Utilizing chitooligosaccharides from shrimp waste biodegradation via recombinant chitinase A: a promising approach for emulsifying hydrocarbon and bioremediation.

BACKGROUND: Hydrocarbon pollution stemming from petrochemical activities is a significant global environmental concern. Bioremediation, employing microbial chitinase-based bioproducts to detoxify or remove contaminants, presents an intriguing solution for addressing hydrocarbon pollution. Chitooligosaccharides, a product of chitin degradation by chitinase enzymes, emerge as key components in this process. Utilizing chitinaceous wastes as a cost-effective substrate, microbial chitinase can be harnessed to produce Chitooligosaccharides. This investigation explores two strategies to enhance chitinase productivity, firstly, statistical optimization by the Plackett Burman design approach to  evaluating the influence of individual physical and chemical parameters on chitinase production, Followed by  response surface methodology (RSM) which delvs  into the interactions among these factors to optimize chitinase production. Second, to further boost chitinase production, we employed heterologous expression of the chitinase-encoding gene in E. coli BL21(DE3) using a suitable vector. Enhancing chitinase activity not only boosts productivity but also augments the production of Chitooligosaccharides, which are found to be used as emulsifiers. RESULTS: In this study, we focused on optimizing the production of chitinase A from S. marcescens using the Plackett Burman design and response surface methods. This approach led to achieving a maximum activity of 78.65 U/mL. Subsequently, we cloned and expressed the gene responsible for chitinase A in E. coli BL21(DE3). The gene sequence, named SmChiA, spans 1692 base pairs, encoding 563 amino acids with a molecular weight of approximately 58 kDa. This sequence has been deposited in the NCBI GenBank under the accession number "OR643436". The purified recombinant chitinase exhibited a remarkable activity of 228.085 U/mL, with optimal conditions at a pH of 5.5 and a temperature of 65 °C. This activity was 2.9 times higher than that of the optimized enzyme. We then employed the recombinant chitinase A to effectively hydrolyze shrimp waste, yielding chitooligosaccharides (COS) at a rate of 33% of the substrate. The structure of the COS was confirmed through NMR and mass spectrometry analyses. Moreover, the COS demonstrated its utility by forming stable emulsions with various hydrocarbons. Its emulsification index remained stable across a wide range of salinity, pH, and temperature conditions. We further observed that the COS facilitated the recovery of motor oil, burned motor oil, and aniline from polluted sand. Gravimetric assessment of residual hydrocarbons showed a correlation with FTIR analyses, indicating the efficacy of COS in remediation efforts. CONCLUSIONS: The recombinant chitinase holds significant promise for the biological conversion of chitinaceous wastes into chitooligosaccharides (COS), which proved its potential in bioremediation efforts targeting hydrocarbon-contaminated sand.

Comparison of rumen and abomasal infusions of an exogenous emulsifier on fatty acid digestibility of lactating dairy cows.

We evaluated the effects of infusing an exogenous emulsifier (polysorbates-C18:1) either into the rumen or abomasum on fatty acid (FA) digestibility and production responses of lactating dairy cows. Nine ruminally cannulated multiparous Holstein cows (170 ± 13.6 d in milk) were assigned to a treatment sequence in replicated 3 × 3 Latin squares with 18-d periods consisting of 7 d of washout and 11 d of infusion. Treatments were abomasal infusions of water carrier only into the rumen and abomasum (control, CON), 30 g/d polysorbate-C18:1 (T80) infused into the rumen (RUM), or 30 g/d T80 infused into the abomasum (ABO). Emulsifiers were dissolved in water and delivered at 6-h intervals (total daily infusion was divided into 4 equal infusions per day). Cows were fed the same diet that contained [% diet dry matter (DM)] 32.2% neutral detergent fiber (NDF), 16.1% crude protein, 26.5% starch, and 3.41% FA (including 1.96% FA from a saturated FA supplement containing 28.0% C16:0 and 54.6% C18:0). Two orthogonal contrasts were evaluated: (1) the overall effect of T80 {CON vs. average of the T80 infusions [1/2 (ABO + RUM)]}, and (2) the effect of ABO versus RUM infusion. Compared with CON, infusing T80 increased the digestibilities of NDF (2.85 percentage units), total (4.35 percentage units), 16-carbon (3.25 percentage units), and 18-carbon FA (4.60 percentage units), and tended to increase DM digestibility and total and 18-carbon FA absorption. Compared with RUM, ABO decreased the intakes of total (28 g/d), 16-carbon (7 g/d), and 18-carbon FA (19 g/d); tended to increase the digestibility of total and 18-carbon FA; and had no effect on the absorption of total, 16-carbon, or 18-carbon FA. Production responses did not change among our treatments. In conclusion, infusing 30 g/d polysorbates-C18:1 increased NDF and total, 16-carbon, and 18-carbon FA digestibility. Compared with RUM, ABO tended to increase the digestibilities of total and 18-carbon FA; however, this may be related to the fact that ABO reduced the intakes of total, 16-carbon, and 18-carbon FA, not necessarily due to better emulsifying action per se. In summary, ABO and RUM both improved FA absorption.

Systematic investigation of factors, such as the impact of emulsifiers, which influence the measurement of skin barrier integrity by in-vitro trans-epidermal water loss (TEWL).

Trans-epidermal water loss (TEWL) has been the most widely used method to assess the integrity of the skin barrier and evaluate the irritation potential or the protective properties of topical products for many years. It detects the amount of water that diffuses across the stratum corneum (SC) to the external environment. As one of the most important functions of the skin is to keep water inside the body, an increase in TEWL is used to indicate the skin's impaired barrier function. So far, a variety of commercial instruments are available to measure the TEWL. Their applications mainly focus on the in-vivo TEWL measurements for dermatological examinations or formulation development. Recently, an in-vitro TEWL probe has also been commercially released enabling preliminary tests with excised skin samples. In our study, we first aimed to optimize the experimental procedures for detecting the in-vitro TEWL of porcine skin. Secondly, different kinds of emulsifiers were applied to the skin, including polyethylene glycol-containing emulsifiers (PEG-ylated emulsifiers), sorbitan esters, cholesterol, and lecithin. Sodium lauryl sulfate (SLS) was used as a positive control, and water as a negative control. Based on the findings, we established a protocol for accurately measuring the in-vitro TEWL values, emphasizing that the temperature of the skin sample should be constantly maintained at 32℃. Subsequently, the influences of emulsifiers on the in-vitro TEWL were analyzed. They indicated a significant skin barrier impairment of PEG-20 cetyl ether, PEG-20 stearyl ether, and SLS on in-vitro skin. Furthermore, we interestingly found that there consistently was an alteration of the TEWL values, even after the application of water to the skin. Our findings are of special interest, as the European Medicines Agency (EMA) recommends the use of in-vitro TEWL to determine skin barrier intactness during Franz cell experiments. Thus, this study provides a validated protocol for measuring the in-vitro TEWL and elucidates the impact of emulsifiers on the skin barrier. It also improves the understanding of tolerable variations of in-vitro TEWL and offers recommendations for its use in research.

Facultative anaerobic conversion of lignocellulose biomass to new bioemulsifier by thermophilic Geobacillus thermodenitrificans NG80-2.

Heavy oil has hindered crude oil exploitation and pollution remediation due to its high density and viscosity. Bioemulsifiers efficiently facilitate the formation and stabilization of oil-in-water emulsions in low concentrations thus eliminating the above bottleneck. Despite their potential benefits, various obstacles had still impeded the practical applications of bioemulsifiers, including high purification costs and poor adaptability to extreme environments such as high temperature and oxygen deficiency. Herein, thermophilic facultative anaerobic Geobacillus thermodenitrificans NG80-2 was proved capable of emulsifying heavy oils and reducing their viscosity. An exocelluar bioemulsifier could be produced by NG80-2 using low-cost lignocellulose components as carbon sources even under anaerobic condition. The purified bioemulsifier was proved to be polysaccharide-protein complexes, and both components contributed to its emulsifying capability. In addition, it displayed excellent stress tolerance over wide ranges of temperatures, salinities, and pHs. Meanwhile, the bioemulsifier significantly improved oil recovery and degradation efficiency. An eps gene cluster for polysaccharide biosynthesis and genes for the covalently bonded proteins was further certificated. Therefore, the bioemulsifier produced by G. thermodenitrificans NG80-2 has immense potential for applications in bioremediation and EOR, and its biosynthesis pathway revealed here provides a theoretical basis for increasing bioemulsifier output.

Effects of an emulsifier blend supplementation on growth performance, nutrient digestibility, intestinal morphology, and muscle fatty acid profile of broiler chickens fed with different levels of energy and protein.

The effects of emulsifier blend (EB) supplementation of diets with various levels of metabolizable energy (ME) and crude protein (CP) on broiler performance, digestibility, gut morphology, and muscle fatty acid profile were investigated over a 42-d period. Diets were arranged factorially (2 × 2 × 3) and consisted of 2 levels of ME (normal [commercially recommended levels] and low [100 kcal/kg reduction in dietary ME]), 2 levels of CP and limiting amino acids (normal [commercially recommended levels] and low [95% of the normal CP level]), and 3 levels of EB supplementation (0, 1, and 2 g/kg of diet). A total of 1,200 one-day-old male broiler chickens (Ross 308) were randomly assigned to 12 treatment groups (5 pens/treatment with 20 birds/pen). Supplemental EB linearly improved (P < 0.05) final body weight, overall average daily gain, and feed conversion ratio, but the magnitude of the responses was greater in low-ME and low-CP treatments, resulting in significant ME × CP × EB interactions. Similarly, the inclusion of EB in the diet, particularly at 2 g/kg, increased the ileal digestibility of crude protein and crude fat, as well as the AMEn value (P < 0.05), but the response was greater at lower ME concentration, indicating significant ME × EB interactions. Additionally, there were CP × EB interactions (P < 0.05) for duodenal villus height and villus height/crypt depth ratio, indicating that the effect of EB on these responses was more marked at lower dietary CP levels. An increase in dietary EB levels was accompanied by a linear increase in the concentration of total saturated fatty acids and a linear decrease (P < 0.05) in the concentrations of total polyunsaturated fatty acids in both breast and thigh meat. In conclusion, the positive effects of EB supplementation, particularly at a dietary inclusion level of 2 g/kg, were clearly evident in broiler chickens fed with low nutrient diets (-100 Kcal/kg ME and/or -5% CP and limiting amino acids) in terms of growth performance, nutrient digestibility, and gut morphology.

Pectin extracted from dragon fruit Peel: An exploration as a natural emulsifier.

As a potential source of pectin, the peel of dragon fruit is a by-product of fruit processing and will bring considerable economic benefits. In this study, pectin (DFP) was extracted from dragon fruit peel by using a hot-acid method, and two commercial pectins were used as controls to correlate structural differences with physical and chemical properties through systematic evaluation. The galacturonic acid (GalA) content, degree of esterification (DM) and molecular weight (Mw) of DFP were 87.02 ± 0.89 %, 37.26 ± 1.37 % and 1181.75 ± 11.21 kDa, respectively. The FTIR and XRD analysis also confirmed that DFP is ultra-low methoxylated pectin and also contains characteristic functional groups naturally present in pectin. Compared to APA140 and LMP, DFP showed excellent emulsification at low concentrations. In particular, the extraordinary antioxidant activity of DFP may be attributed to polyphenols in free or bound form. Overall, DFP can be considered as a promising emulsifier and the results of the study provide an alternative to natural sources of emulsifiers in the food industry.

Replacing PEG-surfactants in self-emulsifying drug delivery systems: Surfactants with polyhydroxy head groups for advanced cytosolic drug delivery.

AIM: Evaluation of different polyhydroxy surfaces in SEDDS to overcome the limitations associated with conventional polyethylene glycol (PEG)-based SEDDS surfaces for intracellular drug delivery. METHODS: Anionic, cationic and non-ionic polyglycerol- (PG-) and alkylpolyglucoside- (APG-) surfactant based SEDDS were developed and compared to conventional PEG-SEDDS. Particular emphasis was placed on the impact of SEDDS surface decoration on size and zeta potential, drug loading and protective effect, mucus diffusion, SEDDS-cell interaction and intracellular delivery of the model drug curcumin. RESULTS: After self-emulsification, SEDDS droplets sizes were within the range of 35-190 nm. SEDDS formulated with high amounts of long PEG-chain surfactants (>10 monomers) a charge-shielding effect was observed. Replacing PEG-surfactants with PG- and an APG-surfactant did not detrimentally affect SEDDS self-emulsification, payloads or the protection of incorporated curcumin towards oxidation. PG- and APG-SEDDS bearing multiple hydroxy functions on the surface demonstrated mucus permeation comparable to PEG-SEDDS. Steric hinderance and charge-shielding of PEG-SEDDS surface substantially reduced cellular uptake up to 50-fold and impeded endosomal escape, yielding in a 20-fold higher association of PEG-SEDDS with lysosomes. In contrast, polyhydroxy-surfaces on SEDDS promoted pronounced cellular internalisation and no lysosomal co-localisation was observed. This improved uptake resulted in an over 3-fold higher inhibition of tumor cell proliferation after cytosolic curcumin delivery. CONCLUSION: The replacement of PEG-surfactants by surfactants with polyhydroxy head groups in SEDDS is a promising approach to overcome the limitations for intracellular drug delivery associated with conventional PEGylated SEDDS surfaces.

Physicochemical and Functional Properties of 2S, 7S, and 11S Enriched Hemp Seed Protein Fractions.

The hemp seed contains protein fractions that could serve as useful ingredients for food product development. However, utilization of hemp seed protein fractions in the food industry can only be successful if there is sufficient information on their levels and functional properties. Therefore, this work provides a comparative evaluation of the structural and functional properties of hemp seed protein isolate (HPI) and fractions that contain 2S, 7S, or 11S proteins. HPI and protein fractions were isolated at pH values of least solubility. Results showed that the dominant protein was 11S, with a yield of 72.70 ± 2.30%, while 7S and 2S had values of 1.29 ± 0.11% and 3.92 ± 0.15%, respectively. The 2S contained significantly (p < 0.05) higher contents of sulfhydryl groups at 3.69 µmol/g when compared to 7S (1.51 µmol/g), 11S (1.55 µmol/g), and HPI (1.97 µmol/g). The in vitro protein digestibility of the 2S (72.54 ± 0.52%) was significantly (p < 0.05) lower than those of the other isolated proteins. The intrinsic fluorescence showed that the 11S had a more rigid structure at pH 3.0, which was lost at higher pH values. We conclude that the 2S fraction has superior solubility, foaming capacity, and emulsifying activity when compared to the 7S, 11S, and HPI.

Effects of Emulsifiers on an In vitro Model of Intestinal Epithelial Tight Junctions and the Transport of Food Allergens.

SCOPE: Certain food emulsifiers may interfere with gut barrier function in ways correlating to increased exposure to allergens. Understanding the consequences of interactions between these food ingredients and the intestinal epithelium is important for evaluating allergen dose exposure characteristics. METHODS AND RESULTS: This study challenged Caco-2 cell monolayers, an in vitro model of human intestinal epithelial tight junctions with synthetic polysorbate-80 or natural lecithin alone, or in combination with known allergens (egg proteins: ovalbumin, ovomucoid, and ovotransferrin; and a synthetic form of galactose-alpha-1,3-galactose [alpha-gal], an allergen of increasing concern). For most doses of individual emulsifiers and allergens, >90% cell viability and <15% cytotoxicity are observed; however, toxicity increased at a 0.5% concentration of emulsifiers. At low cytotoxic concentration (0.2%), only polysorbate-80 treatment reduced monolayer integrity (≈20%) with increased lucifer yellow passage. Dose-related differences in expression of tight junction-associated genes and occludin protein are observed with emulsifier treatments. The transport of all tested allergens across the cell monolayers, excluding ovotransferrin, nearly doubled in the presence of 0.2% polysorbate-80 compared to lecithin and untreated control. CONCLUSION: By modulating paracellular permeability, polysorbate-80 may enhance absorption of allergens in a size-dependent manner.

A hierarchical emulsion system stabilized by soyasaponin emulsion droplets.

Oil/water (O/W) emulsion droplets coated with soyasaponin (Ssa) were used as emulsifiers to prepare emulsions with hierarchical configurations (2.82 µm). Ssa is a natural triterpenoid with amphiphilic properties and an excellent emulsifying activity. Stable O/W emulsions were prepared and characterized using an ultrasonic method at a Ssa concentration of 2.5 wt%. The resultant hierarchical emulsions were further prepared using O/W droplets as emulsifiers. It was observed that the stability of the hierarchical emulsions changed with alterations to the ratio of O/W droplets to the oil phase. As the number of droplets increased, the more the surface area of the hierarchical emulsion was covered. Additional observations included a decreased particle size, increased negative charge and viscoelastic behavior, and enhanced emulsion stability. The emulsion was most stable when the O/W droplet addition was 29%. The addition of O/W droplets continued to increase, and there was an imbalance in the ratio of O/W droplets to the oil phase; the excess O/W droplets induced instability in the emulsion, resulting in a degradation of the emulsion quality. We monitored hierarchical emulsions with different concentrations of emulsifiers for 30 days, and the results indicated that hierarchical emulsions could meet the demand for long-term storage. This provides a new theoretical basis for the construction and application of complex emulsion systems.

Upper digestion fate of citrus pectin-stabilized emulsion: An interfacial behavior perspective.

Citrus pectin can serve as a naturally digestion-resistant emulsifier, although how it achieves this effect is still unknown. In this study, the upper digestion fate of an emulsion stabilized by different concentrations of citrus pectin, and changes in its interfacial properties during digestion, were investigated. Emulsions stabilized by high-concentration citrus pectin (3 %) were relatively stable during digestion and had a lower free fatty acid (FFA) release rate than emulsions stabilized by low-concentration citrus pectin (1 %). At the low concentration, the citrus pectin interface had a thin absorbing layer and was largely replaced by bile salts, while at high concentration the citrus pectin interface possessed a uniform and thick adsorbing layer that resisted the replacement of bile salts and enabled lipase adsorption. This study has improved our understanding of the digestion of emulsion from the interface and will be useful for designing emulsion-based functional foods that can achieve targeted release.

Structure characterization, antioxidant and emulsifying capacities of exopolysaccharide derived from Pantoea alhagi NX-11.

Pantoea alhagi exopolysaccharides (PAPS) have been shown to enhance crop resistance to abiotic stress. However, physicochemical properties and structure of PAPS have not yet been analyzed. In this study, two PAPSs, named PAPS1 and PAPS2, were isolated and purified from the P. alhagi NX-11. The results showed PAPS1 and PAPS2 were composed of glucose, galactose, glucuronic acid, glucosamine and mannose with average molecular weight of 1.326 × 106 Da and 1.959 × 106 Da, respectively. Moreover, the structure of PAPS1 and PAPS2 was investigated by FT-IR and NMR analysis. PAPS1 was identified to have the backbone structure of →4)-ß-D-GlcpA-(1→2)-α-D-Galp-(1→3)-ß-D-Galp-(1→3)-ß-D-GlcpN- (1→3)-α-D-Galp-(1→3)-ß-D-Galp-(1→. PAPS2 had the backbone structure of →4)-ß-D-GlcpA-(1→2)-α-D-Galp-(1→3)-ß-D-Glcp-(1→3)-ß-D-GlcpN-(1→3)-α-D-Galp-(1→3)-α-D-GlcpN-(1→. In addition, PAPS1 and PAPS2 had moderate antioxidant and emulsifying capacities. Overall, the structure analysis of PAPS may point out the direction for the subsequent study of PAPS-mediated microbial and plant interactions, and further exploration of the application of PAPS.

Visualisation of real-time oral biodistribution of fluorescent labeled self-microemulsifying drug delivery system of olmesartan medoxomil using optical imaging method.

In the present study, the biodistribution of self-microemulsifying drug delivery system of hydrophobic olmesartan medoxomil (OM-SMEDDS) was determined by labeling with a fluorescent dye VivoTag®680 XL and Xenolight® DiR. Labeled OM-SMEDDS and control dye solution administered orally to mice; real-time dynamic biodistributions over 7 h were determined by 2D-fluorescent imaging to verify their anatomic location. Fluorescent Emissions by Vivotag 680® XL and Xenolight® DiR labeled OM-SMEDDS emitted 2 to 24 times stronger emission than control dye administered group. To further confirm the results, organs were removed and examined using the same technique at the end of 7 h. VivoTag®680XL and Xenolight® DiR emitted 4 and 1.7 times stronger emission respectively than control dye administered mice in ex-vivo organ imaging studies. This study showed that OM-SMEDDS can be succesfully labeled with fluorescent dye and tracked with optical imaging method for the visualisation of biodistribution of drugs and is also useful for enhanced bioavailability.

Biofunctionality of Enzymatically Derived Peptides from Codfish (Gadus morhua) Frame: Bulk In Vitro Properties, Quantitative Proteomics, and Bioinformatic Prediction.

Protein hydrolysates show great promise as bioactive food and feed ingredients and for valorization of side-streams from e.g., the fish processing industry. We present a novel approach for hydrolysate characterization that utilizes proteomics data for calculation of weighted mean peptide properties (length, molecular weight, and charge) and peptide-level abundance estimation. Using a novel bioinformatic approach for subsequent prediction of biofunctional properties of identified peptides, we are able to provide an unprecedented, in-depth characterization. The study further characterizes bulk emulsifying, foaming, and in vitro antioxidative properties of enzymatic hydrolysates derived from cod frame by application of Alcalase and Neutrase, individually and sequentially, as well as the influence of heat pre-treatment. All hydrolysates displayed comparable or higher emulsifying activity and stability than sodium caseinate. Heat-treatment significantly increased stability but showed a negative effect on the activity and degree of hydrolysis. Lower degrees of hydrolysis resulted in significantly higher chelating activity, while the opposite was observed for radical scavenging activity. Combining peptide abundance with bioinformatic prediction, we identified several peptides that are likely linked to the observed differences in bulk emulsifying properties. The study highlights the prospects of applying proteomics and bioinformatics for hydrolysate characterization and in food protein science.

Effect of emulsifiers on linseed oil emulsion structure, lipolysis and oxidation during in vitro digestion.

Health benefits have been associated with the consumption of omega-3 polyunsaturated fatty acids (PUFA). Linseed oil is rich in long chain omega-3 PUFA, but can generate toxic compounds due to its high susceptibility to oxidation. The nature of the emulsifier can affect both lipolysis and oxidation during digestion since these phenomena occur at the oil-water interface. The objective of this study was to compare the effect of low-molecular weight surfactants (cetyltrimethylammonium bromide (CTAB), Citrem), protein (sodium caseinate, fish gelatin) and polysaccharides (gum arabic, modified starch) on the structure of linseed oil emulsions, lipolysis and formation of reactive oxidation species during in vitro digestion. The emulsion stabilized with Citrem underwent extensive coalescence in the gastric phase, which strongly decreased the extent of lipid digestion and reduced the formation of oxidation markers relative to other emulsions. Emulsions stabilized by proteins and modified starch showed aggregation with partial coalescence in the gastric phase, but protein-stabilized emulsions showed better resistance to oxidation. This study shows that emulsifier properties affect the susceptibility of the emulsion to aggregation and coalescence in the gastrointestinal environment, and strongly influence the extent of lipid digestion and the formation of reactive oxidation products. These findings point out the importance of the choice of the emulsifier to control the lipid digestibility and the protection of sensible lipids thus promoting optimal nutritional properties in omega-3-enriched foods.

Biosurfactant and bioemulsifier as promising molecules produced by Mucor hiemalis isolated from Caatinga soil

BACKGROUND: The present study describes the production of biosurfactant (BS) and emulsifier (BE) by the filamentous fungus Mucor hiemalis UCP 0039, as well as the characterization and stability of the both biomolecules for environmental or industrial applications. RESULTS: Biosurfactants and bioemulsifiers are amphiphilic compounds and are produced as extracellular molecules. The results showed that bioproduct obtained by shaker condition reduced the water surface tension of 72 to 32 mN/m and reached an emulsification index of 96%, while the static cultivation resulted in a biomolecule with a surface tension of 40 mN/m and an emulsification index of 96%, suggesting the production of a biosurfactant and bioemulsifier, respectively. The compounds showed glycolipid nature but the biosurfactant presented cationic charge, while the bioemulsifier, anionic charge. Thus, the results confirmed that M. hiemalis produced two distinct biomolecules under different parameters and in the same culture medium. CONCLUSIONS: It is the first time that biosurfactant and emulsifier production has been described in the same medium and under different physical conditions by Mucor hiemalis. Both biomolecules showed thermal stability, as well as have significant effect on the viscosity of hydrophobic compounds, indicating the excellent potential for environmental safety or industrial applications to improve the efficiency of sustainable and economic technologies.

An overview of fungal biopolymers: bioemulsifiers and biosurfactants compounds production.

Fungal biopolymers have gained considerable attention from the scientific community for various applications due to their biological and physicochemical properties. The wide applications in several areas, especially in the food industry as a bioemulsifier and in the agricultural area as a biosurfactant, have expanded the knowledge on the production of fungal biopolymers to keep up with developments on this subject area. Recent scientific studies have disclosed novel routes, optimized parameters, increased yields, and other related approaches in order to produce and apply fungal bioemulsifiers and biosurfactants. However, there is a need to gather important information in order to provide a way forward. Therefore, this review presents an overview of properties, applications, and perspectives for encouraging further projects and investments in the near future by most categories of investors. The selection of culture media, the definition of cultivation parameters, extraction, recovery, and purification are the initial steps to indicate the conditions for scale-up. Indeed, scale-up is still one of the challenges in this biotechnological field, which could be solved by expanding the tests and operational productions in both pilot and industrial plants.

Low-resolution molecular shape, biochemical characterization and emulsification properties of a halotolerant esterase from Bacillus licheniformis.

Bacterial esterases are highly versatile enzymes, currently widely used in detergents, biosurfactants, bioemulsifiers and as biocatalysts in paper and food industries. Present work describes heterologous expression, purification, and biophysical and biochemical characterization of a halotolerant esterase from Bacillus licheniformis (BlEstA). BlEstA preferentially cleaves pNP-octanoate and both activity and stability of the enzyme increased in the presence of 2 M NaCl, and also with several organic solvents (ethanol, methanol and DMSO). Furthermore, BlEstA has considerable emulsifying properties, particularly with olive oil as substrate. Our studies also show that the enzyme is monomeric in solution and its small-angle X-ray scattering low-resolution molecular envelope fits well its high-resolution homology model.

Hydrophobic ion pair loaded self-emulsifying drug delivery system (SEDDS): A novel oral drug delivery approach of cromolyn sodium for management of bronchial asthma.

The aim of the present study is to develop a self-emulsifying drug delivery system (SEDDS) for the hydrophobic ion pair (HIP) complex of cromolyn sodium (CS), in order to enhance its intestinal absorption and biological activity. Two ion pairing agents (IPAs) were investigated: hexadecyl pyridininum chloride (HPC) and myristyl trimethyl ammonium bromide (MTAB). The optimum binding efficiency for complexation between investigated IPAs and CS was observed at a molar ratio of 1.5:1, where CS binding efficiency was found to be 76.10 ± 2.12 and 91.37 ± 1.73% for MTAB and HPC, respectively. The two prepared complexes exhibited a significant increase in partition coefficient indicating increased lipophilicity. The optimized CS-HIP complex was incorporated into SEDDS formulations. SEDDS formulations F2 (40% oleic acid, 40% BrijTM98, 20% propylene glycol) and F3 (25% oleic acid, 50% BrijTM98, 25% propylene glycol) exhibited nanometric droplet diameters with monodisperse distribution and nearly neutral zeta potential values. Ex vivo intestinal permeation study, using the non-everted gut sac technique, revealed a significantly higher cumulative amount of permeated drug, after 2 h, for F2 and F3 (53.836 and 77.617 µg/cm2, respectively) compared to 8.649 µg/cm2 for plain CS solution. The in vivo evaluation of plain CS solution compared to F2 and F3 was conducted in an ovalbumin sensitization-induced bronchial asthma rat model. Lung function parameters (tidal volume and peak expiratory flow), biochemical parameters (interleukin-5, immunoglobulin-E, myeloperoxidase and airway remodelling parameters) were assessed in addition to histopathological examination. The results indicated the superiority of F3 followed by F2 compared to plain CS solution for prophylaxis of bronchial asthma in rats.

Abomasal infusion of an exogenous emulsifier improves fatty acid digestibility and milk fat yield of lactating dairy cows.

The objective of our study was to determine the effects of abomasal infusion of an emulsifier on fatty acid (FA) digestibility and production responses of lactating dairy cows. Eight rumen-cannulated cows (109 ± 18 d in milk) were randomly assigned to a treatment sequence in replicated 4 × 4 Latin squares with 18-d periods including 7 d of washout and 11 d of infusion with sampling on the last 4 d. Treatments were abomasal infusions of water carrier only (CON) and 3 levels of increasing doses of Tween 80 (polysorbate 80, Tween 80, Sigma-Aldrich, St. Louis, MO) delivering 15 (D-15), 30 (D-30), and 45 (D-45) g/d. The Tween 80 was dissolved in water before infusions, which were delivered at 6-h intervals. Cows were fed the same diet, which contained (% dry matter) 31% neutral detergent fiber, 17% crude protein, 25% starch, and 4% FA (2% dry matter from a saturated fat supplement containing 33% C16:0 and 51% C18:0). Increasing emulsifier infusion dose quadratically increased digestibility of total FA (60.7, 65.3, 70.9, and 66.8%), 16-carbon FA (61.7, 63.9, 70.4, and 66.7%), and 18-carbon FA (59.8, 65.6, 71.1, and 66.6%, respectively). Increasing emulsifier infusion dose quadratically increased absorbed total FA (625, 670, 744, and 658 g/d), 16-carbon FA (151, 157, 197, and 157 g/d, quadratic), and 18-carbon FA (420, 460, 500, and 444 g/d). Increasing emulsifier infusion dose tended to quadratically decrease dry matter intake (29.0, 28.8, 29.6, and 27.6 kg/d). Increasing emulsifier infusion dose quadratically increased milk fat content (3.23, 3.35, 3.45, and 3.35%), milk fat yield (1.54, 1.61, 1.65, and 1.55 kg/d), ECM (45.7, 46.9, 47.5, and 45.3 kg/d), and plasma nonesterified fatty acid concentration (95.6, 98.4, 101.2, and 98.6 µEq/L). On a yield basis, we observed that de novo, mixed, and preformed FA responded quadratically to Tween 80 infusion due to FA yield increasing up to D-30. Treatments had no effect on milk yield (47.9, 48.3, 48.0, and 46.6 kg/d). In conclusion, short-term infusion of an exogenous emulsifier improved FA digestibility and milk fat yield responses when cows were fed a diet containing a saturated fat supplement. Most digestion and production measurements responded quadratically because the highest dose of exogenous emulsifier (45 g/d) decreased dry matter intake and performance.