Integration of Postbiotics in Food Products through Attenuated Probiotics: A Case Study with Lactic Acid Bacteria in Bread.

The study examines the integration of postbiotics in food products through the use of attenuated probiotics, specifically lactic acid bacteria (LAB) in bread. Postbiotics, non-viable microorganisms or their metabolites, offer health benefits similar to probiotics without the risks associated with live bacteria. This research evaluates the regulatory aspects and safety of LAB in sourdough bread production, highlighting their historical and significant use in Europe before 1997. The study includes microbial quantification and Next-Generation Sequencing (NGS) to identify LAB in traditional sourdough, comparing them with historical and current EFSA Qualified Presumption of Safety (QPS) lists. Findings show that the LAB present in sourdough have been extensively and safely used in bread making, supporting their classification as non-novel foods under EU regulations. The stability and consistency of LAB metabolites in sourdough bread are also confirmed, ensuring quality and safety in each batch. The study concludes that LAB in sourdough, when inactivated through bread-making processes, are not considered novel foods, aligning with historical, scientific, and regulatory evidence.

Sourdough Bread Quality: Facts and Factors.

The term "sourdough" denotes a dough composed of flour and water, fermented through the action of yeast and lactic acid bacteria. The utilization of sourdough fermentation technology can enhance the nutritional attributes of bread made from wheat grain. In recent times, sourdough bread has experienced a resurgence, fueled by growing consumer demand for healthier bread options. The market dynamics for sourdough illustrate its rapid expansion and significant role in the contemporary food industry. Sourdough fermentation improves nutritional qualities by altering the structure and function of proteins and starch, enhancing dietary fiber, volatile compound profiles, and antioxidant activity, and reducing FODMAPs. The quality of sourdough bread is influenced by several factors, including fermentation environment, flour particle size, protein quality, starch characteristics, and dietary fiber composition. Moreover, the incorporation of alternative grains (intermediate wheatgrass and legume flour) and non-flour ingredients (fruits, herbs, and dairy products) presents opportunities for creating sourdough bread with unique sensory and nutritional profiles. This review offers updated insights on the quality aspects of sourdough fermentation, the factors that influence the effectiveness of the sourdough fermentation process, sourdough technology with unconventional and non-flour ingredients, and the potential market for frozen sourdough, considering its convenience and extended shelf life.

Evolution-aided improvement of the acid tolerance of Levilactobacillus brevis and its application in sourdough fermentation.

Levilactobacillus brevis is crucial in food fermentation, particularly in sourdough production. However, the cultivation of L. brevis faces a challenge with accumulation of lactic acid, a major inhibitor. We aimed to increase the acid tolerance of L. brevis, an industrial strain for sourdough fermentation. We used the adaptive laboratory evolution (ALE) to obtain lactic acid tolerant strains. The evolved strain's fermentation and metabolite profiles, alongside sensory evaluation, were compared with the parental strain by using various analytical techniques. The ALE approach increased lactic acid tolerance in the evolved strain showing an increased growth rate by 1.1 and 1.9 times higher than the parental strain at pH 4.1 and 6.5, respectively. Comprehensive analyses demonstrated its potential application in sourdough fermentation, promising reduced downstream costs. The evolved strain, free from genetically modified organisms concerns, has great potential for industrial use by exhibiting enhanced growth in acidic conditions without affecting consumers' bread preferences.

Sourdough bread enriched with exopolysaccharides and gazpacho by-products modulates in vitro the microbiota dysbiosis.

Sourdough bread enriched with soluble fiber (by in-situ exopolysaccharides production) and insoluble fiber (by gazpacho by-products addition) showed prebiotic effects an in vitro dynamic colonic fermentation performance with obese volunteer's microbiota. Bifidobacterium population was maintained whereas Lactobacillus increased throughout the colonic sections. Conversely, Enterobacteriaceae and Clostridium groups clearly decreased. Specific bacteria associated with beneficial effects increased in the ascending colon (Lactobacillus fermentum, Lactobacillus paracasei, Bifidobacterium longum and Bifidobacterium adolescentis) whereas Eubacterium eligens, Alistipes senegalensis, Prevotella copri and Eubacterium desmolans increased in the transversal and descending colon. Additionally, Blautia faecis and Ruminococcus albus increased in the transversal colon, and Bifidobacterium longum, Roseburia faecis and Victivallis vadensis in the descending colon. Bifidobacterium and Lactobacillus fermented the in-situ exopolysaccharides and released pectins from gazpacho by-products, as well as cellulosic degraded bacteria. This increased the short and medium chain fatty acids. Acetic acid, as well as butyric acid, increased throughout the colonic tract, which showed greater increases only in the transversal and descending colonic segments. Conversely, propionic acid was slightly affected by the colonic fermentation. These results show that sourdough bread is a useful food matrix for the enrichment of vegetable by-products (or other fibers) in order to formulate products with microbiota modulatory capacities.

Harnessing the Role of Three Lactic Acid Bacteria (LAB) Strains for Type II Sourdough Production and Influence of Sourdoughs on Bread Quality and Maillard Reaction Products.

This study tested the effect of Companilactobacillus paralimentarius E-106, Lactiplantibacillus paraplantarum N-15 and Lactiplantibacillus plantarum SC-9 on the amount of Maillard reaction and aroma profile in bread making with main bread quality parameters. The specific volumes of sourdough and control breads were in the range of 2.97-3.04 cm3/g, and the control II bread had the highest hardness values on all days. The FAST index value was determined to be between 40.48% and 81.22% in all breads. The FAST index value was found to be higher in the control breads than in the sourdough breads. In the volatile compounds analysis, 72 volatile compounds were detected. The variety of volatile compounds in the breads with sourdough addition was higher than the control breads. Among the tested strains, Companilactobacillus paralimentarius E-106 demonstrated superior properties for bread characteristics in comparison to other strains as a type II sourdough starter. In summary, improved aroma profile and decreased Maillard reaction products can be provided by sourdough addition without changing the bread quality, along with meeting consumer demand for less additive use.

Sourdough fermentation for the valorization of sorghum flour: Microbiota characterization and metabolome profiling.

Due to a large adaptability to different cultivation conditions and limited input compared to other cereals, sorghum is considered an emerging crop. Its antioxidant properties, high fiber content and low glycemic index also make it a valuable addition to a healthy diet, nevertheless, the presence of antinutritional factors and the lack of gluten, hamper its use as food ingredient. This study investigated the impact of sourdough fermentation on sorghum nutritional quality. Lactic acid bacteria dominating sorghum flour and sourdough were identified by culture-dependent analysis revealing Lactiplantibacillus plantarum as the dominant species found in the mature sourdough, whereas Weissella cibaria and Weissella paramesenteroides were the species isolated the most after the first refreshment. Among yeasts, Saccharomyces cerevisiae was the most prevalent. Lactic acid bacteria pro-technological and functional performances as starter were evaluated in sorghum type-II sourdoughs through an integrated characterization combining chromatographic and NMR spectroscopic techniques. The metabolic profile of the strains mainly grouped together W. cibaria strains and W. paramesenteroides AI7 which distinguished for the intense proteolysis but also for the presence of compounds particularly interesting from a physiological perspective (allantoin, glutathione, γ-aminobutyric acid and 2-hydroxy-3-methylbutyric acid), whose concentration increased during fermentation in a species or strain specific matter.

Influence of baking time and formulation of part-baked wheat sourdough bread on the physical characteristics, sensory quality, glycaemic index and appetite sensations.

Raw materials and process parameters in bread production can modulate the glycemic index, which on itself has been linked with provision of better hunger satisfaction and maintaining better satiation. The objective of this research was to investigate if using unrefined wheat flour or the addition of intact cereals in formulation or alternating the baking time would have an effect on physical characteristics, sensory quality, glycaemic index and appetite sensations in wheat sourdough bread. In the study, three types of commercial part-baked frozen sourdough bread, baked to the final baking for two different times (long and short baking time) were used. A randomized controlled crossover trial was performed with 10 healthy adults who consumed sufficient quantity of bread to ingest 50 g available carbohydrates. Participants self-reported appetite sensations (desire to eat, hunger, fullness, satisfaction, appetite) on a 10 cm visual analog scale (VAS) scale in a course of 180 min. In addition, bread products were subjected to overall acceptability and different sensory attributes were examined on JAR "just about right" scale. Different bread formulations (refined flour, unrefined wheat flour, cereal flour or intact cereals) and different length of baking time significantly influenced (p < 0.005) physical, textural and sensory features of products. The alternation of aforementioned parameters decreased the glycemic index, but not significantly (p > 0.005). No correlation was found between lower GI, satiety and satiation. Liking score and incremental area under the curve (iAUC) of satiety and satiation were calculated as highest in sourdough bread with added cereals.

Exploring the Nutritional Impact of Sourdough Fermentation: Its Mechanisms and Functional Potential.

Sourdough fermentation is one of the oldest traditional methods in food technology and occurs as a result of fermentation of flour prepared from grains. The nutritional role of sourdough is related to the final composition of fermented foods prepared through sourdough fermentation, and recently, sourdough has become an important application to improve nutrition characteristics of bread. Thanks to lactic acid bacteria (LAB) presented in sourdough microflora and metabolites partially produced by yeasts, technological and important nutritional features of the bread improve and an increase in shelf life is achieved. In addition, sourdough bread has a low glycemic index value, high protein digestibility, high mineral and antioxidant content, and improved dietary fiber composition, making it more attractive for human nutrition compared to regular bread. When the sourdough process is applied, the chemical and physical properties of fibers vary according to the degree of fermentation, revealing the physiological importance of dietary fiber and its importance to humans' large intestine microbiota. Therefore, taking these approach frameworks into consideration, this review highlights the benefits of sourdough fermentation in increasing nutrient availability and contributing positively to support human health.

Wholegrain triticale sourdough: Effects of triticale:Wheat flour ratio and hydration level on bread quality.

Triticale (×Triticosecale) is a hybrid between wheat (Triticum spp.) and rye (Secale cereale), producing higher grain yields than wheat in challenging environments. Triticale grain is also highly nutritious. Thus, the potential of triticale grain for an expanded range of food applications should be explored. Sourdough bread has unique functional and nutritional properties, but understanding the effects of partial substitution of triticale for wheat flour and varying levels of dough moisture content on sourdough quality requires further research. The aim of this study was to evaluate the wholegrain flour of contrasting triticale cultivars in comparison to that of a common wheat cultivar in commercial sourdough breadmaking. Two triticale cultivars (Goanna and Hawkeye, selected from a panel of Australian genotypes) and one wheat cultivar (Scout) were grown in a field trial in northern NSW, Australia. Differences in quantitative texture and color parameters of the dough and sourdough bread resulting from (1) substitution of commercial wholemeal wheat flour with different proportions of wholegrain triticale or wholegrain wheat (Scout) flour (0%, 60%, 70%, 80%, and 90%) and (2) varying the amount of water included in the dough preparation (70%, 80%, 90%, and 100 g water/100 g flour) were determined. Replacement of wholemeal wheat flour with 60% wholegrain Goanna flour (protein content 12.76%; c.f. 11.50% for Hawkeye, 12.40% for Scout), and addition of 100 g water/100 g flour in the dough preparation gave the highest quality sourdough bread based on specific volume, texture, and color parameters and had similar properties to the control made from wheat alone.

Insights into the starch and proteins molecular structure changes of foxtail millet sourdough: Effect of fermentation from grains of cereal to pre-meal.

This study investigated the effects of foxtail millet sourdough fermentation time (0, 8, 16, and 24 h) on the protein structural properties, thermomechanical, fermentation, dynamic rheological, starch granules crystalline regions molecular mobility, and starch microstructural characteristics. The fermentation led to a significant increase in the concentration of free amino acids from protein hydrolysis. Fourier transform infrared spectroscopy (FTIR) revealed changes in protein secondary structure and the presence of functional groups of different bioactive compounds. The result of thermomechanical properties showed a significant increase in the stability (0.70-0.79 min) and anti-retrogradation ability (2.29-3.14 Nm) of lactic acid bacteria (LAB) sourdough compared to the control dough, showing a wider processing applicability with radar profiler index. In contrast, sourdoughs with lower tan δ values had higher elasticity and strength. Scanning electron microscopy showed that the surface of the starch appeared from smooth to uneven with patchy shapes and cavities, which declined the crystallinity from 34.00 % to 21.57 %, 23.64 %, 25.09 %, and 26.34 % respectively. Fermentation changed the To, Tp, Tc, and ΔH of the starch. The results of the study will have great potential for application in the whole grain sourdough industry.

Identification of the Microbiota in Coconut Water, Kefir, Coconut Water Kefir and Coconut Water Kefir-Fermented Sourdough Using Culture-Dependent Techniques and Illumina-MiSeq Sequencing.

The principal objective of this study was to isolate and identify the microorganisms present in commercial kefir grains, a novel kefir-fermented coconut water (CWK) and a novel coconut water kefir-fermented sourdough using phenotypic identification and Sanger sequencing and examine the microbial diversity of CWK and CWK-fermented sourdough throughout the fermentation process using the MiSeq Illumina sequencing method. The phenotypic characterisation based on morphology identified ten isolates of LAB, five AAB and seven yeasts from kefir (K), CWK and CWK-fermented sourdough (CWKS). The results confirm the presence of the LAB species Limosilactobacillus fermentum, Lactobacillus. plantarum, L. fusant, L. reuteri and L. kunkeei; the AAB species Acetobacter aceti, A. lovaniensis and A. pasteurianus; and the yeast species Candida kefyr, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, C. guilliermondii and C. colliculosa. To the best of our knowledge, the identification of Rhodotorula from kefir is being reported for the first time. This study provides important insights into the relative abundances of the microorganisms in CWKS. A decrease in pH and an increase in the titratable acidity for CWK- and CWK-fermented sourdough corresponded to the increase in D- and L-lactic acid production after 96 h of fermentation. Significant reductions in the pHs of CWK and CWKS were observed between 48 and 96 h of fermentation, indicating that the kefir microorganisms were able to sustain highly acidic environments. There was also increased production of L-lactic acid with fermentation, which was almost twice that of D-lactic acid in CWK.

Application of sourdough in gluten-free bakery products.

ABSTRACTSIn recent years, the demand for gluten-free (GF) bakery products has grown rapidly due to the remarkable rising number of celiac patients and the increasing health awareness of GF products. However, GF products generally suffer from defects such as poor sensorial level, low nutritional value, high prices and short shelf life. Sourdough is the important starter culture applied in bakery field, and it has been proven to be ideal for enhancing the overall quality of bakery products. This review aims to systematically reviewed the application of sourdough in GF bakery products and its improvement to GF bakery products in terms of texture, shelf life, nutrition and flavor. Its positive effects derive from the complex metabolic activities of sourdough microorganisms, such as acidification, proteolysis, production of exopolysaccharides (EPS), activation of endogenous enzymes, and production of antibacterial substances. Finally, researchers are encouraged to expand the use of sourdough in GF bakery products to increase the variety of GF products. And the technical and nutritional potential of sourdough should be developed more widely.

Production of γ-aminobutyric acid-enriched sourdough bread using an isolated Pediococcus pentosaceus strain JC30.

A γ-aminobutyric acid (GABA)-producing strain JC30 was isolated from traditional kimchi, which was identified as Pediococcus pentosaceus by 16S rDNA sequencing. P. pentosaceus JC30 was highly tolerant to acid, bile salt, and high temperatures. The survival rate of JC30 in MRS medium (pH 2.5) for 3 h was 60.96 %. Furthermore, the survival rate of JC30 in MRS medium with 3 mg/mL bile salt for 24 h was 86.62 %. The survival rate of JC30 in MRS medium at 56 °C and 58 °C for 10 min was 97.17 % and 78.20 %, respectively. When 2 % v/v JC30 (8.0 log10 CFU/mL) was added to prepare sourdough and the sourdough was then used to make bread, the bread had a higher specific volume (5.13 ± 0.12 mL/g) and GABA content (3.32 ± 0.04 mg/g DW) than the control.

The effects of fermentation time on sourdough bread: An analysis of texture profile, starch digestion rate, and protein hydrolysis rate.

To ensure the best quality bread, it is important to consider the speed of digestion of starch and proteins, as well as how time fermentation and storage time influence the rate of starch digestion and the texture of the bread. This study compared the effect of fermentation time and days of storage on the texture, physicochemical, protein and starch digestibility of sourdough bread. Texture profile analysis showed that the fermentation time in recently baked sourdough bread affects hardness, chewiness, and springiness. The electrophoretic profile showed a decrease in band thickness with increase in fermentation time, consistent with a higher percentage of protein digestion. While fermentation time did not significantly affect rapidly digestible starch (RDS) and slowly digestible starch (SDS), storage time resulted in a decrease in RDS and an increase in SDS. Sourdough breads had higher levels of resistant starch (RS). The digestibility characteristics of protein and starch, as well as texture properties, are significantly influenced by fermentation and storage time. The evidence suggests that sourdough bread has the potential to improve the digestion of protein and to effectively regulate the glycemic response, which is due to its higher levels of SDS and RS.

Casein hydrolysate in naturally-fermented buckwheat sourdough: Effects on fermented and physicochemical characteristics, texture, and bacterial microbial composition.

The effect of a casein hydrolysate (CH) on the fermentation and quality of a naturally-fermented buckwheat sourdough (NFBS) were investigated, through assessing the fermentation characteristics, carbohydrate and protein degradation, texture, and bacterial composition of NFBS. According to the assaying data, CH might both increase the amount of lactic acid bacteria by 2.62 % and shorten the fermentation period by at least 3 h, subsequently leading to enhanced degradation of carbohydrate and protein, accompanied by a softer texture. More importantly, CH increased the relative abundance of lactobacillus in NFBS, making it the dominant bacterial genus and inhibited the growth of spoilage bacteria. In addition, Spearman correlation analysis indicated that the pH value, lactic and acetic acid contents, carbohydrates, protease activity, and these textural indices like hardness, elasticity, and adhesion had a positive/negative correlation with the bacterial composition of NFBS (Spearman correlation coefficient: -0.93-0.95). CH was thus regarded to be helpful to NFBS processing and production mainly by shortening its fermentation time, improving its fermentation performance, causing a finer texture and microstructure, and changing bacterial composition.

The impact of magnetic field-assisted freeze-thaw treatment on the quality of foxtail millet sourdough and steamed bread.

Foxtail millet and sourdough are used to make foxtail millet sourdough steamed bread to improve the flavor and taste. Compared with the conventional freeze-thaw treatment (CFT), the effect of magnetic field-assisted freeze-thaw treatment (MFT) on the storage quality of foxtail millet sourdough and steamed bread is explored. The results showed that compared with CFT, MFT shortened the phase transition time of dough; decreased the water loss rate, the water mobility, and the freezable water content; increased the fermentation volume; stabilized the rheological properties; and minimized the damage of freezing and thawing to the secondary structure and microstructure of the gluten. In addition, an analysis of the specific volume, texture, surface color, and texture structure showed that MFT was beneficial to slowing the deterioration of the steamed bread texture. Finally, MFT effectively inhibited the growth and recrystallization of ice crystals during freezing and thawing, improving the quality of millet dough and steamed bread.

Assessment of Sourdough Fermentation Impact on the Antioxidant and Anti-Inflammatory Potential of Pearl Millet from Burkina Faso.

Millet, a gluten-free cereal, has received attention for its environmental friendliness and higher protein content than other grains. It represents a staple food in many African countries, where fermentation is traditionally used for preserving food products and preparing different cereal-based products. This study aimed to assess the impact of sourdough fermentation on bioactive compounds and antioxidant and anti-inflammatory properties of pearl millet from Burkina Faso. Phenolic compounds were investigated spectrophotometrically and by HPLC-DAD. The antioxidant activity of unfermented (MF) and fermented (FeMF) millet was evaluated in vitro by spectrophotometric and fluorometric assays and ex vivo on oxidized human erythrocytes for hemolysis inhibition. Finally, the potential anti-inflammatory effect of FeMF and MF was evaluated on human adenocarcinoma cell line (HT-29) exposed to TNF-α inflammatory stimulus. Results revealed significantly higher levels of polyphenols, flavonoids, and in vitro antioxidant activity following millet fermentation. Notable differences in phenolic composition between FeMF and MF are observed, with fermentation facilitating the release of bioactive compounds such as gallic acid, quercetin, and rutin. A dose-dependent protection against oxidative hemolysis was observed in both FeMF- and MF-pretreated erythrocytes. Similarly, pretreatment with FeMF significantly reduced the levels of inflammatory markers in TNF-α-treated cells, with effects comparable to those of MF. Fermentation with sourdough represents a simple and low-cost method to improve the bioactive compounds content and in vitro antioxidant activity of millet flour with promising nutraceutical potential.

The effect of sourdough, turnips, and butternut squash on the physicochemical and nutritional properties of Doowina functional food during fermentation.

The dairy-cereal-based food, known as Doowina, is one of the traditional fermented foods in Iran. We aimed to improve the health-promoting properties of Doowina by using turnips, butternut squash, and sourdough as a new functional food with high nutritional value and antioxidant activity. Therefore, the physicochemical, microbial, and sensory properties of samples with nutritional supplements (8% turnip and 8% butternut squash) and different concentrations of sourdough (0, 0.5, and 1%) were studied during 0, 3, 6, and 9 days of fermentation time. The results showed that there was no significant difference (p < .05) in the moisture and ash content between the different formulations of Doowina. There was also no significant difference (p < .05) in the phenolic compound content and antioxidant activity of the Doowina samples during the fermentation period. However, the number of lactic acid bacteria (LAB) increased significantly (p < .05) until the 6th day of fermentation, and the protein content decreased significantly (p < .05) in all samples during the fermentation period. According to the results, the samples with butternut squash and sourdough had the highest total phenolic content, the highest antioxidant activity, the highest linoleic acid content, and the highest sensory rating of all samples.

Contribution of γ-Glutamyl-Cysteine Ligases of Limosilactobacillus reuteri to the Formation of Kokumi-Active γ-Glutamyl Dipeptides in Sourdough Bread.

Kokumi-active γ-glutamyl dipeptides accumulate during sourdough fermentation. γ-Glutamylcysteine ligases (Gcls) of Limosilactobacillus reuteri synthesize γ-glutamyl dipeptides during growth in sourdough. This study aimed to evaluate the contribution of Gcls from strains of L. reuteri in the formation of kokumi-active γ-glutamyl dipeptides in sourdough bread. Among 12 acceptor amino acids, the three Gcls of L. reuteri were the most active to Cys. With the acceptor amino acids Ile, Leu, and Phe, Gcl1 was more active than Gcl2 and Gcl3. Accordingly, Gcl1 contributed to the γ-Glu-Ile synthesis in sourdough fermentation. Proofing and baking strongly influenced the concentration of γ-glutamyl dipeptides in bread. The addition of 10% sourdough increased the content of γ-Glu-Leu and γ-Glu-Phe but not of other γ-glutamyl dipeptides in bread. In conclusion, the accumulation of kokumi γ-glutamyl dipeptides in sourdoughs was attributed to the combined activity of cereal enzymes, γ-glutamyl-cysteine ligases, and other microbial enzymes.

Sourdough bread as nutritional intervention tool for improvement of cognitive dysfunction in diabetic rats.

BACKGROUND: The current research targeted to study the impact of nutritional intervention by two sourdough breads in improvement of cognitive dysfunction in diabetic rats. METHODS: Type-2 diabetes was induced in rats by Streptozotocin-Nicotinamide (STZ-NC). Diabetic rats were fed on balanced diet or balanced diet containing 20% of sourdough bread I or II for a month. Lipid profile, oxidative stress, inflammatory markers and cognitive functions were assessed in all rats. Gene expression of brain-derived neurotrophic factor (BDNF) and nuclear respiratory factor 2 (NRF-2) were assessed in hippocampal tissue, while expression of phosphoenol pyruvate carboxy kinase (PEPCK), and glucose transporter 2 (GLUT2) genes were evaluated in hepatic tissue. Chemical composition and fatty acids profile were evaluated in the prepared sourdough bread. RESULTS: Sourdough bread II showed higher content of phenolic compounds, fat, fiber and carbohydrates. Fatty acids profile revealed that sourdough bread I was higher in saturated fatty acids (16.08%), while sourdough bread sample II was higher in unsaturated fatty acids (79.33%). Sourdough bread I or II feeding rats' showed significant improvement in hyperglycemia, oxidative stress markers, inflammatory markers, lipid profile, liver and kidney functions in association with improvement in cognitive function. Gene expression of BDNF and NRF2 in hippocampal tissue were increased significantly, while hepatic GLUT2 and PEPCK gene expression were down-regulated in diabetic given sourdough bread I or II. CONCLUSION: Sourdough bread II was superior in all the studied parameters. The anti-diabetic effect and protection from cognitive dysfunction of sourdough bread samples may be ascribed to the occurrence of dietary fibers, phenolic compounds, and polyunsaturated fatty acids.