Biomimetic Mineralization of Protein Nanogels for Enzyme Protection

Protein nanogels have found a wide variety of applications, ranging from biocatalysis to drug/protein delivery. However, in practical applications, proteins in nanogels may suffer from enzymic hydrolysis and denaturation. Inspired by the structure and functionalities of the fowl eggshells, biomimetic mineralization of protein nanogels was studied in this research. Protein nanogels with embedded porcine pancreas lipase (PPL) in the cross-linked nanostructures were synthesized through the thiol–disulfide reaction between thiol-functionalized PPL and poly(N-isopropylacrylamide) with pendant pyridyl disulfide groups. The nanogels were further reacted with reduced bovine serum albumin (BSA) and BSA molecules were coated on the nanogels. Mineralization of BSA leads to the synthesis of biomineralized shells on the nanogels. With the growth of CaCO₃ on the shells, the nanogels aggregate into suprastructures. Thermogravimetric analysis, XRD, dynamic light scattering, and TEM were employed to study the mechanism of the biomineralization process and analyze the structures of the mineralized nanogels. The biomineralized shells can effectively protect the PPL molecules from hydrolysis by trypsin; meanwhile, the nanosized channels on the mineralized shells allow the transport of small-molecule substrates across the shells. Bioactivity measurements indicate that PPL in the nanogels maintains more than 80 % bioactivity after biomineralization.     

Towards Photochromic Azobenzene-Based Inhibitors for Tryptophan Synthase

Light regulation of drug molecules has gained growing interest in biochemical and pharmacological research in recent years. In addition, a serious need for novel molecular targets of antibiotics has emerged presently. Herein, the development of a photocontrollable, azobenzene-based antibiotic precursor towards tryptophan synthase (TS), an essential metabolic multienzyme complex in bacteria, is presented. The compound exhibited moderately strong inhibition of TS in its E configuration and five times lower inhibition strength in its Z configuration. A combination of biochemical, crystallographic, and computational analyses was used to characterize the inhibition mode of this compound. Remarkably, binding of the inhibitor to a hitherto-unconsidered cavity results in an unproductive conformation of TS leading to noncompetitive inhibition of tryptophan production. In conclusion, we created a promising lead compound for combatting bacterial diseases, which targets an essential metabolic enzyme, and whose inhibition strength can be controlled with light.     

Synthesis of poly(ω‐pentadecalactone)‐b‐poly(acrylate) diblock copolymers via a combination of enzymatic ring‐opening and RAFT polymerization techniques

Herein the first reported preparation of diblock copolymers of the polyethylene‐like polyester poly(ω‐pentadecalactone) (PPDL) via a combination of enzymatic ring‐opening polymerization (eROP) and reversible addition‐fragmentation chain‐transfer (RAFT) polymerization techniques is described. PPDL was synthesized via eROP using Novozyme 435 as a catalyst and a bifunctional initiator/chain transfer agent (CTA) appropriate for the eROP of ω‐pentadecalactone (PDL) and RAFT polymerization of acrylic and styrenic monomers. Chain growth of the PPDL macro‐CTA was performed to prepare acrylic and styrenic diblock copolymers of PPDL, and demonstrates a facile, metal‐free, and “greener” alternative to preparing acrylic diblock copolymers of polyethylene (PE). Diblock copolymer architecture was substantiated via analysis of ¹H NMR spectroscopic, UV‐GPC chromatographic, DSC onset crystallization (T_c), and MALDI‐ToF mass spectrometric data. © 2016 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2016 , 54, 3326–3335     

Synthesis of Enantiomeric Aminophosphonic Acids and Peptides

Abstract

Synthesis of enantiomeric amino phosphonic acids APA is described by using chiral auxiliary reagent or enzymatic resolution of racemic mixtures of APA phenacyl derivatives. Peptides with APA residue were obtained by application of trimethylsilyl derivatives or condensation in the presence of enzyme-papain     

Synthesis of Main‐Chain Liquid‐Crystalline Polyesters Containing Diphenyl Mesogens by Chemo‐Enzymatic Route †

Main‐chain thermotropic liquid‐crystalline polyesters containing rigid biphenyl mesogens and flexible spacers were synthesized by chemo‐enzymatic route. The enzyme‐catalyzed polymerization showed high regio‐ and chemo‐ selectivity, and is environmentally friendly. The resulting polyesters were characterized with ¹H‐NMR, ¹³C‐NMR, gel permeation chromatography (GPC), differential scanning calorimetry (DSC), and polarized light optical microscopy (POM).     

Simulations of Redox Mediation within Bioactive Hydrogels of Amperometric Biosensors

Highly hydrated bioactive hydrogels containing immobilized oxidoreductase enzymes and immobilized redox mediators were simulated as the biorecognition layer of amperometric biosensors. The linear dynamic range of the amperometric response of mediated biosensors increases and moves to higher concentration brackets with an increase in the concentration of mediator. This informs the design of biosensors that target the same analyte but possesses several independently addressable electrodes modified with hydrogels that contain different concentrations of mediator. Increases in enzyme concentration increase the linear dynamic range but does not alter the sensitivity of amperometric biosensors. Both sensitivity and linear dynamic range of mediated amperometric enzyme biosensors may be “tuned” by varying the concentrations of the enzyme and the mediator. Simulations effectively guide the initial domains of study of complex systems such as implantable biosensors.     

Catalytic Oxidative Cyclocondensation of o‐Aminophenols to 2‐Amino‐3H‐phenoxazin‐3‐ones

The catalytic oxidative cyclocondensation of the o‐aminophenols 1a–f was investigated. The oxidants used were air/laccase, H₂O₂/horseradish peroxidase, H₂O₂/ebselen (3), and TBHP/diphenyl diselenide 4. The products obtained were 2‐amino‐3H‐phenoxazin‐3‐one—questiomycin A, its derivative 2b, and cinnabarinic acid and actinocin (2c,d). Substrates with methyl groups at 4 and 5 positions of benzene ring were converted to different dihydrophenoxazinones 2g,h. Compounds having chlorine atoms at the same positions underwent oxidation to planar phenoxa-zinones 2e,f with elimination of one hydrochloride molecule.     

Photosensitive cholesteric copolymers with spiropyran-containing side groups II. Kinetic features of the photo- and thermo-chromic processes

Kinetic features of the photo- and thermo-chromic behaviour of new cholesteric triple copolymers with photosensitive spiropyran-containing side groups are considered. The kinetics of the back thermal process at different temperatures have been studied; activation energies for this process for the copolymers with different contents of the dye units have been obtained. The photochromic homopolymer and copolymers with a low content of dye units are shown to possess maximum values of the activation energy.     

Enzymatic syntheses of unsaturated polyesters based on isosorbide and isomannide

The search for materials produced from renewable sources aiming at the substitution of petroleum‐based derivates is an area of intense investigation. In this work, the enzymatic copolymerization of isosorbide or isomannide with diethyl adipate and fractions of different unsaturated diesters (diethyl itaconate, diethyl fumarate, diethyl glutaconate, and diethyl hydromuconate) were examined using CAL‐B as catalyst. The polyesters prepared using one‐step syntheses were characterized by SEC, NMR, and MALDI‐TOF MS. In addition, syntheses with linear diols were carried out in bulk to evaluate the reactivity of cyclic diols in producing unsaturated polyesters using enzymatic catalysis, as well as to evaluate the occurrence of addition side reactions on the double bonds. Isosorbide and isomannide yielded unsaturated polymers with values in the order of 4,000‐16,000 when fumarate or glutaconate esters were added in 5 mol % ratio against adipate. In all cases MALDI‐TOF confirmed the presence of unsaturated units. Although these polyesters have unreacted double bonds they are prone to crosslinking and ready to further functionalization, like anchoring bioactive molecules. © 2013 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2013 , 51, 3881–3891