Innovative strategy for polyhydroxyalkanoates recovery from mixed microbial cultures. Effects of aqueous phase and solvent extraction on polymer properties

This work aimed to investigate various methods of polyhydroxyalkanoates (PHA) extraction and to optimize the recovery in the view of reducing solvents’ use and waste. The extraction tests were applied on PHA-rich biomass collected at the end of the accumulation step, conducted both at pilot and lab scale. PHA-rich biomass obtained from the lab scale process was produced from synthetic feedstock (mixture of acetic and propionic acids), while fermented organic waste was used for PHA production at pilot scale. In the extraction tests, different solutions of NaOH, also in combination with a surfactant (sodium dodecyl sulfate, SDS), were used to recover the polymer from the non-polymeric cellular material (NPCM). Different times (3, 6 and 24 h) and concentrations (0.05 – 1 M) were tested, evaluating also the effect of adding SDS (0.1 % w/v). As a benchmark, solvent extraction with chloroform and oxidation with NaClO were also conducted. Finally, extracted samples were characterized through several techniques: DSC, TGA, GC-FID, capillary viscosimetry. Alkaline treatment at short times and low concentrations resulted to be more efficient in terms of purity (85 – 99 % w/w PHA) and recovery (higher than 80 % w/w), than higher concentration. On the other hand, the presence of SDS had dramatic effects on the recovery (lower than 50 % w/w) and also on the molecular weight, which was two folds lower than that obtained from alkaline extraction. Overall, extraction with aqueous phase reagents had no effects on thermal properties, which resulted to be in the range of those reported in literature

Ethylic esters as green solvents for the extraction of intracellular polyhydroxyalkanoates produced by mixed microbial culture

Volatile fatty acids obtained from the fermentation of the organic fraction of municipal solid waste can be used as raw materials for non-toxic ethyl ester (EE) synthesis as well as feedstock for the production of polyhydroxyalkanoates (PHAs). Taking advantage of the concept of an integrated process of a bio-refinery, in the present paper, a systematic investigation on the extraction of intracellular poly(3-hydroxybutyrate-co-3-hydroxyvalerate), produced by mixed microbial culture by using EEs was reported. Among the tested EEs, ethyl acetate (EA) was the best solvent, dissolving the copolymer at the lowest temperature. Then, extraction experiments were carried out by EA at different temperatures on two biomass samples containing PHAs with different average molecular weights. The parallel characterization of the extracted and non-extracted PHAs evidenced that at the lower temperature (100◦C) EA solubilizes preferentially the polymer fractions richer in 3HV comonomers and with the lower molecular weight. By increasing the extraction temperature from 100◦C to 125◦C, an increase of recovery from about 50 to 80 wt% and a molecular weight reduction from 48% to 65% was observed. The results highlighted that the extracted polymer purity is always above 90 wt% and that it is possible to choose the proper extraction condition to maximize the recovery yield at the expense of polymer fractionation and degradation at high temperatures or use milder conditions to maintain the original properties of a polymer

Modelling Mixed Microbial Culture Polyhydroxyalkanoate Accumulation Bioprocess towards Novel Methods for Polymer Production Using Dilute Volatile Fatty Acid Rich Feedstocks

Volatile fatty acid (VFA) rich streams from fermentation of organic residuals and wastewater are suitable feedstocks for mixed microbial culture (MMC) Polyhydroxyalkanoate (PHA) production. However, many such streams have low total VFA concentration (1–10 gCOD/L). PHA accumulation requires a flow-through bioprocess if the VFAs are not concentrated. A flow through bioprocess must balance goals of productivity (highest possible influent flow rates) with goals of substrate utilization efficiency (lowest possible effluent VFA concentration). Towards these goals, dynamics of upshift and downshift respiration kinetics for laboratory and pilot scale MMCs were evaluated. Monod kinetics described a hysteresis between the upshift and downshift responses. Substrate concentrations necessary to stimulate a given substrate uptake rate were significantly higher than the concentrations necessary to sustain the attained substrate uptake rate. A benefit of this hysteresis was explored in Monte Carlo based PHA accumulation bioprocess numerical simulations. Simulations illustrated for a potential to establish continuous flow-through PHA production bioprocesses even at a low (1 gCOD/L) influent total VFA concentration. Process biomass recirculation into an engineered higher substrate concentration mixing zone, due to the constant influent substrate flow, enabled to drive the process to maximal possible PHA production rates without sacrificing substrate utilization efficiency

Measurements of CO2, its stable isotopes, O2/N2, and 222Rn at Bern, Switzerland

A one-year time series of atmospheric CO2 measurements from Bern, Switzerland, is presented. O2/N2 and Ar/N2 ratios as well as stable carbon and oxygen isotopes of CO2 and δ29N2, δ34O2 and δ36Ar were measured periodically during a one year period. Additionally, the 222Rn activity was measured during three months in the winter 2004. Using the correlation from short-term fluctuations of CO2 and 222Rn we estimated a mean CO2 flux density between February 2004 and April 2004 in the region of Bern of 95±39 tC km–2month–1. The continuous observations of carbon dioxide and associated tracers shed light on diurnal and seasonal patterns of the carbon cycle in an urban atmosphere. There is considerable variance in nighttime δ13C and δ18O of source CO2 throughout the year, however, with generally lower values in winter compared to summertime. The O2:CO2 oxidation ratio during the nighttime build-up of CO2 varies between –0.96 and –1.69 mol O2/mol CO2. Furthermore, Ar/N2 measurements showed that artifacts like thermal fractionation at the air intake are relevant for high precision measurements of atmospheric O2

VST - VLT Survey Telescope Integration Status

The VLT Survey Telescope (VST) is a 2.6m aperture, wide field, UV to I facility, to be installed at the European Southern Observatory (ESO) on the Cerro Paranal Chile. VST was primarily intended to complement the observing capabilities of VLT with wide-angle imaging for detecting and pre-characterising sources for further observations with the VLT.Comment: 2 pages, 2 figures, conferenc

A second case with the V374A KCND3 pathogenic variant in an Italian patient with early-onset spinocerebellar ataxia

Background and Objectives To date, approximately 20 heterozygous mainly loss-of-function variants in KCND3 have been associated with spinocerebellar ataxia (SCA) type 19 and 22, a clinically heterogeneous group of neurodegenerative disorders. We aimed at reporting the second patients with the V374A KCND3 mutation from an independent family, confirming its pathogenic role. Methods We describe the clinical history of a patient with SCA and conducted genetic investigations including mitochondrial DNA analysis and exome sequencing. Results This male patient was reported to have unstable gait with tremors at the lower limbs and dysarthric speech since childhood. A neurologic examination also showed dysarthria, nystagmus, action tremor, dysmetria, and weak deep tendon reflexes. He had marked cerebellar atrophy at brain MRI, more evident at vermis. Molecular analysis, including exome sequencing and an in silico panel analysis of genes associated with SCA, revealed the c.1121T>C [p.V374A] mutation in KCND3. Discussion This report consolidates the pathogenicity of the V374A KCND3 mutation and suggests that the ataxic paroxysmal exacerbations are not a key phenotypic feature of this mutation