Homoserine Lactones,Methyl Oligohydroxybutyrates,and Other Extracellular Metabolites of Macroalgae‐Associated Bacteria of the Roseobacter Clade: Identification and Functions

Twenty‐four strains of marine Roseobacter clade bacteria were isolated from macroalgae and investigated for the production of quorum‐sensing autoinducers, N‐acylhomoserine lactones (AHLs). GC/MS analysis of the extracellular metabolites allowed us to evaluate the release of other small molecules as well. Nineteen strains produced AHLs, ranging from 3‐OH‐C10:0‐HSL (homoserine lactone) to (2E,11Z)‐C18:2‐HSL, but no specific phylogenetic or ecological pattern of individual AHL occurrence was observed when cluster analysis was performed. Other identified compounds included indole, tropone, methyl esters of oligomers of 3‐hydroxybutyric acid, and various amides, such as N‐9‐hexadecenoylalanine methyl ester (9‐C16:1‐NAME), a structural analogue of AHLs. Several compounds were tested for their antibacterial and antialgal activity on marine isolates likely to occur in the habitat of the macroalgae. Both AHLs and 9‐C16:1‐NAME showed high antialgal activity against Skeletonema costatum, whereas their antibacterial activity was low.     

Additive Manufacturing of Dense Ceramic Parts via Direct Ink Writing of Aqueous Alumina Suspensions

Additive manufacturing of near‐net‐shaped dense ceramic components has been established via room‐temperature direct writing of highly loaded aqueous alumina suspensions in a layer‐by‐layer fashion. The effect of alumina solid loading on rheology, specimen uniformity, density, microstructure, and mechanical properties was studied. All suspensions contained a polymer binder (~5 vol.%), dispersant, and 51–58 vol.% alumina powder. Rheological measurements indicated all suspensions to be yield‐pseuodoplastic, and both yield stress and viscosity were found to increase with increasing alumina solid loading. Shear rates ranging from 19.5 to 24.2/s, corresponding to viscosities of 9.8 to 17.2 Pa·s, for the 53–56 vol.% alumina suspensions were found to produce the best results for the 1.25‐mm tip employed during writing. All parts were sintered to >98% of true density, with grain sizes ranging from 3.2 to 3.7 μm. The average flexure strength, which ranged from 134 to 157 MPa, was not influenced by the alumina solid loading.     

Protein Engineering of the Progesterone Hydroxylating P450‐Monooxygenase CYP17A1 Alters Its Regioselectivity

The CYP171 enzyme is known to catalyse a key step in the steroidogenesis of mammals. The substrates progesterone and pregnenolone are first hydroxylated at the C17 position, and this is followed by cleavage of the C17?C20 bond to yield important precursors for glucosteroids and androgens. In this study, we focused on the reaction of the bovine CYP17A1 enzyme with progesterone as a substrate. On the basis of a created homology model, active‐site residues were identified and systematically mutated to alanine. In whole‐cell biotransformations, the importance of the N202, R239, G297 and E305 residues for substrate conversion was confirmed. Additionally, mutation of the L206, V366 and V483 residues enhanced the formation of the 16α‐hydroxyprogesterone side product up to 40 % of the total product formation. Furthermore, residue L105 was found not to be involved in this side activity, which contradicts a previous study with the human enzyme.     

Benign route for the modification and characterization of poly(lactic acid) (PLA) scaffolds for medicinal application

Scaffolds fabricated from polymers have imprinted its wide applicability in the field of tissue engineering. The surface of electrospun poly(lactic acid) (PLA) nanofibers was modified to improve their compatibility with living medium. PLA film were treated with alkali solution to introduce carboxyl groups on the surface followed by covalent grafting of gelatin using Xtal Fluoro‐E as coupling agent. The gelatin g‐PLA polymer synthesized via ‘graft‐onto’ method exhibit fascinating properties as studied by contact angle measurement, fourier transformed infrared spectroscopy, thermogravimetric analysis, scanning electron microscopy, water vapor transmission rate(WVTR), swelling studies and differential scanning calorimetry. The fabricated gelatin g‐PLA scaffolds were further characterized to conduct the study on hydrolytic degradation, and extent of biodegradation at ambient temperature. It was observed from the in‐vitro analysis that the gelatin g‐PLA nanofiber (with hemolytic percentage, 0.56 ± 0.13%) was cytocompatible with fibroblast cell and does not impair cell growth. The WVTR obtained for the electrospun mat around 2900 ± 100 g/m². 24 h signifies the optimal moist environment required for tissue engineering especially wound healing. Notably, many of these strategies resulted in porous hydrophilic scaffolds with human cell growth and proliferation for medical applications of various types. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 46056.     

Lipase PS-catalyzed transesterification of citronellyl butyrate and geranyl caproate: Effect of reaction parameters

Pseudomonas sp. lipase PS was immobilized by adsorption and tested for its ability to catalyze the synthesis of citronellyl butyrate and geranyl caproate by transesterification in n-hexane. The reaction parameters investigated were: enzyme load, effect of substrate concentration, added water, temperature, time course, organic solvent, pH memory, and enzyme reuse. Yields as high as 96 and 99% were obtained for citronellyl butyrate and geranyl caproate, respectively, with 300 units (approx. 15% w/w of reactants) of lipase PS. Increasing amounts of terpene alcohol inhibited lipase activity, while excess acyl donor (triacylglycerol) concentration enhanced ester production. Optimal yields were obtained at temperatures from 30–50°C after 24-h incubation time. Yields of 90 and 99% were obtained for citronellyl and geranyl esters, respectively, with 2% added water. Solvents with log P values ≥ 2.5 showed the highest conversion yields. pH 7 and 6–8 seemed to be ideal for citronellyl butyrate and geraniol caproate, respectively. The lipase remained active after reusing 12 times.     

Fluorescent Probe DCVJ Shows High Sensitivity for Characterization of Amyloid β-Peptide Early in the Lag Phase

The aggregation of intrinsically disordered and misfolded proteins in the form of oligomers and fibrils plays a crucial role in a number of neurological and neurodegenerative diseases. Currently, most probes and biophysical techniques that detect and characterize fibrils at high resolution fail to show sensitivity and binding for oligomers. Here, we show that 9-(dicyano-vinyl)julolidine (DCVJ), a class of molecular rotor, binds amyloid beta (Aβ) early aggregates, and we report the kinetics as well as packing of the oligomer formation. The binding of DCVJ to Aβ40 increased its emission intensity with time at 510 nm and produced a second excimer peak at 575 nm. However, DCVJ did not bind to the prefibrillar aggregates of Aβ42, which indicated that the oligomers formed by Aβ40 and Aβ42 were not the same. The F4C F19W mutant of Aβ40, which did not form fibrils, also bound DCVJ, but the emission spectral profile varied from that of the wild-type (WT). Atomic force microscopy images of WT Aβ40, the F4C F19W mutant, and Aβ42 oligomers displayed differences in size and shape, confirming the difference in their DCVJ spectra. The effect of epigallocatechin-3-gallate (EGCG) on the reduction of Aβ42 fibrils was also observed with finer detail than with other techniques. The results of this study show that DCVJ detects early aggregates and provides valuable information regarding the oligomer kinetics, packing, and mechanism of formation.     

Enzymatic Addition of Alcohols to Terpenes by Squalene Hopene Cyclase Variants

Squalene–hopene cyclases (SHCs) catalyze the polycyclization of squalene into a mixture of hopene and hopanol. Recently, amino-acid residues lining the catalytic cavity of the SHC from Alicyclobacillus acidocaldarius were replaced by small and large hydrophobic amino acids. The alteration of leucine 607 to phenylalanine resulted in increased enzymatic activity towards the formation of an intermolecular farnesyl–farnesyl ether product from farnesol. Furthermore, the addition of small-chain alcohols acting as nucleophiles led to the formation of non-natural ether-linked terpenoids and, thus, to significant alteration of the product pattern relative to that obtained with the wild type. It is proposed that the mutation of leucine at position 607 may facilitate premature quenching of the intermediate by small alcohol nucleophiles. This mutagenesis-based study opens the field for further intermolecular bond-forming reactions and the generation of non-natural products.     

Chitosan‐based hydrogels: recent design concepts to tailor properties and functions

Chitosan (CS) has received much attention as a functional biopolymer for designing various hydrogels for biomedical applications. This review provides an overview of the different types of CS‐based hydrogels, the approaches that can be used to fabricate hydrogel matrices with specific features and their applications in controlled drug delivery and tissue engineering. Emphasis is laid on the recent design concepts of hybrid hydrogels based on mixtures of CS and natural or synthetic polymers, interpenetrating polymer networks as well as composite hydrogels prepared by embedding nanoparticles into CS matrices. © 2017 Society of Chemical Industry     

Horizontal Dip‐Spin Casting of aqueous alumina‐polyvinylpyrrolidone suspensions with chopped fiber

A novel ceramic processing method, called Horizontal Dip Spin Casting (HDSC), enabled fabrication of tubular ceramic parts with an aligned chopped fiber phase. HDSC was demonstrated using highly loaded aqueous alumina suspensions with >50 vol.% solids loading and ≤5 vol.% water‐soluble polymer employed as a rheological modifier. Chopped carbon fibers were added to the suspensions to attain maximum loadings of 30 vol.%. During forming, cylindrical foam molds were dipped into the suspension while being rotated radially about the long axis. Simultaneously, a doctor blade was placed at a specified distance from the foam surface to facilitate the flow of the suspension to align the fiber and control the thickness of the material that accrued on the mold. Rheological study of alumina‐PVP suspensions with and without chopped carbon fiber showed that the suspensions exhibited a yield‐pseudoplastic flow behavior. The degree of alignment of the carbon fiber phase in the green bodies was characterized for various suspension formulations, carbon fiber contents and forming speeds. Stereological characterization of green body specimens confirmed the effectiveness of HDSC to attain the desired tubular geometry with considerable fiber alignment for a suspension composition containing ≤20 vol.% chopped fibers.