Aerobic Se(IV) reducing bacteria and their reducing characteristics in estuarine sediment.

Microorganisms play a critical role in the biogeochemical cycling of selenium in natural ecosystems, particularly in reducing selenite (Se(IV)) to element selenium (Se(0)) which reduces its mobility and bioavailability. However, Se(IV)-reducing bacteria and their reducing characteristics in estuarine sediments remain inadequately understood. In this study, the reduction of Se(IV) was confirmed to be microbially driven through the cultivation of a mixture of estuarine sediment and Se(IV) under aerobic conditions. Community analysis indicates that Bacillus was primarily involved in the reduction of Se(IV). A strain with high salt tolerance (7.5 % NaCl) and Se(IV) resistance (up to 200 mM), Bacillus cereus SD1, was isolated from an estuarine sediment. The reduction of Se(IV) occurred concomitantly with the onset of microbial growth, and reduction capacity increased approximately 5-fold by adjusting the pH. In addition, Se(IV) reduction in Bacillus cereus SD1 was significantly inhibited by sulfite, and the key enzyme activity tests revealed the possible presence of a sulfite reductase-mediated Se(IV) reduction pathway. These research findings provide new insights into the bioreducing characteristics and the biogeochemical cycling of selenium in estuarine environments.

Bacterial community in the metal(loid)-contaminated marine vertical sediments of Jinzhou Bay: Impacts and adaptations.

Metal(loid) discharge has led to severe coastal contamination; however, there remains a significant knowledge gap regarding its impact on sediment profiles and depth-resolved bacterial communities. In this study, geochemical measurements (pH, nutrient elements, total and bioavailable metal(loid) content) consistently revealed decreasing nitrogen, phosphorus, and metal(loid) levels with sediment depth, accompanied by reduced alpha diversity. Principal coordinate analysis indicated distinct community compositions with varying sediment depths, suggesting a geochemical influence on diversity. Ecological niche width expanded with depth, favoring specialists over generalists, but both groups decreased in abundance. Taxonomic shifts emerged, particularly in phyla and families, correlated with sediment depth. Microbe-microbe interactions displayed intricate dynamics, with keystone taxa varying by sediment layer. Zinc and arsenic emerged as key factors impacting community diversity and composition using random forest, network analysis, and Mantel tests. Functional predictions revealed shifts in potential phenotypes related to mobile elements, biofilm formation, pathogenicity, N/P/S cycles, and metal(loid) resistance along sediment profiles. Neutral and null models demonstrated a transition from deterministic to stochastic processes with sediment layers. This study provides insights into the interplay between sediment geochemistry and bacterial communities across sediment depths, illuminating the factors shaping these ecosystems.

Divergent adaptation strategies of abundant and rare bacteria to salinity stress and metal stress in polluted Jinzhou Bay.

Understanding how abundant (AT) and rare (RT) taxa adapt to diverse environmental stresses is vital for assessing ecological processes, yet remains understudied. We collected sediment samples from Liaoning Province, China, representing rivers (upstream of wastewater outlet), estuaries (wastewater outlets), and Jinzhou Bay (downstream of wastewater outlets), to comprehensively evaluate AT and RT adaptation strategies to both natural stressors (salinity stress) and anthropogenic stressors (metal stress). Generally, RT displayed higher α- and ß-diversities and taxonomic groups compared to AT. Metal and salinity stresses induced distinct α-diversity responses in AT and RT, while ß-diversity remained consistent. Both subcommunities were dominated by Woeseia genus. Metal stress emerged as the primary driver of diversity and compositional discrepancies in AT and RT. Notably, AT responded more sensitively to salinity stress than RT. Stress increased topological parameters in the biotic network of AT subcommunities while decreasing values in RT subcommunities, concurrently loosening interactions of AT with other taxa and strengthening interactions of RT with others in biotic networks. RT generally exhibited greater diversity of metal resistance genes compared to AT. Greater numbers of genes related to salinity tolerance was observed for the RT than for AT. Compared to AT, RT demonstrated higher diversity of metal resistance genes and a greater abundance of genes associated with salinity tolerance. Additionally, deterministic processes governed AT community assembly, reinforced by salinity stress. However, the opposite trend was observed in the RT, where the importance of stochastic process gradually increased with metal stresses. The study is centered on exploring the adaptation strategies of both AT and RT to environmental stress. It underscores the importance of future research incorporating diverse ecosystems and a range of environmental stressors to draw broader and more reliable conclusions. This comprehensive approach is essential for gaining a thorough understanding of the adaptive mechanisms employed by these microorganisms.

Enhancement of arsenic uptake and accumulation in green microalga Chlamydomonas reinhardtii through heterologous expression of the phosphate transporter DsPht1.

Arsenate (AsV) is a predominant arsenic contaminant in aerobic water. Microalgae have been recently used in the phytoremediation of arsenic-contaminated water. However, the amount of AsV uptake in microalgae is limited, which hinders the application of microalgae in arsenic-contaminated water treatment. Here, we found that the expression of a novel phosphate transporter DsPht1 in Dunaliella salina was highly upregulated after AsV exposure. Fluorescent protein-tagging analysis showed the plasma membrane location of DsPht1. Furthermore, DsPht1 was overexpressed in a model microalga Chlamydomonas reinhardtii. The DsPht1 transgenetic lines accumulated up to 6.4-fold higher total arsenic than the untransformed line, and the AsV amount in total arsenic increased by 8.3-fold. Moreover, the organoarsenic content was also higher in the transgenetic lines. Overall, the DsPht1 transformants generated in this study increased arsenate uptake and transformation, which are promising for the effective phytoremediation of arsenic-contaminated water.

Kinetic modeling and process analysis for photo-production of ß-carotene in Dunaliella salina.

Dunaliella salina is a green microalga with the great potential to generate natural ß-carotene. However, the corresponding mathematical models to guide optimized production of ß-carotene in Dunaliella salina (D. salina) are not yet available. In this study, dynamic models were proposed to simulate effects of environmental factors on cell growth and ß-carotene production in D. salina using online monitoring system. Moreover, the identification model of the parameter variables was established, and an adaptive particle swarm optimization algorithm based on parameter sensitivity analysis was constructed to solve the premature problem of particle swarm algorithm. The proposed kinetic model is characterized by high accuracy and predictability through experimental verification, which indicates its competence for future process design, control, and optimization. Based on the model established in this study, the optimal environmental factors for both ß-carotene production and microalgae growth were identified. The approaches created are potentially useful for microalga Dunaliella salina cultivation and high-value ß-carotene production.

Characterization and RNA-seq transcriptomic analysis of a Scenedesmus obliqnus mutant with enhanced photosynthesis efficiency and lipid productivity.

Microalgae have received significant attention as potential next-generation microbiologic cell factories for biofuels. However, the production of microalgal biofuels is not yet sufficiently cost-effective for commercial applications. To screen higher lipid-producing strains, heavy carbon ion beams are applied to induce a genetic mutant. An RNA-seq technology is used to identify the pathways and genes of importance related to photosynthesis and biofuel production. The deep elucidation of photosynthesis and the fatty acid metabolism pathway involved in lipid yield is valuable information for further optimization studies. This study provided the photosynthetic efficiency and transcriptome profiling of a unicellular microalgae, Scenedesmus obliqnus mutant SO120G, with enhanced lipid production induced by heavy carbon ion beams. The lipid yield (52.5 mg L-1) of SO120G mutant were enhanced 2.4 fold compared with that of the wild strain under the nitrogen deficient condition. In addition, the biomass and growth rate were 57% and 25% higher, respectively, in SO120G than in the wild type, likely owing to an improved maximum quantum efficiency (Fv/Fm) of photosynthesis. As for the major pigment compositions, the content of chlorophyll a and carotenoids was higher in SO120G than in the wild type. The transcriptome data confirmed that a total of 2077 genes with a change of at least twofold were recognized as differential expression genes (DEGs), of which 1060 genes were up-regulated and 1017 genes were down-regulated. Most of the DEGs involved in lipid biosynthesis were up-regulated with the mutant SO120G. The expression of the gene involved in the fatty acid biosynthesis and photosynthesis of SO120G was upregulated, while that related to starch metabolism decreased compared with that of the wild strain. This work demonstrated that heavy-ion irradiation is an promising strategy for quality improvement. In addition, the mutant SO120G was shown to be a potential algal strain for enhanced lipid production. Transcriptome sequencing and annotation of the mutant suggested the possible genes responsible for lipid biosynthesis and photosynthesis, and identified the putative target genes for future genetic manipulation and biotechnological applications.

Quantitative analysis on photon numbers received per cell for triggering ß-carotene accumulation in Dunaliella salina.

Accumulation of ß-carotene in Dunaliella salina is highly dependent on light exposure intensity and duration, but quantitative analysis on photon numbers received per cell for triggering ß-carotene accumulation is not available so far. In this study, experiment results showed that significant ß-carotene accumulation occurred after at least 8 h illumination at 400 µmol photons·m-2·s-1. To quantify the average number of photons received per cell, correlations of light attenuation with light path, biomass concentration, and ß-carotene content were, respectively, established using both Lambert-Beer and Cornet models, and the latter provided better simulation. Using Cornet model, average number of photons received per cell (APRPC) was calculated and proposed as a parameter for ß-carotene accumulation, and constant APRPC was maintained by adjusting average irradiance based on cell concentration and carotenoids content changes during the whole induction period. It was found that once APRPC reached 0.7 µmol photons cell-1, ß-carotene accumulation was triggered, and it was saturated at 9.9 µmol photons cell-1. This study showed that APRPC can be used as an important parameter to precisely simulate and control ß-carotene production by D. salina.

ß-Carotene Production from Dunaliella salina Cultivated with Bicarbonate as Carbon Source.

Bicarbonate has been considered as a better approach for supplying CO2 to microalgae cells microenvironments than gas bubbling owing t°Cost-effectiveness and easy operation. However, the ß-carotene production was too low in Dunaliella salina cultivated with bicarbonate in previous studies. Also, the difference in photosynthetic efficiency between these tw°Carbon sources (bicarbonate and CO2) has seldom been discussed. In this study, the culture conditions, including NaHCO3, Ca2+, Mg2+ and microelement concentrations, were optimized when bicarbonate was used as carbon source. Under optimized condition, a maximum biomass concentration of 0.71 g/l and corresponding ß-carotene content of 4.76% were obtained, with ß-carotene yield of 32.0 mg/l, much higher than previous studies with NaHCO3. Finally, these optimized conditions with bicarbonate were compared with CO2 bubbling by online monitoring. There was a notable difference in Fv/Fm value between cultivations with bicarbonate and CO2, but there was no difference in the Fv/Fm periodic changing patterns. This indicates that the high concentration of NaHCO3 used in this study served as a stress factor for ß-carotene accumulation, although high productivity of biomass was still obtained.

ROS Induce ß-Carotene Biosynthesis Caused by Changes of Photosynthesis Efficiency and Energy Metabolism in Dunaliella salina Under Stress Conditions.

The unicellular alga Dunaliella salina is regarded as a promising cell factory for the commercial production of ß-carotene due to its high yield of carotenoids. However, the underlying mechanism of ß-carotene accumulation is still unclear. In this study, the regulatory mechanism of ß-carotene accumulation in D. salina under stress conditions was investigated. Our results indicated that there is a significant positive correlation between the cellular ROS level and ß-carotene content, and the maximum quantum efficiency (F v /F m ) of PSII is negatively correlated with ß-carotene content under stress conditions. The increase of ROS was found to be coupled with the inhibition of F v /F m of PSII in D. salina under stress conditions. Furthermore, transcriptomic analysis of the cells cultivated with H2O2 supplementation showed that the major differentially expressed genes involved in ß-carotene metabolism were upregulated, whereas the genes involved in photosynthesis were downregulated. These results indicated that ROS induce ß-carotene accumulation in D. salina through fine-tuning genes which were involved in photosynthesis and ß-carotene biosynthesis. Our study provided a better understanding of the regulatory mechanism involved in ß-carotene accumulation in D. salina, which might be useful for overaccumulation of carotenoids and other valuable compounds in other microalgae.

Progress on the development of floating photobioreactor for microalgae cultivation and its application potential.

Microalgae present great potential to replace land crops for the efficient production of large volumes of biomass for food, feed, fuels, and chemicals, as well as to treat wastewater and capture carbon. However, the commercialization of these technologies for bulk commodities requires a great reduction in the current microalgal biomass production cost. The bioreactor is the core of bioprocess engineering and is the premise for the commercial application of certain types of biotechnology. The challenges of phototrophic cultivation are completely different from those of heterotrophic processes because the efficiency of phototrophic cultivation is limited by the energy density of the input sunlight and the inorganic carbon supply. Thus, the development of microalgae cultivation technologies with low manufacturing and operating costs is key to addressing this problem, and floating photobioreactors (PBRs) are a promising solution. PBRs are deployed on the water surface without any land requirements, and wave energy provides free mixing energy. Additionally, the surrounding water can be used to control the culture temperature and to supply nutrients for microalgae growth. In this mini-review, the development of floating PBRs and their recent progress are presented. The effect of the carbon supply approach on the mixing and scaling-up of floating PBRs are critically discussed. The limitations and challenges in commercial applications of floating PBRs are analysed, and the need for future research is proposed. Finally, it is noted that microalgae farming on the ocean is a promising solution for human society to address the challenge of land space exhaustion due to the global population boom.

Large-scale cultivation of Spirulina in a floating horizontal photobioreactor without aeration or an agitation device.

A low-cost floating photobioreactor (PBR) without the use of aeration and/or an agitation device, in which carbon was supplied in the form of bicarbonate and only wave energy was utilized for mixing, was developed in our previous study. Scaling up is a common challenge in the practical application of PBRs and has not yet been demonstrated for this new design. To fill this gap, cultivation of Spirulina platensis was conducted in this study. The results demonstrated that S. platensis had the highest productivity at 0.3 mol L-1 sodium bicarbonate, but the highest carbon utilization (104 ± 2.6%) was obtained at 0.1 mol L-1. Culture of Spirulina aerated with pure oxygen resulted in only minor inhibition of growth, indicating that its productivity will not be significantly reduced even if dissolved oxygen is accumulated to a high level due to intermittent mixing resulting from the use of wave energy. In cultivation using a floating horizontal photobioreactor at the 1.0 m2 scale, the highest biomass concentration of 2.24 ± 0.05 g L-1 was obtained with a culture depth of 5.0 cm and the highest biomass productivity of 18.9 g m-2 day-1 was obtained with a depth of 10.0 cm. This PBR was scaled up to 10 m2 (1000 L) with few challenges; biomass concentration and productivity during ocean testing were little different than those at the 1.0 m2 (100 L) scale. However, the larger PBR had an apparent carbon utilization efficiency of 45.0 ± 2.8%, significantly higher than the 39.4 ± 0.9% obtained at the 1 m2 scale. These results verified the ease of scaling up floating horizontal photobioreactors and showed their great potential in commercial applications.