Protein Surfactant Research Articles

Polymerized whey protein-SDS interactions at their high concentrations

Protein-surfactant interactions have been an ongoing topic of interest for many decades. Applications involving complexes of proteins and surfactants are relevant in food, pharmaceuticals, hygiene, molecular characterization, and other fields. In this study, the interactions of polymerized whey proteins (PWP) and sodium dodecyl sulfate (SDS) at high concentrations are investigated. Different characterization techniques are used, including electrical conductivity, turbidity, isothermal titration calorimetry, dynamic light scattering, dilute solution viscometry, rheology, and surface hydrophobicity to elucidate information on the modes and extent of interactions. Results indicate that PWP-SDS interactions produce highly extended, worm like micelles, with SDS decorating PWP chains and covering non-polar residues. PWP can host SDS up to quite high surfactant to protein ratios (SPR), producing solutions that are highly viscous with shear thickening properties, yet with no networking or gelation. Interestingly, dilution of high viscosity PWP-SDS solutions leads to even smaller size of PWP-SDS molecular complex as compared with PWP without SDS. The current study extends the vision of protein surfactant interactions by examining concentration range beyond that found in literature. The results reveal insights that can help expand studies on other systems and find applications in various fields including coatings, cosmetics, food ingredients, drug transport, and disease treatment.

Fabrication, digestion behavior and β-carotene bioaccessibility of emulsion-filled double-network gel: Effect of corn fiber gum/soy protein isolate ratio and surfactant types

Emulsion fortified with β-carotene was added to corn fiber gum (CFG)/soy protein isolate (SPI) double network gel matrix to obtain emulsion-filled gels (EFG) via dual induction of laccase and glucono-δ-lactone. Protein digestion was accompanied by the release of β-carotene from gel matrix during in vitro digestion. The surfactant types and corn fiber gum/soy protein isolate ratio affected the β-carotene bioaccessibility via changing oil-water interfacial composition and emulsion particle size during in vitro digestion. As compared with Tween-20 EFGs, emulsion droplets released from SPI EFGs was more susceptible to flocculation, followed with coalescence due to proteolysis of interfacial SPI during gastric digestion. The resulting oil droplets with large particle size exhibited lower lipase adsorption, thus reducing the free fatty acid content and β-carotene bioaccessibility. The confocal laser scanning microscope (CLSM) observation confirmed that protein hydrolysate from gel matrix were adsorbed onto the oil-water interface competing with Tween-20 during intestinal digestion. For EFGs with higher CFG content, steric hindrance of CFG molecules and less emulsion release could inhibit droplet flocculation, thus enhancing β-carotene bioaccessibility.

Evolution of the structure and interaction in the surfactant-dependent heat-induced gelation of protein.

The addition of a surfactant and/or an increase in temperature disrupt the native structure of proteins, where high temperature further results in protein gelation. However, in a mixed protein-surfactant system, surfactant concentration and temperature have been observed to exhibit both mutually associative and counter-balancing effects towards heat-induced gelation of protein-surfactant dispersion. This study is conducted on globular bovine serum albumin (BSA) protein and cationic surfactant dodecyl trimethyl ammonium bromide (DTAB), which interact strongly owing to their oppositely charged nature. The findings reveal that the BSA-DTAB suspension undergoes gelation with increasing temperature but only at lower concentrations of DTAB, where the presence of the surfactant facilitates gelation (associative effect). Conversely, as the surfactant concentration increases beyond a critical value, temperature-driven gelation of the BSA-DTAB system is completely inhibited, despite surfactant-induced protein denaturation (counter-balancing effect). To conceptualize these results, we compared them with observations made in a system comprising protein and a similarly charged surfactant, sodium dodecyl sulfate (SDS). It has been further demonstrated that the anionic surfactant (SDS) can restrict protein gelation at much lower concentration compared to the cationic surfactant (DTAB). The evolution of the structure and interaction during gel formation/inhibition has been examined to understand the underlying mechanism guiding these sol-gel transitions. We present a comprehensive phase diagram, encompassing the solution/gel states of the protein-surfactant dispersion, with respect to the dispersion temperature, surfactant concentration, and ionic behavior (anionic or cationic) of the surfactants.

Molecularly informed field theory for estimating critical micelle concentrations of intrinsically disordered protein surfactants.

The critical micelle concentration (CMC) is a crucial parameter in understanding the self-assembly behavior of surfactants. In this study, we combine simulation and experiment to demonstrate the predictive capability of molecularly informed field theories in estimating the CMC of biologically based protein surfactants. Our simulation approach combines the relative entropy coarse-graining of small-scale atomistic simulations with large-scale field-theoretic simulations, allowing us to efficiently compute the free energy of micelle formation necessary for the CMC calculation while preserving chemistry-specific information about the underlying surfactant building blocks. We apply this methodology to a unique intrinsically disordered protein platform capable of a wide variety of tailored sequences that enable tunable micelle self-assembly. The computational predictions of the CMC closely match experimental measurements, demonstrating the potential of molecularly informed field theories as a valuable tool to investigate self-assembly in bio-based macromolecules systematically.

Baseline characteristics from a 3-year longitudinal study to phenotype subjects with COPD: the FOOTPRINTS study

BackgroundFOOTPRINTS® is a prospective, longitudinal, 3-year study assessing the association between biomarkers of inflammation/lung tissue destruction and chronic obstructive pulmonary disease (COPD) severity and progression in ex-smokers with mild-to-severe COPD. Here, we present baseline characteristics and select biomarkers of study subjects.MethodsThe methodology of FOOTPRINTS® has been published previously. The study population included ex-smokers with a range of COPD severities (Global Initiative for Chronic Obstructive Lung Disease [GOLD] stages 1–3), ex-smokers with COPD and alpha-1-antitrypsin deficiency (A1ATD) and a control group of ex-smokers without airflow limitation (EwAL). At study entry, data were collected for: demographics, disease characteristics, history of comorbidities and COPD exacerbations, symptoms, lung function and volume, exercise capacity, soluble biomarkers, and quantitative and qualitative computed tomography. Baseline data are presented with descriptive statistical comparisons for soluble biomarkers in the individual GOLD and A1ATD groups versus EwAL.ResultsIn total, 463 subjects were enrolled. The per-protocol set comprised 456 subjects, mostly male (64.5%). The mean (standard deviation) age was 60.7 (6.9) years. At baseline, increasing pulmonary symptoms, worse lung function, increased residual volume, reduced diffusing capacity of the lung for carbon monoxide (DLco) and greater prevalence of centrilobular emphysema were observed with increasing disease severity amongst GOLD 1–3 subjects. Subjects with A1ATD (n = 19) had similar lung function parameters to GOLD 2–3 subjects, a high residual volume comparable to GOLD 3 subjects, and similar air trapping to GOLD 2 subjects. Compared with EwAL (n = 61), subjects with A1ATD had worse lung function, increased residual volume, reduced DLco, and a greater prevalence of confluent or advanced destructive emphysema. The soluble inflammatory biomarkers white blood cell count, fibrinogen, high-sensitivity C-reactive protein and plasma surfactant protein were higher in GOLD 1–3 groups than in the EwAL group. Interleukin-6 was expressed less often in EwAL subjects compared with subjects in the GOLD and A1ATD groups. Soluble receptor for advanced glycation end product was lowest in GOLD 3 subjects, indicative of more severe emphysema.ConclusionsThese findings provide context for upcoming results from FOOTPRINTS®, which aims to establish correlations between biomarkers and disease progression in a representative COPD population.Trial registration number: NCT02719184, study start date 13/04/2016.

Exploring biosurfactants as a sustainable alternative to chemical surfactants

Surfactants play critical roles in various applications including food, pharmaceuticals, cosmetics, mineral processing, wastewater treatment, etc. However, traditional chemical-based surfactants are often non-biodegradable, toxic, and persistent, leading to environmental pollution and potential health risks. To address these challenges, bio-inspired surfactants have emerged as promising alternatives owing to their well-defined compositions, biodegradability, biocompatibility, and the potential to finely tune their functionality by altering the amino acid sequence. Here, we report a comparative study of traditional foaming agents, MIBC, DPG, and DPM, and a bio-inspired peptide surfactant, AM1, and a protein surfactant, DAMP4. Furthermore, the pH- and metal ion- responsive properties of these two biosurfactants make them an innovative class of smart materials that can further expand the scope of applications. Our findings provide new insights into the potential of applying these new biosurfactants for various applications, representing a sustainable and environmentally friendly alternative approach.

Application of sodium caseinate as protein based coating/stabilizing agent in development and optimization of vitamin D3-loaded nanostructured lipid carriers (NLCs)

Sodium caseinate is a food-grade protein that has several functional properties, such as acting as an emulsifying and stabilizing agent in a colloidal delivery system. Nanostructured lipid carriers (NLCs) have been proven to be highly effective carriers for encapsulating bioactive compounds. In this research, several novel food-grade NLC systems based on sodium caseinate (as a protein surfactant), anhydrous milk fat (as a solid lipid), and vitamin E acetate (as a liquid oil) were developed as an encapsulant for vitamin D3. The D-optimal combined design was employed to optimize the effects of lipid phase ratios and protein concentrations on particle size and loading efficiency. The optimized formulation obtained from the predicted model was composed of 1% w/v caseinate protein and a 73.23/26.78 solid lipid/liquid oil ratio with a desirability function of 0.983. This optimum formulation showed the smallest particle size at 120.16 nm with a narrow size distribution of 0.208 and an encapsulation efficiency of 96.14%. The cytotoxicity study showed that the anticancer effect of the vitamin was improved after incorporation into the NLC structure. The current study suggests that this composition of biopolymeric surfactant and applied lipid matrix could be an effective, health-promoting, and promising carrier system for developing food-grade bioactive delivery systems.

Molecular interaction of di-ester bonded cationic Gemini surfactants with pepsin: in vitro and in silico perspectives

The implications of surfactant–enzyme/protein interactions in a variety of fields, including biotechnology, cosmetics, paints and pharmaceuticals, have attracted a lot of attention in contemporary studies. Herein, we have employed several in vitro and in silico techniques such as excitation and absorption spectroscopies, circular dichroism and FT-IR spectroscopies, density functional and molecular dynamics simulations to understand the interaction behavior of oxy-diester-based green cationic Gemini surfactants, N1,N1,N14,N14-tetramethyl-2,13-dioxo-N1,N14-dialkyl-3,6,12-tetraoxateradecane-1,14-diaminiumdichloride (abbreviated as Cm-E2O2-Cm, where ‘m’ stands for alkyl chain length, m = 12 and 14) with one of the main digestive proteins, pepsin. The spectroscopic techniques confirm the static quenching effect of surfactants on pepsin. The calculated physical parameters (Ksv , Kb and ΔG) and their order reveal the distinguished implications for the surfactants’ chain lengths. The spontaneity of interaction was also confirmed by negative Gibbs free energy change values. The extrinsic spectroscopic study with pyrene as fluorescence probe, FT-IR and CD techniques indicated a potential conformational change in pepsin induced by the Gemini surfactants. DFT, docking and MD simulations provided the theoretical understanding regarding the quantum mechanical environment, location of binding and stability of the protein–surfactant complexation in energy terms. We believe this study will be a humble addition to our existing knowledge in the field of protein–surfactant interactions. Communicated by Ramaswamy H. Sarma

Study on Volumetric, Compressibility and Viscometric Behavior of Cationic Surfactants (CTAB and DTAB) in Aqueous Glycyl Dipeptide: A Thermo-Acoustic Approach

This study aims to understand how glycyl dipeptide affected the compressibility, volumetric behavior and viscometric behavior of the cationic surfactants CTAB (Cetyltrimethylammonium bromide) and DTAB (dodecyltrimethylammonium bromide). Information on solute-solute, solute-solvent, and solvent-solvent interactions has been inferred using the quantification of density (ρ), speed of sound (u) and viscosity in aqueous media containing glycyl dipeptide in the temperature range 293.15-313.15 K at an interval of 5 K. The data from the aforementioned research have been used to enumerate numerous volumetric and compressibility metrics that aid in the collection of information about the interactional behavior of the system under consideration. The study suggests that CTAB interacts strongly compared to DTAB with dipeptide, and it also significantly dehydrates glycyl dipeptide. The difference in water-water interactions caused by the loss of hydrophobic hydration of the surfactant molecules upon the addition of cationic surfactants may be the cause of the variation in determined parameters with surfactant concentration. Consideration of the structural rearrangement of molecules that may occur in the system has been used to explain the results of viscosity and computed factors related to viscosity. The patterns of competitive intermolecular interactions in the ternary (dipeptide + water + surfactant) system have been used to analyze the trends of all the parameters. The study may be helpful to understand the stability and structural changes in protein-surfactant systems mediated through various interactions that may be present in the system.

Self-Assembled Protein–Surfactant Nanoaggregates for Tunable Peptide Bond Hydrolysis by Polyoxometalate Nanoclusters

Self-Assembled Protein–Surfactant Nanoaggregates for Tunable Peptide Bond Hydrolysis by Polyoxometalate Nanoclusters

Surfactant-Assisted Assembly of Dipeptide Forming a Broom-like Structure.

Understanding the influence of surfactants on the assembly of peptides has a considerable practical motivation. In this paper, we systematically study the anionic surfactant-assisted assembly of diphenylalanine (FF). FF forms broom-like structures in a concentration of sodium cholate (NaC) around the CMC, and assembles into linear and unidirectional rods in the presence of low and high surfactant concentrations. FF’s improved hydrogen bonding and controlled assembly rates are appropriate for other anionic surfactants. At this stage, the use of FF as the simplest protein consequence can be helpful in the investigation of further protein–surfactant interactions.

Mechanistic Understanding of Lung Inflammation: Recent Advances and Emerging Techniques.

Acute respiratory distress syndrome (ARDS) is a life-threatening lung injury characterized by an acute inflammatory response in the lung parenchyma. Hence, it is considered as the most appropriate clinical syndrome to study pathogenic mechanisms of lung inflammation. ARDS is associated with increased morbidity and mortality in the intensive care unit (ICU), while no effective pharmacological treatment exists. It is very important therefore to fully characterize the underlying pathobiology and the related mechanisms, in order to develop novel therapeutic approaches. In vivo and in vitro models are important pre-clinical tools in biological and medical research in the mechanistic and pathological understanding of the majority of diseases. In this review, we will present data from selected experimental models of lung injury/acute lung inflammation, which have been based on clinical disorders that can lead to the development of ARDS and related inflammatory lung processes in humans, including ventilation-induced lung injury (VILI), sepsis, ischemia/reperfusion, smoke, acid aspiration, radiation, transfusion-related acute lung injury (TRALI), influenza, Streptococcus (S.) pneumoniae and coronaviruses infection. Data from the corresponding clinical conditions will also be presented. The mechanisms related to lung inflammation that will be covered are oxidative stress, neutrophil extracellular traps, mitogen-activated protein kinase (MAPK) pathways, surfactant, and water and ion channels. Finally, we will present a brief overview of emerging techniques in the field of omics research that have been applied to ARDS research, encompassing genomics, transcriptomics, proteomics, and metabolomics, which may recognize factors to help stratify ICU patients at risk, predict their prognosis, and possibly, serve as more specific therapeutic targets.

Benefit of coconut-based hair oil via hair porosity quantification.

The present study is intended to characterize the surfactant damage suffered by the hair cortex in routine washing and the mechanistic effect of Coconut Based Hair Oils (CBHO) to mitigate the damage. Surfactants which diffuse into the hair structure solubilize protein moieties, leading to an increase in porosity and internal surface area as well as the pore volume. The changes in hair pores occurring in the hair cortex are measured by nitrogen sorption method in line with the Brunauer-Emmett-Teller (BET) theory. Single fiber tensile parameters were measured using Diastron MTT 175. Color protection was measured quantitatively using spectrophotometer as well as visual rating by trained panelists. The pore surface area data clearly show the benefit of introducing coconut-based hair oils (CBHO) into the hair by preventing increase in hair porosity. A statistically significant decrease in break stress and toughness were observed and the same were reversed by the application of CBHO. A pronounced color protection effect was also recorded with the application of CBHO. The porosity reduction effect seen with the use of CBHO is attributed to the CBHO molecules blocking the diffusion pathways in the endocuticle and the matrix part of the cortical cells, limiting protein surfactant interaction resulting in reduced solubilization and loss. Since, the color molecules are likely to be much smaller than the protein moieties, a pronounced color protection effect suggests that the penetrated CBHO molecules form a dense diffusion barrier in the matrix, cell membrane complex (CMC) and the endocuticle regions of hair - which are the main diffusion pathways out of hair. The study confirms the damage repair potential of CBHO and that it works by increasing the hydrophobicity of hair - both on the hair surface and in the cortex.

Deciphering the mechanism of interaction of an ester-functionalized cationic gemini surfactant with bovine serum albumin: A biophysical and molecular modeling study

Protein–surfactant interactions have relevance in the fields of pharmaceuticals, detergency formulation, pharmacology, personal hygiene products, etc. Accordingly, we have analyzed the biomolecular interaction between a recently synthesized C16-E2O2-C16 gemini surfactant with the transport protein, bovine serum albumin (BSA), utilizing surface tension, fluorescence (intrinsic/extrinsic, synchronous, 3-D and resonance Rayleigh scattering (RRS)), circular dichroism (CD), UV–visible, Fourier transform infrared (FT-IR) and computational methods. The critical micelle concentration (CMC) and surface parameters of the gemini surfactant get modified in the presence of BSA. The interaction of the concerned surfactant with BSA occurred via a static quenching mechanism forming a less stable complex at the higher temperature. Based on calculated thermodynamic parameters, the interaction process was found to be spontaneous (∆Gb0 is negative) and that hydrogen bond (HB) and van der Waals (VDW) forces were predominant forces in stabilizing the complex (both ∆Hb0 and ∆Sb0 are negative). FT-IR and CD measurements demonstrated that the secondary structure conformation of BSA was perturbed because of the BSA-C16-E2O2-C16 complexation. The 3-D and RRS fluorescence studies demonstrated that the BSA size enlarged in presence of the gemini and ensued in the increment of scattering effect and suggested some conformational and micro-environmental alterations of Trp/Tyr residues of BSA (the same results were obtained for pyrene extrinsic fluorescence). Molecular docking and site marker displacement investigations recognized that the surfactant C16-E2O2-C16 predominantly binds within BSA’s site I (sub-domain IIA).

Molecular modeling of the interface of an egg yolk protein-based emulsion

Many food emulsions are stabilized by functional egg yolk biomolecules, which act as surfactants at the oil/water interface. Detailed experimental studies on egg yolk emulsifying properties have been largely hindered due to the difficulty in isolating individual chemical species. Therefore, this work presents a molecular model of an oil/water interfacial system where the emulsifier is one of the most surface-active proteins from the egg yolk low-density lipoproteins (LDL), the so-called Apovitellenin I. Dissipative particle dynamics (DPD) was here adopted in order to simulate large systems over long time scales, when compared with full-atom molecular dynamics (MD). Instead of a manual assignment of the DPD simulation parameters, a fully automated coarse-graining procedure was employed. The molecular interactions used in the DPD system were determined by means of a parameter calibration based on matching structural data from atomistic MD simulations. Despite the little availability of experimental data, the model was designed to test the most relevant physical properties of the protein investigated. Protein structural and dynamics properties obtained via MD and DPD were compared highlighting advantages and limits of each molecular technique. Promising results were achieved from DPD simulations of the oil/water interface. The proposed model was able to properly describe the protein surfactant behavior in terms of interfacial tension decrease at increasing protein surface concentration. Moreover, the adsorption time of a free protein molecule was estimated and, finally, an LDL-like particle adsorption mechanism was qualitatively reproduced.

Recyclable optical bioplastics platform for solid state red light harvesting via triplet-triplet annihilation photon upconversion.

Sustainable photonics applications of solid-state triplet-triplet annihilation photon upconversion (TTA-UC) are limited by a small UC spectral window, low UC efficiency in air, and non-recyclability of polymeric materials used. In a step to overcome these issues, we have developed new recyclable TTA-UC bioplastics by encapsulating TTA-UC chromophores liquid inside the semicrystalline gelatin films showing broad-spectrum upconversion (red/far-red to blue) with high UC efficiency in air. For this, we synthesized a new anionic annihilator, sodium-TIPS-anthracene-2-sulfonate (TIPS-AnS), that combined with red/far-red sensitizers (PdTPBP/Os(m-peptpy)2(TFSI)2), a liquid surfactant Triton X-100 reduced (TXr) and protein gelatin (G) formed red/far-red to blue TTA-UC bioplastic films just by air drying of their aqueous solutions. The G-TXr-TIPS-AnS-PdTPBP film showed record red to blue (633 to 478 nm) TTA-UC quantum yield of 8.5% in air. The high UC quantum yield has been obtained due to the fluidity of dispersed TXr containing chromophores and oxygen blockage by gelatin fibers that allowed efficient diffusion of triplet excited chromophores. Further, the G-TXr-TIPS-AnS-Os(m-peptpy)2(TFSI)2 bioplastic film displayed far-red to blue (700-730 nm to 478 nm) TTA-UC, demonstrating broad-spectrum photon harvesting. Finally, we demonstrated the recycling of G-TXr-TIPS-AnS-PdTPBP bioplastics by developing a downstream approach that gives new directions for designing future recyclable photonics bioplastic materials.

On the Effect of pH, Temperature, and Surfactant Structure on Bovine Serum Albumin-Cationic/Anionic/Nonionic Surfactants Interactions in Cacodylate Buffer-Fluorescence Quenching Studies Supported by UV Spectrophotometry and CD Spectroscopy.

Due to the fact that surfactant molecules are known to alter the structure (and consequently the function) of a protein, protein–surfactant interactions are very important in the biological, pharmaceutical, and cosmetic industries. Although there are numerous studies on the interactions of albumins with surfactants, the investigations are often performed at fixed environmental conditions and limited to separate surface-active agents and consequently do not present an appropriate comparison between their different types and structures. In the present paper, the interactions between selected cationic, anionic, and nonionic surfactants, namely hexadecylpyridinium chloride (CPC), hexadecyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), polyethylene glycol sorbitan monolaurate, monopalmitate, and monooleate (TWEEN 20, TWEEN 40, and TWEEN 80, respectively) with bovine serum albumin (BSA) were studied qualitatively and quantitatively in an aqueous solution (10 mM cacodylate buffer; pH 5.0 and 7.0) by steady-state fluorescence spectroscopy supported by UV spectrophotometry and CD spectroscopy. Since in the case of all studied systems, the fluorescence intensity of BSA decreased regularly and significantly under the action of the surfactants added, the fluorescence quenching mechanism was analyzed thoroughly with the use of the Stern–Volmer equation (and its modification) and attributed to the formation of BSA–surfactant complexes. The binding efficiency and mode of interactions were evaluated among others by the determination, comparison, and discussion of the values of binding (association) constants of the newly formed complexes and the corresponding thermodynamic parameters (ΔG, ΔH, ΔS). Furthermore, the influence of the structure of the chosen surfactants (charge of hydrophilic head and length of hydrophobic chain) as well as different environmental conditions (pH, temperature) on the binding mode and the strength of the interaction has been investigated and elucidated.

Interaction between soy protein isolate and surfactant at the interface of antibacterial nanoemulsions loaded with riboflavin tetra butyrate

SummaryRiboflavin tetra butyrate (RTB) is an effective antimicrobial photosensitiser that poses a challenge for long‐term preservation and application due to its hydrophobicity and instability. To overcome this problem, a series of nanoemulsions loaded with photosensitiser RTB was prepared in this study by using soy protein isolate (SPI) as an emulsifier and small molecule surfactant (X) as co‐emulsifier. The interaction mechanism of SPI+X co‐emulsifier and its influence on the stability of emulsion and photosensitiser were explored, and the photodynamic bactericidal effect of the prepared RTB emulsion on S. aureus was studied. SPI and four types of surfactants were present in the form of synergistic (Tween 80, T80) or competition (Span 80, S80 and lecithin, LEC) adsorption or formation of Pickering emulsion (Sodium dodecyl sulphate, SDS) at the oil–water interface. The experimental results revealed that the stable encapsulation ability of RTB emulsions for photosensitisers and their preservation ability can be drastically improved. This work verified that all RTB emulsions could inactive S. aureus under blue light, with an antibacterial ratio of more than 70%. Therefore, the emulsion‐based delivery system effectively improved the water dispersibility and chemical stability of RTB. These results demonstrate that RTB emulsion has the potential to be used as a food‐grade antibacterial emulsion.

Incorporation and Assembly of a Light-Emitting Enzymatic Reaction into Model Protein Condensates.

Eukaryotic cells partition enzymes and other cellular components into distinct subcellular compartments to generate specialized biochemical niches. A subclass of these compartments form in the absence of lipid membranes, via liquid-liquid phase separation of proteins to form biomolecular condensates or "membraneless organelles" such as nucleoli, stress granules, and P-bodies. Because of their propensity to form compartments from simple starting materials, membraneless organelles are an attractive target for engineering new functionalities in both living cells and protocells. In this work, we demonstrate incorporation of a novel enzymatic activity in protein coacervates with the light-generating enzyme, NanoLuc, to produce bioluminescence. Using condensates comprised of the disordered RGG domain of Caenorhabditis elegans LAF-1, we functionalized condensates with enzymatic activity in vitro and show that enzyme localization to coacervates enhances assembly and activity of split enzymes. To build condensates that function as light-emitting reactors, we designed a NanoLuc enzyme flanked by RGG domains. The resulting condensates concentrated NanoLuc by 10-fold over bulk solution and displayed significantly increased reaction rates. We further show that condensate viscosity impacts light emission due to diffusion-limited behavior. Because our model condensates have low viscosities, we predict NanoLuc diffusion-limited behavior in most other condensates and thus propose the condensate-Nanoluc system as a potential strategy for high-throughput screening of condensate targeting drugs. By splitting the NanoLuc enzyme into its constituent components, we demonstrate that NanoLuc activity can be reconstituted via co-condensation. In addition, we demonstrate control of the spatial localization of the enzyme within condensates by targettng NanoLuc to the surface of in vitro condensates. Collectively, this work demonstrates that membraneless organelles can be endowed with localized enzymatic activity and that this activity can be spatially and temporally controlled via biochemical reconstitution and design of protein surfactants.

Nanoemulsions for drug delivery

Emulsions are liquid–liquid dispersions with one liquid phase dispersed in the other liquid phase as small droplets. Nanoemulsions are nano-sized emulsions with sizes ranging from tens to hundreds of nanometers, and have great potential applications in pharmaceutics, foods and cosmetics due to their attractive properties, such as small sizes, high surface area per unit volume, improved dispersion of active hydrophobic components and enhanced absorption. The article provides an overview of nanoemulsions for drug delivery, starting with an introduction of emulsion types, nanoemulsion preparation and nanoemulsion stability. Surfactants play critical roles in producing and stabilizing nanoemulsions. Different types of surfactants are summarized including small molecule surfactants, particle surfactants, phospholipids, peptide and protein surfactants. Then the applications of nanoemulsions as nanomedicine in drug delivery are presented. Finally, clinical applications of nanoemulsions are discussed.