BG-11 Medium Research Articles

Enhancement of dark fermentative hydrogen production in nitrogen-deprived halotolerant unicellular cyanobacterium Aphanothece halophytica by treatment with reducing agents

To enhance H2 production by the halotolerant unicellular cyanobacterium Aphanothece halophytica, effect of various kinds of reducing sugar and reducing agent on H2 production was investigated. The highest H2 production rate of 55.80 ± 0.50 μmol H2 g dry weight−1 h−1 was obtained when the cells were incubated in BG110 medium containing 0.189 mmol C-atom L−1 glucose under dark anaerobic condition. This rate was 1.5 folds higher than that without glucose. Among ten reducing agents tested, β-mercaptoethanol, dithiothreitol, l-cysteine and sodium sulfide had high potential as an effective reducing agent to increase H2 production by A. halophytica. Cells treated with 50 mM sodium sulfide showed the highest H2 accumulation with 4815.59 ± 194.78 μmol H2 g dry weight−1 after 24 h of dark anaerobic incubation. An increase in H2 production was ascribed to an increase of hydrogenase activity and a decrease of O2 production rate. This H2 production yield was approximately 20 folds higher than that without reducing agent. Furthermore, 50 mM sodium sulfide appeared to be non-toxic to A. halophytica cells, since the IC50 of sodium sulfide was higher than 100 mM. The reduced ferredoxin at 1.5 μM-1.5 mM, NADH and NADPH at 0.15–1.5 mM could support in vitro [NiFe]-H2ase activity, demonstrating the ability of these compounds to provide electrons towards [NiFe]-H2ase in A. halophytica.

Formation of Lutein, β-Carotene and Astaxanthin in a Coelastrella sp. Isolate.

In this study, the effect of media composition, N/P ratio and cultivation strategy on the formation of carotenoids in a Coelastrella sp. isolate was investigated. A two-stage process utilizing different media in the vegetative stage, with subsequent re-suspension in medium without nitrate, was employed to enhance the formation of carotenoids. The optimal growth and carotenoid content (β-carotene and lutein) in the vegetative phase were obtained by cultivation in M-8 and BG11 media. Use of a N/P ratio of 37.5 and low light intensity of 40 μmol m−2 s−1 (control conditions) led to optimal biomass production of up to 1.31 g L−1. Low concentrations of astaxanthin (maximum of 0.31 wt. %) were accumulated under stress conditions (nitrogen-deficient medium containing 1.5 % of NaCl and light intensity of 500 μmol m−2 s−1), while β-carotene and lutein (combined maximum of 2.12 wt. %) were produced under non-stress conditions. Lipid analysis revealed that palmitic (C16:0) and oleic (C18:1) constituted the main algal fatty acid chains (50.2 ± 2.1% of the total fatty acids), while esterifiable lipids constituted 17.2 ± 0.5% of the biomass by weight. These results suggest that Coelastrella sp. could also be a promising feedstock for biodiesel production.

First Report of the Endophytic Bacteria Associated with Phormidium sp.

Recent molecular studies on endophytic bacterial diversity have revealed a large richness of species. Associations between endobiotic bacterial-algae interactions have been studied for more than 40 years but were, up to now, never molecularly analyzed within the filamentous Cyanobacteria Phormidium. Therefore, the endophytic bacteria associated with fresh microalgae Phormidium, a group of ubiquitous photosynthetic organisms that play an important role in aquatic ecosystems, has been investigated. To study this partnership, Phormidium sp. was cultured in BG-11 medium using optimal conditions, and after the incubation period, cell biomass was obtained. Total genomic DNA from biomass was extracted and used for endophytic bacteria determination by using the 16S rRNA gene. Sequencing results revealed that a total of seven endophytic bacteria living within the cytoplasm of the host Phormidium sp. have been identified, including six bacteria belonging to three genera, namely Sphingomonas, Sphingopyxis, and Stenotrophobacter and while one bacteria remained unidentified due to low sequence homology in the GenBank database. The results highlighted the importance of endophytic bacteria associated with Phormidium sp. for the first time by using sequence-based identification.

Combining metabolite doping and metabolic engineering to improve 2-phenylethanol production by engineered cyanobacteria.

2-Phenylethanol (2-PE) is a rose-scented aromatic compound, with broad application in cosmetic, pharmaceutical, food and beverage industries. Many plants naturally synthesize 2-PE via Shikimate Pathway, but its extraction is expensive and low-yielding. Consequently, most 2-PE derives from chemical synthesis, which employs petroleum as feedstock and generates unwanted by products and health issues. The need for “green” processes and the increasing public demand for natural products are pushing biotechnological production systems as promising alternatives. So far, several microorganisms have been investigated and engineered for 2-PE biosynthesis, but a few studies have focused on autotrophic microorganisms. Among them, the prokaryotic cyanobacteria can represent ideal microbial factories thanks to their ability to photosynthetically convert CO2 into valuable compounds, their minimal nutritional requirements, high photosynthetic rate and the availability of genetic and bioinformatics tools. An engineered strain of Synechococcus elongatus PCC 7942 for 2-PE production, i.e., p120, was previously published elsewhere. The strain p120 expresses four heterologous genes for the complete 2-PE synthesis pathway. Here, we developed a combined approach of metabolite doping and metabolic engineering to improve the 2-PE production kinetics of the Synechococcus elongatus PCC 7942 p120 strain. Firstly, the growth and 2-PE productivity performances of the p120 recombinant strain were analyzed to highlight potential metabolic constraints. By implementing a BG11 medium doped with L-phenylalanine, we covered the metabolic burden to which the p120 strain is strongly subjected, when the 2-PE pathway expression is induced. Additionally, we further boosted the carbon flow into the Shikimate Pathway by overexpressing the native Shikimate Kinase in the Synechococcus elongatus PCC 7942 p120 strain (i.e., 2PE_aroK). The combination of these different approaches led to a 2-PE yield of 300 mg/gDW and a maximum 2-PE titer of 285 mg/L, 2.4-fold higher than that reported in literature for the p120 recombinant strain and, to our knowledge, the highest recorded for photosynthetic microorganisms, in photoautotrophic growth condition. Finally, this work provides the basis for further optimization of the process aimed at increasing 2-PE productivity and concentration, and could offer new insights about the use of cyanobacteria as appealing microbial cell factories for the synthesis of aromatic compounds.

The Detrimental Effect of High Salinity on the Growth and Microcystins Contamination of Microcystis aeruginosa

The occurrence of cyanobacterial species, especially toxic ones, poses a great threat to coastal and estuary areas. In this study, the toxigenic Microcystis aeruginosa (M. aeruginosa) FACHB-905 was exposed to BG-11 medium with different salinities (1, 4, 7, 10 and 15 ppt) to investigate the physiological responses of this species in terms of oxidative stress, chl a fluorescence and microcystins (MCs) contamination. The results showed that low salinity (≤7 ppt) favored the electron transfer of photosystem II, which promoted the growth and photosynthesis of M. aeruginosa and induced MCs production. However, increased salinity (≥10 ppt) suppressed the growth and photosynthesis of M. aeruginosa and aggravated the oxidative stress of the strain. Salinity of 15 ppt reduced MCs contamination and caused irreversible damage to the photosynthetic system of M. aeruginosa, leading to the lysis and death of algal cells. These results indicated that changes in salinity exerted important regulations on the growth and MCs contamination of the toxic M. aeruginosa, which may provide a reference for the risk assessment of the harmful cyanobacterial species in the coastal and estuary areas.

Mixotrophic Cultivation of a Native Cyanobacterium, Pseudanabaena mucicola GO0704, to Produce Phycobiliprotein and Biodiesel.

Global warming has accelerated in recent decades due to the continuous consumption of petroleum-based fuels. Cyanobacteria-derived biofuels are a promising carbon-neutral alternative to fossil fuels that may help achieve a cleaner environment. Here, we propose an effective strategy based on the large-scale cultivation of a newly isolated cyanobacterial strain to produce phycobiliprotein and biodiesel, thus demonstrating the potential commercial applicability of the isolated microalgal strain. A native cyanobacterium was isolated from Goryeong, Korea, and identified as Pseudanabaena mucicola GO0704 through 16s RNA analysis. The potential exploitation of P. mucicola GO0704 was explored by analyzing several parameters for mixotrophic culture, and optimal growth was achieved through the addition of sodium acetate (1 g/l) to the BG-11 medium. Next, the cultures were scaled up to a stirred-tank bioreactor in mixotrophic conditions to maximize the productivity of biomass and metabolites. The biomass, phycobiliprotein, and fatty acids concentrations in sodium acetate-treated cells were enhanced, and the highest biodiesel productivity (8.1 mg/l/d) was achieved at 96 h. Finally, the properties of the fuel derived from P. mucicola GO0704 were estimated with converted biodiesels according to the composition of fatty acids. Most of the characteristics of the final product, except for the cloud point, were compliant with international biodiesel standards [ASTM 6761 (US) and EN 14214 (Europe)].

Integrated microalgae culture with food processing waste for wastewater remediation and enhanced biomass productivity

Waste generation from food manufacturing facilities poses a serious hazard like environmental degradation, water pollution, and land pollution due to its high nutrient composition. Specifically, solid waste (powder) disposal requires additional energy sources in terms of scientific treatment, structured collection, and disposal packaging according to the safety regulation. Thus, this research discusses the viewpoint of integrating food processing waste as an organic carbon source with BG-11 medium for Chlorella vulgaris (FSP-E) growth. The food processing waste powders investigated in this study were obtained from milk, and biscuit manufacturing facilities. The culture medium was modified by combining both BG-11 and food processing waste powders to identify the optimal algal growth and biochemical content. Compared to the microalgae grown in BG-11 alone (IBG), the combination of biscuit waste and IBG produced higher biomass concentration (44%), with increased lipid (11%), protein (20%), and carbohydrate (57%) contents. Chlorella vulgaris was able to uptake nutrients from the culture medium with combination of food processing waste and IBG thus enhancing its growth. The results obtained also indicate that an integrated culture system using food processing waste and synthetic sources can generate energy out of waste by improving the bio-composition of the microalgae biomass.

Effect of Various Growth Medium on the Physiology and De Novo Lipogenesis of a Freshwater Microalga Scenedesmus rotundus-MG910488 under Autotrophic Condition

The microalga Scenedesmus rotundus, isolated from Jabalpur, Madhya Pradesh, India was designated as Scenedesmus rotundus-MG910488 after morphological and molecular identification. In this study, the effects of various autotrophic growth media on the physiology and lipid accumulation of this microalga were investigated. The cell density, amount of photosynthetic pigments, the productivity of biomass and lipid content and the cell morphology of the microalga were shown to be significantly affected by the variation in growth media. The highest biomass of 754.56 ± 14.80 mg L−1 with biomass productivity of 37.73 ± 0.74 mg L−1 day−1 was achieved when this microalgae was cultivated in the Zarrouk’s medium, whereas the highest lipid content of 33.30 ± 1.21% was observed in the BG-11 medium. The results confirm that the BG-11 is a cost-effective and efficient growth medium for this microalga. It also shows that the ingredients of the growth medium and its concentration influence the growth and synthesis of biomolecules produced by microalga. The biodiesel produced from obtained lipids was qualitatively estimated by Gas Chromatography-Mass Spectroscopy (GC-MS), Nuclear Magnetic Resonance (1H, 13C NMR) and Fourier Transform-Infrared Spectroscopy (FT-IR), which indicate the presence of oleic acid methyl ester, linoleic acid methyl ester and palmitic acid methyl ester as the leading fatty acid methyl esters (FAME) in the samples, which make this strain an ideal feedstock for biodiesel production.

OPTIMIZATION OF CULTURE MEDIA FOR THE GROWTH OF ANABAENA PCC550, ANABAENA PCC 574, AND CYLINDROSPERMUM PCC518, CYLINDROSPERMUM PCC 567

In the present study, Anabaena PCC550, Anabaena PCC574 and Cylindrospermum PCC518, Cylindrospermum PCC 567 have been subjected to 7 different inorganic culture media. In order to identify the best growth medium i.e.; optimized medium, the nutrient requirement of these two algae have been evaluated as prime requisite. The present investigation analyzed the growth of wet biomass of the four microalgae. In order to attain optimal growth of Anabaena and Cylindrospermum species, the 7 culture media employed in the current study were (i) BG11 medium (ii) Knoops medium (iii) Cyanophycean agar medium (iv) Modified Bristols medium (v) Prigsheims Medium (vi) Foggs Medium and (vii) Algal culture medium. Highest growth on 60th day was seen in Cylindrospermum PCC 518 (0.134mg/100ml), Anabaena PCC 574 (0123mg/100ml), Cylindrospermum PCC 567 (0.098mg/100ml), Anabaena PCC 550 (0.094mg/100ml) in Algal culture media which shows luxurient growth when compared to BG11, Foggs Media, Modified Bristols media, Knoops media. While, Prigsheims medium did not show any growth of Anabaena PCC550, Anabaena PCC574 and Cylindrospermum PCC518, Cylindrospermum PCC 567.

Potential use of Chlorella vulgaris KCBAL01 from a freshwater stream receiving treated textile effluent in hexavalent chromium [Cr(VI)] removal in extremely acidic conditions

Remediation of hexavalent chromium with conventional chemical and physical methods is a costly process, while replacing some critical steps in physiochemical remediation with self-sustaining bioremediation agents are expected to be cost-effective and environmentally friendly implementation. In this study, a microalga isolated from a freshwater stream receiving treated textile wastewater was identified up to its molecular level and investigated its ability to tolerate and remove hexavalent chromium from extremely acidic conditions under different temperatures. The ability of microalgae to tolerate and remove Cr(VI) was investigated by growing it in BG11 media with different pH (1, 2, 3 & 7), amended with several concentrations of Cr(VI) and incubated under different temperatures for 96 hrs. Microalga was identified as Chlorella vulgaris and found that the isolated strain has a higher hexavalent chromium removal potential in extremely acidic conditions than in neutral pH conditions at 25 °C. In contrast, its Cr(VI) removal potential is significantly influenced by the pH and temperature of the growth medium. Furthermore, it exhibited a permanent viability loss at extreme acidic conditions (pH 1 − 3) and prolonged exposure to the higher chromium content. The microalga investigated will be a highly useful bioagent in hexavalent chromium remediation in high acidic conditions.

The effect of salinity stress on the growth and lipid accumulation of Scenedesmus quadricauda FACHB-1297 under xylose mixotrophic cultivation

In the commercial production of microalgae biodiesel, high culture cost and low lipid production yields are two major limitations. Finding cheap carbon sources and ways to promote lipid accumulation can achieve low-cost cultivation and efficient lipid production. The impact of NaCl on the growth and lipid accumulation in Scenedesmus quadricauda FACHB-1297 under xylose mixotrophic cultivation was explored in this study. The results showed that when NaCl concentration were 0.88–5.85 g/L, the biomass concentration and lipid content were 2.15–2.68 and 1.19–1.80 times higher than those in BG11 medium, respectively. Under the optimal NaCl concentration of 2.63 g/L, the highest lipid content (39.33%) was achieved, with the biomass concentration of 0.72 g/L, the total nitrogen removal efficiency of 39.00%, and the total phosphorus removal efficiency of nearly 100.00%, except for a higher total sugar content of 19.26% and total protein content of 5.38%. In short, these results indicated that 2.63 g/L NaCl treatment was an efficient way to promote the lipid accumulation of S. quadricauda FACHB-1297 using xylose in papermaking waste liquid to produce biofuel.

Mineral-hydrogel composites for mitigating harmful algal bloom and supplying phosphorous for photo-biorefineries

Harmful algal blooms (HAB) are a major environmental concern in eutrophic aquatic systems. To mitigate HABs and recover the phosphorus that drives algal growth, this study developed hydrogel composites seeded with calcium phosphate and wollastonite particles, which first adsorb phosphate (P) and then precipitate it as calcium phosphate. Using a fast-growing cyanobacterium, Synechococcus elongatus 2973, as a model microalga, we found that the mineral-hydrogel composites reduced dissolved P in BG11 media from 5.1 mg/L to 0.31 mg/L, initially reducing the biomass growth rate by up to 73 % and ultimately reducing the total biomass concentration by 75 %. When applied to municipal wastewater and agricultural run-off, the composites removed 96 % and 91 % of the dissolved P, respectively. Moreover, when the recovered P-enriched composites were reused as a slow-release bio-compatible fertilizer in a photobioreactor, they effectively supported algal growth without blocking light and interfering with photosynthesis. The P-enriched composites could tune the P concentration in the culture medium and significantly promote algal lipid accumulation. This study demonstrates the mineral-hydrogel composites' potential to treat point sources of P pollution and subsequently facilitate photoautotrophic biofuel production as a nutrient, effectively recycling the captured P.

Exploring indigenous freshwater chlorophytes in integrated biophotovoltaic system for simultaneous wastewater treatment, heavy metal biosorption, CO2 biofixation and biodiesel generation

The study explored the combined photosynthetic activities of two green microalgal species, Tetradesmus obliquus and Tetradesmus reginae, on an integrated biophotovoltaic (BPV) platform for simultaneous wastewater treatment, toxic metal biosorption, carbon biofixation, bioelectricity generation and biodiesel production. The experimental setup comprised of a dual-chambered BPV with copper anode surrounded by T. obliquus in BG11 media, and copper cathode with T. reginae in municipal wastewater separated by Nafion 117 membrane. The study reported a maximum power density of 0.344 Wm−2 at a cell potential of 0.415 V with external resistance of 1000 Ω and 0.3268 V maximum open-circuit voltage. The wastewater electrical conductivity and pH increased from 583 ± 22 to 2035 ± 29.31 mS/cm and 7.403 ± 0.174 to 8.263 ± 0.055 respectively, signifying increased photosynthetic and electrochemical activities. Residual nitrogen, phosphorus, chemical oxygen demand, arsenic, cadmium, chromium and lead removal efficiencies by T. reginae were 100%, 80.68%, 71.91%, 47.6%, 88.82%, 71.24% and 92.96%, respectively. T. reginae accumulated maximum biomass of 0.605 ± 0.033 g/L with a CO2 biosequestration rate of 0.166 ± 0.010 gCO2/L/day and 42.40 ± 1.166% lipid content. Methyl palmitate, methyl undecanoate and 13-octadecenoic acid with relative abundances of 37.24%, 24.80% and 12.02%, respectively were confirmed.

Native SodB Overexpression of Synechocystis sp. PCC 6803 Improves Cell Growth Under Alcohol Stresses Whereas Its Gpx2 Overexpression Impacts on Growth Recovery from Alcohol Stressors.

To overcome the limited resistance to alcohol stress, genetically engineered Synechocystis sp. PCC 6803 strains with overexpressions of genes related with the ROS detoxification system (sodB and gpx2, which encode superoxide dismutase and glutathione peroxidase, respectively) were developed. Three engineered strains including a sodB-overexpressing strain (OE + S), a gpx2-overexpressing strain (OE + G), and a sodB/gpx2-overexpressing strain (OE + SG) grew similarly as wild-type control under normal condition. When compared to wild-type control, OE + S and OE + SG strains grew faster for 4days under 2.0% (v/v) ethanol and 0.3% (v/v) n-butanol conditions, as well as having higher chlorophyll a levels. On the other hand, the prominent growth recovery of OE + G and OE + SG was noted within 4days in normal BG11 medium after treating cells with high alcohol stresses for 1h, in particular 15% ethanol and 2.5% n-butanol. Under 4days of recovery from butanol stress, specific levels of intracellular pigments including chlorophyll a and carotenoids were dramatically increased in all modified strains. The overexpression of antioxidant genes then revealed a significant improvement of alcohol tolerance in Synechocystis sp. PCC 6803.

Oilfield-produced water as a medium for the growth of Chlorella pyrenoidosa outdoor in an arid region.

Oilfield-produced wastewater (OPW) provided by the local oil industry was used as a medium (without any pre-treatment) for the outdoor cultivation of microalgaeChlorella pyrenoidosa. The effectiveness of algal growth on the produced water treatment has been investigated. The experimental setups were carried out outdoor, under sunlight radiation, using an open system sited in the desert area. The highest biomass concentration was attaining 1.15 ± 0.07g/L after 21 culture days. FTIR spectroscopy was used to estimate the lipid content in C. pyrenoidosa grown in BG11 and OPW medium. Daytime temperatures fluctuation was between 26 and 31°C. The average insolation was no less than 10h per day with maximum solar irradiation of 1036 ± 30W/m2, measured between 12 and 1 p.m. C. pyrenoidosa was found highly capable of removing COD, NH4 + -N, TN, and TP by 89.67%, 100%, 57.14%, and 75.51%, respectively, throughout the cultivation period. Biosorption of toxic heavy metal pollutants such as Cu, Pb, and Cd was also achieved at rates of approximately 73.39, 72.80, and 48.42%. Overall, the achieved result of C. pyrenoidosa-based process was compared to the actual process using activated carbon.

A Study on the Effect of Nitrate and Phosphate Concentrations on the Production of Mycosporine-Like Amino Acids by Chlorella Vulgaris.

Cyanobacteria can produce compounds absorbing ultraviolet irradiation. Mycosporine like amino acids (MAAs) are some of these important metabolites, which can be potentially considered as a sunscreen agent in the pharmaceutical and cosmetic industry. Different factors have been reported that can affect the biosynthesis of MAA. In this study, the influence of different concentrations of phosphate and nitrate under different environmental conditions on MAA production by Chlorella vulgaris was investigated using an experimental design method, in order to enhance MAAs production in this specious. A 23 full factorial design (FFD) using Design-Expert v7.0.0 software was used to optimize simultaneously all the three factors of nitrate and phosphate concentration and condition of incubation environment on the MAA production by this species of C. vulgaris. Two milliliter of organism stock were grown in 200 mL BG11 medium and after 21 days, the biomasses of all samples were separated. Then, the MAA was extracted from dried biomass using methanol extraction. The extracts were analyzed by reverse-phase high performance liquid chromatography (RP-HPLC). After complete analysis, four samples were then cultured at the optimized conditions and analyzed by liquid chromatohraphy coupled to mass spectrometry (LC/MS). The results showed that this microalga could produce compounds with λmax of 330 nm and a retention time of about 2 min. According to the central composite analysis, phosphate at 0.51 g.L-1 and nitrate at 2.5 g.L-1 can be considered as the optimum concentrations, resulting to the preferable conditions concerning the culture in germinator. Based on LC/MSS analysis, the major compound had a m/z of 332 at the optimum condition. Thus, this species is expected to have the capability of MAA production (maybe Shinorine) or one of its glycosylated derivatives.

Study of temperature and food-grade preservatives affecting the in vitro stability of phycocyanin and phycoerythrin extracted from two Nostoc strains

Cyanobacteria have many bioactive compounds. In the present study, we investigated the degree of purification and free radical scavenging ability of phycocyanin (PC) and phycoerythrin (PE), and compare their stability against selected preservatives at different temperatures with the aim of achieving the best and most stable preservative in increasing shelf life of PC and PE. After collecting and culturing Nostoc sp. strains FSN and ASN in BG-110 medium, the pigments phycocyanin and phycoerythrin were extracted and purified with 56% ammonium sulfate followed by dialysis. The antioxidant activity of pigments was evaluated by DPPH and ABTS assays. Their stability was compared with food-grade preservatives citric acid, sodium chloride, sucrose, and calcium chloride at two temperatures of 5 °C and 35 °C over time period from 3 to 30 days of cultivation. The results showed that the concentration and purity of the pigments increased after the dialysis, the pigments had antioxidant properties and were more stable at 5 °C. In addition, among different preservatives, citric acid caused more stability over time.

Statistical optimization of phytol and polyunsaturated fatty acid production in the Antarctic microalga <italic>Micractinium variabile</italic> KSF0031

Polar microorganisms produce physiologically active substances to adapt to harsh environments, and these substances can be used as biomedical compounds. The green microalga Micractinium variabile KSF0031, which was isolated from Antarctica, produced phytol, a natural antimicrobial agent. Furthermore, several polyunsaturated fatty acids (PUFAs), including omega-3, exhibit antioxidant properties. Here statistical methods (Plackett-Burman design and Box- Behnken design) were used to optimize the culture medium of KSF0031 to improve biomass production, and K2HPO4, MgSO4·7H2O, and ammonium ferric citrate green (AFCg) were selected as significant components of the culture medium. Changes in the concentration of K2HPO4 and MgSO4·7H2O as positive factors and AFCg as a negative factor affected cell growth to a remarkable degree. The biomass production in a 100 L culture using the optimized medium for 24 d at 18°C was improved by 37.5% compared to that obtained using the original BG-11 medium. The quantities of PUFAs and phytol obtained were 13 mg g-1 dry cell weight (DCW) and 10.98 mg g-1 DCW, which represent improved yields of 11.70% and 48.78%, respectively. The results of this study could contribute to an improved production of phytol and fatty acids from Antarctic microalgae in the biomedical industry.

Diversity assessment of photosynthesizers: comparative analysis of pre-cultivated and natural microbiome of sediments from Cerrado biome in Maranhão, Brazil

Photosynthetic microorganisms are important components of most ecosystems and have important roles regarding biogeochemical cycles and the basis of the trophic chain. However, they sometimes are present in low abundance compared to other heterotrophic organisms. The Chapada das Mesas National Park (PNCM) is a Conservation Unit in Brazilian Cerrado biome, which is considered a hotspot for biodiversity conservation and possesses important rivers, waterfalls, and springs with economical and touristic importance. The aim of this study was to perform a comparative analysis of enriched and total microbiome of sediments to understand the impact of pre-cultivation in discovery of underrepresented groups like photosynthesizers. All sediment samples were cultivated in BG-11 medium under illumination to enrich for photosynthetic microorganisms and both the raw samples and the enriched ones were submitted to DNA extraction and sequencing of the V3-V4 hypervariable region of the 16S rRNA gene on the Ion Torrent platform. The reads were analyzed using QIIME2 software and the Phyloseq package. The enrichment allowed detection and identification of many genera of cyanobacteria in the Chapada das Mesas National Park (PNCM), which would probably not be possible without the combination of approaches. A total of 58 groups of photosynthetic microorganisms were classified in the samples from the enrichments and their relative abundance based on amplified 16S rRNA sequences were estimated, highlighting the genus Synechocystis which represented 10.10% of the abundance of the phylum Cyanobacteria and the genus Dunaliella, which represented 45.66% of the abundance of algae as the most abundant groups at the PNCM. In the enrichments, microorganisms from the phyla Proteobacteria (45.2%), Bacteroidetes (18%), and Planctomycetes (3.3%) were also identified, since there are ecological associations between the photosynthetic community and other groups of heterotrophic microorganisms. As for the functional analysis, metabolic functions associated with methanotrophy and methylotrophy, hydrocarbon degradation, phototrophy, and nitrogen fixation were predicted. The results highlight a great diversity of photosynthetic microorganisms in Cerrado and the importance of using a combination of approaches when analyzing target groups which are usually underrepresented such as cyanobacteria and microalgae.

Identification and Characterization of a New Microalga Dysmorphococcus globosus-HI from the Himalayan Region as a Potential Source of Natural Astaxanthin.

Simple SummaryThe natural astaxanthin from algae (marketed only 5%) is the most powerful antioxidant for health compared to the synthetic that shares about 95% of the market. Therefore, there is a huge demand for natural astaxanthin from algae, which can accumulate significantly higher astaxanthin. We have isolated a new algal strain from the Himalayan region, Northern India, which was identified as Dysmorphococcus globosus-HI based on morphological and molecular analysis. Its growth conditions were optimized in the laboratory. Among the seven different tested culture media, MBBM and 3N-BBM provided the maximum growth and astaxanthin production. The highest biomass production (1.14 g L−1) was observed in the modified BBM medium. It is an excellent source of producing natural astaxanthin, to a tune of 391 mg L−1, which is greater than any other known algal species. The productivity of astaxanthin was about 15.6 mg L−1 d−1 under normal conditions, which is higher than the commercially used H. pluvialis species. It is the first report of natural astaxanthin production from D. globosus-HI that has great potential for commercial application.Synthesized astaxanthin (ASX), stereoisomers of 3S,3′R, 3R,3′R, and 3S,3′S, have over 95% market share and have relatively poor antioxidant and bioactivity properties, with persistent issues in terms of biological functions, health benefits, and biosafety if compared to natural ASX. Bioprospecting of new microalgal strains could be vital for a new source of powerful antioxidant (ASX). In this study, a new algal strain was isolated from the Indian foothills of the Himalayas. Its identity was discerned by morphological and DNA barcode studies. It is a unicellular spheroidal cell-shaped alga with 100–200 μm diameter. The isolate has 93.4% similarity to Dysmorphococcus globosus species based on 18S-rDNA phylogenetic analysis and named as D. globosus-HI (HI stands for Himalayan India). Its growth and major cellular components (carotenoids, carbohydrates, protein, lipids, fatty acid profile, and ASX) were optimized using the seven different culture media. The highest biomass (1.14 g L−1) was observed in the MBBM medium, with a specific growth rate (0.087 day−1), division/day (0.125), and cellular yield (6.16 x 106 cells/mL). The highest carotenoids (1.56 mg g−1), lipids (32.5 mg L−1), and carbohydrates (135.62 mg L−1) were recorded in the 3N-BBM medium. The maximum ω3-FAs (17.78%), ω6-FAs (23.11%), and ω9-FAs (7.06%) were observed in MBBM, JW, and BG-11 medium respectively. The highest amount of antioxidant ASX was accumulated in the 3N-BBM medium (391 mg L−1). It is more than any other known algal species used in the production of natural ASX. The optimized biochemical studies on the D. globosus-HI strain should fulfill the increasing demand for natural ASX for commercial application.