Coacervate Phase Research Articles

Diffusive Trends in Concentrated Oppositely-Charged Polyelectrolyte Solutions and Onset of Glassy Dynamics.

We utilize single particle tracking studies to investigate the diffusion of polylysine through concentrated matrices of cationic polylysine and anionic polyglutamic acid with no added salts. These studies show that diffusivity has a strong apparently exponential dependence on concentration in crowded systems that does not appear to be a function of the charge sign. These trends are consistent in both single-phase systems prepared at concentrated conditions and polymer-rich coacervate phases formed from dilute phase-separating systems. The likely origin of this behavior is the onset of glassy dynamics spurred by a decrease in plasticization by water and the large excluded volume associated with charge-bearing species. This effect can be contextualized through free volume-based theories such as the Vrentas-Duda model. Overall, we obtain dynamic behavior that is distinctly different from behavior observed in more dilute systems and warrants further investigation.

Influence of the backbone chemistry and ionic functional groups of five pairs of oppositely charged polyelectrolytes on complex coacervation

Complex coacervation plays an important role in various fields. Here, the influences of the backbone chemistry and ionic functional groups of five pairs of oppositely charged polyelectrolytes on complex coacervation were investigated. These pairs include synthetic polymers with aliphatic hydrocarbon backbones, peptides with amide bonds, and carbohydrates with glycosidic linkages. Despite sharing identical charged groups, specific pairs displayed distinct liquid/liquid and liquid/solid phase separations depending on the polyelectrolyte mixing ratio, buffer, and ionic strength. The coacervate phase boundary broadened in the orders: glycosidic linkages > amide backbone > aliphatic hydrocarbon backbone, and Tris-phosphate > Tris-acetate > Tris-chloride buffers. Coacervates prepared from polyelectrolytes with lower solubilities in water resisted disassembly at high salt concentrations, and their merge rate was slow. These observations suggest that the hydrophobic segments in polyelectrolytes interfere with the formation of complex coacervates; however, following coacervate formation, the hydrophobic segments render the coacervates stable and elastic.

Coacervate Formation in Dilute Aqueous Solutions of Inorganic Molecular Clusters with Simple Divalent Countercations.

We report a complex coacervate formed by a 2.5 nm-diameter, rigid uranyl peroxide molecular cluster (Li68K12(OH)20)[UO2(O2)OH]60, U6060-) and SrCl2 salt in dilute aqueous solutions, including its location in the phase diagram, composition, rheological features, and critical conditions for phase transitions. In this coacervate, the Sr2+ cations are a major building component, and the coacervate phase covers a substantial region of the phase diagram. This coacervate demonstrates features that differ from traditional coacervates formed by oppositely charged long-chain polyelectrolytes, especially in its formation mechanism, dehydration, enhancement of mechanical strength with increasing ionic strength, and the change of salt partition preference into the coacervate and supernatant phases with ionic strength.

Continuous Transformation from Membrane‐Less Coacervates to Membranized Coacervates and Giant Vesicles: Toward Multicompartmental Protocells with Complex (Membrane) Architectures

AbstractThe membranization of membrane‐less coacervates paves the way for the exploitation of complex protocells with regard to structural and cell‐like functional behaviors. However, the controlled transformation from membranized coacervates to vesicles remains a challenge. This can provide stable (multi)phase and (multi)compartmental architectures through the reconfiguration of coacervate droplets in the presence of (bioactive) polymers, bio(macro)molecules and/or nanoobjects. Herein, we present a continuous protocell transformation from membrane‐less coacervates to membranized coacervates and, ultimately, to giant hybrid vesicles. This transformation process is orchestrated by altering the balance of non‐covalent interactions through varying concentrations of an anionic terpolymer, leading to dynamic processes such as spontaneous membranization of terpolymer nanoparticles at the coacervate surface, disassembly of the coacervate phase mediated by the excess anionic charge, and the redistribution of coacervate components in membrane. The diverse protocells during the transformation course provide distinct structural features and molecular permeability. Notably, the introduction of multiphase coacervates in this continuous transformation process signifies advancements toward the creation of synthetic cells with different diffusible compartments. Our findings emphasize the highly controlled continuous structural reorganization of coacervate protocells and represents a novel step toward the development of advanced and sophisticated synthetic protocells with more precise compositions and complex (membrane) structures.

Continuous Transformation from Membrane-Less Coacervates to Membranized Coacervates and Giant Vesicles: Toward Multicompartmental Protocells with Complex (Membrane) Architectures.

The membranization of membrane-less coacervates paves the way for the exploitation of complex protocells with regard to structural and cell-like functional behaviors. However, the controlled transformation from membranized coacervates to vesicles remains a challenge. This can provide stable (multi)phase and (multi)compartmental architectures through the reconfiguration of coacervate droplets in the presence of (bioactive) polymers, bio(macro)molecules and/or nanoobjects. Herein, we present a continuous protocell transformation from membrane-less coacervates to membranized coacervates and, ultimately, to giant hybrid vesicles. This transformation process is orchestrated by altering the balance of non-covalent interactions through varying concentrations of an anionic terpolymer, leading to dynamic processes such as spontaneous membranization of terpolymer nanoparticles at the coacervate surface, disassembly of the coacervate phase mediated by the excess anionic charge, and the redistribution of coacervate components in membrane. The diverse protocells during the transformation course provide distinct structural features and molecular permeability. Notably, the introduction of multiphase coacervates in this continuous transformation process signifies advancements toward the creation of synthetic cells with different diffusible compartments. Our findings emphasize the highly controlled continuous structural reorganization of coacervate protocells and represents a novel step toward the development of advanced and sophisticated synthetic protocells with more precise compositions and complex (membrane) structures.

Advanced coacervation-driven nanoscale polymeric assemblies for biomedical applications

Complex coacervation, a fascinating phenomenon rooted in liquid–liquid phase separation, plays a crucial role in numerous biological systems. This intricate process involves the segregation of a liquid into two distinct phases: a coacervate phase enriched with polymers and a polymer-deficient phase comprising the remaining dilute solution. The potential of coacervates extends beyond their natural occurrence in biological systems, as they possess the capability to encapsulate various types of biomolecules in an aqueous solution, obviating the need for organic solvents. Consequently, considerable efforts have been devoted to designing functional nanoscale coacervate-driven assemblies using both natural and synthetic polymers for a myriad of applications. In this review, we provide a synthesis and discussion of the formation of nanoscale polymeric assemblies driven by complex coacervation. This exploration delves into the fundamental driving forces underpinning the phenomenon and elucidates the diverse fabrication strategies employed. The various biomedical applications of these assemblies are highlighted, with a focus on their roles as drug carriers, gene delivery vehicles, antimicrobial agents, theranostic platforms, mucoadhesives, and nanoreactors. This review aims to contribute to a deeper understanding of coacervation-driven nanoscale assembly systems and their potential impact on the field of biomedical science and technology.

Transforming the Stability, Encapsulation, and Sustained Release Properties of Calcium Alginate Beads through Gel-Confined Coacervation.

Calcium alginate (Ca2+/alginate) gel beads find use in diverse applications, ranging from drug delivery and tissue engineering to bioprocessing, food formulation, and agriculture. Unless modified, however, these gels have limited stability in alkaline media (including phosphate buffers), and their high solute permeability limits their ability to efficiently encapsulate and slowly release water-soluble small molecules. Here, we show how these limitations can be addressed by mixing the alginate solutions used in the bead preparation with the nontoxic anionic polymer polyphosphate (PP). Upon complexing Ca2+ ions, PP undergoes complex coacervation (i.e., liquid/liquid phase separation into a Ca2+/PP-rich coacervate phase and a dilute supernatant phase). At lower PP concentrations, the Ca2+/PP coacervate appears to simply remain dispersed within the beads. Though its presence makes the beads more stable in alkaline media (phosphate-buffered saline and seawater), it has little impact on the bead stiffness, morphology, and (at least in the absence of substantial payload/coacervate association) encapsulation and release properties. When the PP concentrations exceed a critical value, however, Ca2+/PP coacervation within the gelling Ca2+/alginate beads collapses the resulting beads into more compact, interpenetrating polymer networks. Besides their enhanced stability to alkaline environments, these hybrid beads exhibit irregular morphologies with wrinkled and dimpled surface structures and macroscopic (closed) internal pores, and their collapse into these polymer-rich networks also makes them significantly stiffer than their PP-free counterparts. Crucially, these beads also exhibit a much lower solute permeability, which enables highly efficient encapsulation and multiday release of water-soluble small molecules (with the beads encapsulating >90% of the added model payload and sustaining its release over 3-5 d). Collectively, these findings provide a mild and simple (single-step) pathway to generating ionically cross-linked alginate beads with significantly enhanced stability, encapsulation efficiency, and sustained release.

Macromolecular Crowding Effect on Chitosan-Hyaluronic Acid Complexation and the Activity of Encapsulated Catalase.

The associative phase separation of charged biomacromolecules plays a key role in many biophysical events that take place in crowded intracellular environments. Such natural polyelectrolyte complexation and phase separation often occur at nonstoichiometric charge ratios with the incorporation of bioactive proteins, which is not studied as extensively as those complexations at stoichiometric ratios. In this work, we investigated how the addition of a crowding agent (polyethylene glycol, PEG) affected the complexation between chitosan (CS) and hyaluronic acid (HA), especially at nonstoichiometric ratios, and the encapsulation of enzyme (catalase, CAT) by the colloidal complexes. The crowded environment promoted colloidal phase separation at low charge ratios, forming complexes with increased colloidal and dissolution stability, which resulted in a smaller size and polydispersity (PDI). The binding isotherms revealed that the addition of PEG greatly enhanced the ion-pairing strength (with increased ion-pairing equilibrium constant Ka from 4.92 × 104 without PEG to 1.08 × 106 with 200 g/L PEG) and switched the coacervation from endothermic to exothermic, which explained the promoted complexation and phase separation. At the stoichiometric charge ratio, the enhanced CS-HA interaction in crowded media generated a more solid-like coacervate phase with a denser network, slower chain relaxation, and higher modulus. Moreover, both crowding and complex encapsulation enhanced the activity and catalytic efficiency of CAT, represented by a 2-fold increase in catalytic efficiency (Kcat/Km) under 100 g/L PEG crowding and CS-HA complex encapsulation. This is likely due to the lower polarity in the microenvironment surrounding the enzyme molecules. By a systematic investigation of both nonstoichiometric and stoichiometric charge ratios under macromolecular crowding, this work provided new insights into the complexation between natural polyelectrolytes in a scenario closer to an intracellular environment.

Enhanced Catalytic Activity of a de novo Enzyme in a Coacervate Phase

Biomolecular condensates are membraneless organelles that orchestrate various metabolic pathways in living cells. Understanding how these crowded structures regulate enzyme reactions remains yet challenging due to their dynamic and intricate nature. Coacervate microdroplets formed by associative liquid‐liquid phase separation of oppositely charged polyions have emerged as relevant condensate models to study enzyme catalysis. Enzyme reactions within these droplets show altered kinetics, influenced by factors such as enzyme and substrate partitioning, crowding, and interactions with coacervate components; it is often challenging to disentangle the contributions of each. Here, we investigate the peroxidase activity of a de novo enzyme within polysaccharide‐based coacervates. By comparing the reaction kinetics in buffer, in a suspension of coacervates and in the bulk coacervate phase collected after centrifugation of the droplets, we show that the coacervate phase significantly increases the enzyme catalytic efficiency. We demonstrate that the main origin of this enhanced activity lies in macromolecular crowding coupled to changes in the conformational dynamics of the enzyme within the coacervate environment. Altogether, these findings underline the crucial role of the coacervate matrix in enzyme catalysis, beyond simple partitioning effects. The observed boost in enzyme activity within the coacervate phase provides insights for designing biocatalytically active synthetic organelles.

Salt reduction in myofibrillar protein gel via inhomogeneous distribution of sodium-containing encapsulated fish oil coacervate: Mucopenetration ability of sodium carboxymethyl cellulose

The potential application of fish oil microcapsules as salt reduction strategies in low-salt myofibrillar protein (MP) gel was investigated by employing soy protein isolates/carboxymethyl cellulose sodium (SPI-CMC) coacervates enriched with 25 mM sodium chloride and exploring their rheological characteristics, taste perception, and microstructure. The results revealed that the SPI-CMC coacervate phase exhibited the highest sodium content under 25 mM sodium level, albeit with uneven distribution. Notably, the hydrophilic and adhesive properties of CMC to sodium facilitated the in vitro release of sodium during oral digestion, as evidenced by the excellent wettability and mucopenetration ability of CMC. Remarkably, the fish oil microcapsules incorporating SPI-CMC as the wall material, prepared at pH 3.5 with a core-to-wall ratio of 1:1, demonstrated the highest encapsulation efficiency, which was supported by the strong hydrogen bonding. Interestingly, the presence of SPI-CMC coacervates and fish oil microcapsules enhanced the interaction between MPs and strengthened the low-salt MP gel network. Coupled with electronic tongue analysis, the incorporation of fish oil microcapsules slightly exacerbated the non-uniformity of sodium distribution. This ultimately contributed to an enhanced perception of saltiness, richness, and aftertaste in low-salt protein gels. Overall, the incorporation of fish oil microcapsules emerged as an effective salt reduction strategy in low-salt MP gel.

Controlled Supramolecular Polymerization via Bioinspired, Liquid-Liquid Phase Separation of Monomers.

Dynamic supramolecular assemblies, driven by noncovalent interactions, pervade the biological realm. In the synthetic domain, their counterparts, supramolecular polymers, endowed with remarkable self-repair and adaptive traits, are often realized through bioinspired designs. Recently, controlled supramolecular polymerization strategies have emerged, drawing inspiration from protein self-assembly. A burgeoning area of research involves mimicking the liquid-liquid phase separation (LLPS) observed in proteins to create coacervate droplets and recognizing their significance in cellular organization and diverse functions. Herein, we introduce a novel perspective on synthetic coacervates, extending beyond their established role in synthetic biology as dynamic, membraneless phases to enable structural control in synthetic supramolecular polymers. Drawing parallels with the cooperative growth of amyloid fibrils through LLPS, we present metastable coacervate droplets as dormant monomer phases for controlled supramolecular polymerization. This is achieved via a π-conjugated monomer design that combines structural characteristics for both coacervation through its terminal ionic groups and one-dimensional growth via a π-conjugated core. This design leads to a unique temporal LLPS, resulting in a metastable coacervate phase, which subsequently undergoes one-dimensional growth via nucleation within the droplets. In-depth spectroscopic and microscopic characterization provides insights into the temporal evolution of disordered and ordered phases. Furthermore, to modulate the kinetics of liquid-to-solid transformation and to achieve precise control over the structural characteristics of the resulting supramolecular polymers, we invoke seeding in the droplets, showcasing living growth characteristics. Our work thus opens up new avenues in the exciting field of supramolecular polymerization, offering general design principles and controlled synthesis of precision self-assembled structures in confined environments.

Coacervate Phase Evolution and Membrane Formation in Natural Seawater.

Marine organisms produce biological materials through the complex self-assembly of protein condensates in seawater, but our understanding of the mechanisms of microstructure evolution and maturation remains incomplete. Here, we show that critical processing attributes of mussel holdfast proteins can be captured by the design of an amphiphilic, fluorescent polymer (PECHIA) consisting of a polyepichlorohydrin backbone grafted with 1-imidazolium acetonitrile. Aqueous solutions of PECHIA were extruded into seawater, wherein the charge repulsion of PECHIA is screened by high salinity, facilitating interfacial condensation via enhanced "cation-dipole" interactions. Diffusion of seawater into the PECHIA solution caused droplets to form immiscibly within the PECHIA phase (i.e., inverse coacervation). Simultaneously, weakly alkaline seawater catalyzes nitrile cyclization and time-dependent solidification of the PECHIA phase, leading to hierarchically porous membranes analogous to porous architectures in mussel plaques. In contrast to conventional polymer processing technologies, processing of this biomimetic polymer required neither organic solvents nor heating and enabled the template-free production of hollow spheres and fibers over a wide range of salinities.

Tuning of Coacervate Phase Behavior of Polyoxymethylene (4) Lauryl Ether in Aqueous Alcoholic Solution: Investigation of Thermodynamics

In case of non-ionic surfactant, the phase partition behavior (CP) of micellar solution was vary under the influence of various additive mixed systems. The non-ionic surfactant in an aqueous medium shows phase separation above the critical micelle concentration (CMC), There is a change in micellar interactions at cloud point (CP) with increase in temperature. The insertion of foreign material to surfactant solution alters the clouding temperature. In this paper we report how cloud point varies with addition of alcohol as additives. For non-ionic surfactant Polyoxyethylene (4) lauryl ether (Brij-30) the CP temperature was investigated at variable concentrations of surfactant in pure and additive mixed systems. The results we obtained shows that cloud point of pure Brij-30 surfactant decreases with enhancement in surfactant concentration from 1% to 10% (w/v) in a 22% aqueous ethanolic medium. Similarly, the cloud point values of Brij-30 in presence of n-alcohols shows increasing trend in case of propanol and butanol, specifying that their solubilization helps to form swollen water structures, thus favoring micelle hydration, while the presence of alcohols like pentanol, hexanol and heptanol shows a decreasing trend with increasing micelle size. The change in cloud points of Brij-30 in participation of various n-alcohols plays an important role in demonstrating the effect of the nature of additives on the stability of micelle. The foam ability and foam stability data discusses the stability of micelle aggregation. The important information about the thermodynamic entities were calculated using “Phase Separation Model”.

Effect of Polymer Gel Elasticity on Complex Coacervate Phase Behavior.

Gels are key materials in biological systems such as tissues and may control biocondensate formation and structure. To further understand the effects of elastic environments on biomacromolecular assembly, we have investigated the phase behavior and radii of complex coacervate droplets in polyacrylamide (PAM) networks as a function of gel modulus. Poly-l-lysine (PLL) and sodium hyaluronate (HA) complex coacervate phases were prepared in PAM gels with moduli varying from 0.035 to 15.0 kPa. The size of the complex coacervate droplets is reported from bright-field microscopy and confocal fluorescence microscopy. Overall, the complex coacervate droplet volume decreases inversely with the modulus. Fluorescence microscopy is also used to determine the phase behavior and concentration of fluorescently tagged HA in the complex coacervate phases as a function of ionic strength (100-270 mM). We find that the critical ionic strength and complex coacervate stability are nonmonotonic as a function of the network modulus and that the local gel concentration can be used to control phase behavior and complex coacervate droplet size scale. By understanding how elastic environments influence simple electrostatic assembly, we can further understand how biomacromolecules exist in complex, crowded, and elastic cellular environments.

Coexistence of two coacervate phases of polyglycine in water suggested by polymer reference interaction site model theory.

Mixing Gibbs energy and phase equilibria of aqueous solutions of polyglycine were studied theoretically by means of polymer reference interaction site model integral equation theory combined with the Gibbs-Duhem method. In addition to the ordinary liquid-liquid phase separation between dilute and concentrated solutions, the theoretical calculation predicted the coexistence of two coacervate phases, namely, the lower- and higher-density coacervates. The relative thermodynamic stabilities of these two phases change with the polymerization degree of polyglycine. The higher-density coacervate phase was rapidly stabilized by increasing the polymer length, and the lower-density phase became metastable at large polymers. The hydrogen bonds between the peptide chains were strengthened, and water was thermodynamically destabilized in the higher-density coacervate. A possible relation with the formation of amyloid fibril within a liquid droplet is also discussed.

New insights into protein–polysaccharide complex coacervation: Dynamics, molecular parameters, and applications

AbstractFor more than a decade, the discovery of liquid–liquid phase separation within living organisms has prompted colloid scientists to understand the connection between coacervate functionality, phase behavior, and dynamics at a multidisciplinary level. Although the protein–polysaccharide was the first system in which the coacervation phenomenon was discovered and is widely used in food systems, the phase state and relaxation dynamics of protein–polysaccharide complex coacervates (PPCC) have rarely been discussed previously. Consequently, this review aims to unravel the relationship between PPCC dynamics, thermodynamics, molecular architecture, applications, and phase states in past studies. Looking ahead, solving the way molecular architecture spreads to macro‐functionality, that is, establishing the relationship between molecular architecture–dynamics–application, will catalyze novel advancements in PPCC research within the field of foods and biomaterials.

Understanding the Impacts of Molecular and Macromolecular Crowding Agents on Protein-Polymer Complex Coacervates.

Complex coacervation refers to the liquid-liquid phase separation (LLPS) process occurring between charged macromolecules. The study of complex coacervation is of great interest due to its implications in the formation of membraneless organelles (MLOs) in living cells. However, the impacts of the crowded intracellular environment on the behavior and interactions of biomolecules involved in MLO formation are not fully understood. To address this knowledge gap, we investigated the effects of crowding on a model protein-polymer complex coacervate system. Specifically, we examined the influence of sucrose as a molecular crowder and polyethylene glycol (PEG) as a macromolecular crowder. Our results reveal that the presence of crowders led to the formation of larger coacervate droplets that remained stable over a 25-day period. While sucrose had a minimal effect on the physical properties of the coacervates, PEG led to the formation of coacervates with distinct characteristics, including higher density, increased protein and polymer content, and a more compact internal structure. These differences in coacervate properties can be attributed to the effects of crowders on individual macromolecules, such as the conformation of model polymers, and nonspecific interactions among model protein molecules. Moreover, our results show that sucrose and PEG have different partition behaviors: sucrose was present in both the coacervate and dilute phases, while PEG was observed to be excluded from the coacervate phase. Collectively, our findings provide insights into the understanding of crowding effects on complex coacervation, shedding light on the formation and properties of coacervates in the context of MLOs.

Rheological and microstructural properties of heteroprotein complex coacervates formed by lactoferrin and osteopontin

Lactoferrin (LF) and osteopontin (OPN) form heteroprotein complex coacervates (HPCCs) within limited conditions of pH, ionic strength, total protein concentration (TPC), and protein stoichiometry. In this study, it was determined using dynamic oscillatory rheology, that the coacervates exhibit frequency-dependent, viscoelastic behaviour whereby the storage modulus (G′) was less than the loss modulus (G″) for all frequencies studied (≤100 rad/s). By reducing the TPC and/or ionic strength of the system, an increase in the protein concentration and strength of the coacervate was achieved. The G′ and G″ of the coacervates were largely independent of temperature <50 °C, but above this temperature, significant changes occurred due to protein denaturation. Using microscopic techniques, it was also determined that both proteins co-localise throughout the coacervate phase in a highly ordered, structural network of protein and water. The resulting freeze-dried coacervate powder particles had a honeycomb-like appearance due to an intricate network of porous channels formed during the drying process. This physicochemical characterisation of LF-OPN HPCCs has increased scientific understanding of their rheological and microstructural properties which can now be understood, predicted, and controlled for subsequent biochemical analysis.

Compliant Clients: Catechols Exhibit Enhanced Solubility and Stability in Diverse Complex Coacervates.

Polyelectrolyte coacervates, with their greater-than-water density, low interfacial energy, shear thinning viscosity, and ability to undergo structural arrest, mediate the formation of diverse load-bearing macromolecular materials in living organisms as well as in industrial material fabrication. Coacervates, however, have other useful attributes that are challenging to study given the metastability of coacervate colloidal droplets and a lack of suitable analytical methods. We adopt solution electrochemistry and nuclear magnetic resonance measurements to obtain remarkable insights about coacervates as solvent media for low-molecular-weight catechols. When catechols are added to dispersions of coacervated polyelectrolytes, there are two significant consequences: (1) catechols preferentially partition up to 260-fold into the coacervate phase, and (2) coacervates stabilize catechol redox potentials by up to +200 mV relative to the equilibrium solution. The results suggest that the relationship between phase-separated polyelectrolytes and their client molecules is distinct from that existing in aqueous solution and has the potential for insulating many redox-unstable chemicals.

Effect of Triton X-100 on Shiraia laccase production and its application in triclosan biodegradation

Shiraia bambusicola, a bambusicolous fungus exhibits an excellent capability to produce the novel photosensitizer hypocrellin for photodynamic therapy. Recently, Shiraia species attract increasingly more attention as fungal laccase producer due to its extensive application in bioremediation. In this study, the laccase production was enhanced by the addition of surfactants in mycelium culture of Shiraia sp. S9. We sought to explore the mechanism for the surfactant induction and apply the culture to biodegrade a micropollutant triclosan. Triton X-100 was found to induce the burst of reactive oxygen species (ROS), increase membrane permeability and up-regulate laccase-related gene expression, leading to the enhanced laccase production to 9.2 × 104 U/L, about 108-folds over the control. After 2 days of Shiraia mycelium culture under Triton X-100 treatment (0.8%, w/v), the degradation and adsorption rate of triclosan reached 80.71% and 10.21%, respectively. Furthermore, Triton X-100-induced cloud point system was established after the biodegradation, which could further enrich triclosan in the coacervate phase. With the help of a combination of triclosan biodegradation and the cloud point extraction, triclosan was almost completely removed (99.88%) from water phase in the culture. To our knowledge, this is the first report on the biodegradation of triclosan by Shiraia mycelium culture. Our study provided insights into elicitation of surfactants on fungal laccase production and a new strategy for triclosan biodegradation.