Optimization of lactic acid production from apple and tomato pomaces by thermotolerant bacteria.

The production of lactic acid (LA) through biomass fermentation represents a promising alternative to the chemical synthesis. The use of agri-food by-products as fermentable carbohydrate sources can improve process sustainability by reducing waste and valorizing residual biomass. This study assessed the use of apple and tomato pomaces for producing LA through fermentation using thermotolerant bacteria under aerobic and non-sterile conditions. Three bacteria were evaluated and Heyndrickxia coagulans DSM 2314 was selected for its ability to produce LA from hydrolyzates of apple pomace (APH) and tomato pomace (TPH). The fermentation conditions were optimized to maximize LA production from APH, TPH and a mixture of both hydrolyzates. Therefore, LA productions ranged from 36.98 ± 0.41 to 40.72 ± 0.43 g/L, with yields from 0.86 ± 0.02 to 1.01 ± 0.01 g/g. Yeast extract was necessary as a nitrogen source for fermenting APH, while TPH and the mixture of both hydrolyzates did not require any supplementation. Other nitrogen sources, such as wine lees, urea and NH3Cl, were tested for fermenting APH. However, mixing this hydrolyzate with TPH proved to be the most viable alternative. This study demonstrates the potential for valorizing apple and tomato pomaces into LA under feasible fermentation conditions.

Assessment of vine shoots and surplus grape must for succinic acid bioproduction.

Vine shoots and surplus grape must were assessed as feedstocks for succinic acid production with Actinobacillus succinogenes and Basfia succiniproducens. After acidic and enzymatic hydrolysis, vine shoots released 35-40 g/L total sugars. Both bacterial species produced 18-21 g/L succinic acid from this hydrolysate in 120 h. Regarding grape must fermentation, A. succinogenes clearly outperformed B. succiniproducens. Yeast extract (a source of organic nitrogen and vitamins) was the only additional nutrient needed by A. succinogenes to grow on grape must. Under mathematically optimized conditions (145.7 g/L initial sugars and 24.9 g/L yeast extract), A. succinogenes generated 88.9 ± 1.4 g/L succinic acid in 96 h, reaching a succinic acid yield of 0.66 ± 0.01 g/g and a sugar consumption of 96.64 ± 0.30%. Substrate inhibition was not observed in grape musts with 125-150 g/L initial sugars, provided that an adequate amount of yeast extract was available for bacteria. Alternative nitrogen sources to yeast extract (red wine lees, white wine lees, urea, NH4Cl, and choline chloride) were not suitable for A. succinogenes in grape must. KEY POINTS: • Vine shoots and surplus grape must were assessed for succinic acid bioproduction. • Succinic acid bioproduction was 21 g/L with vine shoots and 89 g/L with grape must. • Fermentation was efficient at high sugar loads if organic N supply was adequate.

Revisiting the production of L( +)-lactic acid from vine shoots: bioconversion improvements by employing thermotolerant bacteria.

Vine shoots (Vitis vinifera L.) constitute an abundant lignocellulosic source which is frequently underutilised. Alkaline and acidic pretreatments (with and without washing steps) were compared and optimised to release fermentable sugars from vine shoots. An acidic pretreatment using 1.72% H2SO4 at 134 °C for 17 min (with 10% w/w solid biomass), followed by an enzymatic hydrolysis, offered the most cost-effective results, releasing 40.21 g/L sugars. Three thermotolerant strains, namely, Bacillus coagulans DSM 2314, Geobacillus stearothermophilus DSM 2313, and G. stearothermophilus DSM 494, were assessed to produce lactic acid from vine-shoot hydrolysates under aerobic and non-sterile conditions, without the need of detoxification steps. In addition, wine lees were satisfactorily employed as nitrogen sources for the fermentation, providing similar results to yeast extract and being the only nutrient added to vine-shoot hydrolysates. Under optimal conditions, B. coagulans DSM 2314 produced 29.21 ± 0.23 g/L lactic acid in 24 h, with a sugar consumption of 98.74 ± 0.07% and a yield of 96.38 ± 0.76%, when supplemented with red wine lees. The purity of the isomer L( +) reached 97.59 ± 1.35% of the total lactic acid produced. Although G. stearothermophilus was able to transform the hexoses from vine-shoot hydrolysates into lactic acid, it proved to be inefficient for metabolising pentoses, thus obtaining lower lactic acid values (16-18 g/L).

Biobutanol production from coffee silverskin.

BACKGROUND: Coffee silverskin, a by-product from coffee roasting industries, was evaluated as a feedstock for biobutanol production by acetone-butanol-ethanol fermentation. This lignocellulosic biomass contained approximately 30% total carbohydrates and 30% lignin. Coffee silverskin was subjected to autohydrolysis at 170 °C during 20 min, with a biomass-to-solvent ratio of 20%, and a subsequent enzymatic hydrolysis with commercial enzymes in order to release simple sugars. The fermentability of the hydrolysate was assessed with four solventogenic strains from the genus Clostridium. In addition, fermentation conditions were optimised via response surface methodology to improve butanol concentration in the final broth. RESULTS: The coffee silverskin hydrolysate contained 34.39 ± 2.61 g/L total sugars, which represents a sugar recovery of 34 ± 3%. It was verified that this hydrolysate was fermentable without the need of any detoxification method and that C. beijerinckii CECT 508 was the most efficient strain for butanol production, attaining final values of 4.14 ± 0.21 g/L acetone, 7.02 ± 0.27 g/L butanol and 0.25 ± 0.01 g/L ethanol, consuming 76.5 ± 0.8% sugars and reaching a butanol yield of 0.269 ± 0.008 gB/gS under optimal conditions. CONCLUSIONS: Coffee silverskin could be an adequate feedstock for butanol production in biorefineries. When working with complex matrices like lignocellulosic biomass, it is essential to select an adequate bacterial strain and to optimize its fermentation conditions (such as pH, temperature or CaCO3 concentration).

Biobutanol production from apple pomace: the importance of pretreatment methods on the fermentability of lignocellulosic agro-food wastes.

Apple pomace was studied as a possible raw material for biobutanol production. Five different soft physicochemical pretreatments (autohydrolysis, acids, alkalis, organic solvents and surfactants) were compared in a high-pressure reactor, whose working parameters (temperature, time and reagent concentration) were optimised to maximise the amount of simple sugars released and to minimise inhibitor generation. The pretreated biomass was subsequently subjected to a conventional enzymatic treatment to complete the hydrolysis. A thermal analysis (DSC) of the solid biomass indicated that lignin was mainly degraded during the enzymatic treatment. The hydrolysate obtained with the surfactant polyethylene glycol 6000 (PEG 6000) (1.96% w/w) contained less inhibitors than any other pretreatment, yet providing 42 g/L sugars at relatively mild conditions (100 °C, 5 min), and was readily fermented by Clostridium beijerinckii CECT 508 in 96 h (3.55 g/L acetone, 9.11 g/L butanol, 0.26 g/L ethanol; 0.276 gB/gS yield; 91% sugar consumption). Therefore, it is possible to optimise pretreatment conditions of lignocellulosic apple pomace to reduce inhibitor concentrations in the final hydrolysate and perform successful ABE fermentations without the need of a detoxification stage.

Advances in Microbial Biotechnology for Sustainable Alternatives to Petroleum-Based Plastics: A Comprehensive Review of Polyhydroxyalkanoate Production.

The industrial production of polyhydroxyalkanoates (PHAs) faces several limitations that hinder their competitiveness against traditional plastics, mainly due to high production costs and complex recovery processes. Innovations in microbial biotechnology offer promising solutions to overcome these challenges. The modification of the biosynthetic pathways is one of the main tactics; allowing for direct carbon flux toward PHA formation, increasing polymer accumulation and improving polymer properties. Additionally, techniques have been implemented to expand the range of renewable substrates used in PHA production. These feedstocks are inexpensive and plentiful but require costly and energy-intensive pretreatment. By removing the need for pretreatment and enabling the direct use of these raw materials, microbial biotechnology aims to reduce production costs. Furthermore, improving downstream processes to facilitate the separation of biomass from culture broth and the recovery of PHAs is critical. Genetic modifications that alter cell morphology and allow PHA secretion directly into the culture medium simplify the extraction and purification process, significantly reducing operating costs. These advances in microbial biotechnology not only enhance the efficient and sustainable production of PHAs, but also position these biopolymers as a viable and competitive alternative to petroleum-based plastics, contributing to a circular economy and reducing the dependence on fossil resources.

Fractionation of High-Value Compounds from Hops Using an Optimised Sequential Extraction Procedure.

This study describes the development and optimisation of a process for the extraction and fractionation of high-value compounds from hops. Firstly, the efficacy of ten organic solvents was compared for performing the initial solid-liquid extraction of compounds from hop pellets with subsequent fractionation steps. A methanol-dichloromethane mixture was selected and the extraction variables were optimised in order to maximise the recovery of valuable hop compounds separated into different streams (α- and ß-acids in soft resins, xanthohumol in hard resins, and phenolics in spent solids) after fractionation steps. The optimisation results showed that extraction of hop pellets performed at room temperature with 19.7% (v/v) methanol for 89 min yielded recoveries of 86.57% α-acids and 89.14% ß-acids in soft resins, 78.48% xanthohumol in hard resins and 67.10% phenolics in spent solids. These conditions were successfully validated using six hop varieties. Moreover, the antioxidant properties of all recovered fractions were compared and the soft resins showed the highest antioxidant activities, reaching values of 3.91 ± 0.10 g AAE/100 g for ferric reducing power (FRAP) and 0.10 ± 0.01 mg/mL for 50% of radical scavenging activity (EC50). The optimised sequential extraction could serve as a basis for larger scale-up for industrial production.

Cell Immobilization for Erythritol Production.

Nowadays, commercial erythritol synthesis is performed by free-cell fermentation with fungi in liquid media containing high concentrations of pure carbon sources. Alternative fermentation techniques, such as cell immobilization, could imply an economic and energetic improvement for erythritol-producing factories. The present work describes, for the first time, the feasibility of achieving cell immobilization during erythritol production. Cells of the fungus Moniliella pollinis were successfully immobilized on a cotton cloth which was placed inside a 2-L bioreactor, where they were fed with red grape must supplemented with yeast extract. They produced 47.03 ± 6.16 g/L erythritol in 96 h (yield 0.18 ± 0.04 g/g) over four consecutive fermentation batches. The immobilized cells remained stable and operative during a 456 h period. The erythritol concentration attained was similar (p > 0.05; Tukey HSD test) to the reference value obtained with the use of free cells (41.88 ± 5.18 g/L erythritol) under the same fermentation conditions. The comparable results observed for free and immobilized cells evidences the efficiency of the immobilization system. Therefore, the proposed method for erythritol bioproduction eliminates the need for the continuous preparation of fungal inocula before each fermentation batch, thus reducing the costs of the reagents and energy.

Industrial potato peel as a feedstock for biobutanol production.

Potato peel from a snack factory was assessed as possible feedstock for biobutanol production. This lignocellulosic biomass was subjected to various physicochemical pretreatments (autohydrolysis and hydrolysis with dilute acids, alkalis, organic solvents or surfactants) under different conditions of time, temperature and reagent concentrations, in order to favour the release of sugars and reduce the generation of fermentation inhibitors. Thereafter, the pretreated potato peel was treated enzymatically to complete the hydrolysis. Autohydrolysis at 140 °C and 56 min was the most effective pretreatment, releasing 37.9 ± 2.99 g/L sugars from an aqueous mixture containing 10% (w/w) potato peel (sugar recovery efficiency 55 ± 13%). The fermentability of the hydrolysates was checked with six strains of Clostridium beijerinckii, C. acetobutylicum, C. saccharobutylicum and C. saccaroperbutylacetonicum. C. saccharobutylicum DSM 13864 produced 2.1 g/L acetone, 7.6 g/L butanol and 0.6 g/L ethanol in 96 h (0.186 gB/gS), whereas C. saccharoperbutylacetonicum DSM 2152 generated 1.8 g/L acetone, 8.1 g/L butanol and 1.0 g/L ethanol in 120 h (0.203 gB/gS). Detoxification steps of the hydrolysate before fermentation were not necessary. Potato peel may be an interesting feedstock for biorefineries focused on butanol production.

Yeast screening and cell immobilization on inert supports for ethanol production from cheese whey permeate with high lactose loads.

Eight yeast strains of the genera Saccharomyces and Kluyveromyces were screened to ferment high lactose-load cheese whey permeate (CWP) (>130 g/L lactose) without nutrient supplementation. The fermentation conditions (temperature, pH and time) were optimized to maximize the fermentation performance (ethanol titer, ethanol yield and lactose consumption) for the two preselected strains, K. marxianus DSM 5422 and S. cerevisiae Ethanol Red, using a response surface methodology (RSM). Under optimized conditions, K. marxianus DSM 5422 attained ethanol titers of 6% (v/v) in only 44 h. Moreover, the feasibility of immobilizing this strain on four different inorganic supports (plastic, glass and Tygon silicone Raschig rings and alumina beads) was assessed. Glass Raschig rings and alumina beads showed a more stable performance over time, yielding ethanol titers of 60 g/L during 1,000 hours, which remarkably reduces yeast cultivation costs. Results demonstrate the feasibility of using CWP for successful ethanol production in a simple and economical process, which represents an attractive alternative for waste treatment in dairy industries.

Evaluating the effect of biochar addition on the anaerobic digestion of swine manure: application of Py-GC/MS.

The anaerobic digestion process of swine manure was studied when char was used as supplement for improving performance. The use of pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) was proposed for assessing the organic matter degradation. The assessment on biogas production was carried out using samples of swine manure (SM) supplemented with char in one case and pre-treated by microwave irradiation in the other. This experimental set-up allows for the comparison of the biological degradation observed under these two different configurations and therefore aids in understanding the effect of char particles on the process. Results showed similar performance for both systems, with an average improvement of 39% being obtained in methane production when compared to the single digestion of SM. The analysis of digestate samples by Fourier transform infrared (FTIR) spectroscopy and Py-GC/MS showed improved degradation of proteins, with the Py-GC/MS technique also capable of identifying an increase in microbial-derived material when char was added, therefore highlighting the relevant role of carbon conductive particles on biological systems. Py-GC/MS along with the use of FTIR spectroscopy has proven to be useful tools when evaluating anaerobic digestion.