PHA Production from Cheese Whey and "Scotta": Comparison between a Consortium and a Pure Culture of Leuconostoc mesenteroides.

It is urgent to expand the market of biodegradable alternatives to oil-derived plastics owing to (i) increasingly limited oil availability/accessibility, and (ii) the dramatic impact of traditional plastics on aquatic life, the food chain, all Earth ecosystems, and ultimately, human health. Polyhydroxyalkanoates (PHAs) are promising biodegradable polymers that can be obtained through microbial fermentation of agro-industrial byproducts, e.g., milk and cheese whey. Here, the PHA-accumulating efficiency of a mixed microbial culture (MMC, derived from activated sludges) grown on dairy byproducts (cheese and scotta whey) was measured. Bioreactor tests featuring temperature and pH control showed that both scotta and pre-treated Toma cheese whey could be used for PHA production by MMC, although scotta cheese whey supported higher PHA yield and productivity. The advantages of open MMCs include their plasticity and versatility to fast changing conditions; furthermore, no growth-medium sterilization is needed prior to fermentation. However, the use of pure cultures of efficient PHA producers may support better metabolic performances. Therefore, PHA-producing strains were isolated from a MMC, leading to the satisfactory identification of two bacterial strains, Citrobacter freundii and Leuconostoc spp., whose ability to accumulate PHAs in synthetic media was confirmed. A more detailed investigation by mass spectrometry revealed that the strain was L. mesenteroides. Although the validation of L. mesenteroides potential to produce PHA through fermentation of agro-industrial byproducts requires further investigations, this is the first study reporting PHA production with the Leuconostoc genus.

Rational engineering of the lccß T. versicolor laccase for the mediator-less oxidation of large polycyclic aromatic hydrocarbons.

Laccases are among the most sought-after biocatalyst for many green applications, from biosensors to pollution remedial, because they simply need oxygen from the air to oxidize and degrade a broad range of substrates. However, natural laccases cannot process large and toxic polycyclic aromatic hydrocarbons (PAHs) except in the presence of small molecules, called mediators, which facilitate the reaction but are inconvenient for practical on-field applications. Here we exploited structure-based protein engineering to generate rationally modified fungal laccases with increased ability to process bulky PAHs even in a mediator-less reaction. Computational simulations were used to estimate the impact of mutations in the enzymatic binding pocket on the ability to bind and oxidize a selected set of organic compounds. The most promising mutants were produced and their activity was evaluated by biochemical assays with phenolic and non-phenolic substrates. Mutant laccases engineered with a larger binding pocket showed enhanced activity (up to ~ 300% at pH 3.0) in a wider range of pH values (3.0-8.0) in comparison to the wild type enzyme. In contrast to the natural laccase, these mutants efficiently degraded bulky and harmful triphenylmethane dyes such as Ethyl Green (up to 91.64% after 24 h), even in the absence of mediators, with positive implications for the use of such modified laccases in many green chemistry processes (e.g. wastewater treatment).

Thermal stability and flame resistance of cotton fabrics treated with whey proteins.

It is well described in the literature that whey proteins are able to form coatings, which exhibit high mechanical and oxygen barrier properties, notwithstanding a great water vapour adsorption. These peculiarities have been exploited for applying a novel protein-based finishing treatment to cotton and for assessing the protein effect on the thermal and thermo-oxidative stability and on the flame retardant properties of the cellulosic fabric. Indeed, the deposited whey protein coatings have turned out to significantly affect the thermal degradation of cotton in inert and oxidative atmosphere, and to somehow modify its combustion when a flame has been applied. Furthermore, the influence of the secondary and tertiary structure of these proteins on the morphology of the deposited coating, and thus on the thermal and flame retardant properties of the treated fabrics, has been evaluated by performing a denaturation thermal treatment before the protein application.

Opening Study on the Development of a New Biosensor for Metal Toxicity Based on Pseudomonas fluorescens Pyoverdine.

To date, different kinds of biosensing elements have been used effectively for environmental monitoring. Microbial cells seem to be well-suited for this task: they are cheap, adaptable to variable field conditions and give a measurable response to a broad number of chemicals. Among different pollutants, heavy metals are still a major problem for the environment. A reasonable starting point for the selection of a biorecognition element to develop a biosensor for metals could be that of a microorganism that exhibits good mechanisms to cope with metals. Pseudomonads are characterized by the secretion of siderophores (e.g., pyoverdine), low-molecular weight compounds that chelate Fe3+ during iron starvation. Pyoverdine is easily detected by colorimetric assay, and it is suitable for simple online measurements. In this work, in order to evaluate pyoverdine as a biorecognition element for metal detection, the influence of metal ions (Fe3+, Cu2+, Zn2+), but also of temperature, pH and nutrients, on microbial growth and pyoverdine regulation has been studied in P. fluorescens. Each of these variables has been shown to influence the synthesis of siderophore: for instance, the lower the temperature, the higher the production of pyoverdine. Moreover, the concentration of pyoverdine produced in the presence of metals has been compared with the maximum allowable concentrations indicated in international regulations (e.g., 98/83/EC), and a correlation that could be useful to build a colorimetric biosensor has been observed.