Comparison of flat-sheet and spiral-wound negatively-charged wide-pore ultrafiltration membranes for whey protein concentration

This work compared laboratory-scale flat-sheet and pilot plant-scale spiral-wound wide-pore, negatively-charged ultrafiltration membranes for concentration of whey proteins. By placing a negative charge on the surface of ultrafiltration membranes, a wider pore size could be used to concentrate whey proteins because negatively-charged proteins were rejected by electrostatic repulsion and not simply sized-based sieving. Negatively-charged 100 kDa regenerated cellulose membranes had an 85% higher flux than unmodified 10 kDa membranes, and equivalent protein retention. The pilot plant-scale spiral-wound membranes had 70-fold more area, and a different membrane geometry than the laboratory-scale flat-sheet membranes, yet both membranes were successful in retaining >98% of the whey protein.     

An investigation of the fractionation of whey proteins by two microfiltration membranes with nominal pore size of 0.1 μm

There is growing interest in the fractionation of whey proteins because of the specific properties of individual whey proteins. The objective of this work was to assess the efficiency of two membranes to obtain two fractions, one rich in β‐lactoglobulin (β‐Lg) and the other rich in α‐lactalbumin (α‐La) from a whey‐protein concentrate using microfiltration (MF). Two MF membranes were tested for the fractionation: a flat‐sheet membrane VCWP and a spiral membrane MF‐7002, both with nominal pore sizes of 0.1 μm. The VCWP retained 78% of the proteins in solution, and this membrane was shown to be permeable to both proteins, β‐Lg and α‐La. The retention of protein by MF‐7002 was 65%, and there was a partial retention of lactose. The permeate collected in the MF‐7002 in the concentration stage was over 50%α‐La; although this was present in lower concentrations, it was passed preferentially to the permeate. The results indicated that some aggregation of the proteins may have occurred under the experimental conditions because there was only a partial separation of the proteins under study.     

超滤膜分离技术回收乳清蛋白工艺研究

研究利用超滤膜分离技术,从干酪素乳清废弃液中回收乳清蛋白,通过对不同超滤膜性能的比较,选择最佳的超滤膜材料、工艺流程以及运行参数,并测得分离效果。结果表明:采用PW2540型聚醚砜卷式超滤膜较好,其最佳工艺参数为操作温度35℃,操作压力0.5MPa,且超滤膜透液通量较高,运行稳定。乳清蛋白粉中蛋白质含量72.40%,灰分3.85%。经红外光谱检测证明乳清蛋白粉品质得到较大程度的提高。每吨乳清废弃液中可回收乳清蛋白粉5.13kg,具有较好的经济效益及减排环保效益。     

Selective Enrichment of Proteins Using Formed-In-Place Membranes

A process for selective enrichment of immunoglobulins has been developed using formed-in-place membranes on sintered stainless steel tubes. The process employs both size exclusion and protein charge manipulation around the isoelectric point of proteins to selectively reject or pass components of the stream. Volume recoveries of 95% (20X concentration) could be achieved with a single filtration without loss of fractionation capabilities. The large initial volume reduction allows for small subsequent diafiltration volumes and insures optimization of fractionation while minimizing total processing volume. The process has been used to enrich proteins representing a small percentage of the total protein from a large feed volume. As an example, immunoglobulin G (IgG) from cheese whey was enriched from 8% to 20% of total whey protein with a 90% recovery of the IgG.     

Comparison of the behavior of two nanofiltration membranes for sweet whey demineralization

Nanofiltration is a process used to separate mineral salts from lactose, having previously removed the proteins by ultrafiltration. Both proteins and lactose can be used as raw materials to prepare a variety of products. In this paper, we studied the feasibility of demineralizing sweet whey obtained from the cheese industry of the Comunidad Valenciana (Spain) using membrane technologies. The NF200 membrane showed the highest volumetric flux and solute rejection values, whereas the DS-5 DL membrane showed the lowest values. The volumetric fluxes obtained with the NF200 and DS-5 DL membranes in these experiments with the ultra-filtered whey demonstrated significant differences between membranes. Concerning solute rejection, the highest values were obtained using the NF200 membrane. The chosen parameter to evaluate the demineralization capability was solute flux. In this way, the values obtained for chloride ion were 9.90 and 32.42 g/ (m²·h) for the NF200 and DS-5 DL membranes, respectively, with the highest demineralization rates being achieved with the DS-5 DL membrane.     

Ultrafiltration of whey

Utilization of ultrafiltration for the treatment of whey is increasing in the world dairy industry, due to the progress made in membrane technology itself and a better knowledge of the behaviour of whey components. Pretreatment of whey either by heating accompanied or not with pH adjustment or by ionic strength regulation or by prefiltration has the greatest effect on permeation rate. Preconcentration of whey also improves performances of ultrafiltration equipment. Two new potentialities for whey proteins, made possible by properties of new membranes are also presented.     

Purification of goat beta-lactoglobulin from whey by an ultrafiltration membrane enzymic reactor

This paper presents a novel contribution to the purification of goat beta-lactoglobulin by using an ultrafiltration membrane enzymic reactor. The basis of the purification process was the enzymic hydrolysis of contaminating proteins, alpha-lactalbumin and traces of serum albumin, by pepsin at 40 degrees C and pH 2, conditions under which beta-lactoglobulin is resistant to peptic digestion. Simultaneously, beta-lactoglobulin and peptides were separated by ultrafiltration. beta-Lactoglobulin was retained in the reactor while peptides generated by hydrolysis from alpha-lactalbumin and serum albumin permeated through the membrane. The process was made continuous by the addition of fresh whey to replace the lost permeate. Three mineral membranes with 10, 30 and 50 kDa molecular mass cut-off were tested and the 30 kDa membrane was selected for the continuous process. The simultaneous purification and concentration of beta-lactoglobulin from clarified goats' whey was achieved in a single step. The ultrafiltration membrane enzymic reactor could treat eight reactor volumes of clarified whey. The recovery of beta-lactoglobulin was 74%, its purity was 84% and its concentration 6.6-fold that in the initial clarified whey.     

Selective separation of the major whey proteins using ion exchange membranes

Synthetic microporous membranes with functional groups covalently attached were used to selectively separate β-lactoglobulin, BSA, and α-lactalbumin from rennet whey. The selectivity and membrane performance of strong (quaternary ammonium) and weak (diethylamine) ion-exchange membranes were studied using breakthrough curves, measurement of binding capacity, and protein composition of the elution fraction to determine the binding behavior of each membrane. When the weak and strong anion exchange membranes were saturated with whey, they were both selective primarily for β-lactoglobulin with less than 1% of the eluate consisting of α-lactalbumin or BSA. The binding capacity of a pure β-lactoglobulin solution was in excess of 1.5 mg/cm² of membrane. This binding capacity was reduced to approximately 1.2 mg/cm² when using a rennet whey solution (pH 6.4). This reduction in protein binding capacity can be explained by both the competitive effects of other whey proteins and the effect of ions present in whey. Using binary solution breakthrough curves and rennet whey breakthrough curves, it was shown that α-lactalbumin and BSA were displaced from the strong and weak anion exchange membranes by β-lactoglobulin. Finally, the effect of ionic strength on the binding capacity of individual proteins for each membrane was determined by comparing model protein solutions in milk permeate (pH 6.4) and a 10 mM sodium phosphate buffer (pH 6.4). Binding capacities of β-lactoglobulin, α-lactalbumin, and BSA in milk permeate were reduced by as much as 50%. This reduction in capacity coupled with the low binding capacity of current ion exchange membranes are 2 serious considerations for selectively separating complex and concentrated protein solutions.     

Functional and Biological Properties of Peptides Obtained by Enzymatic Hydrolysis of Whey Proteins

The study of peptides released by enzymatic hydrolysis of whey proteins has been initially focusing on improving their functional properties in food model systems. Our first study showed that peptides 41 to 60 and 21 to 40 from β-lactoglobulin (β-LG) were responsible for improved emulsifying properties of a tryptic hydrolysate of whey protein concentrate (WPC). Further work showed that adding negatively charged peptides from tryptic hydrolysates of WPC could prevent phase separation of dairy-based concentrated liquid infant formula, as a replacement for carrageenan. Hydrolysis of whey proteins using a bacterial enzyme was also successful in improving heat stability of whey proteins in an acidic beverage. Some tryptic peptides demonstrated improvement in the heat stability and in modifying thermal aggregation of whey proteins. Recent research has shown that whey peptides could trigger some physiological functions. Within the scope of this research our work has led to the development of a whey protein enzymatic hydrolysate that has demonstrated antihypertensive properties when orally administered to spontaneously hypertensive rats and human subjects. Our work then focused on the fractionation of hydrolysates by nanofiltration to prepare specific peptidic fractions; however, peptide/peptide and peptide/protein interactions impaired membrane selectivity. The study of those interactions has lead to the demonstration of the occurrence of interactions between β-LG and its hydrophobic fragment 102–105 (opioid peptide), which probably binds in the central cavity of the protein. This latest result suggests that β-LG could be used as a carrier for the protection of bioactive peptides from gastric digestion. Our work therefore has shown that the enzymatic hydrolysis of whey proteins is not only improving their functional properties, but it is also providing powerful technology in the exploitation of their biological properties for functional foods and nutraceutical applications.     

Natural Caprine Whey Oligosaccharides Separated by Membrane Technology and Profile Evaluation by Capillary Electrophoresis

The functional food market is growing rapidly and membrane processing offers several advantages over conventional methods for separation, fractionation and recovery of bioactive components. The aim of the present study was to select a process that could be implemented easily on an industrial scale for the isolation of natural lactose-derived oligosaccharides (OS) from caprine whey, enabling the development of functional foods for clinical and infant nutrition. The most efficient process was the combination of a pre-treatment to eliminate proteins and fat, using an ultrafiltration (UF) membrane of 25-kDa molecular weight cutoff (MWCO), followed by a tighter UF membrane with 1-kDa MWCO. Circa 90 % of the carbohydrates recovered in the final retentate were OS. Capillary electrophoresis was used to evaluate the OS profile in this retentate. The combined membrane-processing system is thus a promising technique for obtaining natural concentrated OS from whey.     

Nanofiltration Concentration Effect on the Efficacy of Lactose Crystallization

Application of nanofiltration membranes to processing sweet whey and skim milk ultrafiltration permeate increased lactose crystal yield by about 10 and 8 %, respectively, at a concentration factor of 3.0. These increases were attributed to depletion of minerals, especially monovalent cations such as sodium and potassium, by the partial demineralization effect of the nanofiltration membrane. These membranes may be incorporated into current industrial processes for producing lactose from whey and milk permeates.     

蛋白含量对牦牛乳清蛋白浓缩物功能特性的影响

通过不同截留分子质量的再生纤维素膜过滤纯化牦牛原乳清液和牦牛甜乳清液,分别制取牦牛原乳清蛋白浓缩物（native whey protein concentrate,NWPC）和牦牛甜乳清蛋白浓缩物（sweet whey protein concentrate,SWPC）,研究蛋白含量不同的乳清蛋白浓缩物（whey protein concentrate,WPC）主要成分（乳糖含量、pH值和总蛋白质含量）和功能特性（溶解性、持水性、持油性、起泡性、乳化性及热稳定性）的特征。结果表明：10 000 Da再生纤维素膜透析得到的牦牛WPC中总蛋白含量达到80%以上,不含乳糖,功能特性（溶解性、持水性、持油性、起泡性、乳化性及热稳定性）均显著高于经3 500 Da卷式膜、5 000 Da再生纤维素膜透析得牦牛WPC,WPC蛋白含量越高,其功能特性越好;不同蛋白含量的牦牛SWPC起泡能力、泡沫稳定性、乳化活性和乳化稳定性均显著（P＜0.05）高于牦牛NWPC。牦牛乳WPC最不稳定温度为85 ℃,高于荷斯坦牛乳WPC的80 ℃,热处理会适当改善牦牛WPC的起泡性能、乳化性能和热稳定性。通过膜牦牛处理获取的高蛋白含量的WPC,功能特性较好,应用广泛,对解决牦牛乳清资源的利用问题、保护环境、提高企业的经济效益起到关键性作用。     

Fermentation of specially formulated milk with single strains of bifidobacteria

A number of strains of bifidobacteria have been used to ferment milk to a yoghurt-like product. The milk used for fermentation was fortified with whey protein and threonine. It was prepared by mixing skimmed milk and cheddar cheese whey which had been concentrated using ultrafiltration membranes which allowed lactose to permeate. A 2 percent inoculum was used, and fermentation was carried out at 37° C overnight. The resulting products resembled voghurt, having good consistency. 'walnutty' aroma (acetaldehyde). and pleasant acidity.     

Surfactant cleaning of ultrafiltration membranes fouled by whey

A polyethersulphone ultrafiltration membrane was prepared for concentration of whey. The membrane was fouled by whey and the effect of different cleaning agents on flux recovery of the fouled membrane was studied. The optimum cleaning procedure for membrane regeneration was elucidated. The results showed that a combination of surfactants (anionic, cationic and nonionic) may be employed as the optimum cleaning agent for maximum flux recovery. The fluorescence studies revealed that the cationic surfactant interact with proteins by breaking the intra‐chain hydrophobic bonding and providing electrostatic repulsion. Changing the alkyl chain from dodecyl to hexadecyl increases the interaction of surfactant–protein. Dodecyltrimethylammonium bromide (DTAB) provided a weak interaction with whey proteins than to tetradecyltrimethylammonium bromide (TTAB) and cetyltrimethylammonium bromide (CTAB). All data obtained in this study support a surfactant–protein interaction in which hydrophobic forces play a dominant role. The nonionic surfactants poly(oxyethylene) isooctyl phenyl ether (TX‐100) and anionic surfactants SDS interact with amino acids in the inner protein structure thus denaturate tertiary protein structure and reduce hydrophobic interaction of proteins by membrane surface.     

Application of a membrane technology to remove bacteriophages from whey

Whey from former cheese batches can be recycled either to increase the yield or to improve texture properties of fat reduced cheeses. However, in the case of the presence of bacteriophages, pasteurization may not be sufficient to eliminate phages in whey. Therefore, in this work, a cross-flow membrane filtration process was designed to separate whey proteins from whey-derived phages. Filtration experiments were carried out using native whey as model filtration medium, three polyethersulfone membranes (100, 300 and 500 kDa) that were studied in detail, and lactococcal phage P008. Filtration performance was characterized by phage retention, total whey protein permeation, and permeation of the major whey proteins α-lactalbumin and β-lactoglobulin. Filtration experiments showed that it is possible to reduce the number of phages in whey by filtration to a level at which subsequent phage multiplication is minimized and, concomitantly, high protein permeation through the membrane is ensured.     

Whey filtration: a review of products,application, and pretreatment with transglutaminase enzyme

In the cheese industry, whey, which is rich in lactose and proteins, is underutilized, causing adverse environmental impacts. The fractionation of its components, typically carried out through filtration membranes, faces operational challenges such as membrane fouling, significant protein loss during the process, and extended operating times. These challenges require attention and specific methods for optimization and to increase efficiency. A promising strategy to enhance industry efficiency and sustainability is the use of enzymatic pre-treatment with the enzyme transglutaminase (TGase). This enzyme plays a crucial role in protein modification, catalyzing covalent cross-links between lysine and glutamine residues, increasing the molecular weight of proteins, facilitating their retention on membranes, and contributing to the improvement of the quality of the final products. The aim of this study is to review the application of the enzyme TGase as a pretreatment in whey protein filtration. The scope involves assessing the enzyme's impact on whey protein properties and its relationship with process performance. It also aims to identify both the optimization of operational parameters and the enhancement of product characteristics. This study demonstrates that the application of TGase leads to improved performance in protein concentration, lactose permeation, and permeate flux rate during the filtration process. It also has the capacity to enhance protein solubility, viscosity, thermal stability, and protein gelation in whey. In this context, it is relevant for enhancing the characteristics of whey, thereby contributing to the production of higher quality final products in the food industry. © 2023 Society of Chemical Industry.     

Membrane Fractionation Processes for Removing 90% to 95% of the Lactose and Sodium from Skim Milk and for Preparing Lactose and Sodium‐Reduced Skim Milk

ABSTRACT: Pilot‐scale microfiltration (MF), microfiltration‐diafiltration (MDF), ultrafiltration (UF), ultrafiltration‐diafiltration (UDF), and nanofilration (NF) membrane fractionation processes were designed and evaluated for removing 90% to 95% of the lactose and sodium from skim milk. The study was designed to evaluate several membrane fractionation schemes as a function of: (1) membrane types with and without diafiltration; (2) fractionation process temperatures ranging from 17 to 45 °C; (3) sources of commercial drinking water used as diafiltrant; and (4) final mass concentration ratios (MCR) ranging from about 2 to 5. MF and MDF membranes provided highest flux values, but were unsatisfactory because they failed to retain all of the whey proteins. UDF fractionation processes removed more than 90% to 95% of the lactose and sodium from skim milk. NF permeate prepared from UDF cumulative permeate contained sodium and other mineral concentrations that would make them unsuitable for use as a diafiltrant for UDF applications. A method was devised for preparing simulated milk permeate (SMP) formulated with calcium, magnesium, and potassium hydroxides, and phosphoric and citric acids for use as UDF diafiltrant or for preparing lactose and sodium reduced skim milk (L‐RSM). MF retentates with MCR values of 4.7 to 5.0 exhibited extremely poor frozen storage stabilities of less than 1 wk at ?20 °C, whereas MCR 1.77 to 2.95 MDF and UDF retentates and skim milk control exhibited frozen storage stabilities of more than 16 wk. L‐RSM exhibited a whiter appearance and a lower viscosity than skim milk, lacked natural milk flavor, and exhibited a metallic off‐flavor.     

PROCESSING WHEY-TYPE BY-PRODUCT LIQUIDS FROM COTTONSEED PROTEIN ISOLATION WITH ULTRAFILTRATION AND REVERSE OSMOSIS MEMBRANES

Cottonseed wheys resulting from protein isolation from cottonseed flour were processed by semi-permeable ultrafiltration (UF) and reverse osmosis (RO) membranes. The UF membrane fractionated the soluble whey constituents by retaining protein and passing through salts, carbohydrates and other non-protein components along with most of the water. The UF membrane effluent was then processed through an RO membrane to recover a secondary product containing the whey materials not retained in the protein product from the UF membrane. The feasibility of recycling effluent from the RO membrane for reuse in subsequent protein extractions was demonstrated. Thus, the threat of water pollution from effluent disposal could be eliminated completely and process water requirements drastically reduced. Spray-dried UF protein concentrates were tested for utilization in protein fortification of breads and noncarbonated beverages and as whipping products. They exhibited commercial potential for use in these food applications. The economics of processing the whey-type liquids by the membrane process under investigation were analyzed. Membrane processing of wheys by each of two alternative whey processing systems proved to be economically attractive.     

膜技术在处理大豆乳清废水中的应用

大豆乳清废水经预处理后,采用超滤膜技术回收含低聚糖废水中的乳清蛋白,再用纳滤膜脱盐、浓缩低聚糖,滤液过反渗透膜即可达到回用或排放要求。探讨了预处理工序的必要性,考察了不同型号膜的运行情况并进行了选择。实验证明,该工艺简单、节能、易操作,污水零排放,且回收产品质量有较大提高,中试数据可供工业化参考与借鉴。     

Evaluation of commercially available, wide-pore ultrafiltration membranes for production of α-lactalbumin-enriched whey protein concentrate

Commercially available, wide-pore ultrafiltration membranes were evaluated for production of α-lactalbumin (α-LA)-enriched whey protein concentrate (WPC). In this study microfiltration was used to produce a prepurified feed that was devoid of casein fines, lipid materials, and aggregated proteins. This prepurified feed was subsequently subjected to a wide-pore ultrafiltration process that produced an α-LA-enriched fraction in the permeate. We evaluated the performance of 3 membrane types and a range of transmembrane pressures. We determined that the optimal process used a polyvinylidene fluoride membrane (molecular weight cut-off of 50 kDa) operated at transmembrane pressure (TMP) of 207 kPa. This membrane type and operating pressure resulted in α-LA purity of 0.63, α-LA:β-LG ratio of 1.41, α-LA yield of 21.27%, and overall flux of 49.46 L/m²·h. The manufacturing cost of the process for a hypothetical plant indicated that α-LA-enriched WPC 80 (i.e., with 80% protein) could be produced at $17.92/kg when the price of whey was considered as an input cost. This price came down to $16.46/kg when the price of whey was not considered as an input cost. The results of this study indicate that production of a commercially viable α-LA-enriched WPC is possible and the process developed can be used to meet worldwide demand for α-LA-enriched whey protein.