Polypyrrole/phytase amperometric biosensors for the determination of phytic acid in standard solutions

Amperometric biosensors based on the physical immobilization of phytase (PhyA) into polypyrrole (PPy) films were prepared in aqueous medium. The PPy/PhyA films were characterized by cyclic voltammetry, and surface and structural characterization techniques, SEM and FTIR. Both voltammetric and amperometric transduction methods were used in order to detect phytic acid in acetate buffer at pH 5.5 at room temperature. The biosensors exhibited a detection limit of 0.15 mmol L⁻¹ and a linear range of phytic acid content from 0.5 to 2.0 mmol L⁻¹, which are adequate values for typical analyses of phytic acids in most seeds, grains, and vegetables.     

pH-Sensitive hydrogels formed by self-assembled amphiphilic triblock copolyelectrolytes

Results are reviewed of a recent extensive investigation of the behavior of self-assembled pH-sensitive triblock copolymers in aqueous solution. The hydrophilic central block was polyacrylic acid and the two hydrophobic end-blocks were statistical copolymers of n-butyl acrylate and acrylic acid containing 50 mol% acrylic acid units. The hydrophobicity of the end blocks could be modified by changing the degree of ionization of the acrylic acid units (α). The relationship between the pH and α was determined. Scattering techniques showed that flower-like micelles are formed that upon increasing concentration connect via bridging into larger aggregates and above a critical concentration into a percolating network. The rheology of the system is controlled by the exchange rate of the end-blocks between micelles and can be fine-tuned by varying the pH. The exchange rate increases exponentially with increasing α. As a consequence the system changes from a quasi-permanent hydrogel at pH < 4.5 to a free flowing liquid at pH > 5.5. The effect of the ionic strength on the structure and the rheology was found to be important only above 0.5 M.     

PLA/metal oxide biocomposites for antimicrobial packaging application

ABSTRACT

The effects of metal oxide addition in polylactic acid (PLA) film were investigated in this study. PLA/Metal Oxide biocomposites were prepared by solvent casting method. The incorporation of metal oxide resulted in no chemical structure change of neat PLA. PLA/ZnO and PLA/MgO exhibited the widest inhibition while PLA/TiO₂ resulted in the smallest inhibition zone. The addition of metal oxides caused fracture and void on the surface of PLA. The addition of metal oxide into PLA films decreased the tensile strength at the concentration higher than 0.1 per hundred resin (phr). Incorporation of metal oxides accelerated the thermal degradation of PLA and tends to increase the biodegradability of PLA.     

2′‐Deoxyribonucleoside Phosphoramidate Triphosphate Analogues as Alternative Substrates for E. coli Polymerase III

Thermostable bacterial polymerases like Taq, Therminator and Vent exo^? are able to perform DNA synthesis by using modified DNA precursors, a property that is exploited in several therapeutic and biotechnological applications. Viral polymerases are also known to accept modified substrates, and this has proven crucial in the development of antiviral therapies. However, non‐thermostable polymerases of bacterial origin, or engineered variants, that have similar substrate tolerance and could be used for synthetic biology purposes remain to be identified. We have identified the α subunit of Escherichia coli polymerase III (Pol III α) as a bacterial polymerase that is able to recognise and process as substrates several pyrophosphate‐modified dATP analogues in place of its natural substrate dATP for template‐directed DNA synthesis. A number of dATP analogues featuring a modified pyrophosphate group were able to serve as substrates during enzymatic DNA synthesis by Pol III α. Features such as the presence of potentially chelating chemical groups and the size and spatial flexibility of the chemical structure seem to be of major importance for the modified leaving group to play its role during the enzymatic reaction. In addition, we could establish that if the pyrophosphate group is altered, deoxynucleotide incorporation proceeds with an efficiency varying with the nature of the nucleobase. Our results represent a great step towards the achievement of a system of artificial DNA synthesis hosted by E. coli and involving the use of altered nucleotide precursors for nucleic acid synthesis.     

Matching relational patterns in nucleic acid sequences

We describe a program that efficiently searches sequence data banks for complex patterns where sites are linked by common relations such as identity, complementarity or span. Its algorithm is closer to those of automatic demonstration than to the finite state machines used in fast pattern matching. The repertory of relations can be enriched at will without rewriting the core of the program. The program is written in Pascal-ISO and runs on a microcomputer.     

Preparation and characterization of ibuprofen-loaded microspheres consisting of poly(3-hydroxybutyrate) and methoxy poly (ethylene glycol)-b-poly (D,L-lactide) blends or poly(3-hydroxybutyrate) and gelatin composites for controlled drug release

Poly-(3-hydroxybutyrate) (P(3HB)) is a biodegradable and biocompatible polymer that has been used to obtain polymer-based drug carriers. However, due to the high crystallinity degree of this polymer, drug release from P(3HB) microspheres frequently occurs at excessive rates. In this study, two strategies for prolonging ibuprofen release from P(3HB)-based microspheres were tested: blending with poly(D,L-lactide)-b-polyethylene glycol (mPEG-PLA); and obtaining composite particles with gelatin (GEL). SEM micrographs showed particles that were spherical and had a rough surface. A slight decrease of the crystallinity degree of P(3HB) was observed only in the DSC thermogram obtained from unloaded-microspheres prepared from 1:1 P(3HB):mPEG-PLA blend. For IBF-loaded microspheres, a reduction of around 10 °C in the melting temperature of P(3HB) was observed, indicating that the crystalline structure of the polymer was affected in the presence of the drug. DSC studies also yielded evidence of the presence of a molecular dispersion coexisting with a crystalline dispersion in the drug in the matrix. Similar results were obtained from X-ray diffractograms. In spite of 1:1 mPEG-PLA:P(3HB) blends having contributed to the reduction of the burst effect, a more controlled drug release was provided by the use of the 3:1 P(3HB):mPEGPLA blend. This result indicated that particle hydration played an important role in the drug release. On the other hand, the preparation of P(3HB):GEL composite microspheres did not allow control of the IBF release.     

Quorum sensing detected by atomic force microscopy imaging of corrals surrounding multicellular arrangement of bacteria

Connectivity of the glycocalyx covering of small communities of Acidithiobacillus ferrooxidans bacteria deposited on hydrophilic mica plates was imaged by atomic force microscopy. When part of the coverage was removed by water rinsing, an insoluble structure formed by corrals surrounding each individual bacterium was observed. A collective ring structure with clustered bacteria (>or=3) was observed, which indicates that the bacteria perceived the neighborhood in order to grow a protective structure that results in smaller production of exopolysaccharides material. The most surprising aspect of these collective corral structures was that they occur at a low bacterial cell density. The deposited layers were also analyzed by confocal Raman microscopy and shown to contain polysaccharides, protein, and glucoronic acid.