Release Of Short-chain Fatty Acids Research Articles

Protective effects of YCHD on the autoimmune hepatitis mice model induced by Ad-CYP2D6 through modulating the Th1/Treg ratio and intestinal flora

BackgroundAutoimmune hepatitis (AIH) is a chronic liver inflammatory disease mediated by autoimmune reactions, the pathogenesis of AIH is probably related to the imbalance of intestinal flora. Yinchenhao decoction (YCHD) has been used to relieve AIH. However, the mechanisms underpinning YCHD’s hepatoprotective effects with the gut microbito have not been fully revealed.ObjectiveTo explore the potential mechanism of YCHD in treating AIH based on changes in the intestinal flora and Th1/Treg ratio in the spleen and hepatic hilar lymph nodes.MethodsThe AIH mice model induced by the adenovirus vectors that overexpress human cytochrome P450 family 2 subfamily D member 6 (Ad-CYP2D6) was established (untreated group). One week after the Ad-CYP2D6 injection, the AIH model mice were treated by administering YCHD by gavage for 14 days (YCHD-treated group). The therapeutic efficacy of YCHD on AIH was evaluated by detecting the histopathological changes of the liver, serum transaminases (ALT and AST), inflammatory factors (TNF-α,IL-17 and IFN-γ), and autoantibodies (including LKM-1 and LC-1). The ratio of Th1 to Treg within the spleen and hepatic hilar lymph nodes of the mice was detected by flow cytometry. The changes in the species and abundance of intestinal flora and intestinal flora metabolites were analyzed via 16S rRNA gene sequencing and gas chromatography-mass spectrometry (GC/MS) to reveal the protective mechanism of YCHD on liver injury.ResultYCHD decreased the transaminase activity (AST and ALT), the content of autoantibodies (LC-1 and LKM-1), and the serum TNF-α, IL-12, and IL-17 levels in AIH mice. The degree of inflammatory infiltration in the YCHD-treated group was significantly less than that in the untreated group. YCHD can effectively reverse the abundance and diversity of intestinal flora in AIH mice and affect the release of short-chain fatty acids (SCFAs), especially butyric acid. Moreover, the flow cytometry results showed that YCHD could also decline the ratio of Th1/Treg, which probably be induced by SCFAs via the G protein-coupled receptor (GPR).ConclusionYCHD may affect the release of SCFAs by regulating the intestinal microbiota, thereby affecting the differentiation of Th1 and Treg, and achieving the effect of alleviating liver damage.

Impact of milking interval and time on milk spontaneous lipolysis and composition in dairy cows

Milk lipolysis is defined as the hydrolysis of triglycerides, the major component of milk fat, resulting in the release of short-chain fatty acids (FA) responsible for rancid flavor and partial glycerides that impair functional properties such as foaming and creaming abilities. Milk lipolysis is a complex phenomenon that depends on both animal parameters and breeding factors. Milk spontaneous lipolysis is known to be higher in milk from evening milkings than from morning milkings. This may be related to the longer length of overnight milking intervals or to the nycthemeral cycle. In this experiment, our objective was thus to study the impact of both milking intervals and time of day on milk spontaneous lipolysis in twice-daily-milking systems with one of 3 milking intervals: Short Day - Long Night (SD-LN, 6.30 a.m. and 4.30 p.m.,); Long Day - Short Night (LD-SN, 6:30 a.m. and 8:30 p.m.,); and Balanced Day and Night (BDN, 6:30 a.m. and 6:30 p.m.,). To achieve this goal, 21 multiparous dairy cows in mid-lactation were used in a 3 × 3 Latin square design over 3 periods. The experiment lasted 5 weeks, corresponding to 3 experimental periods of 6 d alternating with 8 d of milking with conventional hours (morning-evening gap of 10 h). We confirmed that milk spontaneous lipolysis was influenced by milking interval, but not the milking time. Indeed, we observed more lipolysis in SD-LN evening milk (+0.20 mEq/100 g fat) and LD-SN morning milk (+0.22 mEq/100 g fat), both of which corresponded to a 10 h interval between successive milkings. High lipolysis milk came from cows that produced less milk with a higher milk fat content. No significant difference between milkings was observed for BDN. Milk protein, total P and citrate contents increased according to the duration of mammary gland storage of milk (from 10 to 14 h). There was no effect of milking intervals on milk fat globule diameter. The milk Na+/K+ ratio, indicating an opening of tight junctions in the mammary gland, increased only in evening milkings with BDN and LD-SN. In conclusion, we found that the effect of milking intervals on lipolysis is stronger than that of the nycthemeral cycle.

The impact of liquid-pressurized extracts of Spirulina, Chlorella and Phaedactylum tricornutum on in vitro antioxidant, antiinflammatory and bacterial growth effects and gut microbiota modulation

The impact of Spirulina, Chlorella and Phaeodactylum tricornutum (P. tricornutum) microalgal extracts obtained by pressurized liquid extraction (PLE) on antioxidant and anti-inflammatory activities, microbial growth and in vitro gut microbiota composition was evaluated.PLE, compared to conventional extraction, led to a significant (p < 0.05) increase in proteins, carbohydrates, polyphenols, and antioxidant capacities of the three microalgal extracts. Moreover, Spirulina and P. tricornutum extracts significantly (p < 0.05) reduced the in vitro activation of the inflammatory NF-κB pathway. The microalgal extracts had also an inhibitory effect on the pathogenic bacteria while potential beneficial Lactobacillus and Bifidobacterium strains increased growth. The effects of microalgal extracts on specific bacterial groups were analyzed by quantitative PCR technology, and bacterial gene copy numbers were affected by in vitro digestion process and colonic fermentation time. GC–MS results showed that microalgal biomolecules' digestion promoted the release of short-chain fatty acids (SCFAs) during in vitro colonic microbiota fermentation, particularly acetic, butanoic and propanoic, indicating that the biomolecules in microalgae extracts have potential health benefits for human gut.

Comparative characterization of baking lipase substrate specificities using emulsions and the p-nitrophenyl assay

For use in cakes, lipases with suitable substrate specificity are needed to minimize the release of short-chain fatty acids that cause undesirable off-flavors. We analyzed the substrate specificities of 17 lipases using the p-nitrophenyl (PNP)-assay. These results were compared to the reactions of the lipases with the baking fats rapeseed oil, margarine and butter in a new model emulsion. The free fatty acids (FFA) released from this model emulsion were quantitated by gas chromatography. The broad spectrum of lipase specificities seen in the PNP-assay was not apparent in the patterns of FFA released from the model emulsions. There, all lipases released similar percentages of FFA depending on the fat and emulsifier used. The prediction of lipase reactions via the PNP-assay should therefore be critically reevaluated for real food systems such as cakes.

Assessment of the prebiotic potential of globe artichoke by-product through in vitro fermentation by human faecal microbiota

In vitro fermentability of artichoke by-product by pure cultures of lactic bacteria was previously observed. Therefore, prebiotic potential of globe artichoke by-product (Cynara cardunculus L.) was evaluated by in vitro fermentation with human faecal microbiota. Inulin, a well-known prebiotic compound, and sucrose were used as fermentation controls. The inoculated batches were incubated at 37 °C over 72h. Fermentability was evaluated by monitoring bacterial growth, pH, short chain fatty acids (SCFAs) release, lactic acid production and substrate consumption. The most representative gut bacteria were evaluated. Growth of beneficial lactic acid bacteria and bifidobacteria on artichoke by-product was higher than on inulin and similar to sucrose control. Counts of potentially pathogenic bacteria such as coliforms and sulphite-reducing clostridia, where up to 2 log UFC/mL lower than those of beneficial bacteria. After an initial adaptation period, they decreased on inulin and sucrose controls but remained on artichoke by-product. Release of SCFAs was observed on artichoke substrate which showed the highest levels of individual acetic, propionic and butyric acids at 24h or 72h of fermentation, respectively. The lactic acid produced was significantly lower on artichoke substrate than on controls. Human faecal bacteria consumed the 54.6% of artichoke by-product substrate. Therefore, artichoke by-product showed a potential prebiotic effect under the assayed conditions. Further analysis by molecular biology techniques and in vivo studies are needed to determine the specific compounds involved and health-promoting effects in humans. In addition, the possible use of artichoke waste as prebiotic ingredient could contribute to minimize food waste and environmental impact.

Processing treatment of atmospheric- and vacuum-cold plasma improved physical properties, glucose diffusion and fermentability of dietary fibers extracted from de-oiled rice and corn bran

The main objective of this study was to analyze the impact of atmospheric- and vacuum-cold plasma on dietary fibers extracted from rice and corn bran. Cold plasma treatment was observed to be significantly (p < 0.05) changed the physical properties, glucose diffusion and amylolysis kinetic of dietary fibers. Different controlled conditions of plasma treatment were found to led restrained or enhanced release of glucose from dietary fibers. Furthermore, SEM analysis revealed the occurrence of micro-holes in dietary fiber upon cold plasma treatment. However, no changes were observed in the structural and thermal properties of dietary fiber. Cold plasma treatment enhanced the surface area of dietary fibers obtained from brans. Furthermore, growth of probiotics was observed to be enhanced in the fibers obtained from rice and corn brans upon cold plasma treatment. Also, enhanced release of short-chain fatty acids (SCFA) from dietary fibers upon cold plasma treatment documented their improved fermentable properties.

Viability of Lactobacillus delbrueckii in chocolates during storage and in-vitro bioaccessibility of polyphenols and SCFAs

This study evaluated the viability of encapsulated Lactobacillus delbrueckii subsp. bulgaricus in chocolate during storage and in-vitro gastrointestinal transit. Flavonoid contents and short chain fatty acids (SCFAs) production during gastrointestinal transit were also assessed. Encapsulated L. delbrueckii subsp. bulgaricus survived well in chocolates >7 logs both after 120 days of storage at 4 °C and 25 °C, and during in-vitro gastrointestinal transit. The release of SCFAs through in-vitro gastrointestinal digestion and colonic fermentation revealed that probiotic-chocolates could be an excellent source of nutrients for the gut microbiota. Encapsulated probiotic in chocolates with 70% cocoa produced significantly (P < 0.05) more acetic, propionic, isobutyric, butyric and isovaleric acids than that with 45% cocoa. The bioconversion results of a specific polyphenol by L. delbrueckii subsp. bulgaricus exhibited that chocolate polyphenols could be utilized by probiotics for their metabolism. These findings confirmed that chocolate could be successfully fortified with L. delbrueckii subsp. bulgaricus encapsulation to improve health promoting properties of chocolates.

Prebiotic Supplementation During Pregnancy Modifies the Gut Microbiota and Increases Metabolites in Amniotic Fluid, Driving a Tolerogenic Environment In Utero.

The gut microbiota is influenced by environmental factors such as food. Maternal diet during pregnancy modifies the gut microbiota composition and function, leading to the production of specific compounds that are transferred to the fetus and enhance the ontogeny and maturation of the immune system. Prebiotics are fermented by gut bacteria, leading to the release of short-chain fatty acids that can specifically interact with the immune system, inducing a switch toward tolerogenic populations and therefore conferring health benefits. In this study, pregnant BALB/cJRj mice were fed either a control diet or a diet enriched in prebiotics (Galacto-oligosaccharides/Inulin). We hypothesized that galacto-oligosaccharides/inulin supplementation during gestation could modify the maternal microbiota, favoring healthy immune imprinting in the fetus. Galacto-oligosaccharides/inulin supplementation during gestation increases the abundance of Bacteroidetes and decreases that of Firmicutes in the gut microbiota, leading to increased production of fecal acetate, which was found for the first time in amniotic fluid. Prebiotic supplementation increased the abundance of regulatory B and T cells in gestational tissues and in the fetus. Interestingly, these regulatory cells remained later in life. In conclusion, prebiotic supplementation during pregnancy leads to the transmission of specific microbial and immune factors from mother to child, allowing the establishment of tolerogenic immune imprinting in the fetus that may be beneficial for infant health outcomes.

Impact of prebiotics on immune response: from the bench to the clinic.

Several preclinical and clinical studies have shown the immunomodulatory role exerted by prebiotics in regulating the immune response. In this review, we describe the mechanistic and clinical studies that decipher the cell signaling pathways implicated in the process. Prebiotic fibers are conventionally known to serve as substrate for probiotic commensal bacteria that release of short-chain fatty acids in the intestinal tract along with several other metabolites. Subsequently, they then act on the local as well as the systemic immune cells and the gut-associated epithelial cells, primarily through G-protein-coupled receptor-mediated pathways. However, other pathways including histone deacetylase inhibition and inflammasome pathway have also been implicated in regulating the immunomodulatory effect. The prebiotics can also induce a microbiota-independent effect by directly acting on the gut-associated epithelial and innate immune cells through the Toll-like receptors. The cumulative effect results in the maintenance of the epithelial barrier integrity and modulation of innate immunity through secretion of pro- and anti-inflammatory cytokines, switches in macrophage polarization and function, neutrophil recruitment and migration, dendritic cell and regulatory T-cell differentiation. Extending these in vitro and ex vivo observations, some prebiotics have been well investigated, with successful human and animal trials demonstrating the association between gut microbes and immunity biomarkers leading to improvement in health endpoints across populations. This review discusses scientific insights into the association between prebiotics, innate immunity and gut microbiome from in vitro to human oral intervention.

Human colonic in vitro fermentation of water-soluble arabinoxylans from hard and soft wheat alters Bifidobacterium abundance and short-chain fatty acids concentration

The human intestinal microbiome plays an important role in health due to the large number of beneficial effects related to the bacterial profile and the metabolites generated in the intestine. Arabinoxylans are compounds present in different cereals such as wheat and they can modulate the profile and functioning of some beneficial bacteria from human intestinal microbiota. In the present work, a colonic in vitro fermentation with human faecal inoculum was done using arabinoxylans extracted from Argentinian hard and soft wheat as substrates. Molecular size alteration of arabinoxylans were studied during fermentation and Lactobacillus and Bifidobacterium abundance as well as short chain fatty acids concentrations were determined. The arabinoxylans fermentation was proved to induce the growth of Bifidobacterium and the release of short-chain fatty acids. The speed and efficiency of fermentation were different for each of the arabinoxylan extracted from both wheat genotypes, perhaps because of differences in their chemical and physical structures. The consumption of water-extractable arabinoxylans (WE-AX) (either supplemented or enriched) to maintain the balance or modulate in a favorable way the profile of Bifidobacterium can be an important contribution to the human health.

Gastrointestinal digestion of Pickering emulsions stabilised by hydrophobically modified cellulose nanocrystals: Release of short-chain fatty acids

This study aimed to deliver short-chain fatty acids (SCFAs, including propionic and butyric acids) using Pickering emulsions stabilised by hydrophobically modified cellulose nanocrystals (MCNCs). The emulsions (20 wt% oil, 1 wt% MCNCs) were subjected to two in vitro digestion pathways. In the first pathway, the emulsions were used for direct intestinal digestion by bypassing the gastric phase while in the second pathway, the emulsions were subjected to sequential gastrointestinal digestion. Flocculation of emulsion droplets occurred because of charge screening effects by the gastric electrolytes. Such gastric flocculation reduced the droplet surface area, overall lipolysis kinetics and consequently decreased the extent of SCFA release, latter was 40–45% in the gastric-bypassed emulsions and 30–35% in the sequentially-digested emulsions. High proportion of SCFAs remaining after the intestinal digestion (~65%) shows promise in the use of Pickering emulsions for the colon-targeted delivery of SCFAs.

Distinct alterations of gut morphology and microbiota characterize accelerated diabetes onset in nonobese diabetic mice

Distinct alterations of gut morphology and microbiota characterize accelerated diabetes onset in nonobese diabetic mice

Celery cellulose hydrogel as carriers for controlled release of short-chain fatty acid by ultrasound

The feasibility of using celery cellulose hydrogels as carriers was explored for controlled release of short-chain fatty acids (SCFAs) triggered by ultrasound. The hydrogels were prepared with the phase inversion method and further characterized using FT-IR, SEM and XRD techniques. At the optimal cellulose concentration (8.33 and 6.25 mg/mL), the hydrogels (F4 and F5) exhibited the swelling ratio of 185%, and Young’s modulus of the F4 and F5 was lower than that of others. The hydrogels were loaded with SCFAs owing to its hydrophilicity and swelling properties, and the maximum loading capacity of SCFAs achieved nearly 80%. Interestingly, the loaded SCFAs within hydrogel carrier could be readily released if an ultrasound trigger is exerted. Our results indicate that the ultrasound-triggered strategy for the SCFAs delivery system could provide a promising basis to achieve on-demand, reproducible, repeated, and tunable dosing of bioactive molecules.

Prebiotics: Mechanisms and Preventive Effects in Allergy.

Allergic diseases now affect over 30% of individuals in many communities, particularly young children, underscoring the need for effective prevention strategies in early life. These allergic conditions have been linked to environmental and lifestyle changes driving the dysfunction of three interdependent biological systems: microbiota, epithelial barrier and immune system. While this is multifactorial, dietary changes are of particular interest in the altered establishment and maturation of the microbiome, including the associated profile of metabolites that modulate immune development and barrier function. Prebiotics are non-digestible food ingredients that beneficially influence the health of the host by 1) acting as a fermentable substrate for some specific commensal host bacteria leading to the release of short-chain fatty acids in the gut intestinal tract influencing many molecular and cellular processes; 2) acting directly on several compartments and specifically on different patterns of cells (epithelial and immune cells). Nutrients with prebiotic properties are therefore of central interest in allergy prevention for their potential to promote a more tolerogenic environment through these multiple pathways. Both observational studies and experimental models lend further credence to this hypothesis. In this review, we describe both the mechanisms and the therapeutic evidence from preclinical and clinical studies exploring the role of prebiotics in allergy prevention.

Study of antioxidant capacity and metabolization of quebracho and chestnut tannins through in vitro gastrointestinal digestion-fermentation

Quebracho (QUE) and chestnut (CHE) are natural sources of tannins, but there are no connection between QUE, CHE and human health. The study investigated the antioxidant response and metabolization of tannin extracts trough in vitro digestion-fermentation. The FRAP assay pointed a higher reducing capacity of CHE than QUE (6.90 vs. 5.07 mmol Trolox/g), in contrast to a stronger scavenging activity of QUE (8.16 mmol Trolox/g vs 6.70 mmol Trolox/g). The results obtained showed a decrease of the antioxidant capacity of tannins after microbial fermentation, but a high prebiotic activity through release of short-chain fatty acids of both CHE (11.14 mmol/g) and QUE (4.79 mmol/g) was observed. The UPLC-MS investigations on digested and fermented tannins gave an identification and semi-quantification of 18 compounds, including hydrolysable and condensed tannins and their metabolites. The results represent a valid basis for further studies on the potential use of these wood extracts in human diet.

Diet, microorganisms and their metabolites, and colon cancer.

Colorectal cancer is one of the so-called westernized diseases and the second leading cause of cancer death worldwide. On the basis of global epidemiological and scientific studies, evidence suggests that the risk of colorectal cancer is increased by processed and unprocessed meat consumption but suppressed by fibre, and that food composition affects colonic health and cancer risk via its effects on colonic microbial metabolism. The gut microbiota can ferment complex dietary residues that are resistant to digestion by enteric enzymes. This process provides energy for the microbiota but culminates in the release of short-chain fatty acids including butyrate, which are utilized for the metabolic needs of the colon and the body. Butyrate has a remarkable array of colonic health-promoting and antineoplastic properties: it is the preferred energy source for colonocytes, it maintains mucosal integrity and it suppresses inflammation and carcinogenesis through effects on immunity, gene expression and epigenetic modulation. Protein residues and fat-stimulated bile acids are also metabolized by the microbiota to inflammatory and/or carcinogenic metabolites, which increase the risk of neoplastic progression. This Review will discuss the mechanisms behind these microbial metabolite effects, which could be modified by diet to achieve the objective of preventing colorectal cancer in Western societies.

Dietary Fiber and Bacterial SCFA Enhance Oral Tolerance and Protect against Food Allergy through Diverse Cellular Pathways.

The incidence of food allergies in western countries has increased dramatically in recent decades. Tolerance to food antigens relies on mucosal CD103(+) dendritic cells (DCs), which promote differentiation of regulatory T (Treg) cells. We show that high-fiber feeding in mice improved oral tolerance and protected from food allergy. High-fiber feeding reshaped gut microbial ecology and increased the release of short-chain fatty acids (SCFAs), particularly acetate and butyrate. High-fiber feeding enhanced oral tolerance and protected against food allergy by enhancing retinal dehydrogenase activity in CD103(+) DC. This protection depended on vitamin A in the diet. This feeding regimen also boosted IgA production and enhanced T follicular helper and mucosal germinal center responses. Mice lacking GPR43 or GPR109A, receptors for SCFAs, showed exacerbated food allergy and fewer CD103(+) DCs. Dietary elements, including fiber and vitamin A, therefore regulate numerous protective pathways in the gastrointestinal tract, necessary for immune non-responsiveness to food antigens.

The probiotic Propionibacterium freudenreichii as a new adjuvant for TRAIL-based therapy in colorectal cancer.

TNF-Related Apoptosis-Inducing Ligand (TRAIL) is a well-known apoptosis inducer, which activates the extrinsic death pathway. TRAIL is pro-apoptotic on colon cancer cells, while not cytotoxic towards normal healthy cells. However, its clinical use is limited by cell resistance to cell death which occurs in approximately 50% of cancer cells. Short Chain Fatty Acids (SCFA) are also known to specifically induce apoptosis of cancer cells. In accordance, we have shown that food grade dairy propionibacteria induce intrinsic apoptosis of colon cancer cells, via the production and release of SCFA (propionate and acetate) acting on mitochondria. Here, we investigated possible synergistic effect between Propionibacterium freudenreichii and TRAIL. Indeed, we hypothesized that acting on both extrinsic and intrinsic death pathways may exert a synergistic pro-apoptotic effect. Whole transcriptomic analysis demonstrated that propionibacterial supernatant or propionibacterial metabolites (propionate and acetate), in combination with TRAIL, increased pro-apoptotic gene expression (TRAIL-R2/DR5) and decreased anti-apoptotic gene expression (FLIP, XIAP) in HT29 human colon cancer cells. The revealed synergistic pro-apoptotic effect, depending on both death receptors (TRAIL-R1/DR4, TRAIL-R2/DR5) and caspases (caspase-8, -9 and -3) activation, was lethal on cancer cells but not on normal human intestinal epithelial cells (HIEC), and was inhibited by Bcl-2 expression. Finally, milk fermented by P. freudenreichii induced HT29 cells apoptosis and enhanced TRAIL cytotoxic activity, as did P. freudenreichii DMEM culture supernatants or its SCFA metabolites. These results open new perspectives for food grade P. freudenreichii-containing products in order to potentiate TRAIL-based cancer therapy in colorectal cancer.

Esterification of high amylose starch with short chain fatty acids modulates degradation by Bifidobacterium spp.

Large bowel luminal short chain fatty acids (SCFA) are protective against gut diseases such as colorectal cancer, Crohn’s disease and ulcerative colitis. High amylose maize starches acylated with acetic, propionic or butyric were previously shown to deliver SCFA to the gut and increase gut SCFA. This study examines the ability of five Bifidobacterium spp. (B. longum, B. breve, B. infantis, B. adolescentis and B. bifidum), Ruminococcus bromii and Faecalibacterium prausnitzii to degrade starches acylated with SCFA. Release of SCFA from modified starches was not observed, suggesting no preferential hydrolysis of the SCFA ester bond over the glycosidic bond. Acetylated and propionated starches are more readily degraded than either the base or butyrated starches in tested cultures. These observed differences may depend on modification to the starch structure. These structural differences alter the kinetics of starch digestion, irrespective of bacterial type, which may impact current approaches of prebiotic selection.

Enzymatic Interesterification of Butterfat with Rapeseed Oil in a Continuous Packed Bed Reactor

Lipase-catalyzed interesterification of butterfat blended with rapeseed oil (70/30, w/w) was investigated both in batch and in continuous reactions. Six commercially available immobilized lipases were screened in batch experiments, and the lipases, Lipozyme TL IM and Lipozyme RM IM, were chosen for further studies in a continuous packed bed reactor. TL IM gave a fast reaction and had almost reached equilibrium with a residence time of 30 min, whereas RM IM required 60 min. The effect of reaction temperature was more pronounced for RM IM. TL IM showed little effect on the interesterification degree when the temperature was raised from 60 degrees C to 90 degrees C, whereas RM IM had a positive effect when the temperature was increased from 40 degrees C to 80 degrees C. Even though TL IM is an sn-1,3 specific lipase, small changes in the sn-2 position of the triacylglycerol could be seen. The tendency was toward a reduction of the saturated fatty acid C14:0 and C16:0 and an increase of the long-chain saturated and unsaturated fatty acids (C18:0 and C18:1), especially at longer residence times (90 min). In prolonged continuous operation the activity of TL IM was high for the first 5 days, whereafter it dramatically decreased over the next 10 days to an activity level of 40%. In general, the study shows no significant difference for butterfat interesterification in terms of enzyme behavior from normal vegetable oils and fats even though it contains short-chain fatty acids and cholesterol. However, the release of short-chain fatty acids from enzymatic reactions makes the sensory quality unacceptable for direct edible applications.