Low-lactose Milk Products Research Articles

Lactose quantification in UHT milk by high-performance liquid chromatography and cryoscopy (freezing point depression)

Due to the large number of people with lactose maldigestion, the dairy industries have increased production and diversity of low lactose and lactose-free foods. Consequently, the need to control the lactose hydrolysis process has also risen. This study aimed to correlate freezing point depression (cryoscopy) and lactose concentration, quantified by high-performance liquid chromatography (HPLC), in UHT milk. To accomplish this, UHT milk samples were subjected to seven lactose hydrolysis treatments, using lactase enzyme, resulting in different lactose concentrations. All samples were subjected to HPLC analysis and freezing point measurement, using a cryoscope. The results were plotted on a graph and a linear regression was performed. There was a strong correlation between lactose concentration and freezing point (R = 0,9973) and the coefficient of determination (R2) was 0,9946, which means that 99,46% of the variability of the response data is explained by the linear regression model. Therefore, the results point to the feasibility of estimating the lactose concentration in milk during the hydrolysis process for the production of low lactose milk, by cryoscopy, a quick analysis, with lower cost compared to HPLC and that is already among the analyses commonly performed in dairy industries.

Heat-Loving β-Galactosidases from Cultured and Uncultured Microorganisms.

β-galactosidases (EC.3.2.1.23), which hydrolyze lactose to glucose and galactose, have two main applications in the food industry: the production of low-lactose milk and dairy goods for lactose intolerant people, and the generation of galacto-oligosaccharides by transgalactosylation reactions. Due to their thermostability, β-galactosidases from thermophilic microorganisms are very interesting for industrial processes, as high temperatures can increase the initial productivity of the enzyme, provide higher solubility of substrates, and prevent microbial contamination. In the past, it was necessary to cultivate and grow thermophilic microorganisms to discover novel thermozymes, but the development of metagenomic techniques has allowed researchers to access the genomic potential of uncultivated microbes and their enzymes. The present review gives a brief outline of thermophilic β-galactosidases, with a special focus on those obtained through metagenomics. Additionally, the sequences of β-galactosidases found in some public metagenomes from hot springs were studied and compared to other known thermostable β-galactosidases.

Metagenomic approach for the isolation of a thermostable β-galactosidase with high tolerance of galactose and glucose from soil samples of Turpan Basin

Backgroundβ-Galactosidases can be used to produce low-lactose milk and dairy products for lactose intolerant people. Although commercial β-galactosidases have outstanding lactose hydrolysis ability, their thermostability is low, and reaction products have strong inhibition to these enzymes. In addition, the β-galactosidases possessing simultaneously high thermostability and tolerance of galactose and glucose are still seldom reported until now. Therefore, identification of novel β-galactosidases with high thermostability and tolerance to reaction products from unculturable microorganisms accounting for over 99% of microorganisms in the environment via metagenomic strategy is still urgently in demand.ResultsIn the present study, a novel β-galactosidase (Gal308) consisting of 658 amino acids was identified from a metagenomic library from soil samples of Turpan Basin in China by functional screening. After being overexpressed in Escherichia coli and purified to homogeneity, the enzymatic properties of Gal308 with N-terminal fusion tag were investigated. The recombinant enzyme displayed a pH optimum of 6.8 and a temperature optimum of 78°C, and was considerably stable in the temperature range of 40°C - 70°C with almost unchangeable activity after incubation for 60 min. Furthermore, Gal308 displayed a very high tolerance of galactose and glucose, with the highest inhibition constant Ki,gal (238 mM) and Ki,glu (1725 mM) among β-galactosidases. In addition, Gal308 also exhibited high enzymatic activity for its synthetic substrate o-nitrophenyl-β-D-galactopyranoside (ONPG, 185 U/mg) and natural substrate lactose (47.6 U/mg).ConclusionThis study will enrich the source of β-galactosidases, and attract some attentions to β-galactosidases from extreme habitats and metagenomic library. Furthermore, the recombinant Gal308 fused with 156 amino acids exhibits many novel properties including high activity and thermostability at high temperatures, the pH optimum of 6.8, high enzyme activity for lactose, as well as high tolerance of galactose and glucose. These properties make it a good candidate in the production of low-lactose milk and dairy products after further study.

Hydrolysis of milk lactose in a packed bed reactor system using immobilized yeast cells

AbstractBACKGROUND: Hydrolysis of lactose with β‐D‐galactosidase is one of the most promising biotechnological applications in the food industry because of its use in the production of low lactose milk products and whey hydrolysis. To overcome the problem of enzyme extraction from cells due to the intracellular nature of β‐D‐galactosidase and the poor permeability of the cell membrane to lactose, permeabilization of yeast cells was investigated. Permeabilized whole cells have been claimed to have an advantage over more pure enzyme preparations. In view of the advantages of immobilized cell systems over free cell systems, permeabilized cells were immobilized by an entrapment method in calcium alginate gel. A packed bed reactor together with this immobilized cell system has been used for hydrolysis of milk lactose in a continuous system.RESULTS: Different process parameters (temperature, substrate feed rate, biomass load and time‐course) were optimized to maximize lactose hydrolysis. The immobilized yeast cells (300 mg dry wt) resulted in 87.2% hydrolysis of milk lactose at 30 °C and flow rate 7 mL h−1 in a packed bed reactor system.CONCLUSION: This convenient and relatively inexpensive method of immobilization, resulting in high hydrolysis potential in a continuous system, indicates that permeabilized yeast cells have the potential for the production of low lactose milk and milk products. Copyright © 2010 Society of Chemical Industry

Lactose hydrolysis by Lactozym™ immobilized on cellulose beads in batch and fluidized bed modes

Lactozym™ is a commercially available preparation of β-galactosidase from Kluyveromyces fragilis. It has valid generally recognized as safe (GRAS) status for whey hydrolysis and production of low lactose milk. Immobilized β-galactosidase from K. fragilis has been less studied. In this work, Lactozym™ was immobilized on cellulose beads via epichlorohydrin coupling chemistry. The optimized preparation was characterized in terms of its kinetic parameters. The fluidized bed hydrolyzed whey lactose (>90% conversion) in 5 h as compared to 48 h taken by the same enzyme in continuous batch mode. The immobilized enzyme could be reused three times without any change in the performance of the fluidized bed reactor. The fluidized bed could also hydrolyze milk lactose up to 60% within 5 h. The above data show that this enzyme from a GRAS status source can also be used to develop a process for lactose hydrolysis for whey utilization as well as production of low lactose milk.

Production of low-lactose milk by ectopic expression of intestinal lactase in the mouse mammary gland.

We have investigated, in mice, an in vivo method for producing low-lactose milk, based on the creation of transgenic animals carrying a hybrid gene in which the intestinal lactase-phlorizin hydrolase cDNA was placed under the control of the mammary-specific alpha-lactalbumin promoter. Transgenic females expressed lactase protein and activity during lactation at the apical side of mammary alveolar cells. Active lactase was also secreted into milk, anchored in the outer membrane of fat globules. Lactase synthesis in the mammary gland caused a significant decrease in milk lactose (50-85%) without obvious changes in fat and protein concentrations. Sucklings nourished with low-lactose milk developed normally. Hence, these data validate the use of transgenic animals expressing lactase in the mammary gland to produce low-lactose milk in vivo, and they demonstrate that the secretion of an intestinal digestive enzyme into milk can selectively modify its composition.

Microbial technologies in the production of low-lactose dairy foods / Tecnologías microbiológicas para la elaboración de productos lácteos con bajo contenido en lactosa

Low-lactose milk products with 70% or more of the lactose hydrolysed by food grade β-galac tosidase enzymes of yeasts or fungi have become widely accepted for alleviating the symptoms of lactose maldigestion. This condition limits the intake of nutritious dairy foods by large segments of the world's population. Alternative approaches recently proposed for dealing with lactose maldigestion include the supplementation of milk with dormant dairy cultures, treatment of milk with sonicated or permeabilized cultures as food-grade sources of β-galactosidase and the use of cold-active enzymes to hydrolyse lactose in milk under refrigerated storage conditions.