Whey Waste Research Articles

Review: Production and functionality of active peptides from milk

In recent years, research on the production of active peptides obtained from milk and their potential functionality has grown, to a great extent. Bioactive peptides have been defined as specific protein fragments that have a positive impact on body functions or conditions, and they may ultimately have an influence on health. Individual proteins of casein or milk-derived products such as cheese and yogurt have been used as a protein source to study the isolation and activity of peptides with several applications. Currently, the milk whey waste obtained in the production of cheese also represents a protein source from which active peptides could be isolated with potential industrial applications. The active properties of milk peptides and the results found with regard to their physiological effects have led to the classification of peptides as belonging to the group of ingredients of protein nature, appropriate for use in functional foods or pharmaceutical formulations. In this study, the main peptides obtained from milk protein and the past research studies about its production and biological activities will be explained. Second, an analysis will be made on the methods to determinate the biological activities, the separation of bioactive peptides and its structure identification. All of these form the base required to obtain synthetic peptides. Finally, we explain the experimental animal and human trials done in the past years. Nevertheless, more research is required on the design and implementation of equipment for the industrial production and separation of peptides. In addition, different authors suggest that more emphasis should therefore be given to preclinical studies, proving that results are consistent and that effects are demonstrated repeatedly by several research human groups.

Protein Removal from Whey Waste by Foam Fractionation in a Batch Process

The work investigates the separation of proteins from whey waste collected from a local confectionery by the foam fractionation technique in batch mode. The purpose of this work was to evaluate performance criteria of protein separation. The effects of pH, the concentration of initial feed solution, the nitrogen flow rate, the % gas hold up, the bubble diameter, the breaking time of foam, and the optimization of the protein–surfactant ratio (1.5:1) were investigated in detail. Maximum enrichment ratio (48.189), %Rp (96.378) were observed at a gas flow rate of 330 ml/min and pH 5 that is closest to isoelectric point of observed proteins (Bovine serum albumin, β-Lactoglobulin, α-lactalbumin).

Quantitative Characterization of Bovine Serum Albumin, α-Lactalbumin and β-Lactoglobulin in Commercial Whey Sample by RP-LC

A simple, sensitive and precise reverse phase liquid chromatographic method has been developed and validated for quantification of bovine serum albumin (BSA), α-lactalbumin (α-La) and β–lactoglobulin (β-Lg) that are removed from whey waste by foam fractionation method. The data is reproducible over a wide concentration range. This optimized method allowed analysis of BSA, α-La, β-Lg in a mixture within 5 min and could be applied to the analysis of a variety of commercial and laboratory whey products within a short time.

Characterisation and cytomodulatory properties of peptides from Mozzarella di Bufala Campana cheese whey

Bioactive peptides are present in a latent state, encrypted within the amino acid sequence of milk proteins, requiring enzymatic proteolysis for their release. They can be produced by gastrointestinal digestion or food processing, thus they can be present in fermented milks, cheese and also in the by-products of dairy industry such as waste whey. The spectrum of biological activity covered by milk-derived peptides is extremely wide, including antibacterial, immunostimulating, antihypertensive, antithrombotic and opioid actions. However, the characterisation of milk-derived peptides with classical analytical methodologies is severely challenged by the complexity of the milk protein fraction and by the wide dynamic range of relative peptide abundance in both dairy products and by-products. Here we report the characterisation of the peptide fraction released in the whey during the different production stages of Mozzarella di Bufala Campana cheese. The peptide extracts were separated by RP HPLC and analysed by MS in order to identify the peptides produced and to trace the pathway of formation of potential bioactive peptides. The antioxidant properties and the modulatory effect on the cell cycle exerted by the peptide extracts were also studied in CaCo2 cell line. We found that a significant antiproliferative effect on CaCo2 was exerted by Mozzarella di Bufala waste whey peptides.

Analysis of the Mechanism and Regulation of Lactose Transport and Metabolism in Clostridium acetobutylicum ATCC 824

Although the acetone-butanol-ethanol fermentation of Clostridium acetobutylicum is currently uneconomic, the ability of the bacterium to metabolize a wide range of carbohydrates offers the potential for revival based on the use of cheap, low-grade substrates. We have investigated the uptake and metabolism of lactose, the major sugar in industrial whey waste, by C. acetobutylicum ATCC 824. Lactose is taken up via a phosphoenolpyruvate-dependent phosphotransferase system (PTS) comprising both soluble and membrane-associated components, and the resulting phosphorylated derivative is hydrolyzed by a phospho-beta-galactosidase. These activities are induced during growth on lactose but are absent in glucose-grown cells. Analysis of the C. acetobutylicum genome sequence identified a gene system, lacRFEG, encoding a transcriptional regulator of the DeoR family, IIA and IICB components of a lactose PTS, and phospho-beta-galactosidase. During growth in medium containing both glucose and lactose, C. acetobutylicum exhibited a classical diauxic growth, and the lac operon was not expressed until glucose was exhausted from the medium. The presence upstream of lacR of a potential catabolite responsive element (cre) encompassing the transcriptional start site is indicative of the mechanism of carbon catabolite repression characteristic of low-GC gram-positive bacteria. A pathway for the uptake and metabolism of lactose by this industrially important organism is proposed.

Effect of Extraction pH and Temperature on Isoflavone and Saponin Partitioning and Profile During Soy Protein Isolate Production

ABSTRACT: Soy protein isolates (SPIs) are produced industrially using methods using various protein extraction temperatures and pH values. The effects of process temperature (25°C or 60°C) and pH (8.5, 9.5, or 10.5) on isoflavone and group B saponin extraction, partitioning, and profile during bench‐scale SPI production were evaluated. Protein, isoflavone, and saponin extraction increased with increasing temperature and pH. Substantial quantities of isoflavones and saponins remained in the insoluble fraction waste stream. Isoflavones were also lost to the whey waste stream, whereas saponins were not detected in the whey. Neutralization of SPI samples at the time of analytical extraction increased measured isoflavone concentrations for the pH 8.5 process extraction treatments, whereas neutralization substantially increased measured saponin concentrations in SPIs for all process extraction treatments. Malonylglucoside isoflavones were primarily converted to β‐glucoside forms as temperature and pH were increased, and these conditions caused conversion of αg, βg, and βa saponins to saponin V, I, and II forms, respectively. Analytical extraction pH should receive careful consideration when analyzing soy matrices for isoflavones and saponins.

Thermophilic aeration of cattle slurry with whey and/or jam wastes

Thermophilic aeration of cattle slurry and food industrial by-products was studied with the aim to improve hygienic qualities of the slurry so that it could be used as a safe fertiliser for berries to be eaten raw. We also wanted to study if the process would be energetically favourable in an arctic climate. Cattle slurry alone or with whey and/or jam waste was treated. The tests were done in a well heat-insulated reactor with a 10 m 3 volume. Temperature increases up to over 70 °C could be recorded in 19 days even though some processes were carried out in wintertime when the ambient air temperature was less than 0 °C. The heat energy formed was higher than the electrical energy needed to carry out the aeration. The hygienic qualities of the aerated product were good with only minor nitrogen losses. The end product could be useful as a fertiliser and soil improving compound to increase the organic matter content of agricultural soil. Cattle slurry alone was well suited as the raw material if attaining a high temperature was the main goal. A part of slurry could be replaced with food-industrial side products. Whey waste suited better for co-composting than jam waste but the mixture of whey, jam waste, and slurry was optimal for composting.

Production of acetic acid and glycerol from salted and dried whey in a membrane cell recycle bioreactor

The use of salted whey (liquid by-product from the dairy industry containing 7.5% NaCl) as a substrate for either acetic acid or glycerol production was investigated using two yeast strains. One was Kluyveromyces fragilis. The other organism (strain L) can utilize whey lactose that was isolated from waste whey (disposal stream). 8% NaCl, 3% Na 2SO 3 and pH 8.5 at 32°C were the optimum operating conditions for maximum acetic acid production (25.84 g l −1) from salted whey using immobilized cells of yeast strain L. Also, strain L gave a higher yield of acetic acid (0.497 g acetic acid per gram lactose) as compared with K. fragilis (0.322 g acetic acid per gram lactose). 1% Na 2SO 3 at pH 7 and whey supplemented with peptone and malt extract were the optimal conditions for maximum batch glycerol production (13.2 g l −1) from salted whey using immobilized cells of K. fragilis. The extreme values, 18.7 g l −1 for glycerol concentration and 39.78 for the percent yield of glycerol (based on sugar concentration) in a membrane cell recycle bioreactor were higher than those obtained for the immobilized cell batch reactors (13.2 g l −1 and 28, respectively).

Distillery and curd whey wastes as viable alternative sources for biosurfactant production

Biosurfactant production from synthetic medium and industrial waste, viz. distillery and whey wastes was investigated by using an oily sludge isolate Pseudomonas aeruginosa strain BS2. In synthetic medium separately supplemented with glucose and hexadecane as water-soluble and -insoluble carbon sources, respectively, strain BS2 reduced the surface tension of the fermentation broth from 57 to 27 mN/m. The culture produced biosurfactant during the stationary growth phase and its yield was 0.97 g/l. The culture utilized distillery and whey wastes for its growth, as maximum cell counts reached to 54 × 108 and 64 × 109 c.f.u./ml from an initial inoculum size of 1 × 05 c.f.u./ml, respectively, within 48 h of incubation and in these wastes the yields of biosurfactant obtained were 0.91 and 0.92 g/l, respectively. In synthetic medium, distillery and whey wastes, strain BS2 produced a crystalline biosurfactant which belonged to the category of secondary metabolites and its maximum production occurred after the onset of nitrogen-limiting conditions. After recovering biosurfactant from the fermented waste, the chemical oxygen demand (COD) of distillery and whey wastes was significantly reduced by 81 and 87%, respectively. Total acids, nitrogen and phosphate levels in distillery waste were reduced by 90, 92 and 92%, respectively, while in case of whey waste the concentration of these nutrients was reduced by 88, 95 and 93%, respectively. The isolated biosurfactant possessed potent surface active properties, as it effectively reduced the surface tension of water from 72 to 27 mN/m and formed 100% stable emulsions of a variety of water-insoluble compounds such as hydrocarbons, viz. hexadecane, crude oil, kerosene and oily sludge and pesticides, viz. dichlorodiphenyltrichloroethane (DDT) and benzene hexachloride (BHC). The effectiveness of biosurfactant was also evident from its low critical micellar concentration (CMC) which was 0.028 mg/ml.

Stoichiometric analysis ofKluyveromyces fragilis growth on lactose

This paper describes results of the application of simple metabolic model principles to aerobic growth of Kluyveromyces fragilis during the utilisation of a semi-synthetic medium simulating dairy waste (whey). The theoretical yield coefficients (maximum, true and real) of biomass on lactose and oxygen, available electrons and ATP were determined and compared with experimental data. The experimental runs were performed in a B Brown fermenter (15 dm3 of operating capacity). The yeast K fragilis was cultured in semi-synthetic medium containing lactose (40 g dm−3) as a main carbon and energy source and ammonia salts as a main nitrogen source (C/N = 8.74). The yeast growth is closely related to the concentration of oxygen in the media. The experimental yields of biomass on lactose and oxygen reached 94% and 88% of real maximum theoretical values, respectively. Calculated true biomass yield coefficients were closely correlated with the values resulting from the balance analysis of stoichiometric equations. The specific maintenance requirement (mATP) was 14.7 mmole ATP(g DM h)-1. Determination of the elemental composition of the biomass during the course of fermentation resulted in observations of the cellular C/N ratio changes. The conversion of lactose carbon to biomass and CO2 was 74.3%–81.2%. The recovery of ATP reached 79%, and the recovery of oxygen 96.2%. © 2000 Society of Chemical Industry

Production of Bioingredients from Kluyveromyces marxianus Grown on Whey: An Alternative

Whey waste is a major problem for the dairy industry. Finding alternative means to reduce its pollution potential and produce high value-added bioingredients has been attempted by many researchers. Kluyveromyces marxianus var. marxianus is a dairy yeast that produces β-galactosidase, allowing for whey fermentation. Also, K. marxianus has been proposed as a source of: (1) oligonucleotides, used as flavor enhancers in food products; (2) oligosaccharides, used as prebiotics to stimulate the growth of Bifidobacterium sp. in the animal and human intestines; and (3) oligopeptides, immunostimulators added to dairy products that are released in the wort after whey protein proteolysis. Fed-batch fermentation can be used as an alternative process to avoid increases in lactose concentration and prevent the catabolite repression of the respiratory enzyme synthesis during aerobic fermentation, thus allowing for high biomass yields. The relevance of these factors on yeast fermentation of whey is summarized in this critical review.

Production of whey-protein-enriched products.

Whey is the serum liquid portion remaining after removal of fat and casein from milk during the manufacture of cheeses and caseins. Traditionally, whey was regarded in the dairy industry as an undesirable by-product and it was either fed in limited quantities to farm animals or treated as effluent, disposed of as a pollutant into streams and rivers, or irrigated onto pastures. However, environmental awareness led to the imposition of strict controls on the disposal of waste whey which necessitated the construction and operation of costly effluent treatment plants for whey treatment prior to disposal. This, together with a recognition that whey was a potentially valuable source of nutrients, stimulated interest in the development of commercially viable processes to convert liquid whey into viable animal and human food products.

Kinetics of biosurfactant production by Pseudomonas aeruginosa strain BS2 from industrial wastes

Batch kinetic studies were carried out on rhamnolipid biosurfactant production from synthetic medium, industrial wastes viz. distillery and whey waste as substrates. The results indicated that the specific growth rates (μ max) and specific product formation rates (V max) from both the wastes are comparatively better than the synthetic medium, revealing that both the industrial wastes (distillery and whey) can be successfully utilized as substrates for biosurfactant production.

Tegoprens in anaerobic digestion of a mixture of cheese whey, poultry waste, and cattle dung for improved biomethanation

To obtain enriched methane content and improve the anaerobic digestion of a mixture of cattle dung, poultry waste, and cheese whey, the effect of various doses of Tegoprens: T-3012, T-3022, T-5842, T-5843, T-5851, T-5852 has been studied, in bench-scale digesters. Among them, Tegoprens 3022 showed more than a 45% increase in gas production with higher methane content.

Entirely Automated Glucose Monitoring System Based On A Flow Injection Analysis Apparatus

Abstract A process for waste whey treatment, fully based on membrane technologies, has been studied and developed in a pilot plant scale, with the aim to recover chemicals from farm wastes (fats, whey proteins, lactose, salts and water). This process, described in its essential steps, includes a hollow fiber membrane reactor for the hydrolysis of lactose with an immobilized β-galactosidase. The bioreactor performances are studied, in a lab scale, with an entirely automated glucose monitoring system based on a flow injection analysis apparatus. The monitoring system shows interesting performances concerning signal stability, extended linearity (up to 150 mmol l-1) and long term monitoring (up to 3 months without manual operations). The performances of the glucose monitoring system are described.

Utilization of waste whey as a protein source for production of iron proteinate: An antianaemic preparation

Iron proteinates (metalosates) are very effective preparations for the treatment of acute and chronic iron deficiencies in animals. A technology for the production of iron proteinate was elaborated employing waste-whey proteins. The iron content in the preparation could be varied by altering the pH and the amount of precipitating agent (FeSO 4). The formation of iron proteinate was proved by infrared spectra. The absorbance of both CN (1520 cm −1 and 1630 cm −1) and of CO stretchings (1100 cm −1) indicated the presence of chelate bonds typical of an iron-proteinate molecule. The biological activity of the product obtained was tested on piglets. Preliminary results showed a good effect both on the animal body weight and on the blood indices, after oral application.

Saccharomyces cerevisiae cells secreting an Aspergillus niger beta-galactosidase grow on whey permeate.

We describe the construction of a lactose-utilizing Saccharomyces cerevisiae that expresses the cDNA for a secreted, thermostable beta-galactosidase (lacA) from Aspergillus niger. Yeast cells expressing the lacA gene from the yeast ADH1 promotor on a multicopy plasmid secrete up to 40% of the total beta-galactosidase activity into the growth medium. The secreted product is extensively N-glycosylated, and cells expressing the lacA gene grow on whey permeate (4% w/v lactose) with a doubling time of 1.6 hours. Such strains may offer a solution to the increasing problem of waste whey disposal.

Nonpolluting Conversion of Whey Permeate to Food Yeast Protein

By using concentrated whey permeate as substrate, food grade yeast (Kluyveromyces fragilis) was grown continuously in the fermentor to very high cell densities (total broth dry weight of over 140g/L). The high ash content in whey permeate was not inhibitory to cell growth. Cell mass yield remained about constant at pH range 3.9 to 5.1, temperatures between 31 to 37°C, with 6 to 30% whey permeate solids in the feed. Dilution rate (.1 to .3h−1) had little effect on the cell mass yield. At temperatures higher than 38°C, yield decreased. The high cell density fermentation broth was direct-dried without centrifugation and washing and the composition of the direct-dried product was determined. Amino acid profile, vitamin content, and fatty acid composition indicated a quality protein product suitable for food or feed supplementation. Ash content of the product was higher than other dried yeasts because of the high mineral content in whey permeate. Ash content could be significantly reduced by the addition of a small portion of another inexpensive fermentable sugar to the whey permeate feed. With this addition, cell mass yield, productivity, and protein content of the product increased. Because this high cell density, direct-dry process did not require preconcentration of the fermentor broth to dry the product economically, it did not generate a waste stream that required further treatment. This process converts waste whey permeate to useful food protein and essentially eliminates potentially polluting waste streams.

By-products from the food and dairy industries

For the purpose of this paper I intend to include within the term ‘by-products’ all those materials that arise from the food industries which are not consumed by the human population of the United Kingdom. Some of these materials are not acceptable as part of the human diet, or, if acceptable, find no ready market. Other materials are inedible, perhaps by humans and other animals, but can be converted into acceptable food or feed whilst yet others which are edible and acceptable are wasted in processing, storage, transport, distribution and in the kitchen and dining room. It is impossible to give a figure for the annual amount of by-products and waste available but statistical evidence points to approximately 6 million tonnes of food, all of which is edible but not consumed. This figure does not include by-products which are not intended for human consumption such as abattoir wastes, whey and brewery wastes and cereal offals. The total figure is therefore larger and represents a considerable potential economic resource. (It must be observed however, that only a small fraction of these wastes and by-products can be economically utilized. Other speakers are dealing with animal slaughter, fish and cereal wastes and I will therefore give only passing attention to these materials.)About 40 million gallons of cheese whey are produced in the UK each year on the farm. Of this, about 12 million gallons are fed directly to pigs. Although it appears that heavy hog production and cheese making are often associated, disposal of whey on farm remains a problem, since the price fetched by whey may be only a quarter or less than the cost of transport. Disposal on land, where it can be useful as a fertilizer, can lead to pollution problems. Trials carried out by ADAS (Wolverhampton) on both beef and dairy cattle are promising. It has been shown that every kg of barley previously fed to dairy cattle can be replaced by 10 litres of whey at half the cost, making barley feed unnecessary. In diets for steers, calculations have shown that a litre of whey at 6% solids can replace 90g of compound for energy (Peacock, 1977).

Technological Museum of New South Wales

THE annual report of the curator of the Technological Museum of New South Wales for the year ended December 31, 1942, states that the scientific work of the Museum has been placed on a war footing without necessitating any reorganization of laboratory work. Chemical investigations of national importance have included the recovery of by-products from charcoal burning, dyeing of khaki cloth, preparation of coloured flashes and composition of flash powders and smoke screens from motor exhausts, fireproofing paper and 'Cellophane' gas respirators, Australian essential oils as mosquito repellents, rot-proofing of sandbags, strength of laminated plastics, for aircraft, production of lactic acid and lactates from waste whey, and sources of quinine and similar alkaloids in Australia. Work has also been done on synthetic wax, fire-proofing fabrics for camouflage purposes, synthetic rubber from acetylene, production of drugs and natural dyes from Australian plants, camouflage materials, especially paint, wax from sugar-cane, and the relative humidity for controlling the development of moulds likely to attack silk parachutes in Queensland. Botanical investigations into the cultivation of the tung oil tree in Australia, as well as on suitable Australian fibres to relieve the shortage of imported material, and the important research on the cytology of the eucalypts and other genera, were continued. Much time has been devoted to the study of fot-proofing problems in the preservation of jute hessian, canvas and manilla, hemp and cotton rope and to the waterproofing and rot-proofing of canvas duck. Lists of lectures, etc., delivered, papers read before scientific societies, exhibits and publications are included.