Polyhydroxyalkanoates (PHAs) are biodegradable plastics of microbial origin, whose biodegradability and thermochemical properties make them greener alternatives to conventional plastics. Despite their high industrial potential, the PHAs high production costs still hinder their application. Mixed microbial biomass combined with agroindustrial wastes are being used to strategically reduce these costs. However, it is still necessary to optimize the downstream processing, where the extraction process amounts to 30-50 % of the total costs. Conventional processes apply chlorinated solvents to recover PHAs from microbial biomass but cannot be implemented industrially due to environmental regulations. Alternative solvents, with good results of purity and recovery yields, usually have a negative impact on the molecular weight of the final polymer. In this work, the addition of a pre-treatment based on non--114) to extract PHA from mixed microbial biomass selected on fermented agro-industrial wastes was investigated. The best results were obtained with without any prewith chloroform (63%). The extracted polymer was analysed and characterized, revealing a PHA of high purity (>90%) and low molecular weight loss (under 24%). Additionally, a material-focused economic and a carbon footprint analysis were performed and supported the selection of the method as one of the cheapest options and with the lowest carbon footprint.

Acidogenic fermentation (AF) is often applied to wastes to produce short-chain organic acids (SCOAs)—molecules with applications in many industries. Spent coffee grounds (SCGs) are a residue from the coffee industry that is rich in carbohydrates, having the potential to be valorized by this process. However, given the recalcitrant nature of this waste, the addition of a pretreatment step can significantly improve AF. In this work, several pretreatment strategies were applied to SCGs (acidic hydrolysis, basic hydrolysis, hydrothermal, microwave, ultrasounds, and supercritical CO2 extraction), evaluated in terms of sugar and inhibitors release, and used in AF. Despite the low yields of sugar extracted, almost all pretreatments increased SCOAs production. Milder extraction conditions also resulted in lower concentrations of inhibitory compounds and, consequently, in a higher concentration of SCOAs. The best results were obtained with acidic hydrolysis of 5%, leading to a production of 1.33 gSCOAs/L, an increase of 185% compared with untreated SCGs.

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Spent coffee grounds (SCGs) are a promising substrate that can be valorized by biotechnological processes, such as for short-chain organic acid (SCOA) production, but their complex structure implies the application of a pretreatment step to increase their biodegradability. Physicochemical pretreatments are widely studied but have multiple drawbacks. An alternative is the application of biological pretreatments that include using fungi Trametes versicolor and Paecilomyces variotii that naturally can degrade complex substrates such as SCGs. This study intended to compare acidic and basic hydrolysis and supercritical CO2 extraction with the application of these fungi. The highest concentration of SCOAs, 2.52 gCOD/L, was achieved after the acidification of SCGs pretreated with acid hydrolysis, but a very similar result, 2.44 gCOD/L, was obtained after submerged fermentation of SCGs by T. versicolor. This pretreatment also resulted in the best acidification degree, 48%, a very promising result compared to the 13% obtained with the control, untreated SCGs, highlighting the potential of biological pretreatments.