliances.

Illumination promoting biomass production in wastewater treatment for Rubrivivax gelatinosus

In order to improve the reuse degree of Rubrivivax gelatinosus (R. gelatinosus) resource in soybean producing wastewater (SPW) treatment, the impacts of diverse illumination strength on R. gelatinosus biomass and organic contaminant abatement were discussed. It was reported that with the best illumination strength (6,000 lux), the production (3,800 mg/L) of biomass was increased by sixty percent of dark group. COD removal and protein removal rate got 85 and 80%. Influence of diverse illumination strength on R. gelatinosus growth were divided into three aspects, including illumination limitation (0−5,000 lux), illumination saturation (5,000−7,000 lux), illumination inhibition (8,000 lux or higher). Meanwhile, at 6,000 lux, bacteriochlorophyll and carotenoids contents, ATP production were increased by 200, 200, 43.9% severally. Appropriate illumination improved ATP production by increasing the generation of bacteriochlorophyll and carotenoids in photosynthesis. Augment of energy production provoked the biomass production and wastewater treatment. Nevertheless, low or high illumination strength was un-advantage for R. gelatinosus growth due to the illumination deficiency or restrian. Moreover, 6,000 lux shortened the treatment time from 96 h to 48 h.

Extraction and deproteinization process of polysaccharide from purple sweet potato.

Extraction and deproteinization process of polysaccharide from purple sweet potato (PPSP) were optimized via the response surface methodology (RSM). The results indicated that the optimal conditions of extraction in hot water of PPSP were as follows: The extraction temperature was 120℃, the extraction time was 2.5hr, and the solid-liquid ratio was 1∶10 (g/ml). The optimal conditions of Sevage deproteinization were as under the oscillation time was 20min, the deproteinization times was twice, and polysaccharide solution-Sevage reagent ratio was 1:1 (ml/ml). The extraction yield of PPSP was 3.32%, and the protein removal rate was 93.14% insuchacondition.

Purification of xylooligosaccharides from bamboo with non-organic solvent to prepare food grade functional sugars

Food grade xylooligosaccharides (XOS) have a high application value, but most of the reported purification methods of XOS use organic solvents, which has significant problems such as large amount of solvent, residual solvent, and difficulty in solvent recovery. In addition, although there are many reported methods of separation and purification, quantitative analysis is lacking for each specific treatment step, such as sugar loss rate, decolorization rate, protein removal rate, and desalination rate. Therefore, purification of XOS from bamboo with non-organic solvent to prepare food grade functional sugars was presented by this study. What’s more, quantitative analysis and optimization was made for each step of separation, which was of great significance for guiding the actual production. Overall, through activated carbon adsorption, ion exchange and membrane filtration comprehensive treatments, food grade XOS were obtained from bamboo with 7.5% yield, a purity of 92.3%, and a total sugar proportion as high as 98.9%.

Orthogonal Test Design for Optimization of the Extraction of Polysaccharides from Inonotus cuticularis and Their Antioxidant Activities.

Medical fungi polysaccharides belong to a very important species of biological macromolecules, which are the basic substances that effectively maintain and ensure the normal operation of biological life activities. However, research on extraction and biological activity of Inonotus cuticularis polysaccharides has never been reported. In this study, the optimum yield of Inonotus cuticularis polysaccharides was determined by the orthogonal experimental design. The highest yield of 3.10±0.06 % was obtained with extraction temperature of 80 °C, extraction time of 150 min, and water to raw material ratio of 30 mL/g and repeated twice. After deproteinization for 5 times, the protein removal rate reached 70.10±1.75 %, and the content of polysaccharides and protein were 46.64 and 0.42 %. Infrared spectrometer indicated that Inonotus cuticularis polysaccharides are typical β-pyranose with characteristic peaks of polysaccharides. Subsequently, the activities of scavenging free radicals for the deproteinated polysaccharides were studied. When the concentration of Inonotus cuticularis polysaccharides was 0.3 mg/mL, the scavenging activities of the sample on DPPH. , . OH, ABTS.+ and O2 .- reached 83.67±0.27, 65.21±4.82, 43.45±1.36 and 80.28±2.30 %, respectively, and the reducing power reached 0.46±0.01. The IC50 values scavenging DPPH. , . OH, ABTS.+ and O2 .- were 0.139±0.13, 0.162±0.14, 0.317±0.30 and 0.121±0.10 mg/mL, respectively. Results showed that Inonotus cuticularis polysaccharides present potential stronger antioxidant activities, especially .OH scavenging activity and reducing power. Experimental results could provide research basis of Inonotus cuticularis polysaccharides for further exploitation and utilization.

Isolation of lactoferrin and immunoglobulins from dairy whey by an electrodialysis with filtration membrane process

Isolation of high value minor proteins such as lactoferrin and immunoglobulins from crude dairy streams is a major challenge for the dairy industry. Here we investigate an electrodialysis with filtration membrane (EDFM) approach to separate lactoferrin (LF) and immunoglobulins (Ig) from other dairy proteins. A polyvinyl alcohol (PVA) membrane is prepared using phase inversion in a coagulation bath with 80% ethanol to serve as the filtration membrane. A range of treatment times and two electric field strengths (38.5 and 77 V/cm) were then investigated within the electrical cell. The results show that the filtration membrane prepared in-house can offer strong rejection for LF and Ig while allowing a high flux of other proteins. The separation of LF and Ig from a simulated whey solution can be achieved. The use of a partially demineralised whey reduces protein loss, but there is a tradeoff between the energy consumption and the protein removal rate as the extent of this demineralisation is increased.

Domestic wastewater hydrolysis and lipolysis during start-up in anaerobic digesters and microbial fuel cells at moderate temperatures

Raw wastewater hydrolysis rates during start-up in microbial fuel cells (MFCs) and anaerobic digestion (AD) systems, seeded with a mesophilic inoculum from a digester, were compared at moderate temperatures (27.5 ℃ and 8 ℃). Temperature drop affected both the lipids and carbohydrates hydrolysis rates but not necessarily the protein removal rates (temperature-independent rates of MFC), which were significantly influenced from treatment alteration (AD to MFC). MFC showed robust proteolysis at low temperature compared to AD; the latter seems to have a higher potential at warmer conditions. A lipases activity assay showed that although at 27.5 ℃ both AD and MFC are likely to hydrolyse lipids, the latter has a higher lipolysis potential at low temperatures. Preliminary community structure analysis showed that the switch from AD to MFC alters the bacterial community by 15% with the MFC showing higher diversification; temperature decrease, though, alters the community by 40%. Key organisms that appear to be favoured at the MFC set-ups are Geobacteriaceae, taxa likely related to the hydrolytic capacity of this set-up.

Process optimization for extraction and purification of polysaccharides from Cistanche deserticola

In this study, taking Cistanche deserticola in Xinjiang as the experimental material, the optimal process for extracting polysaccharides from C. deserticola with water extraction was studied by using single factor and orthogonal experiment. Its effects on protein removal and polysaccharides retaining were investigated by using Sevag, enzymatic method or combination of these two methods, so as to determine the optimal method for protein removal from polysaccharides of C. deserticola; the decolorization and purification methods such as macroporous resin of AB-8 and activated Carbon were used to determine the optimal process. The results showed that the extraction rate of polysaccharides from C. deserticola was 18.40% during the optimal process of the water extraction as follows: extraction temperature 75 ℃, extraction time 165 min and solid-liquid ratio 1∶55. The protein removal rate can reach 31.40% and polysaccharide retention rate can reach 96.00% under the optimal protein removal process: temperature 50 ℃, time 2 h, and papain dosage 0.2%. The decolorization rate of activated Carbon and macroporous resin called AB-8 was 80.37% and 86.43%, and the recovery rate of polysaccharides was 77.05% and 91.93%, respectively, suggesting that macroporous resin was more suitable for decoloration. Macroporous resin named AB-8 increased the purity of the polysaccharide crude extract from 67.70% to 84.80% under the following conditions: concentration of the sample 4 g·L~(-1), concentration of the eluent 60% ethanol, and the flow rate 1 mL·min~(-1), showing significant purification effect.

Insight into the characteristics, removal, and toxicity of effluent organic matter from a pharmaceutical wastewater treatment plant during catalytic ozonation

Changes in the characteristics, removal efficiency, and toxicity of pharmaceutical effluent organic matter (EfOM) after catalytic ozonation were investigated in this study. After a 90-min treatment with a catalytic ozonation process (COP) in the presence of MnO2 ceramsite, the total organic carbon (TOC), UV254, colority, protein, and humic acid removal rates were 13.24%, 60.83%, 85.42%, 29.36% and 74.19%, respectively. The polysaccharide content increased by 12.73 mg/L during the COP for reaction times between 0 and ~50 min and decreased by 6.97 mg/L between 50 and ~90 min. Furthermore, 64.44% of the total colority was detected in the hydrophobic organic matter (HOM) fraction, and after the COP, and 88.69% of the colority in the HOM was eliminated. Meanwhile, only 59.18% of the colority in the hydrophilic organic matter (HIM) fraction was removed. GC-MS analysis showed that 38 organic pollutant species were completely removed, 8 were partially removed, and 7 were generated. After 90 min of COP treatment, the pharmaceutical EfOM toxicity was effectively reduced based on the higher incubation and lower mortality rates.

A mild extraction and separation procedure of polysaccharide, lipid, chlorophyll and protein from Chlorella spp.

Abstract This study developed a way to separate lipid, polysaccharide, protein and chlorophyll from wet Chlorella spp. (85% moisture content, wt) by using three-phase partitioning (TPP). The optimum conditions for the key steps were researched and defined as (I) cell disruption at the following conditions: biomass concentration 20 g/L, bath temperature 90 °C, duration 2.5 h, enzyme concentration 12 g/L, enzyme hydrolysis time 4 h; and (II) TPP at 40% (NH4)2SO4 (wt, %), sample solution/t-butanol 1: 2 (V/V), room temperature. The extraction yield of lipid and the poysaccharide preservation rate by the optimized procedure were 80.24%, 90.63%, respectively. The protein removal rate was 78.87%. The results provided a promising towards the industrial separation of lipid, polysaccaride, protein and chlorophyll.

The Forms of Fluoride in Antarctic Krill (Euphausia superba) Oil Extracted with Hexane and its Removal with Different Absorbents

ABSTRACTKrill oils (KOs) were prepared using different extraction methods (hexane and supercritical carbon dioxide), different materials (heat-dried krill powder, freeze-dried krill powder, and freeze-dried krill tail meat powder), and different standing times during hexane extraction. The fluoride concentrations in the KOs were positively correlated with the protein concentrations. Absorbents were employed to remove fluoride from KO. Activated clay achieved the highest fluoride removal rate (70.66%), followed by calcium oxide (14.49%), activated carbon (11.97%), calcium chloride (9.32%), activated alumina (0.94%), and chitosan (0.52%). Meanwhile, the protein removal rates of the above absorbents were positively correlated with the fluoride removal rates.

Optimize concentrate process of alkaloid from Leonurus japonicus by ultrafultration-nanofiltration coupling technology

To optimize the concentrate process of alkaloid from Leonurus japonicus by nanofiltration-ultrafiltration coupling technology with response surface methodology. The experiment showed that after ultrafiltration pre-treatment, the total protein removal rate was 94.38% in aqueous extract from L. japonicus, and the nanofiltration technology had obvious advantages over the conventional concentrate process. The optimal concentrate conditions were as follows：molecular weight cut-off 450, pH 3.07, concentration of stachydrine hydrochloride 80.15 mg•L⁻¹, and concentration of the total alkaloid 285.73 mg•L⁻¹. The cut-off rate was 93.37% and 95.85% respectively for stachydrine hydrochloride and the total alkaloid under the optimum conditions, with a relative error of 0.79% and 1.16% respectively. The combination of Box-Behnken design and response surface analysis can well optimize the concentrate process of L. japonicus by nanofiltration, and the results provide the basis for nanofiltration concentrate for heat-sensitive traditional Chinese medicine.