Feed substrates influence biofilm formation on reverse osmosis membranes and their cleaning efficiency

The dairy industry is increasingly using reverse osmosis (RO) membranes for concentration of various fluid feed materials such as whey and ultrafiltration (UF) permeate. This study compared the effect of UF permeate and whey on membrane biofilm formation. A Bacillus sp., previously isolated in our laboratory from a cleaning-resistant membrane biofilm, was used to develop 48-h-old static biofilms on RO membrane pieces, using the different feed substrates (UF permeate, whey, and an alternating whey/UF feed). Biofilms were analyzed for viable counts by the swab technique, and we used scanning electron and atomic force microscopy for microstructure imaging. The membrane cleaning process included 6 sequential steps. We observed differences in the resistance pattern of the 3 types of biofilms to the typical cleaning process. The mean pretreatment counts of the 48-h UF permeate biofilms were 5.39 log cfu/cm², much higher than the whey biofilm counts of 3.44 log, and alternating whey/UF biofilm counts of 4.54 log. After a 6-step cleaning cycle, we found 2.54 log survivors of the Bacillus isolate on UF biofilms, whereas only 1.82 log survivors were found in whey biofilm, and 2.14 log survivors on whey/UF permeate biofilms. In conclusion, the UF permeate biofilms was more resistant to the biofilm cleaning process compared with the whey or whey/UF permeate biofilms. Scanning electron micrographs showed different microstructures of biofilms based on the type of feed. For UF permeate and whey/UF permeate biofilms, bacilli were present in multilayers of cells in aggregates or irregular clusters with foulant layers. In contrast, those in whey biofilms were in monolayers, with a smoother, flatter appearance. Atomic force microscopy analysis indicated that UF permeate biofilms had the greatest surface roughness among the biofilms, reflecting intensified bacterial colonization. The biofilm micro- and nanostructure variations for the 2 feed substrates and their combination may have resulted in differences in their resistance to the cleaning process.     

Resistance of the constitutive microflora of biofilms formed on whey reverse-osmosis membranes to individual cleaning steps of a typical clean-in-place protocol

This experiment evaluates the effectiveness of individual steps of a clean-in-place protocol against the biofilm constitutive microflora isolated from the biofilms developed on whey reverse-osmosis membranes, aged 2 to 14 mo, under industrial processing conditions. The isolates used for the in vitro resistance studies included species of Bacillus, Enterococcus, Streptococcus, Staphylococcus, Micrococcus, Aeromonas, Corynebacterium, Pseudomonas, Klebsiella, and Escherichia. The 6 cleaning steps (alkali, surfactant, acid, enzyme, a second surfactant, and sanitizer treatment) revealed resistance of isolates in both planktonic and biofilm-embedded cell states. The most effective step was the acid treatment, which resulted in 4.54 to 7.90 and 2.09 to 5.02 log reductions of the planktonic and biofilm-embedded cells, respectively. Although the sanitizer step causing a reduction of 4.91 to 8.33 log in the case of planktonic cells, it was less effective against the biofilm-embedded cells, resulting in a reduction of 0.59 to 1.64 log. Bacillus spp. showed the highest resistance in both planktonic, as well as embedded cell states.     

Growth potential and biofilm development of nonstarter bacteria on surfaces exposed to a continuous whey stream

Commercial Cheddar cheese production uses an automated, continuous production system that provides favorable conditions for specific undesirable bacterial subpopulations in certain sections of the processing system. The draining and matting conveyor (DMC) is a large, fully enclosed series of conveyor belts that separates curd and whey on the first drain belt and supports the cheddaring process in subsequent sections. In a previous study, we demonstrated that coliforms increase in the draining section of the DMC (pH 6.0–6.3, 36°C) over a typical 18-h production shift and can lead to detectable coliforms in finished cheese. Sampling at the commercial plant indicated 2 sources of very low levels of coliforms: (1) subpasteurized whey and curd entering the DMC and (2) surfaces in the DMC after sanitation. Mitigation of these sources would require different approaches. The aim of this study was to investigate whether naturally low levels of coliforms in whey could increase in the bulk liquid and attach to different surface materials within 18 h. A laboratory-scale system was created to mimic the conditions of the initial draining section of the DMC and consisted of single-pass, naturally contaminated whey (pH 6.3, 35°C) flowing through a bioreactor (1.11 L/h) containing coupons of surface types found in the DMC (stainless steel and polypropylene). Whey inside the bioreactor chamber and surface coupons were enumerated for bacterial subpopulations on selective media for planktonic and attached bacteria, respectively, at 0, 12, 15, and 18 h. Bacterial isolates were identified by 16S rDNA sequencing. Nonstarter bacteria present in the whey at 0 h included coliforms (Enterobacter), Pseudomonas, and Acinetobacter (0.80, 2.55, and 2.32 log cfu/mL, respectively), with each increasing significantly in whey (6.18, 7.00, and 5.89 log cfu/mL) and on coupons (5.20, 6.85, and 5.29 log cfu/cm², respectively) after 18 h in the continuous flowing system. Scanning electron microscopy confirmed bacterial attachment on both surfaces, with early biofilm development evident on polypropylene coupons by 18 h. Results from this laboratory-scale study demonstrated that naturally low levels of coliforms entering the DMC in the whey could replicate within the conditions of the draining section of the DMC to the levels found in the commercial production environment.     

Preparation and composition of chhana whey powders using membrane processing techniques

Chhana is a traditional Indian product used widely in the confectionery industry. It is produced from cow's milk by a combination of heat and acid coagulation. Chhana whey contains about 6% milk solids yet the vast majority is wasted which leads to pollution problems. This study describes the chemical composition and various options for utilisation of chhana whey using membrane processes. Chhana whey powder containing 956 g kg^?1 total solids, 750 g kg^?1 lactose, 21 g kg^?1 protein. 60 g kg^?1 fat, 65 g kg^?1 ash was produced following concentration of chhana whey by reverse osmosis. Chhana whey protein concentrate powders containing 270, 350, 400 and 580 g kg^?1 protein were produced following ultrafiltration or diafiltration of chhana whey.     

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。     

Efficiency of removal of whey protein from sweet whey using polymeric microfiltration membranes

Our objective was to measure whey protein removal percentage from separated sweet whey using spiral-wound (SW) polymeric microfiltration (MF) membranes using a 3-stage, 3× process at 50°C and to compare the performance of polymeric membranes with ceramic membranes. Pasteurized, separated Cheddar cheese whey (1,080 kg) was microfiltered using a polymeric 0.3-μm polyvinylidene (PVDF) fluoride SW membrane and a 3×, 3-stage MF process. Cheese making and whey processing were replicated 3 times. There was no detectable level of lactoferrin and no intact α- or β-casein detected in the MF permeate from the 0.3-μm SW PVDF membranes used in this study. We found BSA and IgG in both the retentate and permeate. The β-lactoglobulin (β-LG) and α-lactalbumin (α-LA) partitioned between retentate and permeate, but β-LG passage through the membrane was retarded more than α-LA because the ratio of β-LG to α-LA was higher in the MF retentate than either in the sweet whey feed or the MF permeate. About 69% of the crude protein present in the pasteurized separated sweet whey was removed using a 3×, 3-stage, 0.3-μm SW PVDF MF process at 50°C compared with 0.1-μm ceramic graded permeability MF that removed about 85% of crude protein from sweet whey. The polymeric SW membranes used in this study achieve approximately 20% lower yield of whey protein isolate (WPI) and a 50% higher yield of whey protein phospholipid concentrate (WPPC) under the same MF processing conditions as ceramic MF membranes used in the comparison study. Total gross revenue from the sale of WPI plus WPPC produced with polymeric versus ceramic membranes is influenced by both the absolute market price for each product and the ratio of market price of these 2 products. The combination of the market price of WPPC versus WPI and the influence of difference in yield of WPPC and WPI produced with polymeric versus ceramic membranes yielded a price ratio of WPPC versus WPI of 0.556 as the cross over point that determined which membrane type achieves higher total gross revenue return from production of these 2 products from separated sweet whey. A complete economic engineering study comparison of the WPI and WPPC manufacturing costs for polymeric versus ceramic MF membranes is needed to determine the effect of membrane material selection on long-term processing costs, which will affect net revenue and profit when the same quantity of sweet whey is processed under various market price conditions.     

溶液pH对不同超滤膜超滤大豆黄浆水的影响

实验研究溶液pH值对不同超滤膜超滤大豆黄浆水的影响。结果表明,当溶液pH值偏离大豆蛋白等电点时,渗透通量增大,膜衰减系数减小,蛋白质截留率变化不大,总糖透过率略有上升,PES-10膜的适宜pH为11,PS-10膜的适宜pH为6.2。     

Survey of salty and sweet whey composition from various cheese plants in Wisconsin

Salty whey is currently underutilized in the dairy industry because of its high salt content and increased processing and disposal costs. Salty whey accounts for 2 to 5% of the total whey generated during Cheddar and other dry-salted cheese manufacture. Because relatively little information is available on salty whey, this study was conducted to determine the range of compositional components in salty whey from commercial cheese plants. Gross compositional differences in percent protein, salt, solids, and fat between sweet whey and salty whey from various dry-salted cheeses from 8 commercial plants were determined. Differences between individual whey protein compositions were determined using sodium dodecyl sulfate-PAGE. Average total solids, fat, and salt content were significantly greater in the salty whey compared with the corresponding sweet whey. True protein was reduced in salty whey although great variability existed among samples. Individual whey proteins identified included lactoferrin (Lf), BSA, immunoglobulin G, β-lactoglobulin, and α-lactalbumin. Salty whey showed an increase in Lf content and a decrease in α-lactalbumin and β-lactoglobulin content when compared with sweet whey. Salty whey may be a source of Lf, potentially increasing its value to whey processors. However, the compositional assessments showed that commercial salty whey is a highly variable waste stream.     

Determination of the efficiency of removal of whey protein from sweet whey with ceramic microfiltration membranes

Our research objective was to measure percent removal of whey protein from separated sweet whey using 0.1-µm uniform transmembrane pressure ceramic microfiltration (MF) membranes in a sequential batch 3-stage, 3× process at 50°C. Cheddar cheese whey was centrifugally separated to remove fat at 72°C and pasteurized (72°C for 15 s), cooled to 4°C, and held overnight. Separated whey (375 kg) was heated to 50°C with a plate heat exchanger and microfiltered using a pilot-scale ceramic 0.1-µm uniform transmembrane pressure MF system in bleed-and-feed mode at 50°C in a sequential batch 3-stage (2 diafiltration stages) process to produce a 3× MF retentate and MF permeate. Feed, retentate, and permeate samples were analyzed for total nitrogen, noncasein nitrogen, and nonprotein nitrogen using the Kjeldahl method. Sodium dodecyl sulfate-PAGE analysis was also performed on the whey feeds, retentates, and permeates from each stage. A flux of 54 kg/m² per hour was achieved with 0.1-µm ceramic uniform transmembrane pressure microfiltration membranes at 50°C. About 85% of the total nitrogen in the whey feed passed though the membrane into the permeate. No passage of lactoferrin from the sweet whey feed of the MF into the MF permeate was detected. There was some passage of IgG, bovine serum albumen, glycomacropeptide, and casein proteolysis products into the permeate. β-Lactoglobulin was in higher concentration in the retentate than the permeate, indicating that it was partially blocked from passage through the ceramic MF membrane.     

乳清多肽的制备及其发酵饮料的开发

研究了乳清多肽的制备、性质及其发酵饮料的开发,结果表明,碱性蛋白酶比中性蛋白酶水解乳清蛋白的能力强,且更经济,水解最佳条件为加酶量为7000(U/g蛋白)、底物添加量为5%、水解温度为60℃、水解初始pH值为8.5,最大乳清蛋白水解度可达到22.45%。最优酒精发酵条件为接种量5%、初始pH7.5、温度22℃、时间45h。乳清多肽发酵饮料的配方为酸量0.1%,蔗糖量为8%,-β环状糊精量为0.5%。     

酪蛋白与乳清蛋白比例对酸奶凝胶性质的影响

研究了乳中酪蛋白和乳清蛋白比例对凝固型酸奶流变学特性和微观结构的影响,结果表明,固定蛋白质质量分数、降低酪蛋白和乳清蛋白的比例,可以明显提高酸奶凝胶的质量.乳中蛋白质质量分数一致时,酸奶凝胶的硬度、黏度、持水力随着酪蛋白和乳清蛋白比例的减小而增大,凝胶网络结构变得更规则、致密,孔隙更小.在低蛋白质质量分数下,降低乳中酪...     

Effect of whey concentration on protein recovery in fresh ovine ricotta cheese

Ricotta cheese, particularly the ovine type, is a typical Italian dairy product obtained by heat-coagulation of the proteins in whey. The aim of this work was to investigate the influence of whey protein concentration, obtained by ultrafiltration, on yield of fresh ovine ricotta cheese. Ricotta cheeses were obtained by thermocoagulation of mixtures with protein content of 1.56, 3.10, 4.16, and 7.09 g/100 g from the mixing of skim whey and ultrafiltered skim whey. A fat-to-protein ratio of 1.1 (wt/wt) was obtained for all mixtures by adding fresh cream. The initial mixtures, as well as the final ricotta cheeses, were analyzed for their composition and by SDS-PAGE. Protein bands were quantified by QuantityOne software (Bio-Rad, Hercules, CA) and identified by liquid chromatography-tandem mass spectrometry. Significant differences in the composition of the ricotta cheese were observed depending on protein concentration. Particularly, ricotta cheese resulting from the mixture containing 7.09 g/100 g of protein presented higher moisture (72.88 ± 1.50 g/100 g) and protein (10.18 ± 0.45 g/100 g) contents than that prepared from the mixture with 1.56 g/100 g of protein (69.52 ± 1.75 and 6.70 ± 0.85 g/100 g, respectively), and fat content was lower in this sample (12.20 ± 1.60 g/100 g) compared with the other treatments, with mean values between 15.72 and 20.50 g/100 g. Each protein fraction presented a different behavior during thermocoagulation. In particular, the recovery of β-lactoglobulin and α-lactalbumin in the cheese increased as their content increased in the mixtures. It was concluded that concentrating ovine rennet whey improved the extent of heat-induced protein aggregation during the thermal coagulation process. This resulted in a better recovery of each protein fraction in the product, and in a consequent increase of ricotta cheese yield.     

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.     

乳清白酒发酵工艺的研究

利用乳清中的乳糖及丰富的营养物质,将乳清与黑龙江传统白酒酿造原料玉米混合发酵,提高白酒出酒率并改善白酒风味。乳糖水解条件：乳糖酶0．3U／kg,水解温度35℃,水解时间为3h,水解率可达57．23％。酵母菌发酵温度28℃,接菌量0.7％。白酒出酒率从40％提高到61％,口感得到明显改善。     

益生菌发酵乳清饮料的工艺研究

以乳清粉为原料研制出益生菌发酵乳清饮料,优化了包括菌种配比、发酵条件和稳定性在内的工艺与配方。结果表明:嗜酸乳杆菌和干酪乳杆菌配比为1:1、接种量为3%、发酵温度为35℃、发酵时间为8h时,益生菌发酵乳清饮料风味最好。稳定剂的配比为:CMC0.25%、PGA0.15%、果胶0.1%。     

果汁低温浓缩技术研究进展

与传统的多级真空浓缩法相比,低温浓缩技术具有保持果汁风味营养物质、降低能耗、操作简单等优点．目前．果汁低温浓缩技术主要有直接反渗透法、超滤与反渗透相结合的膜联合技术．主要介绍了反渗透、膜联合技术在果汁浓缩中的研究进展。     

Microfiltration of whey proteins adsorbed on yeast cells

Soluble whey proteins (WPs), adsorbed on yeast cells, were recovered by a crossflow microfiltration (MF) technique using a cellulose nitrate membrane with a pore size of 0.45 μm. The crossflow velocity was 1.5 m s^?1 with a transmembrane pressure of 200 kPa at 25 °C. A series of protein rejections occured at various pH values ranging from 2 to 8. WPs adsorbed more on to yeast cells at low pH (pH < 4) than at high pH values, probably because they were positively charged at low pH. It was also shown that permeate flux increased and Modified Membrane Fouling Index values decreased at low pH levels. When the yeast concentration was 50 g L^?1, the flux decreased five times compared with that in the absence of yeast. Protein recovery increased with increasing yeast concentrations. The highest protein recovery was found to be 85% at a yeast concentration of 50 g L^?1 at a steady state flux rate of 10^?6 m s^?1 at 25 °C. When diluted solutions of whey were used, the same rejection of protein, adsorbed on yeast cells, was achieved at ten times lower amounts of yeast cells. This technique not only provides for the recovery of protein but also may give rise to the direct use of yeast cells, which are rich in protein, in the baking industry. WPs absorbed by yeast cells can be used to produce nutritionally rich products in areas where yeasts have been already used.     

Efficient encapsulation of fish oil: Capitalizing on the unique inherent characteristics of whey cream and hydrolyzed whey protein

The effects of protein concentration and of blending a phospholipid-rich whey coproduct, Procream (Salibra 700 Procream, Glanbia Nutritionals), with intact or hydrolyzed whey protein concentrate, on fish oil microencapsulation efficiency and oxidative stability were assessed. Trypsin and protease M, from Aspergillus oryzae, were used to produce 2 unique hydrolysates. All microcapsules had excellent encapsulation efficiencies (>92%) and good physical properties, regardless of protein content and Procream inclusion. Intact α-lactalbumin and β-lactoglobulin and their peptides were involved in stabilizing oil droplets. Disulfide interchange resulted in formation of protein aggregates, which were more pronounced in samples containing Procream. Although all microcapsules had relatively good oxidative stability, most had better stability at 2 versus 0.5% protein. Protease M hydrolysate + Procream microcapsules had the highest stability, regardless of protein content. Results demonstrated that Procream, at a reduced protein inclusion level, can partially replace more expensive whey protein ingredients in microencapsulation, when blended with a select hydrolysate.     

乳清酿酒工艺条件的研究

以乳清蛋白多太水解液为底物,利用混合酵母进行发酵,优化培养条件：接种量8％,起始pH值6,5,温度28℃,乙醇产量可达到3．95％以上,发酵度达60％,风味调配最佳组合为：酸量＝0．1％,糖量＝7．0％,β－环状糊精＝0．6％,多肽乳表酒与去蛋白乳清酒相比,营养成分和风味均得到明显改善。