Halophyte biorefinery for polyhydroxyalkanoates production from Ulva sp. Hydrolysate with Haloferax mediterranei in pneumatically agitated bioreactors and ultrasound harvesting.

The present study tested the outdoor cultivation of Haloferax mediterranei for PHA production from green macroalgae Ulva sp. in pneumatically agitated bioreactors and applied ultrasonic separation for enhanced settling of archaeal cells. Scaled-up cultivation (40 L) yielded maximum biomass productivity of 50.1 ± 0.11 mg·L-1·h-1 with a PHA productivity of 27 ± 0.01 mg·L-1·h-1 and conversion yield of 0.107 g PHA per gram UlvaDW. The maximum mass fraction of PHA achieved in biomass was calculated to be 56% w/w. Ultrasonic harvesting of Hfx. mediterranei cells approached 30% removal at energy inputs around 7.8 kWh·m-3, and indicated no significant aggregation enhancement by Ca2+ addition. Molecular weight analysis showed an increase in Polydispersity Index (PDI) when the corresponding air velocities were increased suggesting that the polymer was more homogeneous at lower mixing velocities. The current study demonstrated scalable processes for PHA production using Ulva sp. feedstock providing new technologies for halophilic biorefinery.

Fighting SARS-CoV-2 with green seaweed Ulva sp. extract: extraction protocol predetermines crude ulvan extract anti-SARS-CoV-2 inhibition properties in in vitro Vero-E6 cells assay.

Due to the global COVID-19 pandemic, there is a need to screen for novel compounds with antiviral activity against SARS-COV-2. Here we compared chemical composition and the in vitro anti- SARS-COV-2 activity of two different Ulva sp. crude ulvan extracts: one obtained by an HCl-based and another one by ammonium oxalate-based (AOx) extraction protocols. The composition of the crude extracts was analyzed and their antiviral activity was assessed in a cytopathic effect reduction assay using Vero E6 cells. We show that the extraction protocols have a significant impact on the chemical composition, anti- SARS-COV-2 activity, and cytotoxicity of these ulvan extracts. The ulvan extract based on the AOx protocol had a higher average molecular weight, higher charge, and 11.3-fold higher antiviral activity than HCl-based extract. Our results strongly suggest that further bioassay-guided investigation into bioactivity of compounds found in Ulva sp. ulvan extracts could lead to the discovery of novel anti-SARS-CoV-2 antivirals.

Polyhydroxyalkanoates and biochar from green macroalgal Ulva sp. biomass subcritical hydrolysates: Process optimization and a priori economic and greenhouse emissions break-even analysis.

Although macroalgae biomass is an emerging sustainable feedstock for biorefineries, the optimum process parameters for their hydrolysis and fermentation are still not known. In the present study, the simultaneous production of polyhydroxyalkanoates (PHA) and biochar from green macroalgae Ulva sp. is examined, applying subcritical water hydrolysis and Haloferax mediterranei fermentation. First, the effects of temperature, treatment time, salinity, and solid load on the biomass and PHA productivity were optimized following the Taguchi method. Hydrolysis at 170 °C, 20 min residence time, 38 g L-1 salinity with a seaweed solid load of 5% led to the maximum PHA yield of 0.104 g g-1Ulva and a biochar yield of 0.194 ± 1.23 g g-1Ulva. Second, the effect of different initial culture densities on the biomass and PHA productivity was studied. An initial culture density of 50 g L-1 led to the maximum volumetric PHA productivity of 0.024 ± 0.002 g L-1 h-1 with a maximum PHA content of 49.38 ± 0.3% w/w Sensitivity analysis shows that within 90% confidence, the annual PHA production from Ulva sp. is 148.14 g PHA m-2 year-1 with an annual biochar production of 42.6 g m-2 year-1. Priori economic and greenhouse gas break-even analyses of the process were done to estimate annual revenues and allowable greenhouse gas emissions. The study illustrates that PHA production from seaweed hydrolysate using extreme halophiles coupled to biochar production could become a benign and promising step in a marine biorefinery.

Energy efficient dewatering of far offshore grown green macroalgae Ulva sp. biomass with pulsed electric fields and mechanical press.

Offshore macroalgae biomass production is a promising, yet challenging, pathway to provide feedstock for biorefineries. In this work, a device and a process for dewatering offshore grown biomass of the green macroalgae Ulva sp. using high-voltage pulsed electric fields (PEF) was developed. Ulva sp. was cultivated attached to fish cages 15 km offshore. Increasing the applied voltage from 250 V to 500 V and invested PEF energy from 9.3 ±â€¯0.4 J g-1 FW to 54.6 ±â€¯0.2Jg-1 FW increased the extracted water from 0.033 ±â€¯0.006 g Water g-1  FW to 0.150 ±â€¯0.031 g Water g-1 FW. The energy consumption to achieve similar moisture content with air convection drying was lower by 78.73 ±â€¯10.41 (JgFW-1) for 250 V and 339.31 ±â€¯48.01 (JgFW-1) for 500 V, pulse duration 50 µs, pulse number 50, pulse repetition frequency 3 Hz. PEF leads to biomass compression of 8.45 ±â€¯1.72% for 250 V protocol and 25.66 ±â€¯2.53% for 500 V protocol. In addition, PEF leads to the reduction of water diffusivity of 18-19% in the treated biomass, reducing air drying kinetics.

Metabolic responses of Ulva compressa to single and combined heavy metals.

Accumulation of metals and metabolic responses were studied for two Cd and Cu concentrations (1 and 10 µM) either alone or as a combination in marine macroalga after 7 days of exposure. Cd accumulated more at a low dose (115 µg of Cd/g DW) but Cu at a high dose (378 µg of Cu/g DW); Cu suppressed Cd accumulation (by 57%). Na and Zn levels were unaffected, but higher metal doses depleted K and Ca levels. Higher metal concentrations strongly stimulated reactive oxygen species and depleted nitric oxide (NO) formation, but differences between the action of Cd and Cu were not extensive. Higher metal doses increased cell wall thickness with a potential relation to NO signal that is visible mainly in the apoplast in those treatments. A higher Cu dose depleted proline, ascorbic acid, and phenol levels more than Cd, whereas Cd elevated nonprotein thiols and ascorbic acid in combined treatments. An eventual role of malic or citric acid in metal chelation was not evident: malic acid level decreased in all treatments. The total content of fatty acids reached 16.7 mg/g DW in control with the quantitative order of PUFAs > SFAs > MUFAs; palmitic, vaccenic, linoleic, and α-linolenic acids were the major compounds. Cu was more toxic for fatty acids than Cd (even at 1 µM); mainly, PUFA levels strongly decreased (from 43% of total acids in control to 28.9% and 5.4% at 1 and 10 µM Cu treatment, respectively). Results are precisely and critically discussed in relation to limited literature focused on macroalgae, and a comparison with microalgae is also provided.

Design of marine macroalgae photobioreactor integrated into building to support seagriculture for biorefinery and bioeconomy.

Seagriculture, which can provide offshore grown macroalgae biomass would play a significant role in bioeconomy. Nevertheless, seagriculture development has been hindered by the lack of laboratory photobioreactors that enable fundamental and pilot scale macroalgae research. In this work, a macroalgae photobioreactor (MPBR) was developed and integrated into the building. The MPBR operation was demonstrated for 6months with cultivation of Cladophora sp., Ulva compressa and Ulva rigida green macroalgae species isolated from 3 sites at the Eastern Mediterranean coast. The growth rate, protein, ash, specific energy density, rhamnose, xylose, arabinose, glucose, galactose and glucuronic acid content of the cultivated species were quantified. The maximum accumulated energy rates were 0.033WhL-1d-1 for Cladophora sp., 0.081WhL-1d-1 for U. compressa and 0.029WhL-1d-1 for U. rigida. This work provides a detailed design of an indoor, urban photobioreactor for cultivation, maintenance and energy balance analysis of macroalgae biomass for biorefinery.