A unique metabolic gene cluster regulates lactose and galactose metabolism in the yeast Candida intermedia.

Lactose assimilation is a relatively rare trait in yeasts, and Kluyveromyces yeast species have long served as model organisms for studying lactose metabolism. Meanwhile, the metabolic strategies of most other lactose-assimilating yeasts remain unknown. In this work, we have elucidated the genetic determinants of the superior lactose-growing yeast Candida intermedia. Through genomic and transcriptomic analyses, we identified three interdependent gene clusters responsible for the metabolism of lactose and its hydrolysis product galactose: the conserved LAC cluster (LAC12, LAC4) for lactose uptake and hydrolysis, the conserved GAL cluster (GAL1, GAL7, and GAL10) for galactose catabolism through the Leloir pathway, and a "GALLAC" cluster containing the transcriptional activator gene LAC9, second copies of GAL1 and GAL10, and a XYL1 gene encoding an aldose reductase involved in carbon overflow metabolism. Bioinformatic analysis suggests that the GALLAC cluster is unique to C. intermedia and has evolved through gene duplication and divergence, and deletion mutant phenotyping proved that the cluster is indispensable for C. intermedia's growth on lactose and galactose. We also show that the regulatory network in C. intermedia, governed by Lac9 and Gal1 from the GALLAC cluster, differs significantly from the galactose and lactose regulons in Saccharomyces cerevisiae, Kluyveromyces lactis, and Candida albicans. Moreover, although lactose and galactose metabolism are closely linked in C. intermedia, our results also point to important regulatory differences.IMPORTANCEThis study paves the way to a better understanding of lactose and galactose metabolism in the non-conventional yeast C. intermedia. Notably, the unique GALLAC cluster represents a new, interesting example of metabolic network rewiring and likely helps to explain how C. intermedia has evolved into an efficient lactose-assimilating yeast. With the Leloir pathway of budding yeasts acting like a model system for understanding the function, evolution, and regulation of eukaryotic metabolism, this work provides new evolutionary insights into yeast metabolic pathways and regulatory networks. In extension, the results will facilitate future development and use of C. intermedia as a cell-factory for conversion of lactose-rich whey into value-added products.

Structure Assessment and Impacts of Lipids' Chemistry on the Structuration of Polyhydroxyalkanoate Biosynthesized by Bacillus licheniformis AZU-A5.

The current study exploited cheese whey supernatant (CWS) as a renewable resource for polyhydroxyalkanoates (PHAs) formation. Structure identification and investigating the influence of lipid membranes' chemistry on PHA structuration were detailed. Approximately 66 bacterial isolates from dairy products companies in Egypt were screened for PHA production, and the bacterial isolate AZU-A5, identified as Bacillus licheniformis strain AZU-A5, showed the highest production PHA 0.93 g L-1) using whey lactose. The highest PHA rate in the individual design was 1.59 g L-1 with a recovery yield of 33.08% (w w-1), while the production rate in the statistical design reached 1.75 g L-1 PHA and 51.60% PHA content. Structurally, the PHA was identified as polyhydroxy-3R-butyrate (R-PH3B). The fibrous texture by SEM highlighted the self-assembly of PHB. The CD analysis of the PHA films suggested favorable hydrophobic interactions between lipids and PHB. Higher lipid contents not only caused a decrement in the CD signal of PHB but also bathochromic effects occurred. The chain length of lipids exerted high impacts on interactions (CD) and structuration of PHB (Δλ). The unsaturation showed little influence on CD and negligible effect on Δλ, while the head group exerted no effect on CD with a considerable impact on Δλ.

Innovative applications of whey protein for sustainable dairy industry: Environmental and technological perspectives-A comprehensive review.

Industrial waste management is critical to maintaining environmental sustainability. The dairy industry (DI), as one of the major consumers of freshwater, generates substantial whey dairy effluent, which is notably rich in organic matter and thus a significant pollutant. The effluent represents environmental risks due to its high biological and chemical oxygen demands. Today, stringent government regulations, environmental laws, and heightened consumer health awareness are compelling industries to responsibly manage and reuse whey waste. Therefore, this study investigates sustainable solutions for efficiently utilizing DI waste. Employing a systematic review approach, the research reveals that innovative technologies enable the creation of renewable, high-quality, value-added food products from dairy byproducts. These innovations offer promising sustainable waste management strategies for the dairy sector, aligning with economic interests. The main objectives of the study deal with, (a) assessing the environmental impact of dairy sector waste, (b) exploring the multifaceted nutritional and health benefits inherent in cheese whey, and (c) investigating diverse biotechnological approaches to fashion value-added, eco-friendly dairy whey-based products for potential integration into various food products, and thus fostering economic sustainability. Finally, the implications of this work span theoretical considerations, practical applications, and outline future research pathways crucial for advancing the sustainable management of dairy waste.

Construction of the advanced flavin mononucleotide producers in the flavinogenic yeast Candida famata.

Flavin mononucleotide (FMN, riboflavin-5'-phosphate) is flavin coenzyme synthesized in all living organisms from riboflavin (vitamin B2 ) after phosphorylation in the reaction catalyzed by riboflavin kinase. FMN has several applications mostly as yellow colorant in food industry due to 200 times better water solubility as compared to riboflavin. Currently, FMN is produced by chemical phosphorylation of riboflavin, however, final product contains up to 25% of flavin impurities. Microbial overproducers of FMN are known, however, they accumulate this coenzyme in glucose medium. Current work shows that the recombinant strains of the flavinogenic yeast Candida famata with overexpressed FMN1 gene coding for riboflavin kinase in the recently isolated by us advanced riboflavin producers due to overexpression of the structural and regulatory genes of riboflavin synthesis and of the putative exporter of riboflavin from the cell, synthesized elevated amounts of FMN in the media not only with glucose but also in lactose and cheese whey. Activation of FMN accumulation on lactose and cheese whey was especially strong in the strains which expressed the gene of transcription activator SEF1 under control of the lactose-induced LAC4 promoter. The accumulation of this coenzyme by the washed cells of the best recombinant strain achieved 540 mg/L in the cheese whey supplemented only with ammonium sulfate during 48 h in shake flask experiments.

Application of enterocin-whey films to reduce Listeria monocytogenes contamination on ripened cheese.

An enterocin whey solution, obtained by growing Enterococcus faecalis L2B21K3 and L3A21K6 in sweet whey - enterocin whey solution (EWS), was incorporated into gelatin/glycerol films that were tested for the control of Listeria monocytogenes. The films containing enterocins produced by either strain (EWS L2 and EWS K6 films) were shown to serve as a suitable matrix for bacteriocin release, preserve the anti-listerial activity for up to 90 days. When applied in cheese, EWS L2 and EWS K6 films were able to reduce L. monocytogenes contamination to undetected levels after 20 or 30 days, respectively, and prevented the migration of this pathogen from the films to cheese. The incorporation of EWS into films did not affect (p < 0.05) moisture content, solubility, permeability (water vapor and limonene), and elongation at break compared to control films (without EWS). However, thickness, swelling index and tensile strength were higher (p < 0.05) in EWS films. These results suggest that active EWS gelatin/glycerol films could be an effective, and safe application to control L. monocytogenes in cheese. In addition, the use of cheese whey as a culture medium for the production of the bacteriocins complemented with the incorporation in films formulation as a packaging material represents an alternative approach to reuse this by-product of cheese production.

Integrated production of hydrogen and methane in a dairy biorefinery using anaerobic digestion: Scale-up, economic and risk analyses.

Anaerobic digestion has emerged as the most appealing waste management strategy in biorefineries. Particularly, recent studies have highlighted the energy advantages of waste co-digestion in industrial biorefineries and the use of two-stage systems. However, there are some concerns about moving the system from laboratory testing to industrial scale. One of them is the high level of investment that is required. Therefore, this study carried out a techno-economic analysis (scale-up and energy production, economic and risk analysis, and factorial design) to assess the feasibility of single- and two-stage systems in the treatment of cheese whey and glycerin for the production of hydrogen and methane. Scenarios (S1 to S9) considered thermophilic and mesophilic single and two-stage systems with different applied organic loading rates (OLRA). The analyses of scale-up and energy production revealed that S3 (a thermophilic single-stage system operated at high OLRA 17.3 kg-COD.m-3.d-1) and S9 (a thermophilic-mesophilic two-stage system operated at high OLRA 134.8 kg-COD.m-3.d-1 and 20.5 kg-COD.m-3.d-1, respectively) were more compact and required lower initial investment compared to other scenarios. The risk analysis performed by a Monte Carlo simulation showed low investment risks (10 and 11%) for S3 and S9, respectively, being the electricity sales price, the key determining factor to define whether the project in the baseline scenario will result in profit or loss. Lastly, the factorial design revealed that while the net present value (NPV) is positively impacted by rising inflation and electricity sales price, it is negatively impacted by rising capitalization rate. Such assessments assist in making decisions regarding which system can be fully implemented, the best market circumstances for the investment, and how market changes may favorably or unfavorably affect the NPV and the internal rate of return (IRR).

A comparison of ultrasonic, ozone, and enzyme pre-treatments on cheese whey degradation for enhancement of anaerobic digestion.

Lactose in cheese whey wastewater (CWW) makes it difficult to degrade under normal conditions. The effect of ultra-sonication (US), ozonation and enzymatic hydrolysis on increasing the bioavailability of organic matter in CWW and biogas production were evaluated. The pre-treatment conditions were: specific energy input varied from 2130 to 8773KJ/KgTS for a sonication time of 4.5-18.5 min, Ozone (O3) dosages ranging from 0.03 to 0.045gO3/gTS were applied for 4-16 min, pH (3.8-7.1), temperature (35°C-55°C), enzyme dosage (0.18-0.52%), was operated from 7.75 to 53 min for enzymatic hydrolysis by ß-galactosidase. The results of the US reported a maximum sCOD solubilisation of 77.15% after 18.5 min of operation, while the corresponding values for ozonation and enzymatic methods were 64.8% at 16 min and 54.79%, respectively. The organic matter degradation rates evaluated in terms of protein and lactose hydrolysis were 68.78%,46.03%; 47.83%,16.15% and 54.22%,86.2%respectively, for US, ozonation and enzymatic methods. The cumulative methane yield for sonicated, ozonised and enzymatically hydrolysed samples were 412.4 ml/g VS, 361.2 ml/g VS and 432.3mlCH4/gVS, respectively. Regardless of the lower COD solubilisation rates attained, enzymatic pre-treatment showed maximum methane generation compared to US and ozonation. This could be attributable to the increased activity of ß-galactosidase in hydrolysing whey lactose. The energy calculations revealed that the pre-conditioning of organic-rich CWW with enzymatic hydrolysis is more effective and efficient, yielding a net energy gain (gross output energy-input energy) of 9166.7 KJ and an energy factor (ratio of output to input energy) of 6.67. The modified Gompertz model well simulated all experimental values.

The effect of adding concentrated cheese whey prepared by ultrafiltration on the quality of Syrian bread.

The objective of this study is to evaluate the effect of adding two types cheese whey (W) contaning 0.9% protein and concentrated cheese whey (CW) containing 7.61% protein on the physical, chemical and sensory properties of Syrian bread. Concentrated whey samples were prepared using a UF membrane unit of 15 KDa at 25 °C and 4 bars. Both W and CW samples were added to wheat flour (72% extraction rate) at dilution level: 0, 25, 50, 75, and 100% levels. The effects of W and CW on dough rheology were evaluated using farinograph and alveograph parameters. The addition of 25% W or 50% CW improved the dough stability in the farinogram, as well as pressure and energy values in the alveogram. Significant increases were observed in ash, minerals (Ca, K, Mg, and P), and protein content (13.8%) when 50% CW was added to the dough. The results of sensory analysis showed that Syrian bread containing 25% whey (W) or 50% whey concentrate (CW) has achieved highest overall acceptability scores than control.

Cheese whey supports high riboflavin synthesis by the engineered strains of the flavinogenic yeast Candida famata.

BACKGROUND: Riboflavin is a precursor of FMN and FAD which act as coenzymes of numerous enzymes. Riboflavin is an important biotechnological commodity with annual market sales exceeding nine billion US dollars. It is used primarily as a component of feed premixes, a food colorant, a component of multivitamin mixtures and medicines. Currently, industrial riboflavin production uses the bacterium, Bacillus subtilis, and the filamentous fungus, Ashbya gossypii, and utilizes glucose and/or oils as carbon substrates. RESULTS: We studied riboflavin biosynthesis in the flavinogenic yeast Candida famata that is a genetically stable riboflavin overproducer. Here it was found that the wild type C. famata is characterized by robust growth on lactose and cheese whey and the engineered strains also overproduce riboflavin on whey. The riboflavin synthesis on whey was close to that obtained on glucose. To further enhance riboflavin production on whey, the gene of the transcription activator SEF1 was expressed under control of the lactose-induced promoter of the native ß-galactosidase gene LAC4. These transformants produced elevated amounts of riboflavin on lactose and especially on whey. The strain with additional overexpression of gene RIB6 involved in conversion of ribulose-5-phosphate to riboflavin precursor had the highest titer of accumulated riboflavin in flasks during cultivation on whey. Activation of riboflavin synthesis was also obtained after overexpression of the GND1 gene that is involved in the synthesis of the riboflavin precursor ribulose-5-phosphate. The best engineered strains accumulated 2.5 g of riboflavin/L on whey supplemented only with (NH4)2SO4 during batch cultivation in bioreactor with high yield (more than 300 mg/g dry cell weight). The use of concentrated whey inhibited growth of wild-type and engineered strains of C. famata, so the mutants tolerant to concentrated whey were isolated. CONCLUSIONS: Our data show that the waste of dairy industry is a promising substrate for riboflavin production by C. famata. Possibilities for using the engineered strains of C. famata to produce high-value commodity (riboflavin) from whey are discussed.

A machine learning proposal method to detect milk tainted with cheese whey.

Cheese whey addition to milk is a type of fraud with high prevalence and severe economic effects, resulting in low yield for dairy products, nutritional reduction of milk and milk-derived products, and even some safety concerns. Nevertheless, methods to detect fraudulent addition of cheese whey to milk are expensive and time consuming, and are thus ineffective as screening methods. The Fourier-transform infrared (FTIR) spectroscopy technique is a promising alternative to identify this type of fraud because a large number of data are generated, and useful information might be extracted to be used by machine learning models. The objective of this work was to evaluate the use of FTIR with machine learning methods, such as classification tree and multilayer perceptron neural networks to detect the addition of cheese whey to milk. A total of 520 samples of raw milk were added with cheese whey in concentrations of 1, 2, 5, 10, 15, 20, 25, and 30%; and 65 samples were used as control. The samples were stored at 7, 20, and 30°C for 0, 24, 48, 72, and 168 h, and analyzed using FTIR equipment. Complementary results of 520 samples of authentic raw milk were used. Selected components (fat, protein, casein, lactose, total solids, and solids nonfat) and freezing point (°C) were predicted using FTIR and then used as input features for the machine learning algorithms. Performance metrics included accuracy as high as 96.2% for CART (classification and regression trees) and 97.8% for multilayer perceptron neural networks, with precision, sensitivity, and specificity above 95% for both methods. The use of milk composition and freezing point predicted using FTIR, associated with machine learning techniques, was highly efficient to differentiate authentic milk from samples added with cheese whey. The results indicate that this is a potential method to be used as a high-performance screening process to detected milk adulterated with cheese whey in milk quality laboratories.

Use of batch anion exchange technique for separation of κ-casein glycomacropeptide from bovine whey fraction.

Bovine κ-casein glycomacropeptide (GMP) is a sialic acid containing glycopeptide, which is considered as a health promoting compound found in cheese whey. The study described in this research communication was undertaken to determine whether GMP with undetectable level of contaminating protein or phenylalanine can be isolated from bovine whey fraction using batch anion exchange technique with chitin as an adsorbent. A soluble whey fraction (SWF) prepared from 1 g whey protein isolate (WPI) was mixed with a slurry of 1 g chitin, and the mixture was incubated at pH 3.0. After incubation, the mixture was filtered, and the residue obtained (containing chitin-GMP complex) was washed with water and eluted stepwise with 0.5 M NaCl and 2.0 M NaCl. Most of GMP (corresponding to 75.8% of total sialic acid recovered) was eluted with 0.5 M NaCl. The recovered GMP accounted for 5.4% dry weight of WPI (or 18.9% dry weight of SWF). Amino acid analysis showed that there was no detectable level of contaminating amino acids including phenylalanine, histidine, arginine and tyrosine in the GMP fraction. It was concluded that the batch anion exchange method with chitin developed in this study can be used for the isolation of high purity GMP from bovine SWF.

Spray-Dried Microcapsules of Cheese Whey and Whey Permeate as a Strategy to Protect Chia Oil from Oxidative Degradation.

Research background: Cheese whey and whey permeate are dairy industry by-products usually sent to effluent treatment or incorrectly disposed in the environment, generating costs for the production of dairy products and environmental problems due to the high organic load. Cheese whey and whey permeate can be reused as wall materials to form chia oil microcapsules, which act as a barrier to prooxidants. This study aims to develop an encapsulation method by spray-drying to protect chia oil using dairy by-products as wall materials. Experimental approach: We evaluated cheese whey, whey permeate and mixtures of m(cheese whey):m(whey permeate)=50, 70 and 80% as encapsulating agents with the spray-drying process. Initially, we characterized the chia oil and encapsulating materials. Chia oil emulsions were prepared using the encapsulating materials and an emulsifier. The stability of the emulsions was evaluated by creaming index, and they were characterized according to size distribution and polydispersity index. Emulsions were encapsulated in a spray dryer with inlet and outlet air temperature at 125 and 105 °C, respectively. After encapsulation, we assessed the oxidative degradation of chia oil over 30 days of storage by determining the peroxide index. Results and conclusions: Emulsions presented creaming index between 51 and 83% in all formulations, and the oxidative stability of microencapsulated chia oil was significantly higher than that of free chia oil after 30 days. Wall material combination affected both encapsulation efficiency and oxidation protection. The cheese whey and whey permeate (8:2) mixture exhibited the highest encapsulation efficiency (70.07%) and ability to protect the chia seed oil. After 30 days, the peroxide value was below the maximum limit considered safe for human consumption. Novelty and scientific contribution: According to these results, dairy by-products can be used for encapsulation of oxidation-sensitive oils. This represents an alternative use for dairy by-products, which otherwise are discarded and can impact the environment due to their high organic load. Our findings suggest that dairy by-products can be effectively used as wall materials to generate value-added products.

Upscale fermenter design for lactic acid production from cheese whey permeate focusing on impeller selection and energy optimization.

This study focusses on the design and scale-up of industrial lactic acid production by fermentation of dairy cheese whey permeate based on standard methodological parameters. The aim was to address the shortcomings of standard scale-up methodologies and provide a framework for fermenter scale-up that enables the accurate estimation of energy consumption by suitable selection of turbine and speed for industrial deployment. Moreover, life cycle assessment (LCA) was carried out to identify the potential impacts and possibilities to reduce the operation associated emissions at an early stage. The findings showed that a 3000 times scale-up strategy assuming constant geometric dimensions and specific energy consumption (P/V w ) resulted in lower impeller speed and energy demand. The Rushton turbine blade (RTB) and LightninA315 four-blade hydrofoil (LA315) were found to have the highest and lowest torque output, respectively, at a similar P/V w of 2.8 kWm-3, with agitation speeds of 1.33 and 2.5 s-1, respectively. RTB demonstrating lower shear damage towards cells (up to 1.33 s-1) was selected because it permits high torque, low-power and acceptable turbulence. The LCA results showed a strong relation between the number of impellers installed and associated emissions suggesting a trade-off between mixing performance and environmental impacts. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-021-05239-6.

Effect of by-products from the dairy industry as alternative inducers of recombinant ß-galactosidase expression.

OBJECTIVE: The aim of the present study was to evaluate the efficiency of lactose derived from cheese whey and cheese whey permeate as inducer of recombinant Kluyveromyces sp. ß-galactosidase enzyme produced in Escherichia coli. Two E. coli strains, BL21(DE3) and Rosetta (DE3), were used in order to produce the recombinant enzyme. Samples were evaluated for enzyme activity, total protein content, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis after induction with isopropyl-ß-D-1-thiogalactoside (IPTG) (0.05 and 1 mM) and lactose, cheese whey, and cheese whey permeate solutions (1, 10, and 20 g/L lactose) at shake-flask cultivation, and whey permeate solution (10 g/L lactose) at bioreactor scale. RESULTS: The highest specific activities obtained with IPTG as inducer (0.05 mM) after 9 h of induction, were 23 and 33 U/mgprotein with BL21(DE3) and Rosetta(DE3) strains, respectively. Inductions performed with lactose and cheese whey permeate (10 and 20 g/L lactose) showed the highest specific activities at the evaluated hours, exhibiting better results than those obtained with IPTG. Specific activity of recombinant ß-galactosidase using whey permeate (10 g/L lactose) showed values of approximately 46 U/mgprotein after 24-h induction at shake-flask study, and approximately 26 U/mgprotein after 16-h induction at bench bioreactor. CONCLUSIONS: The induction with cheese whey permeate was more efficient for recombinant ß-galactosidase expression than the other inducers tested, and thus, represents an alternative form to reduce costs in recombinant protein production.

Batch mesophilic anaerobic co-digestion of spent goat batch mesophilic anaerobic co-digestion of spent goat straw bedding and goat cheese whey: Comparison with the mono-digestion of the two sole substrates.

Spent livestock bedding is a valuable resource for the production of green energy (methane) in rural areas. Comparison and evaluation of batch anaerobic digestion and co-digestion of different mixtures of goat straw bedding (SGSB) and goat cheese whey were carried out. Biochemical methane potential (BMP) tests of the 100% SGSB, 95% SGSB-5% whey, 90% SGSB-10% whey, 85% SGSB-15% whey and 100% whey were found to be 423 ± 7, 354 ± 9, 371 ± 2, 293 ± 1, 274 ± 2 mL CH4 g-1 VS. Two different kinetic models were evaluated. The logistic model revealed a decrease in the maximum methane production rate (Rm) from 34.7 ± 1.5 to 14.1 ± 0.9 mL CH4 g-1 VS·d-1 when the percentage of whey in the mixture increased from 0 to 15% as a consequence of the increased ammonia released during the co-digestion of increased concentrations of whey. The lowest value for the maximum methane production predicted by the model (P) was found for 100% whey (274 ± 10 mL CH4 g-1 VS). A two-substrate model was applied to describe the evident existence of rapid and slowly degradable material. Regarding the hydrolysis kinetic constants predicted by this model, considerable increases in the rapid biodegradation stage (krapid) were observed when comparing to the values found for the slow (kslow) biodegradation stage in all the cases tested. The increases between both constants rose from 5 to 42% when the percentage of whey increased.

In situ biogas upgrading and enhancement of anaerobic digestion of cheese whey by addition of scrap or powder zero-valent iron (ZVI).

Cheese whey is an easily biodegradable substrate with high organic matter that can be anaerobically digested to biogas; however, the process is often inhibited by excess acidification due to the presence of undissociated volatile fatty acids and requires considerable concentration of alkaline buffer. The current study investigates a new approach for biogas upgrading, and increase of total CH4 in conjunction with buffering acidification by using zero-valent iron (powder and scrap metals at concentrations 25, 50, and 100 g/L) in anaerobic granular sludge and cheese whey under mesophilic batch conditions. During the first 2 cycles (total 34 days), a high performance was found in anaerobic bottles with 25 g/L powder zero valent iron (PZVI) and 50 g/L scrap zero valent iron (SZVI) since they had a higher total CH4 production compared to anaerobic bottles free of ZVI, as well as 97% CH4 composition in produced biogas compared to 74% CH4 for anaerobic bottles free of ZVI. Under these conditions, no additional NaOH was added to anaerobic bottles with 25 g/L PZVI and 50 g/L SZVI to increase the pH and at the end of 2nd cycle the concentration of VFAs was substantially lower compared to the anaerobic bottles free of ZVI. However, no positive effects of ZVI in terms of alkaline buffer were found at the 3rd and 4th cycle probably due to ZVI inactivation outer surface layer. Based on the experimental findings (anaerobic bottles: (a) 25 g/L PZVI, (b) 50 g/L SZVI and (c) free of ZVI) an economic comparison for anaerobic digestion of cheese whey by large scale was contacted and pointed out that the best scenario was the anaerobic digestion by addition of 50 g/L SZVI, followed by anaerobic digestion free of ZVI and last was the anaerobic digestion by addition of 25 g/L PZVI. This study highlights a new proof of concept for in-situ biogas upgrading and alleviation of acidification by addition of 50 g/L SZVI or 25 g/L PZVI during anaerobic digestion of cheese whey.

Quality Parameters of Wheat Bread with the Addition of Untreated Cheese Whey.

Τhe present study was carried out to evaluate wheat bread of three different flour compositions prepared by replacing water with untreated cheese whey (WCB). Bread prepared with water was taken as the control (CB). All breads were stored at 24 ± 1 °C for up to 6 days. Microbiological, physicochemical, and sensory analyses were determined as a function of storage time. WCB had lower total viable counts (TVC) (3.81 log cfu/g for CB and 2.78 log cfu/g for WCB on day 2 of storage) and showed delayed mold growth by 1 day (day 4 for CB and day 5 for WCB). WCB also had lower pH (5.91 for CB and 5.71 for WCB on day 0), higher titratable acidity values (TTA) (2.5-5.2 mL NaOH/10 g for CB and 4.5-6.8 mL NaOH/ 10 g for WCB), and higher protein content (PC) (PC 7.68% for CB and 8.88% for WCB). WCB was characterized by a more intense flavor, reduced hardness but similar cohesiveness, springiness, and adhesiveness compared to CB. Based primarily on sensory (appearance/mold formation) data, the shelf life of WCB was 4-5 days compared to 3-4 days for CB stored at 24 ± 1 °C. The proposed use of whey in bread preparation contributes decisively to the environmentally friendly management of whey.

Lactose hydrolysis using ß-galactosidase from Kluyveromyces lactis immobilized with sodium alginate for potential industrial applications.

The present study aimed to evaluate the lactose hydrolysis conditions from "coalho" cheese whey using ß-galactosidase (ß-gal) produced by Kluyveromyces lactis immobilized with sodium alginate. Three sodium alginate-based immobilization systems were evaluated (0.5, 0.7, and 1% w/v) for maximizing the immobilization yield (Y), efficiency (EM), and recovered activity (ar). The lactose hydrolysis capacity of the immobilized form of ß-gal was determined, and simulated environments were used to assess the preservation of the immobilized enzyme in the gastrointestinal tract. The results showed that ß-gal immobilization with 1% (w/v) sodium alginate presented the best results (EM of 66%, Y of 41%, and ar of 65%). The immobilization system maintained the highest pH stability in the range between 5.0 and 7.0, with the highest relative activity obtained under pH 5 conditions. The temperature stability was also favored by immobilization at 50 °C for 30 min was obtained a relative activity of 180.0 ± 1.37%. In 6 h, the immobilized ß-gal was able to hydrolyze 46% of the initial lactose content. For the gastrointestinal simulations, around 40% of the activity was preserved after 2 h. Overall, the results described here are promising for the industrial applications of ß-galactosidase from K. lactis.

Lactic acid production using cheese whey based medium in a stirred tank reactor by a ccpA mutant of Lacticaseibacillus casei.

In lactobacilli, CcpA is known to modulate the expression of genes involved in sugar metabolism, stress response and aerobic adaptation. This study aimed to evaluate a ccpA mutant of Lacticaseibacillus casei BL23 to increase lactic acid production using cheese whey. The ccpA derivative (BL71) showed better growth than the L. casei wild-type in the whey medium. In a stirred tank reactor, at 48 h, lactate production by BL71 was eightfold higher than that by BL23. In batch fermentations, the final values reached were 44.23 g L-1 for BL71 and 27.58 g L-1 for BL23. Due to a decrease in the delay of lactate production in the mutant, lactate productivity increased from 0.17 g (L.h)-1 with BL23 to 0.80 g (L.h)-1 with BL71. We found that CcpA would play additional roles in nitrogen metabolism by the regulation of the proteolytic system. BL71 displayed higher activity of the PepX, PepQ and PrtP enzymes than BL23. Analysis of prtP expression confirmed this deregulation in BL71. Promoter analysis of the prtP gene revealed CcpA binding sites with high identity to the cre consensus sequence and the interaction of CcpA with this promoter was confirmed in vitro. We postulate that deregulation of the proteolytic system in BL71 allows a better exploitation of nitrogen resources in cheese whey, resulting in enhanced fermentation capacity. Therefore, the ccpA gene could be a good target for future technological developments aimed at effective and inexpensive lactate production from dairy industrial wastes.

Valorization of cheese whey using microbial fermentations.

Cheese whey (CW), the liquid resulting from the precipitation and removal of milk casein during cheese-making, and the second cheese whey (SCW) derived from the production of cottage and ricotta cheeses are the main byproducts of dairy industry. The major constituent of CW and SCW is lactose, contributing to the high BOD and COD content. Because of this, CW and SCW are high-polluting agents and their disposal is still a problem for the dairy sector. CW and SCW, however, also consist of lipids, proteins, and minerals, making them useful for production of various compounds. In this paper, microbial processes useful to promote the bioremediation of CW and SCW are discussed, and an overview on the main whey-derived products is provided. Special focus was paid to the production of health-promoting whey drinks, vinegar, and biopolymers, which may be exploited as value-added products in different segments of food and pharmaceutical industries.