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The pulp and paper industry in Sweden annually produce millions of tons of side-stream materials as black liquor, pulp and paper mill sludge, sulfite liquor and lignosulfonate. These lignocellulosic based materials can be more utilized today in biorefinery processes with microorganisms to produce high-value products as lipids, proteins and biofuels. In this work, we used five side-stream materials as carbon source in fermentation with two oleaginous yeasts, Lipomyces starkeyi and Rhodotorula toruloides. We analyzed lipid production, fatty acid profiles, inhibitors, phenolics, free sugars and metals before and after batch fermentation in 2 L bioreactors. Steam-exploded hardwood media was used as reference as it’s known to be a good substrate for the oleaginous yeast species and after fermentation the lipid yield for R. toruloides was 17 % (w/w) and for L. starkeyi 13 % (w/w). The side-stream materials contained less than 30 % of free sugar compared to the reference media and the total lipid yield was thus less than 2.7 %, (w/w). R. toruloides utilized various sugars during fermentation and L. starkeyi mostly consumed glucose and xylose. Both yeast species also showed a possible ability to utilize various phenolics indicating their metabolic potential to convert depolymerized lignin along with wood-based sugars to lipids and proteins. [ABSTRACT FROM AUTHOR]

Research on enhancing the production of lipids, particularly polyunsaturated fatty acids that are considered important for health, has focused on improvement of metabolism as well as heterologous expression of biosynthetic genes in the oleaginous fungus Aspergillus oryzae. To date, the productivity and production yield of free fatty acids have been enhanced by 10-fold to 90-fold via improvements in metabolism and optimization of culture conditions. Moreover, the productivity of ester-type fatty acids present in triacylglycerols could be enhanced via metabolic improvement. Culturing A. oryzae in a liquid medium supplemented with non-ionic surfactants could also lead to the effective release of free fatty acids from the cells. The current review highlights the advancements made in this field so far and discusses the future outlook for research on lipid production using A. oryzae. I hope the contents are useful for researchers in this field to consider the strategy of increasing production of various valuable metabolites as well as lipids in A. oryzae. [ABSTRACT FROM AUTHOR]

Wastewater from different sources with excessive nutrient loading, which is a major contributor to eutrophication, can be a promising medium alternative for microalgae cultivation with effective removal of NH4+-N, PO43--P and organic matter. In this study, three groups of synthetic wastewater (SWW), an ammonium-rich stream (60 to 250 mg L-1 NH4+) containing a specific type of organic matter, such as petroleum effluent (SWW1: 300 mg L-1 glucose and 10 mg L-1 phenol), molasses wastewater (SWW2: 600 mg L-1 glucose), and agro-industrial wastewater (SWW3: no organic matters) were used for growing Chlorella sp. and for investigating its potential for nutrient removal as well as the production of biomass and lipid. The findings showed that Chlorella sp. grown in SWW2 (i.e., the auto/mixotrophic culture with 14 days) served as efficient feedstock for lipid production, and showed the maximum lipid content and yield of 53 % DCW and 188 mg L-1, respectively. Additionally, the maximum removal of 76 % NH4+-N was observed under this condition. Higher removal of NH4+-N with 100 % efficiency was seen in Chlorella sp. under SWW1 and SWW3 conditions. However, under SWW2 the cells had higher NH4+ uptake rate than that under SWW1 and SWW3, resulting in highest biomass productivity in SWW2 grown cells. The spent media after Chlorella sp. cultivation were used for growing the N2-fixing cyanobacterium Fischerella muscicola TISTR 8215 for the production of hydrogen. Among different spent media, F.muscicola TISTR 8215 cultivated in SWW1 spent medium obtained from 7-day Chlorella sp. cultivation had the highest hydrogen production of 200.8 µmol H2 mg-1 chlorophyll a. Overall, the present study employed biorefinery approach using the microalga biomass as feedstock for lipid production with subsequent utilization of spent medium for growing the cyanobacterium for enhanced production of hydrogen. [ABSTRACT FROM AUTHOR]

Summary: Central metabolism produces amino and fatty acids for protein and lipids that establish seed value. Biosynthesis of storage reserves occurs in multiple organelles that exchange central intermediates including two essential metabolites, malate, and pyruvate that are linked by malic enzyme. Malic enzyme can be active in multiple subcellular compartments, partitioning carbon and reducing equivalents for anabolic and catabolic requirements. Prior studies based on isotopic labeling and steady‐state metabolic flux analyses indicated malic enzyme provides carbon for fatty acid biosynthesis in plants, though genetic evidence confirming this role is lacking. We hypothesized that increasing malic enzyme flux would alter carbon partitioning and result in increased lipid levels in soybeans.Homozygous transgenic soybean plants expressing Arabidopsis malic enzyme alleles, targeting the translational products to plastid or outside the plastid during seed development, were verified by transcript and enzyme activity analyses, organelle proteomics, and transient expression assays. Protein, oil, central metabolites, cofactors, and acyl‐acyl carrier protein (ACPs) levels were quantified overdevelopment.Amino and fatty acid levels were altered resulting in an increase in lipids by 0.5–2% of seed biomass (i.e. 2–9% change in oil).Subcellular targeting of a single gene product in central metabolism impacts carbon and reducing equivalent partitioning for seed storage reserves in soybeans. See also the Commentary on this article by Schwender, 239: 1539–1541. [ABSTRACT FROM AUTHOR]

Acne is associated with the excessive production of sebum, a complex mixture of lipids, in the sebaceous glands. The transcription factor Krüppel-like factor 4 (KLF4) plays an important role in skin morphogenesis, but its role in sebum production by sebocytes is not well known. In this study, we investigated the possible action mechanism of KLF4 during calcium-induced lipogenesis in immortalized human sebocytes. Sebocytes were treated with calcium, and lipid production was confirmed by thin-layer chromatography (TLC) and Oil Red O staining. To investigate the effect of KLF4, sebocytes were transduced with the KLF4-overexpressing adenovirus, and then lipid production was evaluated. Calcium treatment resulted in increased sebum production in terms of squalene synthesis in sebocytes. In addition, calcium increased the expression of lipogenic regulators such as sterol-regulatory element binding protein 1 (SREBP1), sterol-regulatory element binding protein 2 (SREBP2), and stearoyl-CoA desaturase (SCD). Similarly, the expression of KLF4 was increased by calcium in sebocytes. To investigate the effect of KLF4, we overexpressed KLF4 in sebocytes using recombinant adenovirus. As a result, KLF4 overexpression increased the expression of SREBP1, SREBP2, and SCD. Parallel to this result, lipid production was also increased by KLF4 overexpression. Chromatin immunoprecipitation revealed the binding of KLF4 to the SREBP1 promoter, indicating that KLF4 may directly regulate the expression of lipogenic regulators. These results suggest that KLF4 is a novel regulator of lipid production in sebocytes. • Calcium promotes differentiation of sebocytes and contributes to lipid production. • Calcium regulates the expression of KLF4 during differentiation of sebocytes. • The transcription factor KLF4 regulates the expression of SREBP1 to facilitate lipid formation. [ABSTRACT FROM AUTHOR]

Currently, microalgae have become a marvelous and resource-friendly alternative source of advantageous bioproducts, such as lipids, carbohydrates, proteins, or other bioactive compounds. Because of the richness of microalgae in these high-value-added metabolites, still, it is an underdeveloped source of sustainable energy and food. There are some hurdles to profitable production, such as culture contamination and costly harvesting techniques. In the current work, a chrysophyte was isolated from dairy wastewater, identified as Poterioochromonas malhamensis based on its morphology and partial 18S rRNA gene sequences. This isolate was used to remediate dairy wastewater (DWW) and to obtain neutral lipids (fatty acids) from microalgae. Microalgal growth was optimized by using different concentrations of DWW, supplemented with all the nutritive requirements for better progression and flourishment. Maximum biomass yield 1.478 g L−1 was achieved by optimized cultural conditions (different concentrations of DWW with BBM media). This strain showed high nitrate and phosphate removal efficiency (87.45% and 88.96%), respectively in 15 days. The experimental results highlighted that the lipid content and the chemical oxygen demand (COD) removal were 31.60% and 88.84%, respectively and the lipid profile of isolated microalga was C16:0, C16:1, C18:0, C18:1, and C18:2 fatty acids. For growth and treatment purposes, 75% DWW with Bold’s Basal Medium (BBM) media showed better results. This is the first report of DWW treatment using the microalga Poterioochromonas malhamensis, as far as we are aware. Its cultivation prevented the spread of pollution of freshwater sources, remedied the DWW, and generated important lipids for industry. [ABSTRACT FROM AUTHOR]

Sucrose non-fermenting 1 (SNF1) protein kinase plays an important role in the utilization of selective carbon sources and regulation of lipid metabolism. In order to further explore the function of SNF1 in regulating lipid accumulation by responding nutritional signals from non-glucose carbon sources, in the present study, the lipid production and SNF1 transcriptional levels of Mucor circinelloides were analyzed and compared on different carbon sources. The results indicated that M. circinelloides could effectively utilize some secondary metabolic carbon sources of monosaccharides and disaccharides for growth and lipids production, such as fructose, maltose and galactose. Snf-β subunit was associated with the regulation of lipid metabolism in response to nutritional signals from different carbon sources. This is the first report on the transcriptional analysis of SNF1 subunits on different carbons metabolism in oleaginous filamentous fungi. This research has suggested that genetic engineering of SNF1 subunits will alter the lipid production of M. circinelloides from alternative carbon sources. [ABSTRACT FROM AUTHOR]

Microalgae can synthesize lipids under controlled settings, making them a viable option as biofuel feedstocks. However, optimizing culture parameters, such as light quality, to maximize lipid yields and achieve commercial viability remains challenging. This study examined the effects of blue and red light on the fatty acid profiles and lipid synthesis in two species with high biotechnological potential for biofuel production: Chlorella vulgaris and Scenedesmus acutus. When grown under blue, red, and white light, these microalgae exhibited unique metabolic responses in terms of biomass increase, lipid content, and fatty acid composition. The expression levels of genes associated with lipid biosynthesis, including glycerol-3-phosphate acyltransferase (GPAT), β-ketoacyl-ACP synthase (KAS), Biotin carboxyl carrier protein-related gene (BCR), Stearoyl-ACP desaturase (SAD), 3-ketoacyl-ACP reductase (KAR), and Beta-carboxyltransferase 1 (BCX1) were analyzed. Blue light significantly increased lipid accumulation in both species, which was correlated with the overexpression of the GPAT and KAS genes. The generation of high-quality biofuel requires polyunsaturated and monounsaturated fatty acids, especially oleic acid (C18:1) and palmitic acid (C16:0), which increased significantly after genetic activation. The increase in oleic acid levels under blue light in C. vulgaris, along with the simultaneous increase in oleic and linoleic acids (C18:2) in S. acutus, demonstrates these metabolic adaptations by the microalgae to optimize lipid synthesis under different light conditions. This study examines how blue and red light affects the growth, lipid accumulation, and gene expression of biofuel-potential microalgae C. vulgaris and S. acutus. We want to optimize light quality for microalgae biofuel precursor generation by evaluating photobiological reactions under regulated light settings. Previously, light quality has been shown to affect photosynthetic efficiency and metabolic pathways, but this study examines gene-level responses to lipid production under blue and red light. The findings show how tailored light conditions can improve lipid biosynthesis gene expression, which directly affects biodiesel fatty acid profiles. [ABSTRACT FROM AUTHOR]

Biodiesel derived from cyanobacterial oils becomes attractive as an efficient renewable energy. The present study aims to optimize growth and lipid production of the halotolerant unicellular cyanobacterium Aphanothece halophytica cultivated in natural seawater. In this study A. halophytica was able to grow in natural seawater when supplemented with low concentration of NaNO3, whereas no growth occurred without supplementation. The specific growth rate of 0.230 day-1 and cell concentration of 25.17 x 106 cells mL-1 were achieved in A. halophytica cultivated in natural seawater supplemented with 17.6 mM NaNO3 and Turk Island salt solution (suitable natural seawater; SNSW) for 14 days. This growth rate was comparable to that of cells grown in normal BG11 plus Turk Island salt solution. The lipid content and fatty acid profiles of A. halophytica varied with changes in NaCl concentrations. The highest lipid content of 50.47 % and lipid productivity of 48.33 mg L-1 day-1 were obtained in cultures supplemented with 1.89 mmol C-atom L-1 glucose and 0.75 M NaCl. The optimal medium pH and cultivation temperature for lipid production was 7.5 and 25-35 °C, respectively. When cultivating A. halophytica in optimized SNSW with various NaCl concentrations, the highest contents of linoleic and linolenic acids, and the lowest contents of palmitic, stearic, and oleic acids were observed with 0.75 M NaCl. In contrast, cultures grown in optimized SNSW with 0.5 M NaCl showed fatty acid methyl ester profiles rich in monounsaturated fatty acids, which are favorable for high-quality biodiesel production. [ABSTRACT FROM AUTHOR]

This article is a Commentary on Morley et al. (2023), 239: 1834–1851. [ABSTRACT FROM AUTHOR]

Lipid extraction from microbial and microalgae biomass requires the separation of oil-rich cells from the production media. This downstream procedure represents a major bottleneck in biodiesel production, increasing the cost of the final product. Flocculation is a rapid and cheap system for removing solid particles from a suspension. This natural characteristic is displayed by some microorganisms due to the presence of lectin-like proteins (called flocculins/adhesins) in the cell wall. In this work, we showed, for the first time, that the heterologous expression of the adhesin Cfl1p endows the oleaginous species Cutaneotrichosporon oleaginosus with the capacity of cell flocculation. We used Helm's test to demonstrate that the acquisition of this trait allows for reducing the time required for the separation of lipid-rich cells from liquid culture by centrifugation without altering the productivity. This improves the lipid production process remarkably by providing a more efficient downstream. [ABSTRACT FROM AUTHOR]

An effective prototype of photobioreactor namely hollow fiber membrane bioreactor is employed to produce algal biomass for biofuel production. The effect of two-stage cultivation system on the biomass productivity and lipid production is studied using freshwater green microalga Tetradesmus obliquus MT188616.1. The hybrid system combines exponential biomass production in hollow fiber membrane photobioreactor (HFMPBr) and a coordinated high-lipid stimulation phase in nitrogen-deprived medium by the shake flask method. This work is proposed to examine the usefulness of HFMPBr module to enhance the microalgal growth rate through the effective mass transfer by standardizing the culture medium re-circulation flow rates (5 – 45 mL min−1) through the hollow fiber membranes. Cultivation is carried out at continuous mode in HFMPBr containing polysulfone fabricated membranes at fixed light intensity of 50 µmol m−2 s−1 and temperature at 28 °C. Biomass productivity and specific growth rate obtained are 0.44 g L−1d−1 and 1.61 µ d−1 with a lipid yield of 0.1 g L− 1. The key operating parameter i.e., liquid flow rate is optimized based on biomass production. The highest biomass concentration is produced at the flow rate of 45 mL min−1. The results showed that HFMPBr module is a better choice for the algal-cultivation to obtain higher biomass yield. The effect of two-stage cultivation system on the growth rate, biomass productivity, and lipid yield was studied in freshwater green microalga Tetradesmus obliquus MT188616.1. The hybrid system combines exponential biomass production in hollow fiber membrane photobioreactor (HFMPBr) and a coordinated high-lipid stimulation phase in nitrogen deprived medium by the shake flask method. The algal cultivation was carried out at continuous mode in HFMPBr containing polysulfone fabricated membranes at fixed light intensity of 50 µmol m−2 s−1 and temperature at 28 °C. Biomass productivity and specific growth rate obtained were 0.44 g L−1d−1 and 1.61 µ d−1 with a lipid yield of 0.1 g L− 1. The key operating parameter i.e., liquid flow rate was optimized based on biomass production. The highest biomass concentration was produced at the highest flow rate of 45 mL min−1, whereas the lowest biomass was at the lowest flow rate of 5 mL min−1. The results showed that HFMPBr module was a better choice for the cultivation of Tetradesmus obliquus MT188616.1 to obtain higher biomass yield. [Display omitted] • Integrated HFMPBr and shake flask approach to attain higher lipid yield of algae. • Optimized salt medium flow rate in HFMPBr for continuous algal growth. • Attained efficient delivery of nutrients via membranes for algal growth and waste removal by maintaining optimal flow rate. • Achieved close to 0.44 gL-1day-1 biomass productivity under 45 mL min-1 flow rate during 10 h light – 14 h dark photoperiod. [ABSTRACT FROM AUTHOR]

Background: Microalgae, with their high adaptability to various stress conditions and rapid growth, are considered excellent biomass resources for lipid production and biodiesel feedstocks. However, lipid yield and productivity of the natural strains are common bottlenecks in their large-scale use for lipid production, which can be overcome by evolving new strains using conventional and advanced mutagenic techniques. It is challenging to generate microalgae strains capable of high lipid synthesis through natural selection. As a result, random mutagenesis is currently considered a viable option in many scenarios. The objective of this study was to explore atmospheric and room temperature plasma (ARTP) as a random mutagenesis technique to obtain high lipid-accumulating mutants of a green microalga for improved biodiesel production. Results: A green microalgal species was isolated from the Chinese Yellow Sea and identified as Parachlorella kessleri (OM758328). The isolated microalga was subsequently mutated by ARTP to obtain high lipid-accumulating mutants. Based on the growth rate and lipid content, 5 mutants (named M1, M2, M4, M5, and M8) were selected from 15 pre-selected mutants. These five mutants varied in their growth rate from 0.33 to 0.68 day-1, with the lipid content varying between 0.25 g/L in M2 to 0.30 g/L in M8 at 10th day of cultivation. Among the mutants, M8 showed the maximum biomass productivity (0.046 g/L/day) and lipid productivity (20.19 mg/L/day), which were 75% and 44% higher than the wild strain, respectively. The triglyceride (TAG) content of M8 was found to be 0.56 g/L at 16th day of cultivation, which was 1.77-fold higher than that of the wild strain. Furthermore, M8 had the highest saturated fatty acids (C16-18) with the lowermost polyunsaturated fatty acid content, which are favorable properties of a biodiesel feedstock according to international standards. Conclusion: The mutant strain of P. kessleri developed by the ARTP technique exhibited significant improvements in biomass productivity, lipid content, and biodiesel quality. Therefore, the biomass of this mutant microalga could be a potential feedstock for biodiesel production. [ABSTRACT FROM AUTHOR]

Background: Schizochytrium sp. is a heterotrophic, oil-producing microorganism that can efficiently produce lipids. However, the industrial production of bulk chemicals using Schizochytrium sp. is still not economically viable due to high-cost culture medium. Replacing glucose with cheap and renewable lignocellulose is a highly promising approach to reduce production costs, but Schizochytrium sp. cannot efficiently metabolize xylose, a major pentose in lignocellulosic biomass. Results: In order to improve the utilization of lignocellulose by Schizochytrium sp., we cloned and functionally characterized the genes encoding enzymes involved in the xylose metabolism. The results showed that the endogenous xylose reductase and xylulose kinase genes possess corresponding functional activities. Additionally, attempts were made to construct a strain of Schizochytrium sp. that can effectively use xylose by using genetic engineering techniques to introduce exogenous xylitol dehydrogenase/xylose isomerase; however, the introduction of heterologous xylitol dehydrogenase did not produce a xylose-utilizing engineered strain, whereas the introduction of xylose isomerase did. The results showed that the engineered strain 308-XI with an exogenous xylose isomerase could consume 8.2 g/L xylose over 60 h of cultivation. Xylose consumption was further elevated to 11.1 g/L when heterologous xylose isomerase and xylulose kinase were overexpressed simultaneously. Furthermore, cultivation of 308-XIXK(S) using lignocellulosic hydrolysates, which contained glucose and xylose, yielded a 22.4 g/L of dry cell weight and 5.3 g/L of total lipid titer, respectively, representing 42.7 and 30.4% increases compared to the wild type. Conclusion: This study shows that engineering of Schizochytrium sp. to efficiently utilize xylose is conducive to improve its utilization of lignocellulose, which can reduce the costs of industrial lipid production. [ABSTRACT FROM AUTHOR]

The pH of skin is very decisive in the life of humans because it helps in enhancing infections that are related to antimicrobials. So far, the skin of humans considered is the main outdoor for several bacterial infections. The acidic Nature (pH) of the skin performs an integral physiologically part in the body and it has been identified as a regulatory component in the stratum corneum's homeostasis and skin barrier permeability. Surveys were done later illustrated that aggregation, skin barrier function& lipid production, epidermal differentiation& desquamation are basically impacted by the pH itself. Multiple epidermal pH figures have also been observed and tested in this study, all within the acidic condition however varying from pH 4.0 to 7.0, and depending on this pH, the 'natural' skin surface pH is expected to be 4.7 to 5.75 or 6. In addition, various studies are available in the latest days that focused primarily on alterations inside the pH of the stratum corneum's deeper layers including the effect of various physiological and pathological variables. Whenever the skin barrier is compromised, pH rises, resulting in a variety of ailments, including dermatitis, irritating contact dermatitis, ichthyosis, acne, and dry and ageing skin. In the current review, we will summarise many autogenous factors and exogenic factors such as skin moisture, sweat, sebum (produced by sebaceous glands), anatomic site, genetic susceptibility, age, detergents, so-called skin health maintenance products, wounds occlusive dressing could bother the pH of the skin. With that, we collected several publications in this review and explored the importance of pH surface and the key factors that influence skin's pH-related skin illness. We'll also go over how to determine the pH of the surface of the skin and also how it would affect skin and wounds. [ABSTRACT FROM AUTHOR]

As an interesting alternative microbial platform for the sustainable synthesis of oleochemical building blocks and biofuels, oleaginous yeasts are increasing in both quantity and diversity. In this study, oleaginous yeast species from northern Thailand were discovered to add to the topology. A total of 127 yeast strains were isolated from 22 forest soil samples collected from mountainous areas. They were identified by an analysis of the D1/D2 domain of the large subunit rRNA (LSU rRNA) gene sequences to be 13 species. The most frequently isolated species were Lipomyces tetrasporus and Lipomyces starkeyi. Based on the cellular lipid content determination, 78 strains of ten yeast species, and two potential new yeast that which accumulated over 20% of dry biomass, were found to be oleaginous yeast strains. Among the oleaginous species detected, Papiliotrema terrestris and Papiliotrema flavescens have never been reported as oleaginous yeast before. In addition, none of the species in the genera Piskurozyma and Hannaella were found to be oleaginous yeast. L. tetrasporus SWU-NGP 2-5 accumulated the highest lipid content of 74.26% dry biomass, whereas Lipomyces mesembrius SWU-NGP 14-6 revealed the highest lipid quantity at 5.20 ± 0.03 g L−1. The fatty acid profiles of the selected oleaginous yeasts varied depending on the strain and suitability for biodiesel production. [ABSTRACT FROM AUTHOR]

Toxic cyanobacterial blooms are recurrent for few decades throughout the globe, due to climate change, atmospheric warming and various anthropogenic activities with severe impacts of potential toxins on various ecosystems finally affecting the entire environment. These cyanobacteria are merely unexplored regarding their biochemical components except toxins. Variable influences and interactions of different factors including nitrogen, carbon, and availability of light are well known to crucially regulate cyanobacterial growth and metabolism. Thus, current research work is motivated for the evaluation and optimization of the effects of the aforementioned vital factors for improvement of biomass and lipid production of a freshwater, toxic strain of Anabaena circinalis. The modelling and optimization of factors such as nitrogen, light intensity and bicarbonate concentration (source of carbon) to maximize growth and lipid production were based on 20 design point experiments by Response Surface Methodology (RSM) and optimized values were further improved and validated by Particle Swarm Optimization (PSO) algorithm. The maximum optima were obtained 1.829 g L-1 and 39.64 % for biomass production and lipid content respectively from PSO optimization with two different sets of optimal values of factors. It shows 0.44 % and 2.77 % higher values of responses than that of RSM optimization. These asynchronous findings pioneered the enhanced lipid accumulation as well as the growth of a toxic cyanobacterium by optimizing interaction effects of culture conditions through various statistical and computational approaches. • Influencing factors were optimized for the improvement of biomass and lipid production. • RSM was used to optimize the system parameters and validated by PSO algorithm. • Maximum optima were obtained as 1.829 g L-1 and 39.64 % for biomass production. • Results shows that 0.44 % and 2.77 % higher values than that of RSM optimization. [ABSTRACT FROM AUTHOR]

This paper studied the bifurcation phenomena that can occur in a continuous bioreactor for lipid-rich biomass production using Rhodococcus opacus that feeds on refinery wastewater. An unstructured model of the bioreactor was developed based on experimentally validated studies reported in the literature. The analysis of the unsteady-state model was carried out both analytically and through numerical simulations. It was shown that the inhibition effect of the biomass growth rate can lead to the occurrence of static bistability in the model. Periodic behavior was also found for some range of model parameters. The effect of model parameters on the productivity of the bioreactor was also investigated. The analysis carried out in this paper allowed for the detection of unsafe operating regions in the bioreactor which, would help in the attenuation of these operational problems in the early stage of process design. [ABSTRACT FROM AUTHOR]

In this work two-stage cultivation of Scenedesmus acuminatus to enhance lipid productivity was investigated. Two two-stage cultivation strategies were employed: 1) S. acuminatus was cultured in nitrogen-limited conditions (0.75 g L−1 NaNO3, stage I), then nitrogen-limited condition (stage II); 2) S. acuminatus was cultured in sulfur-replete conditions (86.80 g L−1 MgSO4, stage I), then nitrogen-limited conditions (stage II). The lengths of stages I and II were varied within a total culture period of 20 days. The highest biomass (12.41 g L−1) and lipid productivity (346.51 mg L−1 day−1) were obtained when stage I lasted 8 days and the cells were then transferred to medium containing 0.3 g L−1 NaNO3 (stage II) for another 12 days; these values were 1.72- and 2.18- times higher, respectively, than those for the control group (single-stage cultivation in control group, 1.5 g L−1 NaNO3). Biochemical analyses indicated that S. acuminatus produced higher biomass and lipid content if stable carbon partitioning model occurred in stage II of culture. Overproduction of carbohydrates or soluble protein after transfer to the second stage was not conducive to the optimal balance of cell growth and lipid content. The findings of this study indicate that the regulation of nitrogen concentration to promote lipid accumulation is more effective than regulation via the sulfur concentration. This two-stage cultivation process is of interest for large-scale cultivation of S. acuminatus for enhanced lipid accumulation. [ABSTRACT FROM AUTHOR]

High salinity, nutrient deficiency, heavy metals, desiccation, temperature fluctuations, and ultraviolet radiations are major abiotic stress factors considered inhospitable to algal growth and development in natural and artificial environments. All these stressful conditions cause effects on algal physiology and thus biochemical functioning. For instance, long-term exposure to hyper/hypo salinity conditions inhibits cell differentiation and reduces growth. Photosynthesis is completely blocked in algae's dehydrated state, resulting in photoinhibition or photodamage. The limitation of nutrients in aquatic environments inhibits primary production via regulating phytoplankton community development and structure. Hence, in response to these stressful conditions, algae develop plenty of cellular, physiological, and morphological defences to survive and thrive. The conserved and generalized defence responses in algae include the production of secondary metabolites, desaturation of membrane lipids, activation of reactive species scavengers, and accumulation of compatible solutes. Moreover, a well-coordinated and timely response to such stresses involves signal perception and transduction mainly via phytohormones that could sustain algae growth under abiotic stress conditions. In addition, the combination of abiotic stresses and plant hormones could further elevate the biosynthesis of metabolites and enhance the ability of algae to tolerate abiotic stresses. This review aims to present different kinds of stressful conditions confronted by algae and their physiological and biochemical responses, the role of phytohormones in combatting these conditions, and, last, the future transgenic approaches for improving abiotic stress tolerance in algae. [ABSTRACT FROM AUTHOR]

Metabolic mechanisms driving rapid lipid production in atmospheric and room temperature plasma-mutagenized Chlorella sorokiniana remain unclear, hindering improvement in lipid productivity through genetic modification. This study investigated two dominant strains, ArX13 and AirX12, mutated by argon and air ARTP, for growth, lipid production, and transcriptomic changes across various culture stages. Results showed that biomass and lipid contents peaked in the late stage, with ArX13 and AirX12 achieving final biomass of 0.71 g/L and 0.70 g/L, and lipid content of 58.57 % g lipid g biomass −1 and 46.90 % g lipid g biomass −1, 2.20 and 1.77 times higher than the control, respectively. The maximum biomass and lipid productivity occurred at the mid-stage, with ArX13 and AirX12 showing biomass productivity of 0.025 g biomass L−1 day−1 and 0.024 g biomass L−1 day−1, and lipid productivity of 17.79 mg lipid L−1 day−1 and 11.40 mg lipid L−1 day−1, respectively. Transcriptomic analysis revealed minimal changes of gene expression during early and late stages, but significant biomass and lipid synthesis during the middle stage. Key genes like PDHB, ATP6A, LSC1 and OGDH were identified in the PPI network throughout the culture cycle, with PDHB remarkably promoting lipid precursor production, up-regulated 8.37 and 4.99-fold in M_ArX13-M_X0 and M_AirX12-M_X0, respectively. In addition, high expression of photosynthesis-related genes (psaL, psaO, psaD, psaK, psbP) contributed the superior lipid production for the argon-based ARTP case, highlighting common and intrinsic mechanisms regulating microalgal lipid accumulation under different ARTP mutations. This study shows that the selection of appropriate ARTP gases can maximize the lipid production of microalgal, and the discovery of some key genes can provide a theoretical basis for the selection of target genes for microalgal research, which is of great significance for promoting the research and development of microalgal lipid engineering. [Display omitted] • Argon source ARTP outperformed air source ARTP in biomass and lipid production. • Genes linked to growth and lipid synthesis are massively up-regulated in mid-culture. • PDHB, ATP6A, LSC1 and OGDH are important key genes throughout the culture cycle. • Photosynthesis genes under argon more effectively promoted lipid and biomass increase. [ABSTRACT FROM AUTHOR]

Oleaginous yeast can produce lipids while degrading phenol in wastewater treatment. In this study, a Plackett-Burman Design (PBD) was adopted to identify key factors of phenol degradation and lipid production using R toruloides 9564T. While temperature, inoculum size, and agitation were significant for both the processes (p < 0.05), pH and incubation were significant for lipid production, and phenol removal, respectively. Results from four factors (pH, temperature, inoculum size, and incubation period) central composite design (CCD) experiment were used to formulate quadratic and genetic algorithm-optimized ANN models. The reduced quadratic model for phenol degradation (R 2 : 0.993) and lipid production (R 2 : 0.958) were marginally inferior to ANN models (R 2 : 0.999, 0.982, respectively) on training sets. Multi-objective optimization with equal importance suggests phenol degradation between 106.4 and 108.76%, and lipid production of 0.864–0.903 g/L, by polynomial and ANN models. Complete phenol degradation (100%) and 3.35-fold increment (0.918 g/L) in lipid production were obtained at pH 6.07, inoculum size 14.68% v/v, at 29.5 °C in 92.17 h experimentally. [Display omitted] • PB design identified effective variables for phenol degradation and lipid synthesis. • RSM-CCD shows R2 values of 0.99 and 0.96 for phenol degradation and lipid synthesis. • RSM-CCD revealed a 3.5-fold increase in lipid synthesis under optimal conditions. • ANN-GA validate RSM-CCD data on phenol degradation and lipid synthesis. • ANN-GA found R2 values of 0.99 and 0.95 for phenol degradation and lipid synthesis. [ABSTRACT FROM AUTHOR]

Microalgae biomasses offer important benefits regarding macromolecules that serve as promising raw materials for sustainable production. In the present study, the microalgae Arthrospira platensis DHR 20 was cultivated in horizontal photobioreactors (HPBR), with and without temperature control, in batch mode (6 to 7 days), with anaerobically digested cattle wastewater (ACWW) as substrate. High dry biomass concentrations were observed (6.3–7.15 g L−1). Volumetric protein, carbohydrate, and lipid productivities were 0.299, 0.135, and 0.108 g L−1 day−1, respectively. Promising lipid productivities per area were estimated between 22.257 and 39.446 L ha−1 year−1. High CO2 bio-fixation rates were recorded (875.6–1051 mg L−1 day−1), indicating the relevant potential of the studied microalgae to mitigate atmospheric pollution. Carbon concentrations in biomass ranged between 41.8 and 43.6%. ACWW bioremediation was satisfactory, with BOD5 and COD removal efficiencies of 72.2–82.6% and 63.3–73.6%. Maximum values of 100, 95.5, 92.4, 80, 98, and 94% were achieved concerning the removal of NH4+, NO3−, Pt, SO42−, Zn, and Cu, respectively. Total and thermotolerant coliform removals reached 99–99.7% and 99.7–99.9%. This microalgae-mediated process is, thus, promising for ACWW bioremediation and valuation, producing a microalgae biomass rich in macromolecules that can be used to obtain friendly bio-based products and bioenergy. [ABSTRACT FROM AUTHOR]

Microbial fermentation offers a sustainable source of fuels, commodity chemicals, and pharmaceuticals, yet strain performance is influenced greatly by the growth media selected. Specifically, trace metals (e.g., iron, copper, manganese, zinc, and others) are critical for proper growth and enzymatic function within microorganisms yet are non-standardized across media formulation. In this work, the effect of trace metal supplementation on the lipid production profile of Yarrowia lipolytica was explored using tube scale fermentation followed by biomass and lipid characterization. Addition of iron (II) to the chemically defined Yeast Synthetic Complete (YSC) medium increased final optical density nearly twofold and lipid production threefold, while addition of copper (II) had no impact. Additionally, dose-responsive changes in lipid distribution were observed, with the percent of oleic acid increasing and stearic acid decreasing as initial iron concentration increased. These changes were reversible with subsequent iron-selective chelation. Use of rich Yeast Peptone Dextrose (YPD) medium enabled further increases in the production of two specialty oleochemicals ultimately reaching 63 and 47% of the lipid pool as α-linolenic acid and cyclopropane fatty acid, respectively, compared to YSC medium. Selective removal of iron (II) natively present in YPD medium decreased this oleochemical production, ultimately aligning the lipid profile with that of non-supplemented YSC medium. These results provide further insight into the proposed mechanisms for iron regulation in yeasts especially as these productions strains contain a mutant allele of the iron regulator, mga2. The work presented here also suggests a non-genetic method for control of the lipid profile in Y. lipolytica for use in diverse applications. Key points: • Iron supplementation increases cell density and lipid titer in Yarrowia lipolytica. • Iron addition reversibly alters lipid portfolio increasing linolenic acid. • Removal of iron from YPD media provides a link to enhanced oleochemical production. [ABSTRACT FROM AUTHOR]

The most common method of treating palm oil mill effluent (POME) is by using various types of bacteria communities. However, the utilization of microalgae in consuming the high nutrient content in wastewater offer additional benefit, particularly for CO2 sequestration. In this study, we proposed to evaluate the performance of multicultural bacteria obtained from municipal wastewater and Chlorella sp. for batch treatment of POME at different COD concentrations (ranges between 600 to 1,300 mg/L), microalgae species (C. vulgaris and C. pyrenoidosa) and speed of agitation (0 and 150 rpm). The results showed that between the bacteria and microalgae that are involved in POME treatment, microalgae give high removal of colour (93%) and ammoniacal nitrogen (95%). As for the COD and phosphate removal, both microorganisms show comparable performances. It was observed that C. pyrenoidosa was able to remove more colour compared to C. vulgaris where higher lipid yield production was obtained (47.6%). However, there is no significant impact of agitation on pollutant removal. This study also reveals that co-cultivation of different microalgae species does not affect the efficiency of the system. This study provides an important insight into developing an efficient and environmentally friendly method to treat wastewater by incorporating green technology in the treatment system. [ABSTRACT FROM AUTHOR]

Cerium is the most abundant rare earth metal in the earth's crust, and it has deleterious effects on aquatic ecosystems from fertilizer runoff. Scenedesmus obliquus is an oil‐rich microalga that grows rapidly and is sensitive to many kinds of toxins. Given that microalgae are useful indicators of eutrophication and toxic stress, it was found that lower concentrations of cerium (0.50–5.00 mg·L−1) stimulated algal growth and increased chlorophyll a content, whereas higher concentrations (above 50.00 mg·L−1) had an inhibitory effect on algal growth and chlorophyll a content. The algal growth rate and chlorophyll a content peaked at a cerium concentration of 5.00 mg·L−1. Both the donor and acceptor sides of photosystem II (PSII) reaction centers were sensitive to cerium‐induced stress. Specifically, high concentrations of cerium damaged the oxygen evolving complex and PSII reaction center and suppressed electron transport at the donor and receptor side of the reaction center, influencing the absorption, transfer, and application of light energy in S. obliquus XJ002. In addition, we established a simple method to quantify the intracellular lipid content of S. obliquus XJ002, and the optimum staining conditions for Nile red were as follows: volume percentage of dimethyl sulfoxide was 2%, the concentration of Nile red was 2.0 µg·mL−1, and the staining time of Nile red was 5 min. The addition of cerium resulted in a significant increase in the total lipid content of XJ002. When the concentration of cerium was 50 mg·L−1, the total lipid content was 16.26% higher than the control group. This information will enhance our ability to utilize microelement fertilizer in biomass accumulation programs and will help to further reveal the key regulatory factors in the lipid metabolism, and would lay the foundation for promoting the research of microalgae bioenergy. [ABSTRACT FROM AUTHOR]

Rhodosporidium toruloides is a useful oleaginous yeast, but lipids production is affected by various factors including nutrients in the culture medium. Herein, the R-ZL2 high-yield mutant strain was used to investigate the effects of different carbon sources (sucrose, glucose, xylose), nitrogen sources (ammonium sulphate, ammonium nitrate), and C/N ratio on lipids production capacity, get the following conclusion (1) Compared with glucose and xylose, sucrose was a superior carbon source for lipids production; (2) When using ammonium sulphate as the nitrogen source, a C/N ratio of 200:1 achieved the highest biomass, lipids production and lipids content (10.7 g/L, 6.32 g/L and 59%, respectively), and lipids produced under different C/N conditions have potential for biodiesel production (except for C/N = 40 and C/N = 80); (3) When using ammonium nitrate as the nitrogen source, a C/N ratio of 200:1 achieved the highest biomass, lipids production and lipids content (12.1 g/L, 8.25 g/L and 65%, respectively), and lipids produced under different C/N ratio conditions have potential for biodiesel production. Thus, a combination of sucrose and ammonium nitrate was optimal for the lipid accumulation in R-ZL2. The findings will lay a foundation for further improving lipids yields. [ABSTRACT FROM AUTHOR]

Biodiesel, as a renewable and eco-friendly energy source that can be produced through algae oil esterification, has recently received much attention. Maximization of algal biomass and lipid content is crucial for commercial biodiesel production. In this study, Chlorella sp. PG96, a microalgal strain isolated from urban wastewater, was identified considering its morphological and molecular characteristics. Fractional factorial design (2 11 - 7) was employed to screen medium and environmental factors for achieving high lipid productivity. The effects of eleven factors including light intensity, light spectrum, aeration rate, temperature, salinity, NaHCO 3 , CO 2 , NaNO 3 , NH 4 Cl, MgSO 4.7H 2 O, and K 2 HPO 4 and their interactions on growth characteristics of Chlorella sp. PG96 (biomass and lipid production) were statistically assessed. Based on the experimental results, lipid productivity was at its maximum (54.19 ± 8.40 mg lipid L−1 day−1) under a combination of high levels of all factors. The analysis also showed that physical parameters of light intensity and temperature were more effective on algal growth compared to nutritional parameters. Furthermore, nitrogen source of ammonium and carbon source of bicarbonate played more significant roles in biomass and lipid production, compared with nitrate and CO 2 , respectively. Although the effect of sulfur limitation on cellular growth was similar to phosphorus deficiency, S-limitation had a greater impact on lipid accumulation. The interaction between NaHCO 3 and NH 4 Cl was the most prominent interaction affecting all responses. It is concluded that Chlorella sp. PG96 at a high level of light intensity and temperature (22500 Lux and 32 °C, respectively) can be a prospective candidate for biodiesel production. [Display omitted] • Chlorella sp. PG96 was found to be a suitable candidate for biodiesel production. • FFD (2 11 - 7) successfully determined combined effects of 11 factors on algal growth. • Maximum lipid productivity was obtained at high levels of all factors. • Light intensity and temperature had the most impact on biomass and lipid. • Interaction between NaHCO 3 and NH 4 Cl was the most effective term for all responses. [ABSTRACT FROM AUTHOR]

In the global effort to reduce CO 2 emissions, the concurrent enhancement of pollutant degradation and reductions in fossil fuel consumption are pivotal aspects of microalgae-mediated wastewater treatment. Clarifying the degradation mechanisms of bacteria and microalgae during pollutant treatment, as well as regulatory biolipid production, could enhance process sustainability. The synergistic and inhibitory relationships between microalgae and bacteria are introduced in this paper. The different stimulators that can regulate microalgal biolipid accumulation are also reviewed. Wastewater treatment technologies that utilize microalgae and bacteria in laboratories and open ponds are described to outline their application in treating heavy metal-containing wastewater, animal husbandry wastewater, pharmaceutical wastewater, and textile dye wastewater. Finally, the major requirements to scale up the cascade utilization of biomass and energy recovery are summarized to improve the development of biological wastewater treatment. [Display omitted] • The synergistic and antagonistic interactions between bacteria and microalgae are summarized. • Different stimulators that could regulate microalgal biolipid accumulation are also reviewed. • Wastewater treatment technologies that utilize microalgae and bacteria in the laboratory and open ponds are described. • The trends in microalgae-bacterial consortium in multistage energy production are proposed. [ABSTRACT FROM AUTHOR]

The biochemical treatment of cellulosic ethanol wastewater (CEW) is considered to be an ideal green process. To screen out the best oleaginous yeastfor the utilization of cellulosic ethanol wastewater, four oleaginous yeasts (Trichosporon cutaneum, Rhorosporidium toruloides, Cryptococcus albidus and T. coremiiforme) were compared to assess their abilities for lipid production in terms of biomass production, lipid content and lipid yield. Furthermore, thechemical oxygen demand (COD) conversion rate, COD degradation and fatty acid composition were calculated to analyze the effect of wastewaters treatment. According to the fermentation results, the highest biomass and lipid yield of T. cutaneum in CEW were 20.945 and 1.56 g/L, respectively, while the R. toruloides reached the highest lipid content (17.32%). The maximum conversion rates of T. cutaneum are 73.64 and 6.06%, respectively, in terms of conversion yield of biomass/COD and lipids/COD. The content of fatty acids showed that after six days' fermentation, T. coremiiforme obtained the highest unsaturated fatty acid content, the content of C18:1 and C18:2 was 57.64%. This study suggests that T. cutaneum has great potential for lipid production and wastewaters treatment from cellulosic ethanol fermentation. Four oleaginous yeasts were used for lipid production. Cellulosic ethanol fermentation wastewater as growth medium for lipid production. Biomass and lipid yield of Trichosporon cutaneum showed potential for lipid production. Chemical oxygen demand conversion rate and chemical oxygen demand degradation of Trichosporon cutaneum showed potential for wastewater treatment. [ABSTRACT FROM AUTHOR]

Most of the oleaginous microorganisms cannot assimilate xylose in the presence of glucose, which is the major bottleneck in the bioconversion of lignocellulose to biodiesel. Our present study revealed that overexpression of xylose isomerase (XI) gene xylA or xylulokinase (XK) gene xks1 increased the xylose consumption by 25 to 37% and enhanced the lipid content by 8 to 28% during co-fermentation of glucose and xylose. In xylA overexpressing strain Mc-XI, the activity of XI was 1.8-fold higher and the mRNA level of xylA at 24 h and 48 h was 11- and 13-fold higher than that of the control, respectively. In xks1 overexpressing strain Mc-XK, the mRNA level of xks1 was 4- to 11-fold of that of the control strain and the highest XK activity of 950 nmol min−1 mg−1 at 72 h which was 2-fold higher than that of the control. Additionally, expression of a translational fusion of xylA and xks1 further enhanced the xylose utilization rate by 45%. Our results indicated that overexpression of xylA and/or xks1 is a promising strategy to improve the xylose and glucose co-utilization, alleviate the glucose repression, and produce lipid from lignocellulosic biomass in the oleaginous fungus M. circinelloides. Key points: • Overexpressing xylA or xks1 increased the xylose consumption and the lipid content. • The xylose isomerase activity and the xylA mRNA level were enhanced in strain Mc-XI. • Co-expression of xylA and xks1 further enhanced the xylose utilization rate by 45%. [ABSTRACT FROM AUTHOR]

Thraustochytrid strains belonging to the genus Aurantiochytrium accumulate significant amounts of lipids including polyunsaturated fatty acids and carotenoids and, therefore, are expected to be used for industrial production of various valuable materials. Although various efforts such as chemical mutagenesis and homologous gene recombination have been made to improve lipid productivity of Aurantiochytrium species, low specificity and efficiency in the conventional methods hinder the research progress. Here, we attempted to apply a genome editing technology, the CRISPR-Cas9 system as an alternative molecular breeding technique for Aurantiochytrium species to accelerate the metabolic engineering. The efficiency of specific gene knock-in by the homologous recombination increased more than 10-folds by combining the CRISPR-Cas9 system. As a result of disrupting the genes associated with β-oxidation of fatty acids by the improved method, the genome edited strains with higher fatty acid productivity were isolated, demonstrating for the first time that the CRISPR-Cas9 system was effective for molecular breeding of the strains in the genus Aurantiochytrium to improve lipid productivity. [ABSTRACT FROM AUTHOR]

• First report of lipids production in the marine diatom Chaeotceros affinis. • Palm oil mill effluent (POME) was sole source of N, P and other trace elements. • POME growth media were compared with a standard control medium. • Biomass grown in a vitamins-free POME medium exceeded 48 % in lipids content. • Lipid productivity exceeded 16 mg L–1 d –1 in a 18-day batch culture. Biomass and lipid production by the marine diatom Chaetoceros affinis were characterized under continuous light with aeration. Media based on palm oil mill effluent (POME; 10, 20 and 30 % v/v in distilled water) were used together with a standard control medium. The maximum biomass concentration on day 12 of batch cultures in control medium was 821 ± 71 mg L–1. Under identical conditions, in the best POME medium (20 % POME v/v in distilled water with other inorganic components), the biomass concentration was reduced by ∼11 % to 734 ± 66 mg L–1. The lipid content of the biomass grown in the control medium was 50.8 ± 4.5 % by dry weight, but was a little lower (48.9 ± 4.1 % by dry wt) in the above specified best POME medium. In the best POME medium, oleic acid was the major fatty acid (72.3 ± 5.2 % by weight) in the total lipids extracted from the biomass and monounsaturated fatty acids were the main type of fatty acids (74.6 ± 5.2 %). POME levels of >20 % in the medium suppressed both biomass and lipid production relative to the medium with 20 % POME. [ABSTRACT FROM AUTHOR]

Research on lipid production in microalgae, especially to target biofuels, has been widely published in recent decades. This literature is mostly focused on the optimization of culture conditions, metabolic engineering, biorefinery and downstream processes. At the lab scale, lipid production is very promising concerning biofuels production based on microalgae due to their high lipid content. However, the accumulation of knowledge associated with the development of processes and technologies for the production of biomass and associated lipids has so far not achieved the hoped-for commercial success due to still too expensive operating costs compared to fossil fuels. This review aims to present the influence of various abiotic factors on the accumulation of lipids and lipid profile in microalgae, such as the depletion of the culture medium, temperature and light, but also most innovative like the induction by hormones. Other optimization procedures and the use of wastewater to improve the cost-viability are discussed in perspective. [ABSTRACT FROM AUTHOR]

The crucial need for alternative oil sources due to increasing energy demand has motivated research to find alternatives resources. In the current study, different fungal groups were isolated from diverse Egyptian localities for oil production screening as well as sugarcane bagasse management. A total of 499 fungal isolates were recovered from soil and dung samples; one isolate was identified as Mortierella wolfii AH12 using morphological and molecular techniques and was demonstrated to produce high lipid concentrations. Taguchi design was used to obtain higher lipid productivity. Accordingly, dry biomass, total lipids, and lipid percentage were 3.81 g l−1, 1.57 g l−1, and 41.2%, respectively, under the optimized culture conditions: 40 g glucose, 5 g peptone, pH 5 at 30 0°C for 10 days. Non-pretreated sugarcane bagasse was not only utilized as an alternative carbon source, but it was also an effective source of nutrients for mold sprouting and lipid biosynthesis using solid-state fermentation (SSF) technique. Promising fatty acid profile was deduced, where palmitic (PA) and oleic acid (OA) were predominant. Moreover, unsaturated fatty acids (USFAs) as linoleic acid (LA), Gamma linolenic acid (GLA), Dihomo gamma linolenic acid (DGLA), arachidonic acid (ARA) were present in low quantities according to GC–MS analysis. The study offers a promising result that serves in the management of agricultural waste, converting it into lipids with a promising profile which could be considered as a precursor for biodiesel. [ABSTRACT FROM AUTHOR]

Background: Converting wastewater sludge to lipid is considered as one of the best strategies of sludge management. The current problem of lipid production from wastewater sludge is the low yield (0.10–0.16 g lipid/g dry sludge) due to the low availability of easily uptaken materials (such as soluble monosaccharide and oligosaccharide) in sludge to oleaginous microorganism (Rhodotorula glutinis, Trichosporon oleaginosus, Lipomyces starkeyi). Pretreatments are efficient methods to improve sludge bioavailability. This study is aimed to achieve high lipid production from sludge and high sludge reduction. Results: In this study, it was observed that the soluble chemical oxygen demand (SCOD) had significantly increased after different pretreatment. The SCOD in the supernatant was increased from 32.64 to 180.25 mg/L, 924.16 mg/L, 1029.89 mg/L and 3708.31 mg/L after acidic (pH 2 for 2 h), alkaline (pH 12 for 2 h), microwave irradiation (15 min with 5 min interval), and ultrasonication (30 min at 450 W and 20 kHz frequency with 5 s on and 2 s off mode) pretreatment, respectively. Pretreatments have also increased the release of total nitrogen (TN) and total phosphorus (TP) from solids. The sludge after different pretreatments were used as a medium for lipid production, and the highest lipid content (36.67% g/g) was obtained in the fermentation with ultrasonication pretreatment sludge, and the sludge reduction was 63.10%. For other pretreatments, the lipid content and sludge reduction were 18.42% and 32.63% in acid pretreatment case, 21.08% and 36.44% in alkaline pretreatment case, and 26.31% and 43.03% in microwave pretreatment case, respectively. Conclusion: It was found that ultrasonication pretreatment was the most efficient way to increase the sludge biodegradability (SCOD) and to release TN and TP from solid phase to liquid phase. Pretreated sludge for lipid production achieved significant improvement in lipid yield and sludge reduction. Lipids produced from pretreated sludge were transesterified to biodiesel and the analysis showed that biodiesel had a similar composition as commercial biodiesel. The study reveals that pretreatment on sludge is a promising method for enhancing biological sludge management efficiency. [ABSTRACT FROM AUTHOR]

Blastobotrys raffinosifermentans is an ascomycetous yeast with biotechnological applications, recently shown to be an oleaginous yeast accumulating lipids under nitrogen limitation. Diacylglycerol acyltransferases (DGATs) act in the lipid storage pathway, in the last step of triacylglycerol biosynthesis. Two DGAT families are widespread in eukaryotes. We first checked that B. raffinosifermentans strain LS3 possessed both types of DGAT, and we then overexpressed the native DGAT-encoding genes, DGA1 and DGA2 , separately or together. DGA2 (from the DGAT1 family) overexpression was sufficient to increase lipid content significantly in LS3, to up to 26.5% of dry cell weight (DCW), 1.6 times the lipid content of the parental strain (16.90% of DCW) in glucose medium under nitrogen limitation. By contrast, DGA1 (of the DGAT2 type) overexpression led to a large increase (up to 140-fold) in the amount of the corresponding transcript, but had no effect on overall lipid content relative to the parental strain. Analysis of the expression of the native genes over time in the parental strain revealed that DGA2 transcript levels quadrupled between 8 and 24 h in the N-limited lipogenic medium, whereas DGA1 transcript levels remained stable. This survey highlights the predominant role of the DGAT1 family in lipid accumulation and demonstrates the suitability of B. raffinosifermentans for engineering for lipid production. [ABSTRACT FROM AUTHOR]

Comparative profilings of cell growth and lipid production in the morphologically engineered strain (Δags1) and the wild type (WT) of Aspergillus oryzae BCC7051 were implemented. Using various nitrogen sources, a discrimination in cell morphology between the two strains was found, of which the Δags1 culture exhibited mycelial growth as small pellets in contrast to the WT. Of them, sodium nitrate and potassium nitrate were optimal for lipid production of the WT and Δags1 strains, respectively, which the highest lipid concentrations of 7.2 and 7.9 g L−1 were obtained in the respective cultures. The mathematical models of the growth kinetics and lipid phenotypes of both fungal strains were developed, enabling to distinguish three lipid-producing stages, including low lipid-producing, lipid accumulation, and lipid turnover stages. The model validation showed good performances in all nitrogen sources tested for the WT, but only NaNO3 and mixed yeast extract/NH4Cl were fitted well for the Δags1. The difference in the period of lipid-producing stages between the WT and Δags1 indicated the metabolic alterations of A. oryzae by the defect of a gene involved in the cell wall biosynthesis, which exhibited benefits for bioprocessing practices in addition to the high productivities of biomass and lipid. These findings would further permit the manipulation in the metabolic hub of the fungal production platform for other industrial purposes. [ABSTRACT FROM AUTHOR]

Lignocellulosic biomass shows high potential as a renewable feedstock for use in biodiesel production via microbial fermentation. Yarrowia lipolytica, an emerging oleaginous yeast, has been engineered to efficiently convert xylose, the second most abundant sugar in lignocellulosic biomass, into lipids for lignocellulosic biodiesel production. Yet, the lipid yield from xylose or lignocellulosic biomass remains far lower than that from glucose. Here we developed an efficient xylose‐utilizing Y. lipolytica strain, expressing an isomerase‐based pathway, to achieve high‐yield lipid production from lignocellulosic biomass. The newly developed xylose‐utilizing Y. lipolytica, YSXID, produced 12.01 g/L lipids with a maximum yield of 0.16 g/g, the highest ever reported, from lignocellulosic hydrolysates. Consequently, this study shows the potential of isomerase‐based xylose‐utilizing Y. lipolytica for economical and sustainable production of biodiesel and oleochemicals from lignocellulosic biomass. [ABSTRACT FROM AUTHOR]

In this work, we propose and describe a novel diatom species, Fistulifera alcalina sp. nov., isolated from Port Mayaca on Lake Okeechobee, Florida (USA). This new diatom species represents a distinct lineage within the genus Fistulifera, which includes F. iranensis (Hustedt) Lange-Bertalot, F. saprophila (Lange-Bertalot & Bonik) Lange-Bertalot, F. pelliculosa (Kützing) Lange-Bertalot, and F. solaris S. Mayama, M. Matsumoto, K. Nemoto & T. Tanaka. Morphometric and molecular phylogenetic analyses of the 18S rRNA and rbcL genes strongly support Fistulifera alcalina as a new species. Fistulifera alcalina sp. nov. is cultured in media with moderate salinity and high bicarbonate concentrations in contrast to the other species of Fistulifera which show ecological preferences for high salinity and eutrophic or oligotrophic waters. This is also the first record of Fistulifera in the southeastern United States (Florida, USA). [ABSTRACT FROM AUTHOR]

Oleaginous fungi have recently gained increasing attention among different microorganisms due to their ability for lipid production for the preparation of biofuel. In the present study, a locally isolated fungus E45, identified genetically as Aspergillus sp. strain EM2018, was found to produce 25.2% of the total lipids content of its dry cell weight (DCW). Optimization of culture conditions was performed and lipid accumulation increased by about 2.4 fold (from 25.2% to 60.1% of DCW) when the fungus was grown for seven days in the potato dextrose (50 g/L) liquid medium at pH 5.0, incubation temperature at 30 °C and inoculum size of 2 × 106 spore/mL. Supplementation of the medium with yeast extract and NaNO3 at a concentration of 0.05% as organic and inorganic nitrogen sources, respectively, increased lipid production (53.3% lipid/dry biomass). Gas chromatography analysis of fungal lipids revealed the presence of saturated (mainly palmitic acid C16:0 (33%) and lignoceric acid C24:0 (15%)) and unsaturated fatty acids in different proportions (mainly linoleic acid C18:2 (24.4%), oleica cid C18:1 (14%) and arachidonic C20:4 (7.4%). These findings suggest this new oleaginous fungus as a promising feedstock for various industrial applications and for the preparation of biodiesel. [ABSTRACT FROM AUTHOR]

Orange peel waste (OPW), the primary byproduct of the juice extraction process, is annually generated in massive amounts (21 Mton), and its aqueous extraction in biorefining operations yields a liquid fraction, referred to as orange peel extract (OPE). Although OPE contains significant amounts of easily assimilable carbohydrates, such as fructose, glucose, and sucrose, no investigations have been conducted yet to assess its possible use in biodiesel production by oleaginous yeasts. Consequently, the objective of the present study was to assess whether OPE might act as the basis of a liquid medium for microbial lipid production. A screening conducted with 18 strains of oleaginous yeasts in shaken flask on the OPE-based medium showed that Rhodosporidium toruloides NRRL 1091 and Cryptococcus laurentii UCD 68-201 gave the best results in terms of lipid production (5.8 and 4.5 g L−1, respectively) and accumulation (77 and 47% on a dry matter basis, respectively). The subsequent scale transfer of the process to a 3-L STR operated in batch mode halved the time required to reach the lipid peak with the ensuing increase in volumetric productivities in R. toruloides NRRL 1091 (3646 mg L−1 day−1) and C. laurentii UCD 68-201 (2970.7 mg L−1 day−1). The biodiesel yields from the lipids of the former and the latter strain were 36.9 and 31.9%, respectively. Based on multivariate analysis of fatty acid methyl ester compositions, the lipids from the former and the latter strain were highly resembling those of Jatropha and palm oils, two commonly used feedstocks for biodiesel manufacturing. [ABSTRACT FROM AUTHOR]

Navicula incerta is a benthic diatom with potential use in nutrition and health for humans and aquaculture. Therefore, it is important to know its optimal growth conditions and biochemical composition. In this study, the effects of nitrate and silicate concentration on the growth kinetics and biochemical composition of N. incerta under laboratory conditions were measured. F/2 media was used as the control, and different concentrations of nitrogen (N/4, N/8 and N/16) and silicates (Si/4, Si/8 and Si/16) were evaluated. We measured cell concentration, chlorophyll-a and conducted proximal chemical analyses. It was found that different concentrations of nutrients affected the diatom's growth kinetics and affected the concentrations of ash, chlorophyll, protein, lipids and carbohydrates. The highest concentration of lipids was obtained in the limiting treatment of nitrogen N/8 (27.09%), while the lowest value was found with silicate Si/8 media (16.97%). Carbohydrates increased compared to the control, with the N/16 treatment presenting the maximal concentration (23.31%). Treatments with reduced nitrate (N/8 and N/16) demonstrated the lowest concentrations of protein (18.75 and 12.44%, respectively), while in reduced silicate treatments, no statistical differences (P = 0.05) were observed. Therefore, media limited nitrogen and silicates affected the growth kinetics and proximal chemical composition of N. incerta. The growth of this species using the N/8 medium is a suitable method for increasing lipid concentration in N. incerta. [ABSTRACT FROM AUTHOR]

Background: Without sufficient alternatives to crude oil, as demand continues to rise, the global economy will undergo a drastic decline as oil prices explode. Dependence on crude oil and growing environmental impairment must eventually be overcome by creating a sustainable and profitable alternative based on renewable and accessible feedstock. One of the promising solutions for the current and near-future is the substitution of fossil fuels with sustainable liquid feedstock for biofuel production. Among the different renewable liquid feedstock's studied, wastewater is the least explored one for biodiesel production. Sago wastewater is the byproduct of the cassava processing industry and has starch content ranging from 4 to 7%. The present investigation was aimed to produce microbial lipids from oleaginous yeast, Candida tropicalis ASY2 for use as biodiesel feedstock and simultaneously decontaminate the sago processing wastewater for reuse. Initial screening of oleaginous yeast to find an efficient amylolytic with maximum lipid productivity resulted in a potent oleaginous yeast strain, C. tropicalis ASY2, that utilizes SWW as a substrate. Shake flask experiments are conducted over a fermentation time of 240 h to determine a suitable fatty acid composition using GC-FID for biodiesel production with simultaneous removal of SWW pollutants using ASY2. Results: The maximum biomass of 0.021 g L−1 h−1 and lipid productivity of 0.010 g L−1 h−1 was recorded in SWW with lipid content of 49%. The yeast strain degraded cyanide in SWW (79%) and also removed chemical oxygen demand (COD), biological oxygen demand (BOD), nitrate (NO3), ammoniacal (NH4), and phosphate (PO4) ions (84%, 92%, 100%, 98%, and 85%, respectively). GC-FID analysis of fatty acid methyl esters (FAME) revealed high oleic acid content (41.33%), which is one of the primary fatty acids for biodiesel production. Conclusions: It is evident that the present study provides an innovative and ecologically sustainable technology that generates valuable fuel, biodiesel using SWW as a substrate and decontaminates for reuse. [ABSTRACT FROM AUTHOR]

Background: Oleaginous yeasts can be grown on different carbon sources, including lignocellulosic hydrolysate containing a mixture of glucose and xylose. However, not all yeast strains can utilize both the sugars for lipogenesis. Therefore, in this study, efforts were made to isolate dual sugar-utilizing oleaginous yeasts from different sources. Results: A total of eleven isolates were obtained, which were screened for their ability to utilize various carbohydrates for lipogenesis. One promising yeast isolate Trichosporon mycotoxinivorans S2 was selected based on its capability to use a mixture of glucose and xylose and produce 44.86 ± 4.03% lipids, as well as its tolerance to fermentation inhibitors. In order to identify an inexpensive source of sugars, nondetoxified paddy straw hydrolysate (saccharified with cellulase), supplemented with 0.05% yeast extract, 0.18% peptone, and 0.04% MgSO4 was used for growth of the yeast, resulting in a yield of 5.17 g L−1 lipids with conversion productivity of 0.06 g L−1 h−1. Optimization of the levels of yeast extract, peptone, and MgSO4 for maximizing lipid production using Box-Behnken design led to an increase in lipid yield by 41.59%. FAME analysis of single cell oil revealed oleic acid (30.84%), palmitic acid (18.28%), and stearic acid (17.64%) as the major fatty acids. Conclusion: The fatty acid profile illustrates the potential of T. mycotoxinivorans S2 to produce single cell oil as a feedstock for biodiesel. Therefore, the present study also indicated the potential of selected yeast to develop a zero-waste process for the complete valorization of paddy straw hydrolysate without detoxification. [ABSTRACT FROM AUTHOR]

Thiamine is a vitamin that functions as a cofactor for key enzymes in carbon and energy metabolism in all living cells. While most plants, fungi, and bacteria can synthesize thiamine de novo, the oleaginous yeast Yarrowia lipolytica cannot. In this study, we used proteomics together with physiological characterization to elucidate key metabolic processes influenced and regulated by thiamine availability and to identify the genetic basis of thiamine auxotrophy in Y. lipolytica. Specifically, we found that thiamine depletion results in decreased protein abundance for the lipid biosynthesis pathway and energy metabolism (i.e., ATP synthase), leading to the negligible growth and poor sugar assimilation observed in our study. Using comparative genomics, we identified the missing 4-amino-5-hydroxymethyl-2- methylpyrimidine phosphate synthase (THI13) gene for the de novo thiamine biosynthesis in Y. lipolytica and discovered an exceptional promoter, P3, that exhibits strong activation and tight repression by low and high thiamine concentrations, respectively. Capitalizing on the strength of our thiamine-regulated promoter (P3) to express the missing gene from Saccharomyces cerevisiae (scTHI13), we engineered a thiamine-prototrophic Y. lipolytica strain. By comparing this engineered strain to the wild-type strain, we revealed the tight relationship between thiamine availability and lipid biosynthesis and demonstrated enhanced lipid production with thiamine supplementation in the engineered thiamine-prototrophic Y. lipolytica strain. [ABSTRACT FROM AUTHOR]

Using the effluent of municipal wastewater treatment plant (WWTP) to cultivate lipid-producing microalgae is likely to reduce the production cost of algae-derived biodiesel. However, the toxicity of antibiotics present in the effluent may affect the growth of microalgae and lipid production. This study investigated the effects of erythromycin (ERY) and tetracycline (TC) on the growth and lipid production of Tetradesmus (Scenedesmus) obliquus cultivated in the effluent of municipal WWTP. The results indicated that ERY was more toxic to T. obliquus compared to TC. The critical concentrations of ERY resulting in S. obliquus algae growth inhibition and increased lipid accumulation were 100 and 10 μg L−1, respectively. Maximum lipid accumulation was observed at an ERY concentration of 50 μg L−1, while lipid accumulation was significantly decreased at ERY concentrations greater than 250 μg L−1. Interestingly, the contents of polyunsaturated fatty acids in T. obliquus exposed to ERY were significantly increased, while the contents of total saturated fatty acids and monounsaturated fatty acids were significantly decreased. TC had no effect on the growth or lipid production when the concentration reached 500 μg L−1. Overall, the presence of ERY and TC in the municipal WWTP effluent does not largely affect the growth or lipid production of T. obliquus. [ABSTRACT FROM AUTHOR]

Microalgal lipids have been considered as one of the most promising feedstocks for biodiesel production. In order to solve the contradiction of algae growth and lipid accumulation, Chlorella vulgaris was co-cultured with Mesorhizobium sangaii under nitrogen deficiency conditions. The biomass and lipid production of C. vulgaris–bacteria co-culture with initial ratio of algae/bacteria = 40:1 were significantly improved compared with the pure algae culture. The maximum biomass, lipid content and productivity of algae in the co-cultures at 40:1 ratio were 1.89 mg L−1, 51.2%, and 96.77 mg L−1 day−1, respectively, which were 1.5, 2.2, and 3.3 times higher than those of the pure algae culture. Furthermore, the proportion of unsaturated fatty acids and C18:1 fatty acid of the consortium system was also significantly increased. Our study clearly suggests that co-cultivation of algae–bacteria can effectively contribute to the quality and quantity of microalgal bio-oil and shows promising applications for production of algal biomass and biodiesel. [ABSTRACT FROM AUTHOR]

Thiamine is a vitamin that functions as a cofactor for key enzymes in carbon and energy metabolism for all living cells. While most plants, fungi and bacteria can synthesize thiamine de novo, the oleaginous yeast, Yarrowia lipolytica, cannot. In this study, we used proteomics together with physiological characterization to understand key metabolic processes influenced and regulated by thiamine availability and identified the genetic basis of thiamine auxotrophy in Y. lipolytica. Specifically, we found thiamine depletion results in decreased protein abundance of the lipid biosynthesis pathways and energy metabolism (i.e., ATP synthase), attributing to the negligible growth and poor sugar assimilation observed in our study. Using comparative genomics, we identified the missing 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate synthase (THI13) in the de novo thiamine biosynthesis of Y. lipolytica, and discovered an exceptional promoter, P3, that exhibits strong activation or tight repression by low and high thiamine concentrations, respectively. Capitalizing on the strength of our thiamine-regulated promoter (P3) to express the missing gene from Saccharomyces cerevisiae (scTHI13), we engineered the thiamine-prototrophic Y. lipolytica. By comparing this engineered strain to the wildtype, we unveiled the tight relationship linking thiamine availability to lipid biosynthesis and demonstrated enhanced lipid production with thiamine supplementation in the engineered thiamine-prototrophic Y. lipolytica. Thiamine plays a crucial role as an essential cofactor for enzymes in carbon and energy metabolism of all living cells. Thiamine deficiency has detrimental consequences on cellular health. Yarrowia lipolytica, a non-conventional oleaginous yeast with broad biotechnological applications, is a native thiamine auxotroph, whose effects on cellular metabolism are not well understood. Therefore, Y. lipolytica is an ideal eukaryotic host to study thiamine metabolism, especially as mammalian cells are also thiamine-auxotrophic and thiamine deficiency is implicated in several human diseases. This study elucidates the fundamentals of thiamine deficiency on cellular metabolism of Y. lipolytica and identifies genes and novel thiamine- regulated elements that eliminate thiamine auxotrophy in Y. lipolytica. Furthermore, discovery of thiamine-regulated elements enables development of thiamine biosensors with useful applications in synthetic biology and metabolic engineering. [ABSTRACT FROM AUTHOR]

[Display omitted] • Combined micro-current and salt stresses promoted lipid production of Chlorella vulgaris. • Periodic micro-current enhanced algal growth and carbon flow distribution to lipid production. • Increased activities of antioxidant and lipid-regulation enzymes contributed to lipid synthesis. • Current and salinity successively favored fatty acid composition for biodiesel production. Two-stage cultivation of Chlorella with periodic micro-current coupled with NaCl stress was investigated to analyze the performance and contribution of increased lipid production. In the growth stage, periodic micro-current of 3 mA resulted in an escalation of 290.06 mg/L of Chlorella's biomass by 61.25 % in comparison to the control. During the induction stage, under a NaCl concentration of 8 g/L, the highest lipid content and yield attained were 42.11 % and 199.32 mg/L, revealing an increment of 53.13 % and 39.11 % respectively when compared to the control. Additionally, the polyunsaturated fatty acids content (PUFA) decreased from 30.11 % to 23.09 %, while the saturated fatty acids content (SFA) increased from 31.86 % to 42.39 %. Chlorella demonstrated adaptability to the alterations in the cultivation environment by an initial upsurge and later a reduction in SOD and CAT enzyme activities, and concurrently facilitated lipid synthesis by similar changes in the activities of ACCase and DGAT. The above cultivation enabled a synergistic effect of swift growth and efficient lipid accumulation in Chlorella , providing groundwork for large-scale biofuel production. [ABSTRACT FROM AUTHOR]