Genome-scale metabolic modelling of extremophiles and its applications in astrobiological environments.

Metabolic modelling approaches have become the powerful tools in modern biology. These mathematical models are widely used to predict metabolic phenotypes of the organisms or communities of interest, and to identify metabolic targets in metabolic engineering. Apart from a broad range of industrial applications, the possibility of using metabolic modelling in the contexts of astrobiology are poorly explored. In this mini-review, we consolidated the concepts and related applications of applying metabolic modelling in studying organisms in space-related environments, specifically the extremophilic microbes. We recapitulated the current state of the art in metabolic modelling approaches and their advantages in the astrobiological context. Our review encompassed the applications of metabolic modelling in the theoretical investigation of the origin of life within prebiotic environments, as well as the compilation of existing uses of genome-scale metabolic models of extremophiles. Furthermore, we emphasize the current challenges associated with applying this technique in extreme environments, and conclude this review by discussing the potential implementation of metabolic models to explore theoretically optimal metabolic networks under various space conditions. Through this mini-review, our aim is to highlight the potential of metabolic modelling in advancing the study of astrobiology.

Inhibition of dengue virus infection in vitro by fucoidan and polysaccharide extract from marine alga Sargassum spp.

Dengue virus (DENV) infection poses a global health threat, leading to severe conditions with the potential for critical outcomes. Currently, there are no specific drugs available whereas the vaccine does not offer comprehensive protection across all DENV serotypes. Therefore, the development of potential antiviral agents is necessary to reduce the severity risk and interrupt the transmission circuit. The search for effective antiviral agents against DENV has predominantly focused on natural resources, particularly those demonstrating diverse biological activities and high safety profiles. Cyanobacteria and algae including Leptolyngbya sp., Spirulina sp., Chlorella sp., and Sargassum spp., which are prevalent species in Thailand, have been reported for their diverse biological activities and high safety profiles. However, their anti-DENV activity has not been documented. In this study, the screening assay was performed to compare the antiviral activity against DENV of crude polysaccharide and ethanolic extracts derived from 4 species of cyanobacteria and algae in Vero cells. Polysaccharide extracts from Sargassum spp. were the most effective in inhibiting DENV-2 infection under co-infection conditions, where the virus was exposed to the extract at the time of infection. Treatment of the extract significantly reduced the ability of DENV to bind to the host cells to 47.87 ± 3.88 % while treatment upon virus binding step had no antiviral effect suggesting the underlaying mechanism of the extract on interfering virus binding step. Fucoidan, a key bioactive substance in Sargassum polysaccharide, showed to reduce DENV-2 infection to 26.59 ± 5.01 %, 20.46 ± 6.58 % under the co-infection condition in Vero and A549 cells, respectively. In accompanied with Sargassum polysaccharide, fucoidan disturbed the virus binding to the host cells. These findings warrant further development and exploration of the Sargassum-derived polysaccharide, fucoidan, as a promising candidate for combating DENV infections.

Zero-waste biorefining co-products from ultrasonically assisted deep eutectic solvent-pretreated Chlorella biomass: Sustainable production of biodiesel and bio-fertilizer.

Microalgal biomass is gaining increasing attention to produce high-value co-products. This study proposes integrating Chlorella microalgal biomass into a zero-waste biorefining system, aiming to produce biodiesel and biofertilizer. It investigates optimal conditions for ultrasound-assisted deep eutectic solvent (DES) pretreatment and lipid recovery to enhance the extraction of lipids. Optimal DES pretreatment was identified as a 1.6:1 acetic acid-to-choline chloride molar ratio, 0.36 g biomass loading, and 2.50 min of pretreatment. Lipid recovery succeeded with a 10-minute extraction time and a 1:3 methanol-to-butanol volume ratio. These conditions yielded biodiesel-quality lipids at 139.52 mg/g microalgal biomass with superior fuel characteristics. The de-oiled microalgal biomass residue exhibited promise as a lettuce biofertilizer, enhancing photosynthetic pigments but potentially reducing yields by 40 %. The study also notes changes in rhizosphere microbial communities, indicating both stimulatory and inhibitory effects on beneficial microbes. This study has the potential to enhance sustainability in energy, agriculture, and the environment.

Unlocking microalgal host-exploring dark-growing microalgae transformation for sustainable high-value phytochemical production.

Microalgae have emerged as a promising, next-generation sustainable resource with versatile applications, particularly as expression platforms and green cell factories. They possess the ability to overcome the limitations of terrestrial plants, such non-arable land, water scarcity, time-intensive growth, and seasonal changes. However, the heterologous expression of interested genes in microalgae under heterotrophic cultivation (dark mode) remains a niche area within the field of engineering technologies. In this study, the green microalga, Chlorella sorokiniana AARL G015 was chosen as a potential candidate due to its remarkable capacity for rapid growth in complete darkness, its ability to utilize diverse carbon sources, and its potential for wastewater treatment in a circular bioeconomy model. The aims of this study were to advance microalgal genetic engineering via dark cultivation, thereby positioning the strain as promising dark-host for expressing heterologous genes to produce high-value phytochemicals and ingredients for food and feed. To facilitate comprehensive screening based on resistance, eleven common antibiotics were tested under heterotrophic condition. As the most effective selectable markers for this strain, G418, hygromycin, and streptomycin exhibited growth inhibition rates of 98%, 93%, and 92%, respectively, ensuring robust long-term transgenic growth. Successful transformation was achieved through microalgal cell cocultivation with Agrobacterium under complete darkness verified through the expression of green fluorescence protein and ß-glucuronidase. In summary, this study pioneers an alternative dark-host microalgal platform, using, Chlorella, under dark mode, presenting an easy protocol for heterologous gene transformation for microalgal host, devoid of the need for expensive equipment and light for industrial production. Furthermore, the developed genetic transformation methodology presents a sustainable way for production of high-value nutrients, dietary supplements, nutraceuticals, proteins and pharmaceuticals using heterotrophic microalgae as an innovative host system.

Dual-bioaugmentation strategy to enhance the formation of algal-bacteria symbiosis biofloc in aquaculture wastewater supplemented with agricultural wastes as an alternative nutrient sources and biomass support materials.

This study performs an integrated evaluation of the formation and distribution of algal-bacterial bioflocs in aquaculture wastewater supplemented with agricultural waste, together with an assessment of their behavior in the microbial community and of the water quality of the system in which a new bioaugmentation strategy was applied. Results indicated that the dual bioaugmentation strategy via the consortium addition of bacteria and microalgae had the highest formation performance, providing the most compact biofloc structure (0.59 g/L), excellent settleability (71.91%), and a large particle diameter (4.25 mm). The fed-batch supplementation of molasses and rice bran, in terms of changes in the values of COD, NH4+, NO3-, and PO43-, stimulated the formation of biofloc through algal-bacterial bioflocs and microbe-rice bran complexes within a well-established microbial community. These findings provide new insight into the influence of bioaugmentation on the formation of an innovative algal-bacterial biofloc.

Lipid Profile, Antioxidant and Antihypertensive Activity, and Computational Molecular Docking of Diatom Fatty Acids as ACE Inhibitors.

Diatoms, as single cell eukaryotic microalgae, are rich sources of lipids, which have either beneficial or detrimental effects on the prevention and treatment of many diseases. Gas chromatography-mass spectrometry (GC-MS) identified diatom lipids with high levels of essential fatty acids (EFAs), especially polyunsaturated FAs (PUFAs) containing both omega-3 and omega-6. Nutritional values of FAs indicated possible applications in the pharmaceutical, nutraceutical, and functional food industries. Diatom FAs showed antioxidative potential on harmful radicals by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) scavenging, with high inhibition of the angiotensin-converting enzyme (ACE) that causes cardiovascular disease (CVD) and hypertension. A computational molecular docking simulation confirmed the inhibition mechanisms of FAs on ACE, with comparable levels of binding free energy to chemically synthesized ACE drugs. Findings suggested that diatom lipids showed potential for use as alternative ACE inhibitors or food supplement for CVD prevention.

Naturally Occurring Functional Ingredient from Filamentous Thermophilic Cyanobacterium Leptolyngbya sp. KC45: Phytochemical Characterizations and Their Multiple Bioactivities.

Cyanobacteria are rich in phytochemicals, which have beneficial impacts on the prevention of many diseases. This study aimed to comprehensively characterize phytochemicals and evaluate multifunctional bioactivities in the ethanolic extract of the cyanobacterium Leptolyngbya sp. KC45. Results found that the extract mainly contained chlorophylls, carotenoids, phenolics, and flavonoids. Through LC-ESI-QTOF-MS/MS analysis, 38 phenolic compounds with promising bioactivities were discovered, and a higher diversity of flavonoids was found among the phenolic compounds identified. The extract effectively absorbed the harmful UV rays and showed high antioxidant activity on DPPH, ABTS, and PFRAP. The extract yielded high-efficiency inhibitory effects on enzymes (tyrosinase, collagenase, ACE, and α-glucosidase) related to diseases. Interestingly, the extract showed a strong cytotoxic effect on cancer cells (skin A375, lung A549, and colon Caco-2), but had a much smaller effect on normal cells, indicating a satisfactory level of safety for the extract. More importantly, the combination of the DNA ladder assay and the TUNEL assay proved the appearance of DNA fragmentation in cancer cells after a 48 h treatment with the extract, confirming the apoptosis mechanisms. Our findings suggest that cyanobacterium extract could be potentially used as a functional ingredient for various industrial applications in foods, cosmetics, pharmaceuticals, and nutraceuticals.

Enhanced production of microalgal biomass and lipid as an environmentally friendly biodiesel feedstock through actinomycete co-culture in biogas digestate effluent.

In this study, an innovative approach to enhance the production of microalgal biomass and lipid as a promising sustainable feedstock for biodiesel was proposed using an actinomycetes co-culture with microalgae in the biogas digestate effluent (BDE) that can be employed as an environmentally friendly and cost-effective strategy. Among tested actinomycete isolates, Piscicocus intestinalis WA3 produced indole-3-acetic acid and siderophores as algal growth promoting agents and showed effective lipid accumulation with satisfying fatty acids composition. During co-cultivation of P. intestinalis WA3 with microalga Tetradesmus obliquus AARL G022 in the BDE, biomass production, chlorophyll a content, and lipid productivity were significantly increased by 1.30 folds, 1.39 folds, and 1.55 folds, respectively, compared to microalgae monoculture. The accumulated lipids contained long-chain fatty acids with better fuel properties that could potentially be used as biodiesel feedstock. The overall results evidenced that actinomycete co-culture would contribute greatly to the cost-effective production of environmental-friendly microbial-based biofuel.