Demulsification of oil-rich emulsion from enzyme-assisted aqueous extraction of extruded soybean flakes

Extraction of soybean oil from flaked and extruded soybeans using enzyme-assisted aqueous extraction processing (EAEP) is a promising alternative to conventional hexane extraction. The efficiencies of four proteases releasing oil from extruded material were compared. Protex 51FP, Protex 6L and Protex 7L each extracted 90% of the total oil available while Protex 50FP gave similar extraction yield as the control (without enzyme treatment). During EAEP, however, a stable emulsion forms that must be broken in order to recover free soybean oil. The potential of various proteases and phospholipases to destabilize the emulsion was determined. Two enzymes, a phospholipase A2 (LysoMax) and a protease (Protex 51FP) were selected to determine the effect of enzyme concentration on demulsification. Although at a 2% concentration (w/w, enzyme/(cream+free oil)), each enzyme tested was effective in totally destabilizing the cream; the protease released significantly more free oil than did the phospholipase at concentrations less than 2%. At 0.2% concentration, 88 and 48% of free oil were obtained with the protease and phospholipase, respectively. Reducing the pH of the cream also destabilized the cream with maximum demulsification at the isoelectric point of soy proteins. These results provide destabilization strategies for the oil-rich emulsion formed during aqueous extraction processing of extruded flakes and significantly contribute to the development of this environmentally-friendly technology.     

Efficiency of pretreatments for optimal enzymatic saccharification of soybean fiber

The effectiveness of several pretreatments [high-power ultrasound, sulfuric acid (H₂SO₄), sodium hydroxide (NaOH), and ammonium hydroxide (NH₃OH)] to enhance glucose production from insoluble fractions recovered from enzyme-assisted aqueous extraction processing of extruded full-fat soybean flakes (FFSF) was investigated. Sonication of the insoluble fraction at 144 μm_{pp (peak-to-peak)} for 30 and 60 s did not improve the saccharification yield. The solid fractions recovered after pretreatment with H₂SO₄ [1% (w/w), 90 °C, 1.5 h], NaOH [15% (w/w), 65 °C, 17 h], and NH₃OH [15% (w/w), 65 °C, 17 h] showed significant lignin degradation, i.e., 81.9%, 71.2%, and 75.4%, respectively, when compared to the control (7.4%). NH₃OH pretreatment resulted in the highest saccharification yield (63%) after 48 h of enzymatic saccharification. A treatment combining the extraction and saccharification steps and applied directly to the extruded FFSF, where oil extraction yield and saccharification yield reached 98% and 43%, respectively, was identified.     

Destabilization of the emulsion formed during the enzyme-assisted aqueous extraction of oil from soybean flour

Soybean oil extraction from soybean flour by a neutral metallo-endopeptidase enzyme-assisted aqueous extraction process, optimized for effectiveness in reducing oil content of the solid residue, yields a small fraction of the oil as free oil whereas most is emulsified in a cream layer. Analysis of the cream showed the presence of peptides/proteins and phospholipids, either of which could serve as emulsifiers. Several demulsification treatments targeting these components – protease addition, phospholipase addition, and acidification (pH 4.5) – were evaluated for effectiveness against the targeted emulsifiers and in conversion of the cream to free oil.Acidification increased the oil yield from 2% to 83%. After a two-stage enzymatic demulsification process with an alkaline endopeptidase, the oil recovery increased to 95%. A lysophospholipase A₁ treatment at pH 4.5 provided complete conversion of emulsified to free oil. The phospholipids (PL) present in the original and post-treatment cream were quantified using phosphorus-31 nuclear magnetic resonance spectroscopy. PL contained phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidic acid (PA) and their lyso-derivatives. Protein size distribution, PL proportions and fatty acid composition changed after the demulsification treatments. Recycle of the endopeptidase is feasible as more than 90% of its activity was retained.     

Development of an optimal formulation for oxidative stability of walnut-beverage emulsions based on gum arabic and xanthan gum using response surface methodology

The susceptibility of lipids to oxidation is one of the most fundamental problems in oil-in-water emulsions. A response surface methodology 5-level-3-factor central-composite rotatable design was applied to study the effects of key formula ingredients including walnut oil (WO, 3-6%, w/w), gum arabic (GA, 5-10%, w/w) and xanthan gum (XG, 0.05-0.15%, w/w) on lipid oxidation in walnut-beverage emulsions. During 30 days’ storage, the oxidation process was monitored by evaluating the peroxide value, anisidine value and total oxidation (Totox) value in different emulsion formulations. Use of XG as a stabilizer at high concentrations considerably inhibited the oxidation of WO in the prepared emulsions. The experimental data were satisfactorily fitted to quadratic models using multiple regression analysis. The optimum conditions to obtain the minimum peroxide (0.923 mequiv. O₂/kg oil), anisidine (0.500) and Totox (2.347) values are met when a walnut-beverage emulsion is formulated with 3% WO, 10% GA and 0.12% XG.     

Effect of Soy Soluble Polysaccharide on the Stability of Soy-Stabilised Emulsions During In Vitro Protein Digestion

This study investigated physicochemical properties of soy soluble polysaccharide (SSP) and pectinase-hydrolysed soy soluble polysaccharide (PH-SSP) from okara, the residue from soy milk production, and their influences when used as a fibre source in oil-in-water (o/w) emulsions. Although pectinase hydrolysed only the carbohydrate fraction in SSP, it resulted in the self-association of PH-SSP to the large-size aggregates. When PH-SSP was added to liquid emulsion containing 3.33% (w/v) rice bran oil and 3.75% (w/v) heated soy protein, it regulated the contents of protein in serum phase, sediment phase and at oil–water interface. The types and contents of soy proteins in the serum phase and sediment phase could be manipulated by pre-heating of soy proteins at 80 °C for 30 min and the addition of PH-SSP. The presence of PH-SSP (0–6% w/v) induced different distribution of proteins to the sediment phase and subsequent in vitro protein digestion in the emulsion. Overall, this study proposed the means to design the distributions of proteins in different phases of o/w emulsion for different degrees of oil release, emulsion stability and protein-polysaccharide coacervation during the course of in vitro peptic and tryptic digestion.     

生物破乳剂产生菌的筛选及其方法研究

针对生物破乳剂产生菌筛选难的问题, 采用显色法、溶血细胞测试法、表面张力测定法和排油圈法从6种不同菌源对生物破乳菌产生菌进行了筛选。通过试验筛选得到了17株生物破乳剂产生菌, 其中24h内破乳率高于70%的破乳菌有5株; 油田含油污泥、采油废水生物处理污泥和污水处理厂剩余污泥是筛选破乳菌的较好的菌源; 显色法、溶血圈法存在检测范围的局限性; 表面张力测定法和排油圈法是最为简易和准确的生物表面活性剂产生菌的筛选方法, 采用模型乳状液对生物破乳剂产生菌进行筛选最为直接和准确, 但工作量大、所需时间长, 因此在筛选高效破乳菌时, 建议采用表面张力、排油圈法进行初筛, 而后通过模型乳状液破乳进行验证。     

生物破乳剂产生菌的筛选及其方法研究

针对生物破乳剂产生菌筛选难的问题,采用显色法、溶血细胞测试法、表面张力测定法和排油圈法从6种不同菌源对生物破乳菌产生茵进行了筛选.通过试验筛选得到了17株生物破乳剂产生茵,其中24h内破乳率高于70%的破乳菌有5株;油田含油污泥、采油废水生物处理污泥和污水处理厂剩余污泥是筛选破乳菌的较好的菌源:显色法、溶血圈法存在检测范围的局限性;表面张力测定法和排油圈法是最为简易和准确的生物表面活性剂产生茵的筛选方法,采用模型乳状液对生物破乳剂产生菌进行筛选最为直接和准确,但工作量大、所需时间长,因此在筛选高效破乳菌时,建议采用表面张力、排油圈法进行初筛,而后通过模型乳状液破乳进行验证.     

Changes in Colloidal Properties of Oil in Water Emulsions Stabilized with Sodium Caseinate Observed by Acoustic and Electroacoustic Spectroscopy

Sodium caseinate is a commonly used emulsifier in foods, as it adsorbs on the surface of oil droplets and stabilizes them via electrostatic and steric stabilization, forming a polyelectrolyte layer at the interface. Since the protein interface is affected by varying environmental conditions such as pH, ionic strength, concentration of unadsorbed polymers, these emulsions are prone to a variety of destabilization mechanisms. The objective of the present work was to observe the destabilization of sodium caseinate stabilized oil in water emulsions using electroacoustic spectroscopy. This technique can be utilized for the characterization of concentrated colloidal systems in situ, without dilution. The electroacoustic and ultrasonic properties of soy oil in water emulsions were determined for sodium caseinate stabilized emulsions under conditions known to cause destabilization. Ultrasonic attenuation and electrophoretic mobility (ζ-potential) could clearly follow the changes occurring in the emulsion droplets, under minimal sample disruption. This is critical for these systems in a very fragile, metastable state. The emulsions were stable to the addition of high methoxyl pectin (HMP) up to 0.1% HMP. Addition of free sodium caseinate induced depletion flocculation, causing a decrease in the attenuation and electrophoretic mobility measured. The presence of HMP limited depletion interactions. Acidification of the emulsion droplets resulted in a clear sol–gel transition, as shown by a steep increase in the particle size and a decrease in attenuation. Again, destabilization was limited by HMP addition. It was concluded that ultrasonics and electroacoustics are suitable techniques to understand the details of the destabilization processes occurring to food emulsions, measured in situ.     

Promising perspectives for ruminal protection of polyunsaturated fatty acids through polyphenol-oxidase-mediated crosslinking of interfacial protein in emulsions

Previously, polyunsaturated fatty acids (PUFA) from linseed oil were effectively protected (>80%) against biohydrogenation through polyphenol-oxidase-mediated protein crosslinking of an emulsion, prepared with polyphenol oxidase (PPO) extract from potato tuber peelings. However, until now, emulsions of only 2 wt% oil have been successfully protected, which implies serious limitations both from a research perspective (e.g. in vivo trials) as well as for further upscaling toward practical applications. Therefore, the aim of this study was to increase the oil/PPO ratio. In the original protocol, the PPO extract served both an emulsifying function as well as a crosslinking function. Here, it was first evaluated whether alternative protein sources could replace the emulsifying function of the PPO extract, with addition of PPO extract and 4-methylcatechol (4MC) to induce crosslinking after emulsion preparation. This approach was then further used to evaluate protection of emulsions with higher oil content. Five candidate emulsifiers (soy glycinin, gelatin, whey protein isolate (WPI), bovine serum albumin and sodium caseinate) were used to prepare 10 wt% oil emulsions, which were diluted five times (w/w) with PPO extract (experiment 1). As a positive control, 2 wt% oil emulsions were prepared directly with PPO extract according to the original protocol. Further, emulsions of 2, 4, 6, 8 and 10 wt% oil were prepared, with 80 wt% PPO extract (experiment 2), or with 90, 80, 70, 60 and 50 wt% PPO extract, respectively (experiment 3) starting from WPI-stabilized emulsions. Enzymatic crosslinking was induced by 24-h incubation with 4MC. Ruminal protection efficiency was evaluated by 24-h in vitro batch simulation of the rumen metabolism. In experiment 1, protection efficiencies were equal or higher than the control (85.5% to 92.5% v. 81.3%). In both experiments 2 and 3, high protection efficiencies (>80%) were achieved, except for emulsions containing 10 wt% oil emulsions (<50% protection), which showed oiling-off after enzymatic crosslinking. This study demonstrated that alternative emulsifier proteins can be used in combination with PPO extract to protect emulsified PUFA-rich oils against ruminal biohydrogenation. By applying the new protocol, 6.5 times less PPO extract was required.     

Hydrophobically modified alginate for emulsion of oil in water

Dodecanol was covalently coupled to sodium alginate (NaAlg) via ester functions using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC-HCl) as a coupling reagent to provide an amphiphilic dodecanol alginate (DA) for subsequent use in oil-in-water (O/W) emulsion application. The structure of DA was confirmed by FT-IR spectrometry. The stability of the emulsions prepared with different concentrations (0.3-1.2 wt%) of DA or 1.0 wt% NaAlg was evaluated by measuring droplet size, microstructure, viscosity and creaming. The results showed that the emulsions containing 1.0 wt% NaAlg, 0.3 and 0.5 wt% DA were unstable and the emulsions containing 0.8-1.2 wt% DA presented better stability during storage.     

Integration of Gas Enhanced Oil Recovery in Multiphase Fermentations for the Microbial Production of Fuels and Chemicals

In multiphase fermentations where the product forms a second liquid phase or where solvents are added for product extraction, turbulent conditions disperse the oil phase as droplets. Surface‐active components (SACs) present in the fermentation broth can stabilize the product droplets thus forming an emulsion. Breaking this emulsion increases process complexity and consequently the production cost. In previous works, it has been proposed to promote demulsification of oil/supernatant emulsions in an off‐line batch bubble column operating at low gas flow rate. The aim of this study is to test the performance of this recovery method integrated to a fermentation, allowing for continuous removal of the oil phase. A 500 mL bubble column is successfully integrated with a 2 L reactor during 24 h without affecting cell growth or cell viability. However, higher levels of surfactants and emulsion stability are measured in the integrated system compared to a base case, reducing its capacity for oil recovery. This is related to release of SACs due to cellular stress when circulating through the recovery column. Therefore, it is concluded that the gas bubble‐induced oil recovery method allows for oil separation and cell recycling without compromising fermentation performance; however, tuning of the column parameters considering increased levels of SACs due to cellular stress is required for improving oil recovery.     

Chitosan-based microcapsules containing grapefruit seed extract grafted onto cellulose fibers by a non-toxic procedure

A novel non-toxic procedure is described for the grafting of chitosan-based microcapsules containing grapefruit seed oil extract onto cellulose. The cellulose was previously UV-irradiated and then functionalized from an aqueous emulsion of the chitosan with the essential oil. The novel materials are readily attained with durable fragrance and enhanced antimicrobial properties. The incorporation of chitosan as determined from the elemental analyses data was 16.08 ± 0.29 mg/g of sample. Scanning electron microscopy (SEM) and gas chromatography-mass spectroscopy (GC-MS) provided further evidence for the successful attachment of chitosan microcapsules containing the essential oil to the treated cellulose fibers. The materials thus produced displayed 100% inhibition of Escherichia coli and Staphylococcus epidermidis up to 48 h of incubation. Inhibition of bacteria by the essential oil was also evaluated at several concentrations.     

Quantitative studies of phagocytosis by alveolar macrophages

The initial rate of phagocytosis by rabbit alveolar macrophages of paraffin oil emulsions stabilized with albumin was markedly increased by prior incubation of the emulsion with serum. The active component(s) of serum was non-dialyzable and heat-labile and was absent from zymosan-treated serum. Magnesium and calcium were both required for the maximal rate of phagocytosis. At 4 °C or in the presence of 1 mM N-ethylmaleimide, the rate of phagocytosis was less than 2% of the control (37° C) rate. The initial rates of phagocytosis of this emulsion by alveolar macrophages from rabbits injected with Freund's adjuvant were not demonstrably different from those observed with macrophages from normal rabbits. Per of mg of cell protein, polymorphonuclear leukocytes ingested serum-treated emulsion more rapidly than did macrophages, but per cell the rates were not different.     

Antifreeze Emulsions Produced from Linoleic Acid and Water Using Enzymatically Modified Gelatin

An enzymatically modified gelatin with covalently attached leucine dodecyl ester, referred to as EMG-12, was used as a surfactant to prepare emulsions with different properties by changing the surfactant concentration, oil volume fraction, and pH in the water phase. The emulsions generally resisted the freezing of their constituent bulk water at approximately ?10°C, but similar emulsions produced with soy protein isolate, casein, or Tween-80 as control agents were less resistant. The freezing (or unfreezing) of the bulk water in these emulsions depended on the kind of agent used, not on the emulsion properties such as average area of the oil/water interface, stability against coalescence, and stability against creaming. The emulsion produced with EMG-12, like that produced with polyglycerol stearate, tended to maintain its unfrozen state even in the presence of silver iodide crystals added as heterogeneous ice-nuclei. The significance of producing such an antifreeze emulsion is discussed from the standpoint of cryopreservation of cold-sensitive food and biological systems.     

Comparison between waste frying oil and paraffin as carbon source in the production of biodemulsifier by Dietzia sp. S-JS-1

In order to lower the production cost, waste frying oils were used in the biosynthesis of demulsifier by Dietzia sp. S-JS-1, which was isolated from petroleum contaminated soil. After 7 days of cultivation, the biomass concentration of the most suitable waste frying oil (WFO II) culture reached 3.78 g/L, which was 2.4 times the concentration of paraffin culture. The biodemulsifier produced with WFO II culture broke the emulsions more efficiently than that produced with paraffin culture, given the same volume ratio of carbon source in the culture medium and the same cultivation conditions. It achieved 88.3% of oil separation ratio in W/O emulsion and 76.4% of water separation ratio in O/W emulsion within 5 h. With the aid of thin layer chromatography (TLC) and Fourier transform infrared (FTIR) spectrometry, biodemulsifiers produced from both paraffin and WFO II were identified as a mixture of lipopeptide homologues. The subtle variation in the distribution of these homologues and high biomass concentration of WFO II cultures may account for the afore-mentioned good demulsification performance.     

Ecofriendly demulsification of water in oil emulsions by an efficient biodemulsifier producing bacterium isolated from oil contaminated environment

Objective

Water in oil emulsions increase oil processing costs and cause damage to refinery equipment which necessitates demulsification. Since chemical demulsifiers cause environmental pollution, biodemulsifiers have been paid more attention. This study aims to identify biodemulsifier-producing bacteria from petroleum contaminated environments.

Results

As a result, several biodemulsifier producing strains were found that Stenotrophomonas sp. strain HS7 (accession number: MF445088) which produced a cell associated biodemulsifier showed the highest demulsifying ratio, 98.57% for water in kerosene and 66.28% for water in crude oil emulsion after 48 h. 35 °C, pH 7, 48 h incubation and ammonium nitrate as nitrogen source were optimum conditions for biodemulsifier production. Furthermore, it was found that hydrophobic carbon sources like as liquid paraffin is not preferred as the sole carbon source while a combination of various carbon sources including liquid paraffin will increase demulsification efficiency of the biodemulsifier.

Conclusions

The appropriate potential of this biodemulsifier strengthens the possibility of its application in industries especially petroleum industry.

     

Emulsifier Dependent <Emphasis Type="Italic">in vitro</Emphasis> Digestion and Bioaccessibility of β-Carotene Loaded in Oil-in-Water Emulsions

This study was performed to examine the effect of emulsifiers used to coat emulsion droplets containing β-carotene on the behavior of lipid digestion and bioaccessibility. Different emulsifiers (whey protein isolate, soy protein isolate, sodium caseinate, Tween 20, and soy lecithin) were used to prepare emulsions with similar sized droplets (200–400 nm). Protein-stabilized emulsions showed a similar behavior of digestion, and morphological change in the simulated gastrointestinal conditions. Soy lecithin-stabilized emulsions showed the lowest rate and extent of lipid digestion probably due to the low emulsifying capability of soy lecithin, showing coalesced droplets occurring after exposure to the gastric phase. Tween 20-stabilized emulsions had a lower rate and extent of lipid digestion than that of protein-stabilized emulsions, even though Tween 20-stabilized emulsions had a more stable structure to resistant to aggregation in gastric phase. Even though the difference in the digestion rate and extent, β-carotene bioaccessibility was not significantly different among emulsions stabilized by different emulsifiers at p?<?0.05.     

Dietzia sp.S-JS-1利用煎炸废油生产生物破乳剂及其性能研究

以前期研究中筛选得到的破乳剂产生菌Dietzia sp.S-JS-1为研究对象,采用煎炸废油为培养碳源,考察菌株的生物量和表面张力,研究处理方式、温度、乳状液pH对破乳剂在两种模型乳状液W/O型(water in oil)和O/W型(oilin water)中破乳性能的影响,并初步分析生物破乳剂成分。结果表明:菌株最大生物量为2.6 g/L,其产生的破乳剂能够将纯水表面张力从72.0 mN/m降低到32.5 mN/m。冻融对破乳剂效果的影响小于高温灭菌;破乳剂经冷冻干燥处理后的破乳效果明显好于烘干处理;破乳剂在35℃～75℃时具有较好的破乳效果,脱水率均在75%以上;破乳剂在W/O型乳状液中的效果随着pH变大而逐渐增加,pH=10时的脱水率高达99.8%,而在O/W型乳状液中,pH=7时的脱水率最高,为90%左右。薄层色谱结果表明S-JS-1利用煎炸油生产的生物破乳剂可能含有5种脂肽类物质。     

Microwave-assisted extraction of human hair proteins

The effectiveness of microwave-assisted extraction of proteins from human hair samples was evaluated. Extractions were performed from 2-mg hair samples in an extraction solution consisting of 25 mM Tris-HCl (pH 8.5), 2.6 M thiourea, 5 M urea, and 5% mercaptoethanol. During extraction, samples were exposed to microwave radiation (600 W) for a specified incubation period (5-120 min). The extraction efficiency of samples that had been incubated for 60 min was similar to that of samples that had been heated at 50 °C for 24 h using the conventional Shindai method.