Enhancement of phycocyanin productivity and thermostability from Arthrospira platensis using organic acids.

Intracellular hyperaccumulation of phycocyanin (PC) and its high susceptibility to degradation at higher temperatures are major challenging problems associated with its production from cyanobacteria. The present study evaluated different concentrations of organic acids (1, 2, and 3 mM) (citric acid, acetic acid, succinic acid, fumaric acid, and oxalic acid) under fed-batch mode on the biomass and phycobiliproteins' production from Arthrospira platensis. Besides they were evaluated at 2.5-7.5 mM as preservative to stabilize PC at high temperatures. The incorporation of 3 mM of succinic acid into the cultivation medium enhanced the biomass and PC productivity to 164.05 and 26.70 mg L-1 day-1, which was ~ 2- and threefold higher than control, respectively. The produced PC in this treatment was food-grade with a 2.2 purity ratio. The use of organic acids also enhanced the thermal stability of PC. Citric acid (7.5 mM) markedly promoted the half-life values of PC to 189.44 min compared to 71.84 min in the control. The thermodynamic analysis confirmed higher thermostability of PC in the presence of organic acids and indicated the endothermic and non-spontaneity of the thermal denaturation process. The findings of the present study confirmed that organic acids could be utilized as cost effective and sustainable compounds for promoting not only phycobiliproteins' production but also the thermostability of PC for potential application in food industry.

Low-cost biosorption of Fe(II) and Fe(III) from single and binary solutions using Ulva lactuca-derived cellulose nanocrystals-graphene oxide composite film.

The marine algal biomass of Ulva lactuca was utilized for the extraction of cellulose and the development of cellulose nanocrystals/graphene oxide film. Cellulose nanocrystals with 50-150 nm were produced by H2SO4 hydrolysis of the algal cellulose. The adsorption efficiency of the nanocomposite film for Fe(II) and Fe(III) ions was successfully evaluated using Box-Behnken design. The maximum removal for Fe(II) (64.15%) could be attained at pH 5.13, adsorbent dosage 7.93 g L-1 and Fe(II) concentration 15.39 mg L-1, while the biosorption of Fe(III) was 69.92% at pH 5.0, adsorbent dosage 2 g L-1, and Fe(III) concentration 15.0 mg L-1. However, in the binary system, the removal efficiency of Fe(II) was enhanced to 95.48% at Fe(II):Fe(III) ratio of 1:1, while the Fe(III) removal was increased to 79.17% at ratio 1:2. The pseudo-second-order kinetics exhibited better fitting to the experimental results of Fe(II) and Fe(III) adsorption in both single and binary systems. The intra-particle diffusion was prominent during the biosorption, but the effect of the external mass transfer was significant. The Langmuir, Freundlich, Langmuir-Freundlich, Temkin, and Dubinin-Radushkevich isotherms showed satisfactory fitting to the experimental data, but they differ in priority based on iron state and pH. The adsorption of Fe(II) in the presence of Fe(III) in a mixture was best represented by the extended Langmuir model, while the extended Langmuir-Freundlich model best fitted the adsorption of Fe(III). The FT-IR analysis indicated that physisorption through electrostatic interaction/complexation is the predominant mechanism for the adsorption of iron using the nanocomposite film.

Hormesis effects of phenol on growth and cellular metabolites of Chlorella sp. under different nutritional conditions using response surface methodology.

The present study investigated the effects of different phenol concentrations (200 - 1000 mg L-1) towards Chlorella sp. under different culture conditions (light vs. dark) and NaNO3 concentrations (0 - 0.1 g L-1) using central composite design. Phenol induced hormesis effects on the algal growth and cellular metabolites. Nitrate was identified as a crucial factor for promoting the uptake of phenol by Chlorella cells, while light was a limiting factor for growth, but the phyco-toxicity of phenol was decreased in the dark. The pigment contents were generally increased in the treated cells to protect against the oxidative phenol stress. The incorporation of 200 mg L-1 phenol and 0.05 g L-1 NaNO3 to the illuminated cells markedly promoted biomass and lipid contents to 0.22 g L-1 and 26.26% w/w, which was 44 and 112% higher than the phenol-less control, respectively. Under the same conditions, the increase of phenol concentration to 600 mg L-1, the protein contents were increased to 18.59% w/w. Conversely, the algal cells were able to accumulate more than 60% w/w of soluble carbohydrates under dark conditions at 600 mg L-1 of phenol. Nitrate replete conditions stimulated lipid accumulation at the expense of protein biosynthesis. Furthermore, most of the treatments showed an increase of H2O2 and malonaldehyde contents, especially for the illuminated cells. However, catalase activity tended to increase under dark conditions, especially at low phenol and nitrate concentrations. This study is valuable in indicating the effects of phenol on microalgae by exploiting response surface methodology, which can be applied as a powerful tool in growth monitoring and toxicity assessment.

Fungicidal activities and molecular docking of the marine alga Ulva lactuca and Punica granatum peel extracts on Alternaria tomato spot disease.

In this study, we utilized pomegranate peel and marine algae Ulva lactuca (U. lactuca) as rich and sustained sources of bioactive compounds to combat tomato-black spot disease. n-Hexane extracts from the peel of pomegranate (Punica granatum) (PPE) and the marine algal biomass U. lactuca (ULE) were used alone and in combinations to verify their impact against Alternaria alternata (A. alternata). The applied extracts exhibited severe destructive effects on both fungal growth and structure such as mycelia malformation, underdeveloped conidia, cell wall deformation, and shrinkage. Moreover, increased deformations and protrusions, and notch-like structures, were noticed in A. alternata mycelia treated with mixed extracts (PPE and ULE) compared to all other treatments. The protein and reduced sugar contents in tomato fruits were significantly increased in the infected fruits with A. alternata. The highest enzyme activities of pectinase, cellulase, catalase (CAT), and ascorbate peroxidase (APX) were recorded in infected tomatoes in comparison with the healthy ones. Molecular docking studies showed that each extract is rich with bioactive compounds that have a promising inhibition effect on A. alternata cellulases. Pomegranate and Ulva extract showed promising antifungal activity against A. alternata which revealed their feasibility and applicability as biocontrol agents in postharvest disease management and food preservation against fungal pathogens.

Biosorption of ketoprofen and diclofenac by living cells of the green microalgae Chlorella sp.

There is a growing interest for the removal of different pharmaceuticals from water owing to their toxicity to various organisms. The present study investigated the use of living cells of the green alga Chlorella sp. in the short-term adsorption of ketoprofen (KET) and diclofenac (DIF) from aqueous solutions. The bioremoval efficiency of both KET and DIF was highly dependent on various parameters such as time, pH, algal dosage, and drug concentration. The adsorption efficiencies of both KET and DIC were maximized at pH 6. The biosorption of KET was better described by pseudo-first-order kinetics, while DIC obeyed the pseudo-second-order model. The maximum adsorption capacities of KET and DIF were attained as 0.328 and 0.429 mg g-1, respectively. The equilibrium data of the investigated drugs showed a better fit to the Freundlich model than the Langmuir model. The Elovich and Temkin models indicated that the algal surface was heterogeneous with different binding energies, while the intraparticle diffusion model assumed a boundary layer effect. Additionally, the Dubinin-Radushkevich isotherm indicated that the adsorption process was predominantly physisorption. FT-IR analysis revealed that H-bonding and n-π interactions were prominent in the biosorption process of the investigated pharmaceuticals on the surface of microalgae. The results of the present study showed that microalgae living cells could be applied as an eco-friendly and cost-effective biosorbent for the removal of KET and DIF at low concentrations.

Environmental risk analysis of pharmaceuticals on freshwater phytoplankton assemblage: effects on alpha, beta, and taxonomic diversity.

Antibiotics and nonsteroidal anti-inflammatory drugs (NSAIDs) have a wide range of bioactivities and are released into the ecosystem in large amounts. Heretofore, little information is available regarding their potential risk to the phytoplankton assemblage. Different alpha, taxonomic, and beta diversity measures were investigated and linked to the spatial variation of nine drugs. Distance-based redundancy analysis (dbRDA) indicated that pharmaceutical pollution had adverse effects on both phytoplankton diversity and taxonomic structure leading to the existence of congeneric taxa. However, different phytoplankton groups respond differently to different pharmaceuticals and Cyanoprokaryotes was suggested as the most sensitive group. According to the EC50 value and the detected concentration for each drug, a hazard index (Hq) was calculated for each polluted site to investigate environmental risk analysis. Increasing Hq values exhibited negative effects on phytoplankton diversity. Phytoplankton community was characterized by high beta diversity values, which suggested that microalgae were able to disperse and select suitable environmental conditions. High beta diversity values were driven by species difference rather than species replacement due to the disappearance of most sensitive taxa from highly polluted sites. Additionally, microalgae were classified into different morpho-functional groups (FGs), and principal component analysis (PCA) indicated that different FGs had different responses to pharmaceutical pollution. A laboratory toxicity experiment was also conducted to identify the negative effects of short-term exposure to low doses of paracetamol and ciprofloxacin.

Upgrading the antioxidant properties of fucoidan and alginate from Cystoseira trinodis by fungal fermentation or enzymatic pretreatment of the seaweed biomass.

The seaweed Cystoseira trinodis was fermented by different fungi prior to extraction of fucoidan and alginate to enhance their antioxidative potential. All the investigated fungi were able to produce fucoidanase (1.05-3.41 U/ml) and alginate lyase (7.27-18.59 U/mL). Different fungal species induced a reduction in the molecular weight (MW) of fucoidan and alginate in comparison to the unfermented control. The MW of fucoidan reduced by 41-81.5%, while the MW of alginate was reduced by 28-75%, depending on the fungal species. Significant increases in the fucose and sulphate contents of fucoidan and mannuronic/guluronic acid ratio of alginate were induced by fungal fermentation. Fungal pretreatment enhanced the ferric reducing antioxidant power, total antioxidant capacity and hydroxyl radical scavenging activity of both fucoidan and alginate. Additionally, enzymatic pretreatment of the macroalgal biomass assisted in the recovery of fucoidan and alginate with low molecular weight and enhanced antioxidative potential.

Statistical Optimization of Culture Variables for Enhancing Agarase Production by Dendryphiella arenaria Utilizing Palisada perforata (Rhodophyta) and Enzymatic Saccharification of the Macroalgal Biomass.

Agarase is a promising biocatalyst for several industrial applications. Agarase production was evaluated by the marine fungus Dendryphiella arenaria utilizing Palisada perforata as a basal substrate in semi-solid state fermentation. Seaweed biomass, glucose, and sucrose were the most significant parameters affecting agarase production, and their levels were further optimized using Box-Behnken design. The maximum agarase activity was 7.69 U/mL. Agarase showed a degree of thermostability with half-life of 99 min at 40 °C, and declining to 44.72 min at 80 °C. Thermodynamics suggested an important process of protein aggregation during thermal inactivation. Additionally, the enzymatic saccharification of the seaweed biomass using crude agarase was optimized with respect to biomass particle size, solid/liquid ratio, and enzyme loadings. The amount of biosugars obtained after optimization was 26.15 ± 1.43 mg/g. To the best of our knowledge, this is the first report on optimization of agarase in D. arenaria.

Optimization of alginate alkaline extraction technology from Sargassum latifolium and its potential antioxidant and emulsifying properties.

Alginate was recovered from Sargassum latifolium biomass using different conditions of alkali treatment. Box-Behnken experimental design was evaluated to study the influence of alkali:alga ratio, temperature and time on alginate yield, and its molecular weight (MW) and mannuronic/guluronic acid ratio (M/G). The second-order polynomial equations were analyzed by appropriate statistical methods. Extraction temperature and time were the most important factors during alginate alkaline extraction. MW and M/G ratio played an important role in controlling the reducing power of alginate. Increasing pH of the alginate solutions enhanced its reducing capacity, while thermal treatment showed a negative effect. Additionally, alginate exhibited good emulsion stabilizing capacities with diverse hydrophobic compounds. Emulsifying activity was less sensitive to temperature, ionic strength and more stable at acidic pH.