Protein Recovery in Aqueous Extraction Processing of Soybeans Using Isoelectric Precipitation and Nanofiltration

Isoelectric precipitation and whey nanofiltration were evaluated in recovering protein from skim fractions produced by enzyme-assisted aqueous extraction processing (EAEP) of extruded full-fat soybean flakes. Countercurrent two-stage EAEP was performed at 1:6 solids-to-liquid ratio, 50 °C, pH 9.0, and 120 rpm for 1 h to extract oil and protein from soybeans. Two protein recovery strategies were applied to skim fractions produced by different extraction treatments: Treatment 1 using 0.5% protease (wt/g extruded flakes) in both extraction stages; Treatment 2 using 0.5% protease only in the 2nd extraction stage; and Treatment 3 using no enzyme in either extraction stage. Protein recovery by using isoelectric precipitation was inversely related to the extent of hydrolysis with recoveries of 27, 61, and 87% of skim proteins from Treatments 1, 2, and 3, respectively. Overall protein recoveries of 26, 54, and 57% of the original protein in the extruded full-fat flakes were achieved when combining extraction treatments and isoelectric precipitation. Nanofiltering isoelectric wheys (500-Da membrane) achieved protein retentate yields of 96.3, 94.5, and 91.8% (1.9–2.8 concentration factor) with permeate fluxes up to 1.35 kg/h m². About 97, 98, and 99% of skim protein were recovered by isoelectric precipitation and whey nanofiltration for Treatments 1, 2, and 3, respectively. Overall protein recoveries of 93, 87, and 65% of the protein in the extruded flakes were achieved for Treatments 1, 2, and 3, respectively. Although high protein retentions were achieved, very low permeate fluxes were observed for whey nanofiltration.     

Two-Stage Countercurrent Enzyme-Assisted Aqueous Extraction Processing of Oil and Protein from Soybeans

Enzyme-assisted aqueous extraction processing (EAEP) is an increasingly viable alternative to hexane extraction of soybean oil. Although considered an environmentally friendly technology where edible oil and protein can be simultaneously recovered, this process employs much water and produces a significant amount of protein-rich aqueous effluent (skim). In standard EAEP, highest oil, protein and solids yields are achieved with a single extraction stage using 1:10 solids-to-liquid ratio (extruded flakes/water), 0.5% protease (wt/g extruded flakes), pH 9.0, and 50 °C for 1 h. To reduce the amount of water used, two-stage countercurrent EAEP was evaluated for extracting oil, protein and solids from soybeans using a solids-to-liquid ratio of 1:5–1:6 (extruded flakes/water). Two-stage countercurrent EAEP achieved higher oil, protein and solids extraction yields than using standard EAEP with only one-half the usual amount of water. Oil, protein and solids yields up to 98 and 96%, 92 and 87%, and 80 and 77% were obtained when using two-stage countercurrent EAEP (1:5–1:6) and standard single-stage EAEP (1:10), respectively. Recycling the second skim obtained in two-stage countercurrent EAEP enabled reuse of the enzyme, with or without inactivation, in the first extraction stage producing protein with different degrees of hydrolysis and the same extraction efficiency. Slightly higher oil, protein and solids extraction yields were obtained using unheated skim compared to heated skim. These advances make the two-stage countercurrent EAEP attractive as the front-end of a soybean biorefinery.     

Scale-up of Enzyme-Assisted Aqueous Extraction Processing of Soybeans

The effects of scaling-up enzyme-assisted aqueous extraction process (EAEP) using 2 kg of flaked and extruded soybeans as well as the effects of different extrusion and extraction conditions were evaluated. Standard single-stage EAEP at 1:10 solids-to-liquid ratio (SLR) was used to evaluate the effects of different extruder screw speeds and whether or not collets were extruded directly into water. Increasing extruder screw speed from 40 to 90 rpm improved oil extraction yield from 85 to 95%. Oil, protein, and solids extraction yields of 97, 86, and 78% were obtained when extruding directly into water and 95, 84, and 77% when not extruding into water. When not extruding into water, standard single-stage EAEP (1:10 SLR) yielded 95, 84, and 77% of total oil, protein, and solids extraction, respectively, and two-stage countercurrent EAEP (1:6 SLR) yielded 99, 94, and 83% total oil, protein, and solids extraction, respectively. These yields were similar to those previously obtained in the laboratory (0.08 kg soybeans), but higher oil contents were observed in the skim fractions produced at pilot-plant scale for both processes. Modifying processing parameters improved the oil distribution among the fractions, increasing oil yield in the cream fraction (from 76 to 86%) and reducing oil yield in the skim fraction (from 23 to 12%). Steady-state oil extraction was achieved after two 2-stage extractions. Two-stage countercurrent EAEP is particularly attractive due to reduced water usage compared to conventional single-stage extraction.     

Integrated Countercurrent Two-Stage Extraction and Cream Demulsification in Enzyme-Assisted Aqueous Extraction of Soybeans

Countercurrent two-stage extraction and cream demulsification were fully integrated and demonstrated on laboratory scale (2 kg soybeans) wherein the enzyme used for demulsifying the cream was used in the extraction steps of enzyme-assisted aqueous extraction processing (EAEP). Protease enzyme (Protex 6L) entered the integrated EAEP process in the demulsification step and the skim, which contained the enzyme, resulting from breaking the cream emulsion was recycled upstream into the second extraction stage and then to the first extraction stage. Oil, protein and solids extraction yields of 96.1 ± 1.4%, 89.3 ± 1.0%, and 81.2 ± 2.0%, respectively, were achieved with steady-state operation of integrated EAEP. Higher degrees of protein hydrolysis (DH) were obtained when using the integrated process compared with the process when not recycling the enzyme. Higher extents of hydrolysis probably increased emulsion formation thereby affecting lipid distribution among the fractions. Overall free oil recovery was reduced due to more oil shifting to the skim fraction.     

Properties of Soy Protein Produced by Countercurrent,Two-Stage,Enzyme-Assisted Aqueous Extraction

Enzyme-assisted aqueous extraction processing (EAEP) is an environmentally friendly technology where oil and protein can be simultaneously extracted from soybeans by using water and protease. Countercurrent, two-stage, EAEP was performed at a 1:6 solids-to-liquid ratio, 50 °C, pH 9.0, and 120 rpm for 1 h to extract oil and protein from soybeans. The skim fractions were produced by three methods: (1) by treating with 0.5 % protease (wt/g extruded flakes) in both extraction stages; (2) by treating with 0.5 % protease in the 2nd extraction stage only; and (3) by using the same two-stage extraction procedure without enzymes in either extraction stages. Countercurrent, two-stage, protein extraction of air-desolventized, hexane-defatted, soybean flakes was used as a control. Solubility profiles of the skim proteins were the typical U-shaped curves with the lowest solubility at the isoelectric point of soy protein (pH 4.5). The use of the enzyme slightly improved solubility of the recovered protein with hydrolyzed proteins having higher solubilities at acid pH. Emulsification and foaming properties were generally reduced by the use of enzyme during EAEP extractions. The skims produced with protease-extracted (hydrolyzed) proteins gave gels with lower hardness than did unhydrolyzed proteins when heated at 80 °C. The essential amino acid compositions and protein digestibilities were not adversely affected by either extrusion or extraction treatments.     

Enzyme-Assisted Aqueous Extraction of Oil and Protein from Soybeans and Cream De-emulsification

The effects of two commercial endoproteases (Protex 6L and Protex 7L, Genencor Division of Danisco, Rochester, NY, USA) on the oil and protein extraction yields from extruded soybean flakes during enzyme-assisted aqueous extraction processing (EAEP) were evaluated. Oil and protein were distributed in three fractions generated by the EAEP: cream + free oil, skim and insolubles. Protex 6L was more effective for extracting free oil, protein and total solids than Protex 7L. Oil and protein extraction yields of 96 and 85%, respectively, were obtained using 0.5% Protex 6L. Enzymatic and pH treatments were evaluated to de-emulsify the oil-rich cream. Cream de-emulsification generated three fractions: free oil, an intermediate residual cream layer and an oil-lean second skim. Total cream de-emulsification was obtained when using 2.5% Protex 6L and pH 4.5. The extrusion treatment was particularly important for reducing trypsin inhibitor activity (TIA) in the protein-rich skim fraction. TIA reductions of 69 and 45% were obtained for EAEP skim (the predominant protein fraction) from extruded flakes and ground flakes, respectively. Protex 6L gave higher degrees of protein hydrolysis (most of the polypeptides being between 1,000 and 10,000 Da) than Protex 7L. Raffinose was not detected in the skim, while stachyose was eliminated by α-galactosidase treatment.     

Flaking as a Pretreatment for Enzyme-Assisted Aqueous Extraction Processing of Soybeans

Soybean moisture content (7.2–12.8%) and conditioning temperature (51–79 °C) during flaking were evaluated to determine their effects on oil and protein extraction and oil distribution among fractions produced in enzyme-assisted aqueous extraction processing (EAEP). Extractions were performed by using two-stage countercurrent EAEP at a 1:6 solids-to-liquid ratio with 0.5% protease (wt/g extruded flakes) at pH 9.0 and 50 °C for 1 h. Oil extraction improved when using soybeans with moisture contents ranging from 8.0 to 12.0% for flaking but was not affected by conditioning temperature. Oil extraction was reduced when moving away from 10% moisture with the lowest values at 7.2 and 12.8% moisture. Free oil extraction increased as soybean moisture content increased from 7.2 to 12.8% although total oil extraction was reduced at 12.8% moisture. Higher (79 °C) and lower conditioning temperatures (51 °C) improved free oil extraction and reduced cream emulsion formation. Skim oil content was not significantly affected by soybean moisture content and the conditioning temperature, although an undesirable high oil content in the skim was observed at 8% moisture and at 55 °C. The cream with a high oil yield was easily demulsified compared with cream containing a low oil yield (95 vs. 76.5% de-emulsification). Due to differences in cream stability, similar oil recoveries (78–80%) were obtained for treatments yielding creams with either low or high oil yields. Mean protein extraction of 95% was achieved for all treatments and was not significantly affected by soybean moisture content at flaking or conditioning temperature.     

Pilot-Plant Proof-of-Concept for Integrated, Countercurrent, Two-Stage, Enzyme-Assisted Aqueous Extraction of Soybeans

Proof-of-concept for integrated, countercurrent, two-stage, enzyme-assisted aqueous extraction processing of soybeans was demonstrated on a pilot-plant scale (75 kg extruded flaked soybeans) where the protease used to demulsify the cream was recycled into upstream extraction stages. Oil, protein, and solids extraction yields of 98.0 ± 0.5%, 96.5 ± 0.4%, and 86.8 ± 0.5% were achieved by using the integrated countercurrent process. A three-phase horizontal decanter centrifuge efficiently separated the solids from the two liquid fractions (skim and cream). Fine separation between the two liquid fractions was important to reducing the volume of skim contaminating the cream fraction, thereby reducing the amount of enzyme used for cream demulsification and subsequent extraction. We were able to reduce enzyme use when moving from the laboratory to the pilot-plant scale, which reduced the degree of protein hydrolysis and improved cream demulsification. Enzyme-catalyzed cream demulsification was 91.6% efficient and 93.0% free oil recovery from cream was achieved by using the integrated approach.     

Separating Oil from Aqueous Extraction Fractions of Soybean

Previous research has shown that enzyme-assisted aqueous extraction processing (EAEP) extracts 88–90% of the total soybean oil from extruded full-fat soy flakes into the aqueous media, which is distributed as cream (oil-in-water emulsion), skim, and free oil. In the present work, a simple separatory funnel procedure was effective in separating aqueous skim, cream and free oil fractions allowing mass balances and extraction and recovery efficiencies to be determined. The procedure was used to separate and compare liquid fractions extracted from full-fat soy flour and extruded full-fat soy flakes. EAEP extracted more oil from the extruded full-fat soy flakes, and yielded more free oil from the resulting cream compared to unextruded full-fat soy flour. Dry matter partitioning between fractions was similar for the two procedures. Mean oil droplet sizes in the cream and skim fractions were larger for EAEP of extruded flakes compared to non-enzymatic AEP of unextruded flour (45 vs. 20 μm for cream; 13 vs. 5 μm for skim) making the emulsions from EAEP of extruded flakes less stable. All major soy protein subunits were present in the cream fractions, as well as other fractions, from both processes. The cream could be broken using phospholipase treatments and 70–80% of total oil in the extruded full-fat flakes was recovered using EAEP and a phospholipase de-emulsification procedure.     

Characteristics of Oil and Skim in Enzyme-Assisted Aqueous Extraction of Soybeans

The economic viability of enzyme-assisted aqueous extraction processing (EAEP) of soybeans depends on properties and potential applications of all fractions (skim and insolubles as well as oil). EAEP oil contained lower free fatty acid, phosphorus, and tocopherol contents, similar unsaponifiable matter levels, and higher degrees of oxidation (peroxide and p-anisidine values) than hexane-extracted oil. The phospholipid profile of EAEP fractions was mainly composed of phosphatidic acid, followed by phosphatidylcholine, phosphatidylinositol, and phosphatidylethanolamine. Most of phospholipids were present in the skim, except for phosphatidic acid, which was the major phospholipid in the cream fraction. Skim and cream contained 55 and 3 % of the soluble carbohydrates in the original extruded flakes, respectively. Soluble carbohydrates of the skim were mainly composed of stachyose (5.8 ± 0.8 mg/mL) and sucrose (9.9 ± 0.8 mg/mL), which were hydrolyzed into glucose, galactose, and fructose after addition of α-galactosidase. Skim and cream peptides contained <20 kDa MW molecules. About 71 % of the skim peptides were <20 kDa MW, with 49 % being <1.35 kDa MW, 22 % being 17–1.35 kDa MW, and 29 % being 44–670 kDa MW. Skim protein and carbohydrate contents make this fraction suitable for replacing water in ethanol fermentations, thereby improving the fermentation rate/production and the nutritional quality of distiller’s dried grains with solubles.     

Aqueous Extraction of Oil and Protein from Soybeans with Subcritical Water

Aqueous extraction using subcritical water is an environmentally friendly alternative to extracting oil and protein from oilseeds with flammable organic solvents. The effects of solids-to-liquid ratio (1:3.3–1:11.7), temperature (66–234 °C), and extraction time (13–47 min) were evaluated on the extraction of oil and protein from soybean flakes and from extruded soybeans flakes with subcritical water. A central composite design (2³) with three center points and six axial points was used. Subcritical water extractions were carried out in a 1-L high-pressure batch reactor with constant stirring (300 rpm) at 0.03–3.86 MPa. In general, oil extraction was greater for extruded soybean flakes than with soybean flakes. More complete oil extraction for extruded soybean flakes was achieved at around 150 °C and extraction was not affected by solids-to-liquid ratios over the range tested, while oil extraction from soybean flakes was more complete at 66 °C and low solids-to-liquid ratio (1:11.7). Protein extraction yields from flakes were generally greater than from extruded flakes. Protein extraction yields from extruded flakes increased as temperature increased and solids-to-liquid ratio decreased, while greater protein extraction yields from soybean flakes were achieved when using low temperatures and low solids-to-liquid ratio.     

Retention of cadmium ions from aqueous solutions by poly(ammonium acrylate) enhanced ultrafiltration

Cadmium removal from aqueous solution by polyelectrolyte enhanced ultrafiltration (PEUF) with relatively low transmembrane pressure was investigated at varying conditions of polyelectrolyte and cadmium concentrations, transmembrane pressure, ionic strength and pH. The poly(ammonium acrylate), with two average molecular weights (8000 and 15 000 Da) were used as polyelectrolyte. Flux declines during ultrafiltration of polyelectrolyte solutions. An effort has been made to evaluate these resistances independently at different operating conditions. The hydraulic membrane resistance is higher for processing solutions of PAmA₈ than solutions of PAmA₁₅. The study of ionic strength effect demonstrates that it decreases the retention of cadmium ions and increases the permeate flux. More than 99% of cadmium was retained for a NaNO₃ feed concentration less than 5 × 10⁻² mol L⁻¹. The pH effect study on the cadmium recovery revealed a maximum retention around 98% for pH 4.     

Critical flux in cross-flow ultrafiltration of protein solutions

The effect of solute size relative to membrane pore size on the critical flux during the ultrafiltration of protein solutions was investigated using the constant pressure method. Hydrophilic regenerated cellulose membranes with a cut-off of 10, 30 and 100 kg mol⁻¹, model proteins and skimmed milk solutions were used. The critical flux mainly increased with the pore size of the ultrafiltration membrane. The lowest critical fluxes, 40-50 L m⁻²h⁻¹, were obtained with the retentive 10 kg mol⁻¹ cut-off membrane. This membrane had a very low permeability and, thus, the critical fluxes were achieved at high transmembrane pressures (TMP): 1.7-2.3 bar. With the 100 kg mol⁻¹ cut-off membrane critical fluxes were obtained at 0.2 bar TMP, which were around 100 L m⁻² h⁻¹, slightly declining with increasing protein molar mass. In skimmed milk experiments the permeate flux decreased when the protein molecules were enzymatically split to peptides. A critical flux for skimmed milk solution could not be found unless the protein concentration was diluted to 0.3-w% or lower. The results with model proteins were then compared to those obtained with skimmed milk resulting in β-lactoglobulin being the worst foulant.     

Refining of biodiesel by ceramic membrane separation

A ceramic membrane separation process for biodiesel refining was developed to reduce the considerable usage of water needed in the conventional water washing process. Crude biodiesel produced by refined palm oil was micro-filtered by ceramic membranes of the pore size of 0.6, 0.2 and 0.1 μm to remove the residual soap and free glycerol, at the transmembrane pressure of 0.15 MPa and temperature of 60 °C. The flux through membrane maintained at 300 L m^− 2 h^− 1 when the volumetric concentrated ratio reached 4. The content of potassium, sodium, calcium and magnesium in the whole permeate was 1.40, 1.78, 0.81 and 0.20 mg/kg respectively, as determined by inductively coupled plasma-atomic emission spectroscopy. These values are lower than the EN 14538 specifications. The residual free glycerol in the permeate was estimated by water extraction, its value was 0.0108 wt.%. This ceramic membrane technology was a potential environmental process for the refining of biodiesel.     

酶-膜法提取纯化荔枝核中氨基酸

采用酶-膜法提取纯化荔枝核中氨基酸。使用X JT9503中性蛋白酶酶解辅助提取荔枝核中氨基酸,实验确定酶解的优化条件为:酶用量600 U/g、酶解温度50℃、pH=6.5、酶解时间90 m in,氨基酸得率为1.16%。采用水提法和酸解提取法氨基酸得率分别仅为0.41%和0.92%。实验表明,酶解提取法要优于水提法和酸解提取法。使用截留相对分子质量50 000的陶瓷超滤膜纯化酶解提取液,实验表明,膜通量随操作压力和料液温度升高而增加。在操作压力0.22 MPa,料液温度30℃的条件下,膜的平均通量为75.63 L/(m2.h),氨基酸的截留率仅为10.3%,蛋白质的截留率为98.1%,多糖的截留率为85.2%,氨基酸能够与蛋白质、多糖等大分子实现有效分离。     

Fabrication and characterization of hydrophobic PVDF hollow fiber membranes for desalination through direct contact membrane distillation

The mixture of inorganic salt LiCl and soluble polymer polyethylene glycol (PEG) 1500 as non-solvent additive was introduced to fabricate hydrophobic hollow fiber membrane of polyvinylidene fluoride (PVDF) by phase inversion process, using N,N-dimethylacetamide (DMAc) as solvent and tap water as the coagulation medium. Compared with other three membranes from PVDF/DMAc, PVDF/DMAc/LiCl and PVDF/DMAc/PEG 1500 dope solution, it can be observed obviously by scanning electron microscope (SEM) that the membrane spun from PVDF/DMAc/LiCl/PEG 1500 dope had longer finger-like cavities, ultra-thin skins, narrow pore size distribution and porous network sponge-like structure owing to the synergistic effect of LiCl and PEG 1500. Besides, the membrane also exhibited high porosity and good hydrophobicity. During the desalination process of 3.5 wt% sodium chloride solution through direct contact membrane distillation (DCMD), the permeate flux achieved 40.5 kg/m² h and the rejection of NaCl maintained 99.99% with the feed solution at 81.8 °C and the cold distillate water at 20.0 °C, this performance is comparable or even higher than most of the previous reports. Furthermore, a 200 h continuously desalination experiment showed that the membrane had stable permeate flux and solute rejection, indicating that the as-spun PVDF hollow fiber membrane may be of great potential to be utilized in the DCMD process.     

Ethanol Production from Soybean Fiber,a Co-product of Aqueous Oil Extraction,Using a Soaking in Aqueous Ammonia Pretreatment

The effectiveness of soaking in aqueous ammonia (SAA) as a pretreatment method for the conversion of soybean fiber to ethanol via simultaneous saccharification and fermentation (SSF) was investigated. Insoluble fiber is a co-product from oil and protein extraction using two-stage, countercurrent, enzyme-assisted, aqueous extraction processing of full-fat soybean flakes (FFSF) and extruded FFSF. The fiber fractions were soaked in 15 wt% aqueous ammonia at 1:10 solid-to-liquid ratio. The effects of operating variables, including treatment times (6, 12, and 24 h), treatment temperatures (60 and 80 °C), and cellulase loadings (15 and 60 FPU/g-glucan) on the degree of enzymatic hydrolysis were determined. The best SAA conditions were 80 °C for 12 h followed by an enzyme loading of 15 FPU/g-glucan, which produced a 152-mg/g glucose yield after 48 h of hydrolysis. This was 8.7 times the amount produced from the same fiber not pretreated with SAA. The glucose yield increased to 381 mg/g when fiber obtained from extruded FFSF was submitted to SAA. SAA (80 °C, 12 h) on extruded fiber subjected to SSF increased ethanol yield from 0.06 g of ethanol/g [40% of theoretical yield] (for non SAA pretreated fiber) to 0.25 g of ethanol/g [92% of theoretical yield]. The combination of extrusion and SAA was an efficient means for converting the fiber-rich soybean fraction into ethanol.     

Application of ultrafiltration integrated with coagulation for improved NOM removal

Alum coagulation followed by ultrafiltration was studied in order to improve water quality for surface water treatment. The influence of pH (from 5 to 10) and coagulant dose (25, 40 and 50 g alum/m³ equal to 1.795, 2.872 and 3.59 g Al/m³) on the investigated process performance was analyzed. Two ultrafiltration membranes (cut-off 30 kD) made of regenerated cellulose and polyethersulphone were used. The experiments showed that even the lowest dose of alum significantly increased the quality of permeate. The maximum NOM separation was observed at pH 6 and a further increase of solution reaction resulted in a worsening of the permeate quality.     

Enzymatic Hydrolysis of Cassava Starch into Maltose Syrup in a Continuous Membrane Reactor

This study deals with the use of a membrane reactor for the enzymatic conversion of cassava starch to maltose. The enzymes used were Maltogenase and Promozyme (Novo Nordisk). Maltogenase activity was unaffected after a 5 h incubation period at 65°C, but Promozyme was markedly heat-unstable even at 37°C. Batch hydrolysis of liquefied cassava starch (30% w/w) by Maltogenase and Promozyme resulted in a maximum degree of starch conversion to maltose of 72% (≈254 g dm⁻³ maltose). The conversion degree fell by 11% when no debranching enzyme was used. The residence time distribution of the ultrafiltration reactor (UFR) was that of an ideal continuously stirred tank reactor. Rejection of Maltogenase by Carbosep M4 membranes (MWCO: 50 kDa) was not total. The overall enzyme activity loss after a 5 h diafiltration period was 28%, however about half this loss appeared to be due to enzyme denaturation inside the reactor. During saccharification trials conducted in the UFR at a starch concentration of 30% (w/w), severe membrane fouling occurred. The average permeate fluxes obtained were 14 and 23 dm³ h⁻¹ m⁻² at constant transmembrane pressures of 100 and 200 kPa respectively. When the reactor was operated at a space-time of 4·2 h, the degree of starch conversion to maltose in the permeate rapidly stabilized around 55–56%. © 1997 SCI.     

Extraction and Separation of α-lactalbumin and β-Lactoglobulin from Skim Milk by Microfiltration and Ultrafiltration at High Shear Rates: A Feasibility Study

Abstract

This paper presents a two-stage membrane filtration process for extracting and separating α-Lactalbumin (α-La) and β-Lactoglobulin (β-Lg), from UHT skim milk, using dynamic filtration. The 1st stage separates casein micelles in retentate from whey proteins in the permeate with rotating 0.2 µm pores ceramic membrane disks. Casein micelles rejection was excellent, while α-La and β-Lg transmissions remained between 80 and 90%. The permeate flux at 40°C ranged from 105 to 40 Lh^?1m^?2 at a volume reduction ratio of VRR = 4. The 2nd stage consisted of ultrafiltration of the previous permeate with a metal disk rotating at 2000 rpm near a fixed 50 kDa PES membrane, in order to concentrate β-Lg in retentate, while collecting α-La in the permeate. The flux dropped from 270 Lh^?1m^?2 at VRR = 1, and remained nearly constant at 200 Lh^?1m^?2 until a VRR of 3.3. α-La transmission increased with VRR to reach 23% at VRR = 3.3, while β-Lg transmission decayed at increasing VRR to 3%, to give a maximum selectivity of 8.