Gouda Cheese with Modified Content of β-Casein as a Source of Peptides with ACE- and DPP-IV-Inhibiting Bioactivity: A Study Based on In Silico and In Vitro Protocol

In silico and in vitro methods were used to analyze ACE- and DPP-IV-inhibiting potential of Gouda cheese with a modified content of β-casein. Firstly, the BIOPEP-UWM database was used to predict the presence of ACE and DPP-IV inhibitors in casein sequences. Then, the following Gouda cheeses were produced: with decreased, increased, and normative content of β-casein after 1 and 60 days of ripening each (six variants in total). Finally, determination of the ACE/DPP-IV-inhibitory activity and the identification of peptides in respective Gouda-derived water-soluble extracts were carried out. The identification analyses were supported with in silico calculations, i.e., heatmaps and quantitative parameters. All Gouda variants exhibited comparable ACE inhibition, whereas DPP-IV inhibition was more diversified among the samples. The samples derived from Gouda with the increased content of β-casein (both stages of ripening) had the highest DPP-IV-inhibiting potency compared to the same samples measured for ACE inhibition. Regardless of the results concerning ACE and DPP-IV inhibition among the cheese samples, the heatmap showed that the latter bioactivity was predominant in all Gouda variants, presumably because it was based on the qualitative approach (i.e., peptide presence in the sample). Our heatmap did not include the bioactivity of a single peptide as well as its quantity in the sample. In turn, the quantitative parameters showed that the best sources of ACE/DPP-IV inhibitors were all Gouda-derived extracts obtained after 60 days of the ripening. Although our protocol was efficient in showing some regularities among Gouda cheese variants, in vivo studies are recommended for more extensive investigations of this subject.

Metataxonomic Profiling of Native and Starter Microbiota During Ripening of Gouda Cheese Made With Listeria monocytogenes-Contaminated Unpasteurized Milk

Unpasteurized milk is used to produce aged artisanal cheeses, which presents a safety concern due to possible contamination with foodborne pathogens, especially Listeria monocytogenes. The objective of this study was to examine the composition of the bacterial community in unpasteurized milk used to prepare Gouda cheese artificially contaminated with L. monocytogenes (~1 log CFU/ml) and assess the community dynamics and their potential interaction with L. monocytogenes during a 90-day ripening period using targeted 16S rRNA sequencing. The diversity of bacterial taxa in three batches of unpasteurized milk was not significantly different, and the microbiomes were dominated by species of Lactococcus, Streptomyces, Staphylococcus, and Pseudomonas. The highest relative abundances were observed for Pseudomonas fluorescens (31.84–78.80%) and unidentified operational taxonomic units (OTUs) of Pseudomonas (7.56–45.27%). After manufacture, both with and without L. monocytogenes-contaminated unpasteurized milk, Gouda cheese was dominated by starter culture bacteria (including Lactococcus lactis subsp. cremoris, lactis, lactis bv. diacetylactis, and Streptococcus thermophilus), in addition to unassigned members in the taxa L. lactis and Streptococcus. During ripening there was an overall decrease in L. lactis abundance and an increase in the number of taxa with relative abundances >0.1%. After 90-day ripening, a total of 82 and 81 taxa were identified in the Gouda cheese with and without L. monocytogenes, respectively. Of the identified taxa after ripening, 31 (Gouda cheese with L. monocytogenes) and 56 (Gouda cheese without L. monocytogenes) taxa had relative abundances >0.1%; 31 were shared between the two types of Gouda cheese, and 25 were unique to the Gouda cheese without added L. monocytogenes. No unique taxa were identified in the Gouda cheese with the added L. monocytogenes. This study provides information on the dynamics of the bacterial community in Gouda cheese during ripening, both with and without the addition of L. monocytogenes.

Reduction in Spoilage Microbiota and Cyclopiazonic Acid Mycotoxin with Chestnut Extract Enriched Chitosan Packaging: Stability of Inoculated Gouda Cheese

Active chitosan-based films, blended with fibrous chestnut (Castanea sativa Mill.) tannin-rich extract were used to pack Gouda cheese that has been contaminated with spoilage microflora Pseudomonas fluorescens, Escherichia coli, and fungi Penicillium commune. A comprehensive experimental plan including active chitosan-based films with (i) chestnut extract (CE), (ii) tannic acid (TA), and (iii) without additives was applied to evaluate the film′s effect on induced microbiological spoilage reduction and chemical indices of commercial Gouda cheese during 37 days while stored at 4 °C and 25 °C, respectively. The cheese underwent microbiology analysis and chemical assessments of ultra-high-performance liquid chromatography (UHPLC) (cyclopiazonic acid), pH, and moisture content. The biopackaging used for packing cheese was characterized by mechanical properties before food packaging and analyzed with the same chemical analysis. The cheese microbiology showed that the bacterial counts were most efficiently decreased by the film without additives. However, active films with CE and TA were more effective as they did not break down around the cheese and showed protective properties against mycotoxin, moisture loss, and pH changes. Films themselves, when next to high-fat content food, changed their pH to less acidic, acted as absorbers, and degraded without plant-derived additives.

Investigation of the antimicrobial properties of nanoclay and chitosan based nanocomposite on the microbial characteristics of Gouda cheese

Background and Objectives: In recent years, active packaging has been introduced as a new method to better preserve food. Chitosan and nanoclay have been used for preparation of an active nanocomposite with respect to their antimicrobial properties to investigate its effects on the microbial limitation in Gouda cheese. Materials and Methods: Nanoclay film, chitosan film, chitosan-based nanocomposites and nanoclay-based nanocompos- ites were prepared and their antimicrobial properties were evaluated to the microbial limitations of Gouda cheese con- sist of coliforms, Escherichia coli, Salmonella spp., coagulase-positive Staphylococcus, mold and yeast by agar diffusion method. Results: The results indicated, the best antimicrobial effect belonged to nanocomposite film with the composition of chitosan 3 wt% by adding nanoclay 1 wt%, which can prevent microbial characteristics of Gouda cheese. Conclusion: The chitosan and nanoclay nanocomposite had excellent antibacterial activity and performed well against microbial limitations (coliforms, E. coli, Salmonella spp., coagulase-positive Staphylococcus, mold and yeast) of Gouda cheese. Therefore, the nanocomposite may be possibly used as a surface coating in addition to Gouda cheese as well as sim- ilar cheeses and other food to enhance microbial characteristics and extend shelf life.

Essential Oil Based PVP-CMC-BC-GG Functional Hydrogel Sachet for ‘Cheese’: Its Shelf Life Confirmed with Anthocyanin (Isolated from Red Cabbage) Bio Stickers

‘Gouda cheese’ is one of the most popular varieties of cheese eaten worldwide. The preservation problem of gouda arises due to microbial contamination and infestation. Therefore, essential oil (EO) based PVP-CMC-BC-GG hydrogel film was prepared to solve the problem and to extend the shelf-life of ‘Gouda cheese’. Anthocyanin (isolated from red cabbage) based pH stickers are integrated into the packaging system to recognize the spoilage of ‘cheese’. EOs (clove and/or cinnamon) are added to PVP-CMC-BC-GG hydrogel film to improve its antimicrobial, physical, mechanical, and thermal properties as well as shelf-life of cheese. The films are assessed based on their physical, structural, and functional properties, real-time assessment on cheese, and biodegradability. The results revealed that although the addition of oils to the PVP-CMC-BC-GG hydrogel films enhanced its mechanical, hydrophobic, and antimicrobial properties, the biodegradability of PVP-CMC-BC-GG films declined with the addition of EOs. The thermal properties remained the same irrespective of the addition of EOs. The shelf life of cheese was extended for more than 10–12 days, inside the PVP-CMC-BC-GG hydrogel sachet compared to the conventional PE packaging system. Hence the use of the PVP-CMC-BC-GG sachet (containing EO or without EO) is recommended for cheese packaging along with the use of PVP-CMC-BC-GG anthocyanin bio stickers for monitoring the quality of cheese.