Enzymatic Hydrolysis Of Lactose Research Articles

Formulation of hypotonic electrolyte re-hydration whey drinks from paneer and cheese whey

In the present study, formulation of electrolyte drink from paneer and cheese whey was done by enzymatic hydrolysis of lactose to simple sugars and adjusting the salt levels to meet the World Health Organization requirements (245 mOsm/L) for Oral Rehydration Salt. Complete hydrolysis of lactose was achieved in whey using Lactozyme enzyme (3000L) used at level of 2 ml/L and incubated at 40°C for 2 h at neutral pH. Lactose hydrolyzed whey was diluted (1:2.3, v/v) using potable quality demineralised water (Total dissolved solids: 60 ppm) to achieve reduction in osmolarity of 75 mosm/l. The salt concentration was adjusted by adding 3.25 g sodium chloride and 0.8 g potassium chloride for paneer whey (per L) and 2.5 g sodium chloride, 0.9 g of potassium chloride and 1.5 g of trisodium citrate for cheese whey (per L) to meet sodium, potassium and citrate levels in the resultant whey drink.

Mini review on role of β-galactosidase in lactose intolerance

This review mainly focuses on the role and properties of β-galactosidase in lactose intolerance and its industrial application. β-Galactosidase, hydrolyses the lactose into glucose and galactose and it is most commonly used in food based technology, particularly in the dairy manufacturing industry. This catalyst mainly focus for the improvement of new and novel products with hydrolyzed lactose, which can be appropriate for the lactose-intolerant persons, to improve the technological, texture and scientific properties of non-fermented dairy products. β-Galactosidase derived from the group of saccharides which is a converting enzymes in the family of hydrolases. They are broadly distributed in the several biological living systems. The enzymatic hydrolysis of lactose is also preferred in food based technology due to the low soluble range of lactose. The concentration lactose was found to be high in fermented dairy products such as ice cream, butter, cheese curd, yogurt, etc., can prompt extreme lactose crystallization bringing about items through a coarse, abrasive surface. Lactose hydrolysis in dairy products enhances adaptability also, richness altogether. These products are extra edible. Also for this purpose, the utilization of β-galactosidase enzyme prior to the condensing operation can reduce the lactose content to a point where lactose was no longer a problem industrial application of β-galactosidase. In Industries, due to the positive and constructive effect on intestinal bacterial microflora, different types of applications are possible in β-galactosidase enzyme.

Technological aspects of lactose-hydrolyzed milk powder

Few reports describe the effect of lactose hydrolysis on the properties of milk powder during production and storage. Hence, the aim of this study was to evaluate the effects of five different levels of enzymatic lactose hydrolysis during the production and storage of milk powder. As the lactose hydrolysis rate increased, adhesion to the drying chamber also increased, due to higher levels of particle agglomeration. Additionally, more brown powder was obtained when the lactose hydrolysis rate was increased, which in turn negatively affected rehydration ability. Using Raman spectroscopy, crystallization of the lactose residues in various samples was assessed over 6weeks of accelerated aging at a room temperature environment with 75.5% of air moisture. Products with 25% or greater lactose hydrolysis showed no signs of crystallization, in contrast to the non-hydrolyzed sample.

Investigation of lactose hydrolysis by crude extract of Trichoderma viride ATCC 32098

Abstract Objective Aim of this study was to find the optimum pH, temperature and incubation conditions for efficient lactose hydrolysis by beta-galactosidase produced by Trichoderma viride ATCC 32098. In addition, enzymatic lactose hydrolysis in milk, whey and lactose solutions were studied and compared. Material and methods Trichoderma viride ATCC 32098 was incubated at 30°C for 8 days on a rotary shaker. Lactose hydrolysis was calculated based on the amount of glucose measured. Hydrolysis was carried out in lactose solution, milk and whey. Results Maximum efficiency was observed at pH 5.0 and at 60°C. Agitation increases lactose hydrolysis slightly. When enzymatic lactose hydrolysis in milk, whey and lactose solutions were studied, it was observed that after 26 h, hydrolyses in lactose solution and whey were almost 100% and hydrolysis in milk was 92%. Conclusion It can be concluded that the enzyme produced from T. viride ATCC 32098 in this study is a good alternative for use in lactose hydrolysis.

β-galactosidase Production by Aspergillus niger ATCC 9142 Using Inexpensive Substrates in Solid-State Fermentation: Optimization by Orthogonal Arrays Design.

Background:Enzymatic hydrolysis of lactose is one of the most important biotechnological processes in the food industry, which is accomplished by enzyme β-galactosidase (β-gal, β-D-galactoside galactohydrolase, EC 3.2.1.23), trivial called lactase. Orthogonal arrays design is an appropriate option for the optimization of biotechnological processes for the production of microbial enzymes.Methods:Design of experimental (DOE) methodology using Taguchi orthogonal array (OA) was employed to screen the most significant levels of parameters, including the solid substrates (wheat straw, rice straw, and peanut pod), the carbon/nitrogen (C/N) ratios, the incubation time, and the inducer. The level of β-gal production was measured by a photometric enzyme activity assay using the artificial substrate ortho-Nitrophenyl-β-D-galactopyranoside.Results:The results showed that C/N ratio (0.2% [w/v], incubation time (144 hour), and solid substrate (wheat straw) were the best conditions determined by the design of experiments using the Taguchi approach.Conclusion:Our finding showed that the use of rice straw and peanut pod, as solid-state substrates, led to 2.041-folds increase in the production of the enzyme, as compared to rice straw. In addition, the presence of an inducer did not have any significant impact on the enzyme production levels.

Raman-Based Feedback Control of the Enzymatic Hydrolysis of Lactose

The development of real-time monitoring and an improved control system was performed according to a process analytical technology initiative focusing on the benefits of the pharmaceutical industry. As an extension of the results of pioneering Raman-based closed-loop feedback control of crystallization and synthesis, a bioprocess, namely, the enzymatic hydrolysis of lactose, was investigated in this work. For this purpose, an effective control system was developed to maintain the lactose concentration at a level of 20 g/L. Real-time evaluation of Raman spectra was accomplished using classical least-squares (CLS) and partial least-squares (PLS) multivariate data analysis methods. Formation of oligosaccharides during the hydrolysis of lactose causes inaccuracy in the CLS evaluation, and thus, optimization of lactose hydrolysis was needed to decrease oligosaccharide formation. After optimization, control of lactose hydrolysis could be achieved. To improve real-time evaluation, the PLS method was used, resulti...

Hyper Production of Β‐Galactosidase From Newly Isolated Strain of Aspergillus nidulans

AbstractSeveral Aspergillus species were isolated from soil samples collected from local dairy farms and screened for the production of β‐galactosidase. The potent isolate was identified as Aspergillus nidulans on the basis of microscopic and phenotypic characteristics. The production of β‐galactosidase from A. nidulans was optimized in batch fermentation using one‐variable‐at‐a‐time approach. Evaluation of process parameters indicated that A. nidulans produced high titer of β‐galactosidase at 35°C after 10.0 days of fermentation. It was found that A. nidulans is capable of releasing 60.0 U mg−1 of β‐galactosidase at pH 3.0. Among various carbon sources xylose (0.5%) showed maximum enzyme production. Glucose (1.0%), tri‐sodium citrate (0.7%), and potassium dihydrogen phosphate (0.3%) indicated positive affect by inducing the culture for maximum synthesis of β‐galactosidase. Various organic and inorganic nitrogen sources were also tested and it was noticed that peptone (0.5%), yeast extract (0.5%), ammonium sulfate (0.2%), and ammonium nitrate (0.2%) improved the production of β‐galactosidase. These results suggested the feasibility of A. nidulans for the large scale production of β‐galactosidase.Practical ApplicationsBeta galactosidase has potential applications in food processing industry and enzymatic hydrolysis of lactose is best process as compared to high temperature and acids treatment as it retains maximum quality of product. This enzyme is used to obtain the hydrolyzed lactose from milk and milk whey for utilization in bakery products, animal feeds and as a sugar source for several fermentation products. This enzyme is also used in ice creams and condensed dairy products to avoid lactose crystallization. The syrup produced by lactose conversion using enzyme hydrolysis is used in dairy and soft drink industries. In pharmaceutical industry, it is used to produce lactose‐free milk for lactose intolerant consumers who are unable to digest the milk lactose.

HIDRÓLISE DA LACTOSE E PRODUÇÃO DE LEITE EM PÓ: ASPECTOS TECNOLÓGICOS

A indústria de alimentos tem como desafio e oportunidade desenvolver novos produtos com reduzido ou baixo teor de lactose, a fim de atender uma crescente massa de portadores de má digestão da lactose. A secagem dos produtos com lactose hidrolisada é um desafio tecnológico devido a higroscopicidade dos mesmos, influenciando na produtividade, no entupimento das câmaras e na conservação dos pós. O objetivo deste estudo foi avaliar o efeito de diferentes níveis de hidrólise enzimática da lactose (0%, 25%, 50%, 75% e > 99%), na produção e estocagem de leite em pó integral utilizando como ferramentas de caracterização dos produtos fabricados em planta piloto a análise de composição, o controle do grau de hidrólise e as isotermas de sorção. Todos os resultados indicam que a hidrólise da lactose afeta a produção do leite em pó por aumentar a adesão no spray dryer e a absorção de umidade durante a estocagem.

Modeling lactose hydrolysis for efficiency and selectivity: Toward the preservation of sialyloligosaccharides in bovine colostrum whey permeate

Enzymatic hydrolysis of lactose has been shown to improve the efficiency and selectivity of membrane-based separations toward the recovery of bioactive oligosaccharides. Achieving maximum lactose hydrolysis requires intrinsic process optimization for each specific substrate, but the effects of those processing conditions on the target oligosaccharides are not well understood. Response surface methodology was used to investigate the effects of pH (3.25–8.25), temperature (35–55°C), reaction time (6 to 58 min), and amount of enzyme (0.05–0.25%) on the efficiency of lactose hydrolysis by β-galactosidase and on the preservation of biologically important sialyloligosaccharides (3′-siallylactose, 6′-siallylactose, and 6′-sialyl-N-acetyllactosamine) naturally present in bovine colostrum whey permeate. A central composite rotatable design was used. In general, β-galactosidase activity was favored at pH values ranging from 3.25 to 5.75, with other operational parameters having a less pronounced effect. A pH of 4.5 allowed for the use of a shorter reaction time (19 min), lower temperature (40°C), and reduced amount of enzyme (0.1%), but complete hydrolysis at a higher pH (5.75) required greater values for these operational parameters. The total amount of sialyloligosaccharides was not significantly altered by the reaction parameters evaluated, suggesting specificity of β-galactosidase from Aspergillus oryzae toward lactose as well as the stability of the oligosaccharides at pH, temperature, and reaction time evaluated.

Kluyveromyces lactis β-galactosidase immobilization in calcium alginate spheres and gelatin for hydrolysis of cheese whey lactose

ABSTRACT: One of the greatest challenges for dairy industries is the correct destination of all the whey generated during cheese making, considering its high impact, the large volume created, and its technological potential. Enzymatic hydrolysis of cheese whey lactose is a biotechnological alternative. However, one of the limiting factors of its use is the relatively high cost of the enzymes, which could be lowered with the immobilization of these biocatalysts. Considering this context, the objective of this research was to evaluate the commercial Kluyveromyces lactis β-galactosidase enzyme immobilized in calcium alginate spheres and gelatin, using glutaraldehyde and concanavalin A (ConA) as modifying agents in the hydrolysis of cheese whey lactose process. Results have shown that the enzyme encapsulation complexed with ConA in alginate-gelatin spheres, without glutaraldehyde in the immobilization support, has significantly increased the hydrolysis of lactose rate, achieving a maximum conversion of 72%.

Expression of a functional cold active β-galactosidase from Planococcus sp-L4 in Pichia pastoris

Lactase deficiency problem discourages many adults from consuming milk as a major source of micro- and macronutrients. Enzymatic hydrolysis of lactose is an ideal solution for this problem but such processing adds significant costs. In this study, a cold active β-galactosidase from Planococcus sp-L4 (bgal) was optimized for expression of recombinant “BGalP” in Pichia pastoris. As a result of codon optimization, the codon adaptation index was improved from 0.58 to 0.85 after replacing rare codons. After transformation of two P. pastoris strains (KM71H and GS115), the activity of BGalP enzyme was measured in the culture supernatants using ortho-Nitrophenyl-β-galactoside (ONPG). Maximal activity was recorded as 3.7U/ml on day 11 in KM71H clone #2 which was 20% higher than the best GS115 clone. Activity measurements under different conditions indicated optimal activity at pH 6.5. It was active at temperatures ranging from 0 to 55°C with deactivation occurring at or above 60°C. Protein analysis of the crude ultra-filtrate showed the enzyme was ∼75kDa and was the major constituent (85%) of the sample. This enzyme have the potential to find utility for the breakdown of lactose in chilled milk and subsequently can be deactivated by pasteurization. The use of BGalP would minimize energy consumption thus decreasing cost and also help to preserve the nutritional elements of the milk.

HIDRÓLISE ENZIMÁTICA DA LACTOSE DE PERMEADO DE SORO

O permeado de soro é o resíduo do processo de concentração das proteínas do soro pelo método de ultrafiltração. Contém nutrientes importantes, como lactose, minerais e traços de proteínas e lipídeos. É um resíduo sem fim industrial estabelecido que causa sérios danos ao meio ambiente. Para que o seu aproveitamento seja integral e para permitir seu consumo por pessoas intolerantes ao açúcar do leite é preciso que a lactose seja hidrolisada. A hidrólise enzimática pela lactase (β-galactosidase) do fungo Kluyveromyces lactis é um método seguro e que não compromete a integridade dos demais nutrientes, permitindo assim o posterior uso do permeado como matériaprima. Este trabalho objetivou realizar ensaios de hidrólise enzimática da lactose de formulações de permeado de soro, em concentração de 0,2%, 0,7% e 1% nos tempos 30, 60 e 90 minutos com o pH do meio de 6,3 e temperatura de 37 ºC. As reações foram acompanhadas por cromatografia líquida de alta eficiência que mostrou que a partir da concentração enzimática de 0,7% no tempo de 30 minutos, as formulações se tornaram seguras para o consumo de intolerantes à lactose, de acordo com níveis mínimos estabelecidos pela legislação.

Improved Saccharomyces Cerevisiae Growth on Cheese Whey by Controlling Enzymatic Lactose Hydrolysis

Whey generated in cheese manufacturing poses serious environmental issues that limit process profitability. The innovation in the dairy sector recognizes the bio-refinery as a key to successful handling of whey disposal and economic rise. Cheese whey valorisation is a complex process involving multiple technologies that might lead to value-added products (biomass, fine or bulk chemicals). This work focuses on the optimization of a fermentation process using whey as growth medium and carbon source. Lactose, which is abundant in whey, is a valuable carbon source. However, the microorganism more widely used in industrial fermentation processes, the yeast Saccharomyces cerevisiae, is not a lactose-fermenting yeast. We set up an innovative biotechnological process for the production on large scale of a not-genetically modified yeast biomass that can be used in different contexts, such as bread making, production of probiotics, nutraceuticals, bio-active molecules. In order to use the cheese whey as raw material for the cultivation of S. cerevisiae and to overcome the limitations in the use of lactose we used and externally added the enzyme β-galactosidase. The careful optimization of the amount of added enzyme allowed the gradual release by hydrolysis and the simultaneous consumption of glucose and galactose with a consequent decrease of ethanol and an increase of the biomass produced.

Evaluation of the cold-active Pseudoalteromonas haloplanktis β-galactosidase enzyme for lactose hydrolysis in whey permeate as primary step of d-tagatose production

d-Tagatose is an innovative natural low-calorie bulk sweetener with a broad potential for low-calorie and low-glycaemic foods and drinks. Production of this healthy sweetener is realized through enzymatic d-galactose isomerization. d-Galactose needs to be produced in situ due to its limited availability. Whey permeate contains a substantial amount of lactose, which is an interesting source for d-galactose production through enzymatic lactose hydrolysis. In this context, the cold-active β-galactosidase from the psychrophile Pseudoalteromonas haloplanktis was studied. Optimal parameters for efficient lactose hydrolysis in whey permeate have been deduced, viz. optimal incubation temperature, pH and lactose concentration. Hydrolysis efficiencies above 96.0% were realized within 24h at 23°C and pH 7.0 in whey permeate with a maximum dry matter content of 10.0% (w/w). In addition, the effect of the presence of d-glucose and d-galactose was investigated up to concentrations of 100gl−1. d-Glucose inhibited lactose hydrolysis more strongly compared to d-galactose. Also, the operational stability of the cold-active β-galactosidase was studied. Hydrolysis efficiencies above 90.0% were maintained during 7 subsequent hydrolysis cycles.

Permeabilization of Kluyveromyces marxianus with Mild Detergent for Whey Lactose Hydrolysis and Augmentation of Mixed Culture

Cheese whey is a by-product of cheese-manufacturing industries, and the utilization of whey is a challenging problem either to use it or dispose it, because only few microorganisms can metabolize the whey lactose. Enzymatic hydrolysis of whey lactose to glucose and galactose by β-galactosidase is the approach for biotechnological application. Kluyveromyces marxianus cells were permeabilized with non-toxic, biodegradable, anionic detergent N-lauroyl sarcosine (N-LS) for the enzyme activity. The permeabilization process parameters (N-LS concentration, solvent volume, temperature and incubation time) were optimized. The maximum β-galactosidase activity of 1,220 IU/g dry weight was obtained using permeabilized cells under optimized conditions. Moreover, viability of the permeabilized cells was also evaluated, which showed that cells were alive; however, viability was reduced by two log cycles. The permeabilized cells were evaluated for whey lactose hydrolysis. The maximum lactose hydrolysis of 91% was observed with 600 mg (dry cell weight/100 mL) in whey powder (5% w/v) solution at 180-min incubation, pH 6.5 and 30 °C. Further, the hydrolyzed whey was evaluated for amelioration of growth of non-lactose-consuming yeast Saccharomyces cerevisiae. S. cerevisiae was able to grow in hydrolyzed whey simultaneously with K. marxianus. The study confirmed that N-LS could be used to permeabilize K. marxianus cells to make available the enzyme activity.

Recovery of Whey Proteins and Enzymatic Hydrolysis of Lactose Derived from Casein Whey Using a Tangential Flow Ultrafiltration Module

In this study, ultrafiltration (UF) of pretreated casein whey was carried out in a cross-flow module fitted with 5 kDa molecular weight cut-off polyethersulfone membrane to recover whey proteins in the retentate and lactose in the permeate. Effects of processing conditions, like transmembrane pressure and pH on permeate flux and rejection were investigated and reported. The polarised layer resistance was found to increase with time during UF even in this high shear device. The lactose concentration in the permeate was measured using dinitro salicylic acid method. Enzymatic kinetic study for lactose hydrolysis was carried out at three different temperatures ranging from 30 to 50 °C using β-galactosidase enzyme. The glucose formed during lactose hydrolysis was analyzed using glucose oxidase–peroxidase method. Kinetics of enzymatic hydrolysis of lactose solution was found to follow Michaelis–Menten model and the model parameters were estimated by Lineweaver–Burk plot. The hydrolysis rate was found to be maximum (with Vmax = 5.5091 mmol/L/min) at 30 °C.

Galactooligosaccharides formation during enzymatic hydrolysis of lactose: Towards a prebiotic-enriched milk

The formation of galactooligosaccharides (GOS) in skim milk during treatment with several commercial β-galactosidases (Bacillus circulans, Kluyveromyces lactis and Aspergillus oryzae) was analysed in detail, at 4 and 40°C. The maximum GOS concentration was obtained at a lactose conversion of approximately 40–50% with B. circulans and A. oryzae β-galactosidases, and at 95% lactose depletion for K. lactis β-galactosidase. Using an enzyme dosage of 0.1% (v/v), the maximum GOS concentration with K. lactis β-galactosidase was achieved in 1 and 5h at 40 and 4°C, respectively. With this enzyme, it was possible to obtain a treated milk with 7.0g/L GOS − the human milk oligosaccharides (HMOs) concentration is between 5 and 15g/L – and with a low content of residual lactose (2.1g/L, compared with 44–46g/L in the initial milk sample). The major GOS synthesised by this enzyme were 6-galactobiose [Gal-β(1→6)-Gal], allolactose [Gal-β(1→6)-Glc] and 6′-O-β-galactosyl-lactose [Gal-β(1→6)-Gal-β(1→4)-Glc].

Presence of mono-, di- and galactooligosaccharides in commercial lactose-free UHT dairy products

In this work a study of mono-, di- and galactooligosaccharide (GOS) content as well as the determination of thermal indicators of different commercial lactose-free UHT (LF-UHT) milks and dairy drinks has been carried out. Moreover, GOS formation during hydrolysis of lactose in a commercial UHT milk using Lactozym® pure 6500 was also studied. Presence of GOS was detected in all analyzed samples ranging from 950 to 4350mg/L in LF-UHT milks and from 600 to 2260mg/L in LF-UHT dairy drinks. Their GOS contents correlate roughly with the remaining lactose content of samples. During enzymatic hydrolysis of lactose in commercial UHT milk, the GOS formed reached a maximum about 10,000mg/L when 75–90% of lactose was hydrolyzed and then gradually decreased to values considerably lower than 5000mg/L when over 99% of the lactose was hydrolyzed. Therefore, GOS formation during low lactose milk manufacture might be improved by controlling enzymatic lactose hydrolysis so that it would be possible to elaborate low-lactose milk products able to satisfy the physiological requirements of the majority of intolerant groups and with a GOS content high enough to confer beneficial effect as prebiotic.

Quantification of lactose using ion-pair RP-HPLC during enzymatic lactose hydrolysis of skim milk

The correct labelling of dairy foods as “lactose-free” requires a suitably sensitive and valid analytical method for the quantification of lactose in complex food matrices. Thus, an ion-pair RP-HPLC method for the simultaneous determination of lactose, glucose and galactose in original skim milk was investigated. The samples derived from an enzymatic lactose hydrolysis approach (0.5L) using the commercial β-galactosidase Godo-YNL2. After derivatisation with p-aminobenzoic acid and sodium cyanoborohydride, the samples were injected on a RP-C18 column. Tetrabutylammonium hydrogen sulphate was used as the ion-pair reagent in the eluent system. The sugars were quantified using photometric- (UV; 303nm) and fluorescence-detection (λex 313nm, λem 358nm). The overall run time was 27min. The limits of detection (LOD) were estimated at 2mgL−1 (UV detection) and at 0.13mgL−1 (fluorescence detection). The limits of quantification were 6mgL−1 (UV detection) and 0.45mgL−1 (fluorescence detection). Thus, this analytical method is suitable for sensitive lactose quantification in milk systems of less than 10mgL−1.

Production of Galacto-Oligosaccharides from Lactose and whey Permeate by β-Glycosidase from Sporotrichum Thermophile ATCC 34628

ABSTRACTβ-Glycosidases catalyze not only the hydrolysis reactions, but also the transglycosylation ones, producing different oligosaccharides. The production of galactooligosaccharides using intracellular β'-glycosidase from Sporotrichum thermophile ATCC 34628 and toluene-treated cell suspensions was investigated. Enzymatic hydrolysis of lactose and synthesis of galactooligosaccharides (Gal-OS) by intracellular p-glycosidase reached an yield of 5.9% when 20% (w/v) lactose was used as substrate at 50 °C and pH 6.0. The production of Gal-OS by toluene-treated cells of Sporotrichum thermophile ATCC 34628 revealed an increasing of the trisaccharides content from 4% after 2 hours of incubation up to 13.5% after 72 hours at a lactose concentration of 20% (w/v). When whey permeate (10%, w/v) was used as substrate for the transglycosylation reaction, mainly trisaccharides were produced. The results from TLC and HPLC analyses suggested a transfer of glucose to lactose and formation of glucosyllactose, because of the lack of glucose on the chromatograms as a product of lactose hydrolysis at 22h of incubation. The produced Gal-OS were purified and their potential role as prebiotics and growth factors for Bifidobacteria strains was established.