Enzymatic Polymerization Research Articles

Peculiar Properties of Template-Assisted Aniline Polymerization in a Buffer Solution Using Laccase and a Laccase-Mediator System as Compared with Chemical Polymerization.

The conventional chemical polymerization of aniline has been described in multiple publications, while enzymatic polymerization has been poorly explored. A comparative study of the template-assisted enzymatic and chemical polymerization of aniline in a buffer solution of sodium dodecylbenzenesulfonate micelles was performed for the first time. The high-redox potential laccase from the fungus Trametes hirsuta was used as a catalyst and air oxygen served as an oxidant. Potentiometric and spectral methods have shown that oligomeric/polymeric products of the enzymatic polymerization of aniline are synthesized in the conducting emeraldine salt form immediately after the reaction is initiated by the enzyme. The use of the laccase-mediator system enabled a higher rate of enzymatic polymerization and a higher yield of final products. Potassium octocyanomolybdate (IV) served as a redox mediator. The products of the enzymatic polymerization of aniline were studied by the ATR-FTIR, MALDI-TOF and atomic force microscopy methods. The chemical oxidative polymerization of aniline under the same conditions resulted in forming a non-conducting dark brown product.

Chemoenzymatic cascade reaction for the production of furan-based polyesters with different thermal performance

The oxidation products of 5-hydroxymethylfurfural (HMF) have been classified as favorable bio-source building blocks and employed to the production of polymers. Herein, a chemoenzymatic cascade reaction was proposed for the production of furan-based polyesters. Ruthenium lignosulphonate (Ru-LigS) was prepared using a facile hydrothermal process for HMF oxidation. The synergistic effect between ruthenium and sodium lignin sulfonate promoted this process, resulting in obvious improvement in catalytic performance. After separation, two furan-based polyesters were prepared through enzymatic polymerization by using two HMF oxidation products as the monomers. The minor difference in structure afforded a significant difference in the thermal properties of these two polyesters. Self-polymerized polyester derived from 5-hydroxymethyl-2-furancarbonxylic acid (HMFCA) was amorphous along with a glass transition temperature as low as −43.9 °C, demonstrating excellent cold resistance performance. Instead, the polyester polymerized by 2,5-furandicarboxylic acid (FDCA) and 2,5-bis(hydroxymethyl)furan (BHMF) was crystalline and possessed good thermodynamic properties. This study presented sustainable chemical conversion processes for both oxidizing HMF using highly active biomacromolecule catalysts and producing valuable furan-based polyesters.

Engineered polysaccharide alpha‐1,3‐glucan in highly filled thermoplastic polyurethane systems

AbstractThis study explores the use of α‐1,3‐glucan, which can be produced through an enzymatic polymerization process, as additive for thermoplastic polyurethane composite systems. Most importantly, highly filled thermoplastic polyurethane composites can be achieved with up to 80% by weight polysaccharide content. Analysis of mechanical properties demonstrates that strength and modulus can be significantly improved, while the plasticizer content allows for adjustments of compound flexibility. The addition of 60% α‐1,3‐glucan increased the flexural modulus of the thermoplastic polyurethane by 497% while adding 80% α‐1,3‐glucan increased the flexural modulus by 3193%. Citrate esters can be incorporated as plasticizer to improve processibility and flexibility of the compounds. An increase in impact strength of 55% was achieved when 10% of tributyl citrate was added to the 80% α‐1,3‐glucan composite. Based on the compatibility and property enhancements, the use of these engineered polysaccharides may provide additional options for producing sustainable composite materials with high renewable content and unique properties.

Enzymatic polymerization of enantiomeric L−3,4-dihydroxyphenylalanine into films with enhanced rigidity and stability

L−3,4-dihydroxyphenylalanine is an important molecule in the adhesion of mussels, and as an oxidative precursor of natural melanin, it plays an important role in living system. Here, we investigate the effect of the molecular chirality of 3,4-dihydroxyphenylalanine on the properties of the self-assembled films by tyrosinase-induced oxidative polymerization. The kinetics and morphology of pure enantiomers are completely altered upon their co-assembly, allowing the fabrication of layer-to-layer stacked nanostructures and films with improved structural and thermal stability. The different molecular arrangements and self-assembly mechanisms of the L+D-racemic mixtures, whose oxidation products have increased binding energy, resulting in stronger intermolecular forces, which significantly increases the elastic modulus. This study provides a simple pathway for the fabrication of biomimetic polymeric materials with enhanced physicochemical properties by controlling the chirality of monomers.

Hydrogelation from Self-Assembled and Scaled-Down Chitin Nanofibers by the Modification of Highly Polar Substituents.

Chitin nanofibers (ChNFs) with a bundle structure were fabricated via regenerative self-assembly at the nanoscale from a chitin ion gel with an ionic liquid using methanol. Furthermore, the bundles were disentangled by partial deacetylation under alkaline conditions, followed by cationization and electrostatic repulsion in aqueous acetic acid to obtain thinner nanofibers called scaled-down ChNFs. This review presents a method for hydrogelation from self-assembled and scaled-down ChNFs by modifying the highly polar substituents on ChNFs. The modification was carried out by the reaction of amino groups on ChNFs, which were generated by partial deacetylation, with reactive substituent candidates such as poly(2-oxazoline)s with electrophilic living propagating ends and mono- and oligosaccharides with hemiacetallic reducing ends. The substituents contributed to the formation of network structures from ChNFs in highly polar dispersed media, such as water, to produce hydrogels. Moreover, after the modification of the maltooligosaccharide primers on ChNFs, glucan phosphorylase-catalyzed enzymatic polymerization was performed from the primer chain ends to elongate the amylosic graft chains on ChNFs. The amylosic graft chains formed double helices between ChNFs, which acted as physical crosslinking points to construct network structures, giving rise to hydrogels.

Preparation of fabric-based electronic device via surface topological modification and enzymatic polymerization for personal thermal management, subtle motion detection, and ultraviolet protection

With the increasingly severe problem of population aging, human motion detection and personal thermal management have attracted widespread attention in the field of smart wearable electronics. However, the current multifunctional flexible devices face problems, such as lower stability and poor comfort, which greatly hinder their further development. In this work, an improved fabrication strategy combining in situ enzymatic polymerization and surface topological modification is proposed to facile and efficiently prepare poly(3,4-ethylenedioxythiophene)/polyacrylamide (PEDOT/PAAm) smart fabric. To increase the conductivity and the bonding force between fabric and PEDOT particles, the PAAm was used to pre-construct a microgel layer on the surface of cotton fabric to make PEDOT uniformly coated on the fabric, thus forming a highly electrically conductive interconnection network. The resulting fabric samples show outstanding duel-driven thermal performance (88.6 °C for 2 sun, 84.5 °C for 10 V), satisfactory detection capability for subtle and rapid motion (drops of water and paper pill, bending of fingers), and remarkable ultraviolet protection performance (24.6 times than the UPF value of unmodified fabric sample). All in all, multifunctional fabric-based devices with remarkable performance exhibited great application potential in rehabilitation therapy and personal home care, especially for people with disabilities and older people.

Preparation of multifunctional composite materials based on PEDOT via enzymatic cascade polymerization

Bioenzyme-catalyzed conductive polymer polymerization is an ecological and environmental protection method that aligns with the current sustainable development plan. Hence, a new strategy of in situ assembly of poly (3,4-ethylenedioxythiophene) (PEDOT) on the surface of fabrics via free radical polymerization initiated by the cascade reaction of glucose oxidase (GOx) and horseradish peroxidase (HRP) was introduced. With excellent photothermal conversion capability and sensitive humidity response, PEDOT endows the fabric with remarkable photothermal performance, humidity sensing performance, and UV resistance capability. It is worth noting that the PEDOT-coated fabric can reach a fantastic 54.2 °C within 20 s under 2 sun irradiation intensity and finally reach 98.4 °C. In addition, it can maintain stable and fast responses in repeated and long-term operations. Moreover, the PEDOT-coated fabric has a fast response and long-term working ability in high humidity. This work provides a new environmental protection and sustainability idea for designing and developing conductive polymer-based high-performance composites.

Synthesis of Functionalized Triblock Copolyesters Derived from Lactic Acid and Macrolactones for Bone Tissue Regeneration.

Synthetic and functional grafts are a great alternative to conventional grafts. They can provide a physical support and the precise signaling for cells to heal damaged tissues. In this study, a novel RGD peptide end-functionalized poly(ethylene glycol)-b-poly(lactic acid)-b-poly(globalide)-b-poly(lactic acid)-b-poly(ethylene glycol) (RGD-PEG-PLA-PGl-PLA-PEG-RGD) is synthetized and used to prepare functional scaffolds. The PGl inner block is obtained by enzymatic ring-opening polymerization of globalide. The outer PLA blocks are obtained by ring-opening polymerization of both, l-lactide or a racemic mixture, initiated by the α-ω-telechelic polymacrolactone. The presence of PGl inner block enhances the toughness of PLA-based scaffolds, with an increase of the elongation at break up to 300% when the longer block of PGl is used. PLA-PGl-PLA copolymer is coupled with α-ω-telechelic PEG diacids by esterification reaction. PEGylation provides hydrophilic scaffolds as the contact angle is reduced from 114° to 74.8°. That difference improves the contact between the scaffolds and the culture media. Moreover, the scaffolds are functionalized with RGD peptides at the surface significantly enhancing the adhesion and proliferation of bone marrow-derived primary mesenchymal stem cells and MC3T3-E1 cell lines in vitro. These results place this multifunctional polymer as a great candidate for the preparation of temporary grafts.

Effect of enzymatic polymerization on the thermal stability of flavonoids

The mechanisms of the enzymatic polymerization of flavonoids, including quercetin, rutin and catechins, have been fairly well researched, but the properties of polymeric forms of flavonoids still require in-depth analysis. The products of enzymatic polymerization are oligomeric flavonoids. The antioxidant and antimicrobial properties of many oligomeric flavonoids have been described in the literature. However, data on the thermal properties of oligomeric flavonoids are lacking and the supplementing of these deficiencies is a scientific novelty of this work. The aim of this study is to determine the effect of enzymatic polymerization on thermal stability of oligomeric flavonoids. As part of the work, oligomeric quercetin, rutin and catechin were prepared by the enzymatic polymerization reaction with laccase and horseradish peroxidase enzymes. The oligomeric structure of the flavonoids was confirmed by spectroscopic (Fourier transform infrared spectroscopy (FTIR) and UV–visible spectroscopy (UV–Vis)) as well as chromatographic (ultra-performance liquid chromatography (UPLC) and gel permeation chromatography (GPC)) methods. Thermal properties of oligomeric flavonoids were investigated using differential scanning calorimetry (DSC) and thermogravimetry (TG). Moreover, the antioxidant and antimicrobial properties of oligomeric forms of selected polyphenols were investigated. Based on DSC analysis, it was found that the common feature of all oligomeric flavonoids was a higher final oxidation temperature and a higher oxidation enthalpy than the reference flavonoids. Thermogravimetric analysis showed that oligomeric poly(flavonoids)-laccase had better thermal stability, which correlated with higher molar mass.

Fungal-Lactobacteria Consortia and Enzymatic Catalysis for Polylactic Acid Production.

Polylactic acid (PLA) is the main biobased plastic manufactured on an industrial scale. This polymer is synthetized by chemical methods, and there is a strong demand for the implementation of clean technologies. This work focuses on the microbial fermentation of agro-industrial waste rich in starch for the production of lactic acid (LA) in a consolidated bioprocess, followed by the enzymatic synthesis of PLA. Lactic acid bacteria (LAB) and the fungus Rhizopus oryzae were evaluated as natural LA producers in pure cultures or in fungal-lactobacteria co-cultures formed by an LAB and a fungus selected for its metabolic capacity to degrade starch and to form consortia with LAB. Microbial interaction was analyzed by scanning electron microscopy and biofilm production was quantified. The results show that the fungus Talaromyces amestolkiae and Lactiplantibacillus plantarum M9MG6-B2 establish a cooperative relationship to exploit the sugars from polysaccharides provided as carbon sources. Addition of the quorum sensing molecule dodecanol induced LA metabolism of the consortium and resulted in improved cooperation, producing 99% of the maximum theoretical yield of LA production from glucose and 65% from starch. Finally, l-PLA oligomers (up to 19-LA units) and polymers (greater than 5 kDa) were synthetized by LA polycondensation and enzymatic ring-opening polymerization catalyzed by the non-commercial lipase OPEr, naturally produced by the fungus Ophiostoma piceae.

An Antibacterial Polypeptide Coating Prepared by In Situ Enzymatic Polymerization for Preventing Delayed Infection of Implants.

Delayed implant-associated infection is an important challenge, as the treatment involves a high risk of implant replacement. Mussel-inspired antimicrobial coatings can be applied to coat a variety of implants in a facile way, but the adhesive 3,4-dihydroxyphenylalanine (DOPA) group is prone to oxidation. Therefore, an antibacterial polypeptide copolymer poly(Phe7-stat-Lys10)-b-polyTyr3 was designed to prepare the implant coating upon tyrosinase-induced enzymatic polymerization for preventing implant-associated infections. Both poly(Phe7-stat-Lys10) and polyTyr3 blocks have specific functions: the former provides intrinsic antibacterial activity with a low risk to induce antimicrobial resistance, and the latter is attachable to the surface of implants to rapidly generate an antibacterial coating by in situ injection of polypeptide copolymer since tyrosine could be oxidized to DOPA under catalyzation of skin tyrosinase. This polypeptide coating with excellent antibacterial effect and desirable biofilm inhibition activity is promising for broad applications in a multitude of biomedical materials to combat delayed infections.

Crystallinity-tuned ultrasoft polymeric DNA networks for controlled release of anticancer drugs.

Despite the vast interest in utilizing rolling circle amplification (RCA)-based DNA networks for bioapplications, precise control of the mechanical and physicochemical properties is highly challenging. To address this concern, we aimed to develop ultrasoft self-supporting polymerized DNA networks (pDNets) of variable crystallinities to manipulate sequence-mediated drug release efficiency. A controlled ratio of the inorganic magnesium pyrophosphate (MgPPi) crystal to the organic polymeric DNA resulted in the synthesis of pDNets of various nanoporosities. The number of crystal microstructures influencing drug localization and release pattern and the tunable mechanical properties influencing injectability and structural stability under physiological conditions were investigated. The pDNets exhibited ultrasoft properties with Young's moduli of 0.06-0.54Pa; approximately 9-fold differences in mechanical properties were obtained by varying the degree of crystallinity. With functional DNA sequences, the developed platforms showed pH stimuli-responsive drug release profiles of the dynamic DNA structures and aptamer-specific cell target adhesion efficiency. Analyses of controlled delivery of anticancer therapeutics in vitro and in vivo revealed crystallinity-dependent antitumor efficacy without side effects. This strategy provides an effective one-pot enzymatic polymerization methodology and a favorable microenvironment for a three-dimensional DNA network based on demand-localized drug delivery.

Metabolite-induced in vivo fabrication of substrate-free organic bioelectronics.

Interfacing electronics with neural tissue is crucial for understanding complex biological functions, but conventional bioelectronics consist of rigid electrodes fundamentally incompatible with living systems. The difference between static solid-state electronics and dynamic biological matter makes seamless integration of the two challenging. To address this incompatibility, we developed a method to dynamically create soft substrate-free conducting materials within the biological environment. We demonstrate in vivo electrode formation in zebrafish and leech models, using endogenous metabolites to trigger enzymatic polymerization of organic precursors within an injectable gel, thereby forming conducting polymer gels with long-range conductivity. This approach can be used to target specific biological substructures and is suitable for nerve stimulation, paving the way for fully integrated, in vivo-fabricated electronics within the nervous system.

Tyrosine-Derived Polymers as Potential Biomaterials: Synthesis Strategies, Properties, and Applications.

Peptide-based polymers are evolving as promising materials for various biomedical applications. Among peptide-based polymers, polytyrosine (PTyr)-based and l-tyrosine (Tyr)-derived polymers are unique, due to their excellent biocompatibility, degradability, and functional as well as engineering properties. To date, different polymerization techniques (ring-opening polymerization, enzymatic polymerization, condensation polymerization, solution-interfacial polymerization, and electropolymerization) have been used to synthesize various PTyr-based and Tyr-derived polymers. Even though the synthesis starts from Tyr, different synthesis routes yield different polymers (polypeptides, polyarylates, polyurethanes, polycarbonates, polyiminocarbonate, and polyphosphates) with unique functional characteristics, and these polymers have been successfully used for various biomedical applications in the past decades. This Review comprehensively describes the synthesis approaches, classification, and properties of various PTyr-based and Tyr-derived polymers employed in drug delivery, tissue engineering, and biosensing applications.

Vine-Twining Inclusion Behavior of Amylose towards Hydrophobic Polyester, Poly(β-propiolactone), in Glucan Phosphorylase-Catalyzed Enzymatic Polymerization.

This study investigates inclusion behavior of amylose towards, poly(β-propiolactone) (PPL), that is a hydrophobic polyester, via the vine-twining process in glucan phosphorylase (GP, isolated from thermophilic bacteria, Aquifex aeolicus VF5)-catalyzed enzymatic polymerization. As a result of poor dispersibility of PPL in sodium acetate buffer, the enzymatically produced amylose by GP catalysis incompletely included PPL in the buffer media under the general vine-twining polymerization conditions. Alternatively, we employed an ethyl acetate-sodium acetate buffer emulsion system with dispersing PPL as the media for vine-twining polymerization. Accordingly, the GP (from thermophilic bacteria)-catalyzed enzymatic polymerization of an α-d-glucose 1-phosphate monomer from a maltoheptaose primer was performed at 50 °C for 48 h in the prepared emulsion to efficiently form the inclusion complex. The powder X-ray diffraction profile of the precipitated product suggested that the amylose-PPL inclusion complex was mostly produced in the above system. The 1H NMR spectrum of the product also supported the inclusion complex structure, where a calculation based on an integrated ratio of signals indicated an almost perfect inclusion of PPL in the amylosic cavity. The prevention of crystallization of PPL in the product was suggested by IR analysis, because it was surrounded by the amylosic chains due to the inclusion complex structure.

Overcoming challenges in the synthesis of a lignin-carbohydrate complex (LCC) model: Mitsunobu versus Appel product

Arabinoxylan ferulate representing a macromolecular LCC model valid for annual plants is synthesized under Mitsunobu conditions. The content of ferulic acid ester is tuned by the reaction conditions achieving degree of substitution values from 0.09 to 0.45. Utilization of the chloride-free solvent N-methyl-2-pyrrolidone allows the design of pure Mitsunobu products without occurrence of deoxychloro moieties arising from Appel type reaction. 2D NMR experiments reveal nature-identical structure of ferulate moieties present at position 5 of the arabinose side chain. Enzymatic dehydrogenation polymerization of coniferyl alcohol on ferulate anchor groups under homogeneous conditions lead to beta -O-4, beta -5, and Hibbert ketone structures identified by Py-GC-MS. The results are valuable to study structure-property relationships within the formation of natural and non-native lignins.Graphical abstract

Fibrin-Rhamnogalacturonan I Composite Gel for Therapeutic Enzyme Delivery to Intestinal Tumors.

Therapy of colorectal cancer with protein drugs, including targeted therapy using monoclonal antibodies, requires the preservation of the drug's structure and activity in the gastrointestinal tract or bloodstream. Here, we confirmed experimentally the fundamental possibility of creating composite protein-polysaccharide hydrogels based on non-degrading rhamnogalacturonan I (RG) and fibrin as a delivery vehicle for antitumor RNase binase. The method is based on enzymatic polymerization of fibrin in the presence of RG with the inclusion of liposomes, containing an encapsulated enzyme drug, into the gel network. The proposed method for fabricating a gel matrix does not require the use of cytotoxic chemical cross-linking agents and divalent cations, and contains completely biocompatible and biodegradable components. The process proceeds under physiological conditions, excluding the effect of high temperatures, organic solvents and ultrasound on protein components. Immobilization of therapeutic enzyme binase in the carrier matrix by encapsulating it in liposomes made from uncharged lipid made it possible to achieve its prolonged release with preservation of activity for a long time. The release time of binase from the composite carrier can be regulated by variation of the fibrin and RG concentration.

Influence of the enzymatic and the chemical oxidative polymerization of trifluoromethyl‐substituted aromatic diamine on thermal and photophysical properties

AbstractSemi‐conductive polymers have been used in a few area applications such as coatings, accumulators, solar cells, rechargeable batteries, ion sensors, photoreceptors, light‐emitting diodes and electrochromic instruments. Poly(4‐(trifluoromethyl)benzene‐1,2‐diamine)s of Poly(TFMBDA)‐O and Poly(TFMBDA)‐E were synthesized via chemical oxidative and enzymatic polymerization methods, respectively, using 4‐(trifluoromethyl)benzene‐1,2‐diamine (TFMBDA) as monomer. FT‐IR, UV–vis and 1H‐NMR and 13C‐NMR instruments were utilized in order to confirm the structures of all compounds. The Mw values of the polymers and their PDI values were found to be between 5000–7000 Da and 1.045–1.115, respectively, from GPC measurements. The glass temperature of phenazine‐type structure of Poly(TFMBDA)‐O was higher compared to that of Poly(TFMBDA)‐E, which was composed of phenyl units linked with –NH bridges. As soon as Poly(TFMBDA)‐O and Poly(TFMBDA)‐E were excited by UV light, they had a quantum efficiency of 6.3% and 13.7%, respectively, in DMF for violet photoluminescence (PL) emission depending on photoluminescence measurement. SEM images revealed that Poly(TFMBDA)‐O consisted of approximately 1–2 μm of uniform micro‐spheres and a granular surface with different pore diameters. The enzymatic oxidation with HRP afforded Poly(TFMBDA)‐E micro‐rods of about 5–10 μm‐long shape in their SEM images.

Harnessing the power of polyol-based polyesters for biomedical innovations: synthesis, properties, and biodegradation.

Polyesters based on polyols have emerged as promising biomaterials for various biomedical applications, such as tissue engineering, drug delivery systems, and regenerative medicine, due to their biocompatibility, biodegradability, and versatile physicochemical properties. This review article provides an overview of the synthesis methods, performance, and biodegradation mechanisms of polyol-based polyesters, highlighting their potential for use in a wide range of biomedical applications. The synthesis techniques, such as simple polycondensation and enzymatic polymerization, allow for the fine-tuning of polyester structure and molecular weight, thereby enabling the tailoring of material properties to specific application requirements. The physicochemical properties of polyol-based polyesters, such as hydrophilicity, crystallinity, and mechanical properties, can be altered by incorporating different polyols. The article highlights the influence of various factors, such as molecular weight, crosslinking density, and degradation medium, on the biodegradation behavior of these materials, and the importance of understanding these factors for controlling degradation rates. Future research directions include the development of novel polyesters with improved properties, optimization of degradation rates, and exploration of advanced processing techniques for fabricating scaffolds and drug delivery systems. Overall, polyol-based polyesters hold significant potential in the field of biomedical applications, paving the way for groundbreaking advancements and innovative solutions that could revolutionize patient care and treatment outcomes.

Self-decorating cells via surface-initiated enzymatic controlled radical polymerization.

Through the innovative use of surface-displayed horseradish peroxidase, this work explores the enzymatic catalysis of both bioRAFT polymerization and bioATRP to prompt polymer synthesis on the surface of Saccharomyces cerevisiae cells, with bioATRP outperforming bioRAFT polymerization. The resulting surface modification of living yeast cells with synthetic polymers allows for a significant change in yeast phenotype, including growth profile, aggregation characteristics, and conjugation of non-native enzymes to the clickable polymers on the cell surface, opening new avenues in bioorthogonal cell-surface engineering.