Flow cytometry: a tool for understanding the behaviour of polyhydroxyalkanoate accumulators.

The use of mixed microbial cultures (MMCs) is seen as an attractive strategy for polyhydroxyalkanoate (PHA) production. In order to optimize the MMC-PHA production process, tools are required to improve our understanding of the physiological state of the PHA-storing microorganisms within the MMC. In the present study, we explored the use of flow cytometry to analyse the metabolic state and polyhydroxybutyrate (PHB) content of the microorganisms from an MMC-PHA production process. A sequencing batch reactor under a feast and famine regime was used to enrich an MMC with PHB-storing microorganisms. Interestingly, once the PHB-storing microorganisms are selected, the level of PHB accumulation depends largely on the metabolic state of these microorganisms and not exclusively on the consortium composition. These results demonstrate that flow cytometry is a powerful tool to help to understand the PHA storage response of an MMC-PHA production process. KEY POINTS: • Flow cytometry allows to measure PHB content and metabolic activity over time. • Microorganisms showing high PHB content also have high metabolic activity. • PHB producers with low metabolic activity show low PHB content.

Enzymatic esterification of oleic acid by Candida rugosa lipase immobilized onto biochar.

The immobilization of Candida rugosa lipase (CRL) onto biochar was studied in a series of batch experiments. CRL sorption behavior was evaluated as a function of pH, enzyme concentration, temperature and ionic strength. As the immobilized lipase was used for the catalytic esterification of oleic acid, its resistance to solvents and thermal stability were evaluated. CRL adsorption increased by increasing temperature, and with higher pH, reaching a maximum at pH 7.0. Immobilization increased lipase stability at 40 °C by more than 80% when compared to the free enzyme. Moreover, immobilized CRL showed high stability in the presence of tert-butanol, which prevents premature deactivation of the enzyme caused by alcohols during the reaction. Immobilization of CRL increased the oleic acid conversion rate. Our results suggest that biochar is a highly promising material for the immobilization of CRL lipase for the catalytic production of esters.

Pectin-Chitosan Hydrogel Beads for Delivery of Functional Food Ingredients.

A common challenge in hydrogel-based delivery systems is the premature release of low molecular weight encapsulates through diffusion or swelling and reduced cell viability caused by the low pH in gastric conditions. A second biopolymer, such as chitosan, can be incorporated to overcome this. Chitosan is usually associated with colonic drug delivery systems. We intended to formulate chitosan-coated pectin beads for use in delaying premature release of the encapsulate under gastric conditions but allowing release through disintegration under intestinal conditions. The latter is of utmost importance in delivering most functional food ingredients. Therefore, this study investigated the impact of formulation and process conditions on the size, sphericity, and dissolution behavior of chitosan-coated hydrogel beads prepared by interfacial coacervation. The size and sphericity of the beads depend on the formulation and range from approximately 3 to 5 mm and 0.82 to 0.95, respectively. Process conditions during electro-dripping may be modulated to tailor bead size. Depending on the voltage, bead size ranged from 1.5 to 4 mm. Confocal laser scanning microscopy and scanning electron microscopy confirmed chitosan shell formation around the pectin bead. Chitosan-coated beads maintained their size and shape in simulated gastric fluid but experienced structural damage in simulated intestinal fluid. Therefore, they represent a novel delivery system for functional food ingredients.

Life cycle comparison of waste-to-energy alternatives for municipal waste treatment in Chilean Patagonia.

The energy system in the Region of Aysén, Chile, is characterized by a strong dependence on fossil fuels, which account for up to 51% of the installed capacity. Although the implementation of waste-to-energy concepts in municipal waste management systems could support the establishment of a more fossil-independent energy system for the region, previous studies have concluded that energy recovery systems are not suitable from an economic perspective in Chile. Therefore, this work intends to evaluate these technical options from an environmental perspective, using life cycle assessment as a tool for a comparative analysis, considering Coyhaique city as a case study. Three technical alternatives were evaluated: (i) landfill gas recovery and flaring without energy recovery; (ii) landfill gas recovery and energy use; and (iii) the implementation of an anaerobic digestion system for the organic waste fraction coupled with energy recovery from the biogas produced. Mass and energy balances of the three analyzed alternatives have been modeled. The comparative LCA considered global warming potential, abiotic depletion and ozone layer depletion as impact categories, as well as required raw energy and produced energy as comparative regional-specific indicators. According to the results, the use of the recovered landfill gas as an energy source can be identified as the most environmentally appropriate solution for Coyhaique, especially when taking into consideration the global impact categories.

Is biodegradable waste a porous environment? A review.

This article presents a review of the porous physical characteristics, phenomena and simulation models so far investigated and applied in the management of biodegradable wastes (BW), summarising the main properties of porous media and the dynamics of fluids within its voids. The aim is to highlight how the description of biodegradable wastes as porous media and the use of porous media models can facilitate the development of new sustainable and affordable technologies for BW recycling. However, it is pointed out how the lack of physical experimental data and of tailored modelling tools has so far hampered the use of this approach. Therefore, it is suggested that a simpler way to design and implement modelling tools simulating BW treatment technologies is by modifying available models designed originally for other porous media, such as soil and rock.

Life cycle assessment of solid waste management strategies in a chlor-alkali production facility.

The waste management of a chlor-alkali and calcium chloride industrial facility from southern Chile was the object of this study. The main solid waste materials generated in these processes are brine sediments and calcium chloride sediments, respectively. Both residues are mixed in the liquid phase and filtered in a press filter, obtaining a final low humidity solid waste, called 'mixed sediments', which is disposed of in an industrial landfill as non-hazardous waste. The aim of the present study was to compare by means of LCA, the current waste management option of the studied chlor-alkali facility, namely landfill disposal, with two new possible options: the reuse of the mixed sediments as mineral additive in compost and the use of brine sediments as an unconventional sorbent for the removal of heavy metals from wastewater. The functional unit was defined as 1 tonne of waste being managed. To perform this evaluation, software SimaPro 7.0 was used, selecting the Ecoindicator 99 and CML 2000 methodologies for impact evaluation. The obtained results indicate that the use of brine sediments as a novel material for the removal of heavy metals from wastewater (scenario 3) presented environmental benefits when compared with the waste management option of sediments landfilling (scenario 1). The avoided environmental loads, generated by the substitution of activated granular carbon and the removal of Cu and Zn from wastewater in the treatment process generated positive environmental impacts, enhancing the environmental performance of scenario 3.

Development and thermochemical characterization of an antioxidant material based on polyhydroxybutyrate electrospun microfibers.

The use of antioxidants such as curcumin (Cur) or quercetin (Que) in biomedical and biotechnological applications has been studied owing to their capability to prevent oxidative stress and inhibit free radicals. Using polyhydroxybutyrate (PHB) electrospun fibers is presented as a proper option to encapsulate curcumin and quercetin due to its biocompatibility and biodegradability characteristics. Electrospun fibers were obtained dissolving commercial PHB in chloroform:N,N-dimethylformamide (DMF) (4:1) at 7% m/V, and adding two different concentrations of antioxidant (Cur, and Que) 1%m/m, and 7% m/m. These polymeric solutions were electrospun at different conditions and the obtained fibers were characterized by scanning electron microscopy (SEM), thermogravimetric (TGA) analysis, and Fourier transform infrared spectroscopy (FT-IR). The curcumin and quercetin releases into phosphate buffer saline (PBS) at pH 7.4 were obtained in vitro and measured by spectrophotometry. Antioxidant activities were measured by spectrophotometry in a microplate reader using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Fibers obtained with different formulations presented a chemical composition in accordance with PHB according to FTIR spectra, the diameters fluctuate between 0.761 ± 0.123 and 1.803 ± 0.557 µm, with qualities over 0.95 according to their morphology, and the melting temperature resulted near 178 °C according to the bibliography. The crystallinity of fibers decreases while curcumin or quercetin concentration increases for the studied interval, indeed, quercetin showed a higher impact on the relative crystallinity of fibers. Antioxidant activity of active compounds is maintained after encapsulation in PHB electrospun fibers, and quercetin resulted in near four times antioxidant activity compared to curcumin according to DPPH analysis.

Genome-Wide Metabolic Reconstruction of the Synthesis of Polyhydroxyalkanoates from Sugars and Fatty Acids by Burkholderia Sensu Lato Species.

Burkholderia sensu lato (s.l.) species have a versatile metabolism. The aims of this review are the genomic reconstruction of the metabolic pathways involved in the synthesis of polyhydroxyalkanoates (PHAs) by Burkholderia s.l. genera, and the characterization of the PHA synthases and the pha genes organization. The reports of the PHA synthesis from different substrates by Burkholderia s.l. strains were reviewed. Genome-guided metabolic reconstruction involving the conversion of sugars and fatty acids into PHAs by 37 Burkholderia s.l. species was performed. Sugars are metabolized via the Entner-Doudoroff (ED), pentose-phosphate (PP), and lower Embden-Meyerhoff-Parnas (EMP) pathways, which produce reducing power through NAD(P)H synthesis and PHA precursors. Fatty acid substrates are metabolized via ß-oxidation and de novo synthesis of fatty acids into PHAs. The analysis of 194 Burkholderia s.l. genomes revealed that all strains have the phaC, phaA, and phaB genes for PHA synthesis, wherein the phaC gene is generally present in ≥2 copies. PHA synthases were classified into four phylogenetic groups belonging to class I II and III PHA synthases and one outlier group. The reconstruction of PHAs synthesis revealed a high level of gene redundancy probably reflecting complex regulatory layers that provide fine tuning according to diverse substrates and physiological conditions.

Biotechnological processes for biodiesel production using alternative oils.

As biodiesel (fatty acid methyl ester (FAME)) is mainly produced from edible vegetable oils, crop soils are used for its production, increasing deforestation and producing a fuel more expensive than diesel. The use of waste lipids such as waste frying oils, waste fats, and soapstock has been proposed as low-cost alternative feedstocks. Non-edible oils such as jatropha, pongamia, and rubber seed oil are also economically attractive. In addition, microalgae, bacteria, yeast, and fungi with 20% or higher lipid content are oleaginous microorganisms known as single cell oil and have been proposed as feedstocks for FAME production. Alternative feedstocks are characterized by their elevated acid value due to the high level of free fatty acid (FFA) content, causing undesirable saponification reactions when an alkaline catalyst is used in the transesterification reaction. The production of soap consumes the conventional catalyst, diminishing FAME production yield and simultaneously preventing the effective separation of the produced FAME from the glycerin phase. These problems could be solved using biological catalysts, such as lipases or whole-cell catalysts, avoiding soap production as the FFAs are esterified to FAME. In addition, by-product glycerol can be easily recovered, and the purification of FAME is simplified using biological catalysts.

Applications of Electrospun Nanofibers with Antioxidant Properties: A Review.

Antioxidants can be encapsulated to enhance their solubility or bioavailability or to protect them from external factors. Electrospinning has proven to be an excellent option for applications in nanotechnology, as electrospun nanofibers can provide the necessary environment for antioxidant encapsulation. Forty-nine papers related to antioxidants loaded onto electrospun nanofibers were categorized and reviewed to identify applications and new trends. Medical and food fields were commonly proposed for the newly obtained composites. Among the polymers used as a matrix for the electrospinning process, synthetic poly (lactic acid) and polycaprolactone were the most widely used. In addition, natural compounds and extracts were identified as antioxidants that help to inhibit free radical and oxidative damage in tissues and foods. The most recurrent active compounds used were tannic acid (polyphenol), quercetin (flavonoid), curcumin (polyphenol), and vitamin B6 (pyridoxine). The incorporation of active compounds in nanofibers often improves their bioavailability, giving them increased stability, changing the mechanical properties of polymers, enhancing nanofiber biocompatibility, and offering novel properties for the required field. Although most of the polymers used were synthetic, natural polymers such as silk fibroin, chitosan, cellulose, pullulan, polyhydroxybutyrate, and zein have proven to be proper matrices for this purpose.