Giuseppe Olivieri

A novel approach to single-cell protein (SCP) production via hydrogen-oxidizing bacteria (HOB) is introduced to minimize explosion risks in hydrogen-based bioprocesses while assessing its implications for renewable-energy-based protein production systems. An alternative process is proposed where hydrogen (H2) and oxygen (O2) are spatially separated into two compartments: bioreactor and regenerator. Hydrogen is combined with an alternative electron carrier in a bioreactor, while oxygen is directed to a second compartment where the electron carrier is regenerated and energy is recovered. A thermodynamic analysis was performed to estimate yields and energetic efficiencies with a 13% margin of biological variability. Alternative processes based on HNO3 as electron carrier showed the highest potential when reduced to N2, NO, or N2O at low pH. Energy demand for SCP production was estimated to be 5.26, 5.82, and 5.52 MJ/molprotein, respectively. This was comparable to the 5.47 MJ/molprotein when using O2 as electron acceptor. The estimated yields for HNO3 reduction to N2 were in accordance with empirical biological yields. This showed that HOB-SCP energetic efficiency could be competitive with that of the conventional single-compartment aerobic method, offering a comparable energy demand, and a more integrated process design while eliminating explosion risks. This perspective on energetic efficiency is crucial for feasibility and techno-economic analysis of hydrogen-based SCP as a renewable-energy-driven route for sustainable protein production.

The demand for natural antioxidants to be used in food industry is increasing, as synthetic antioxidants are toxic and have high production costs. Specifically, food processing and preservation require antioxidants resistant to thermal sterilization processes. In this study, twenty-five strains among microalgae and cyanobacteria were screened as antioxidants producers. The species Enallax sp., Synechococcus bigranulatus and Galdieria sulphuraria showed the highest content of chlorophyll a and total carotenoids. In vitro stability and antioxidant activity of the ethanolic extracts were performed. The results revealed that pigments present in the extracts, obtained from the previously mentioned species, were stable at room temperature and exhibited in vitro free radical scavenging potential with IC₅₀ values of 0.099 ± 0.001, 0.048 ± 0.001 and 0.13 ± 0.02 mg mL^-1, respectively. Biocompatibility assay showed that the extracts were not toxic on immortalized cell lines. The antioxidant activity was also tested on a cell-based model by measuring intracellular ROS levels after sodium arsenite treatment. Noteworthy, extracts were able to exert the same protective effect, before and after the pasteurization process. Results clearly indicate the feasibility of obtaining biologically active and thermostable antioxidants from microalgae. Green solvents can be used to obtain thermo-resistant antioxidants from cyanobacteria and microalgae which can be used in the food industry. Thus, the substitution of synthetic pigments with natural ones is now practicable. Graphical abstract

Microalgae biomass production rate in short light-path photobioreactors potentially can be improved by mixing-induced flashing light regimes. A cascade photobioreactor features a thin liquid layer flowing down a sloping, wavy-bottomed surface where liquid flow exhibits peculiar local recirculation hydrodynamics, potentially conducive to an ordered flashing light regime. This article presents a model-based analysis of the frequency distribution of perceived irradiance in said wavy-bottomed photobioreactor. The model combines a Lagrangian description of the motion of individual cells, in turn derived from the hydrodynamic parameters of the photobioreactor extracted from an experimentally validated Computational Fluid Dynamic model, with a simplified description of the irradiance field across the culture thickness, down to the spectral analysis of perceived irradiance. The main finding of the work is that the wavy bottomed photobioreactor provides a ‘robust’ spectral excitation to the circulating microalgae up to 3 Hz frequency, while in flat panels and bubble columns excitation decays evenly at a 24 db/octave rate. This analysis paves the way to improving the light flashing performance of the wavy-bottomed photobioreactor with respect to geometry (cavity size and installation inclination) and operation (flow rate).

Saccharification of lignocellulosic biomass is a fundamental step in the biorefinery of second generation feedstock. The physicochemical and enzymatic processes for the depolymerization of biomass into simple sugars has been achieved through numerous studies in several disciplines. The present review discusses the development of technologies for enzymatic saccharification in industrial processes. The kinetics of cellulolytic enzymes involved in polysaccharide hydrolysis has been discussed as the starting point for the design of the most promising bioreactor configurations. The main process configurations—proposed so far—for biomass saccharification have been analyzed. Attention was paid to bioreactor configurations, operating modes and possible integrations of this operation within the biorefinery. The focus is on minimizing the effects of product inhibition on enzymes, maximizing yields and concentration of sugars in the hydrolysate, and reducing the impact of enzyme cost on the whole process. The last part of the review is focused on an emerging process based on the catalytic action of laccase applied to lignin depolymerization as an alternative to the consolidated physicochemical pretreatments. The laccases-based oxidative process has been discussed in terms of characteristics that can affect the development of a bioreactor unit where laccases or a laccase-mediator system can be used for biomass delignification.

Microalgae started receiving attention as producers of third generation of biofuel, but they are rich in many bioactive compounds. Indeed, they produce many molecules endowed with benefic effects on human health which are highly requested in the market. Thus, it would be important to fractionate algal biomass into its several high-value compounds: this represents the basis of the microalgal biorefinery approach. Usually, conventional extraction methods have been used to extract a single class of molecules, with many side effects on the environment and on human health. The development of a green downstream platform could help in obtaining different class of molecules with high purity along with low environmental impact. This review is focused on technical advances that have been performed, from classic methods to the newest and green ones. Indeed, it is fundamental to set up new procedures that do not affect the biological activity of the extracted molecules. A comparative analysis has been performed among the conventional methods and the new extraction techniques, i.e., switchable solvents and microwave-assisted and compressed fluid extractions.

The increasing impact of plastic materials on the environment is a growing global concern. In regards to this circumstance, it is a major challenge to find new sources for the production of bioplastics. Poly-β-hydroxybutyrate (PHB) is characterized by interesting features that draw attention for research and commercial ventures. Indeed, PHB is eco-friendly, biodegradable, and biocompatible. Bacterial fermentation processes are a known route to produce PHB. However, the production of PHB through the chemoheterotrophic bacterial system is very expensive due to the high costs of the carbon source for the growth of the organism. On the contrary, the production of PHB through the photoautotrophic cyanobacterium system is considered an attractive alternative for a low-cost PHB production because of the inexpensive feedstock (CO₂ and light). This paper regards the evaluation of four independent strategies to improve the PHB production by cyanobacteria: (i) the design of the medium; (ii) the genetic engineering to improve the PHB accumulation; (iii) the development of robust models as a tool to identify the bottleneck(s) of the PHB production to maximize the production; and (iv) the continuous operation mode in a photobioreactor for PHB production. The synergic effect of these strategies could address the design of the optimal PHB production process by cyanobacteria. A further limitation for the commercial production of PHB via the biotechnological route are the high costs related to the recovery of PHB granules. Therefore, a further challenge is to select a low-cost and environmentally friendly process to recover PHB from cyanobacteria.

The setup of an economic and sustainable method to increase the production and commercialization of products from microalgae, beyond niche markets, is a challenge. Here, a cascade approach has been designed to optimize the recovery of high valuable bioproducts starting from the wet biomass of Galdieria phlegrea. This unicellular thermo-acidophilic red alga can accumulate high-value compounds and can live under conditions considered hostile to most other species. Extractions were performed in two sequential steps: a conventional high-pressure procedure to recover phycocyanins and a solvent extraction to obtain fatty acids. Phycocyanins were purified to the highest purification grade reported so far and were active as antioxidants on a cell-based model. Fatty acids isolated from the residual biomass contained high amount of PUFAs, more than those recovered from the raw biomass. Thus, a simple, economic, and high effective procedure was set up to isolate phycocyanin at high purity levels and PUFAs.