Carotenoid Synthesis Research Articles

Optimizing pH and Light for Enhanced Carotenoid Synthesis and Antioxidant Properties in Sub-Aerial Cyanobacteria.

Carotenoid, natural pigments, synthesized by plants and microbes are now much favored in global markets due to the awareness of their putative health benefits, and a wide array of commercial applications. There is a diversity of natural and synthetic carotenoid, but only a few of them are commercially produced, including carotenes (β-carotene and lycopene) and xanthophylls (astaxanthin, canthaxanthin, lutein, zeaxanthin, and capsanthin). However, for commercial production, plants and algae are more favored than cyanobacteria because of their much less carotenoid synthesis than land plants; although they are well known for producing commercially important carotenoid. But with advances in optimization of their carotenoid production, cyanobacteria can be used as a potential source of carotenoid production in the future allowing us to exploit its various applications. Hence, this study investigated the effects of pH and light conditions on carotenoid production in the sub-aerial cyanobacterium Desertifilum dzianense ON358232.1. The results revealed that the highest carotenoid synthesis occurred under alkaline conditions (pH 9) and red-light exposure, significantly increasing compared to the control (pH 7.2, white light). UV-Vis and FTIR analyses confirmed the presence of β-carotene as the primary carotenoid, demonstrating strong antioxidant potential. The study's findings highlight the optimal environmental parameters for enhancing carotenoid yield, which can be applied for industrial and pharmaceutical uses due to their antioxidant properties.

Exploring the lipids, carotenoids, and vitamins content of Rhodotorula glutinis with selenium supplementation under lipid accumulating and growth proliferation conditions

BackgroundRhodotorula glutinis, a specific type of yeast, has been recognised as a superior resource for generating selenium-enriched biomass that possesses exceptional nutritional and functional attributes. The purpose of this investigation was to assess the effect of sodium selenite at different concentrations on lipid and carotenoid synthesis, as well as the growth of R. glutinis.MethodsThe lipid’s fatty acid composition was determined using gas chromatography (GC). The vitamins were detected by high-performance liquid chromatography (HPLC). Transmission electron microscopy was used to detect the structural modification of yeast cells caused by the addition of sodium selenite to the growth medium, as well as the accumulation of elemental selenium in the yeast cells.ResultsThe yeast cells demonstrated the ability to endure high concentrations of sodium selenite under lipid accumulation (LAM) and growth-promoting (YPD) conditions. 25.0 mM and 30.0 mM, respectively, were published as the IC50 values for the LAM and YPD conditions. In both growth media, 1 mM sodium selenite boosted lipid synthesis. Lipid accumulation increased 26% in LAM to 11.4 g/l and 18% in YPD to 4.3 g/l. Adding 1 mM and 3 mM sodium selenite to YPD medium increased total and cellular carotenoids by 22.8% (646.7 µg/L and 32.12 µg/g) and 48.7% (783.3 µg/L and 36.43 µg/g), respectively. Palmitic acid was identified as the most abundant fatty acid in all treatments, followed by oleic acid and linoleic acid. The concentrations of water soluble vitamins (WSV) and fat soluble vitamins (FSV) were generally significantly increased after supplementation with 1.0 mM sodium selenite. TEM examination revealed a significant reduction in lipid bodies accumulation in the yeast cells when sodium selenite was added to lipid-promoting environments. This decline is accompanied by an augmentation in the formation of peroxisomes, indicating that selenium has a direct impact on the degradation of fatty acids. In addition, autophagy appears to be the primary mechanism by which selenium ions are detoxified. Additionally, intracellular organelles disintegrate, cytoplasmic vacuolization occurs, and the cell wall and plasma membrane rupture, resulting in the discharge of cytoplasmic contents, when a high concentration of sodium selenite (20.0 mM) is added. Also, the presence of numerous electron-dense granules suggests an intracellular selenium-detoxification pathway.ConclusionThis study proposes the use of YPD with 1 mM sodium selenite to cultivate selenium-enriched biomass from R. glutinis. This approach leads to heightened lipid levels with higher accumulation of oleic, linoleic and linolenic acids, carotenoids, and vitamins. Hence, this biomass has the potential to be a valuable additive for animal, fish, and poultry feed. Furthermore, explain certain potential factors that indicate the impact of selenium in reducing the accumulation of lipid droplets in R. glutinis during lipogenesis, as detected through TEM examination.

Increasing carotenoid production in Xanthophyllomyces dendrorhous/Phaffia rhodozyma: SREBP pathway activation and promoter engineering

The yeast Xanthophyllomyces dendrorhous synthesizes astaxanthin, a high-value carotenoid with biotechnological relevance in the nutraceutical and aquaculture industries. However, enhancing carotenoid production through strain engineering remains an ongoing challenge. Recent studies have demonstrated that carotenogenesis in X. dendrorhous is regulated by the SREBP pathway, which includes the transcription factor Sre1, particularly in the mevalonate pathway that also produces precursors used for ergosterol synthesis. In this study, we explored a novel approach to enhance carotenoid synthesis by replacing the native crtE promoter, which drives geranylgeranyl pyrophosphate synthesis (the step where carotenogenesis diverges from ergosterol biosynthesis), with the promoter of the HMGS gene, which encodes 3-hydroxy-3-methylglutaryl-CoA synthase from the mevalonate pathway. The impact of this substitution was evaluated in two mutant strains that already overproduce carotenoids due to the presence of an active Sre1 transcription factor: CBS.cyp61-, which does not produce ergosterol and strain CBS.SRE1N.FLAG, which constitutively expresses the active form of Sre1. Wild-type strain CBS6938 was used as a control. Our results showed that this modification increased the crtE transcript levels more than threefold and fourfold in CBS.cyp61−.pHMGS/crtE and CBS.SRE1N.FLAG.pHMGS/crtE, respectively, resulting in 1.43-fold and 1.22-fold increases in carotenoid production. In contrast, this modification did not produce significant changes in the wild-type strain, which lacks the active Sre1 transcription factor under the same culture conditions. This study highlights the potential of promoter substitution strategies involving genes regulated by Sre1 to enhance carotenoid production, specifically in strains where the SREBP pathway is activated, offering a promising avenue for strain improvement in industrial applications.

Defense Mechanisms of Xylopia aromatica (Lam.) Mart. in the Dry Season in the Brazilian Savanna

Water availability and light during the dry and rainy seasons in the Cerrado may influence plants’ stomatal movement and the entry of CO2 for organic synthesis, which is the main electron drain. A lower stomatal conductance may contribute to the energy accumulated in the chloroplasts being directed towards the synthesis of compounds, which contributes to the activity of antioxidant enzymes to neutralize reactive oxygen species. Xylopia aromatica is a characteristic Cerrado species, and it is often recommended for recovering degraded areas. This study aimed to investigate the influence of the dry and rainy seasons on the metabolic adjustments of Xylopia aromatica in a portion of the Brazilian savanna in the state of São Paulo. In the rainy season, better photosynthetic performance led to greater investment in essential oil production. In the dry season, the plants may direct part of their reducing sugars to the syntheses of carotenoids and anthocyanins, which may help the antioxidant enzymes to neutralize reactive oxygen species. Carotenoids assist in the dissipation of photosystem energy, which has the potential to cause oxidative stress. During this season, lower stomatal conductance prevented excessive water loss. These results suggest the acclimatization of this species to the conditions of the Brazilian savanna.

Proteomic analysis identified proteins that are differentially expressed in the flavonoid and carotenoid biosynthetic pathways of Camellia Nitidissima flowers

BackgroundCamellia nitidissima Chi is a popular ornamental plant because of its golden flowers, which contain flavonoids and carotenoids. To understand the regulatory mechanism of golden color formation, the metabolites of C. nitidissima petals at five different developmental stages were detected, a proteome map of petals was first constructed via tandem mass tag (TMT) analysis, and the accuracy of the sequencing data was validated via parallel reaction monitoring (PRM).ResultsNineteen color components were detected, and most of these components were carotenoids that gradually accumulated, while some metabolites were flavonoids that were gradually depleted. A total of 97,647 spectra were obtained, and 6,789 quantifiable proteins were identified. Then, 1,319 differentially expressed proteins (DEPs) were found, 55 of which belong to the flavonoid and carotenoid pathways, as revealed by pairwise comparisons of protein expression levels across the five developmental stages. Notably, most DEPs involved in the synthesis of flavonoids, such as phenylalanine ammonium lyase and 4-coumarate-CoA ligase, were downregulated during petal development, whereas DEPs involved in carotenoid synthesis, such as phytoene synthase, 1-deoxy-D-xylulose-5-phosphate synthase, and β-cyclase, tended to be upregulated. Furthermore, protein‒protein interaction (PPI) network analysis revealed that these 55 DEPs formed two distinct PPI networks closely tied to the flavonoid and carotenoid synthesis pathways. Phytoene synthase and chalcone synthase exhibited extensive interactions with numerous other proteins and displayed high connectivity within the PPI networks, suggesting their pivotal biological functions in flavonoid and carotenoid biosynthesis.ConclusionProteomic data on the flavonoid and carotenoid biosynthesis pathways were obtained, and the regulatory roles of the DEPs were analyzed, which provided a theoretical basis for further understanding the golden color formation mechanism of C. nitidissima.

Ultraviolet-diethyl sulfate composite mutagenesis of Rhodosporidium toruloides and culture optimization to improve carotenoid production

Rhodosporidium toruloides is a carotenoid-producing oleaginous yeast, and its carotenoids are of considerable value to food, chemical, and medical industries. In this study, to improve carotenoid production capacity, the parent R. toruloides strain (CK) was compound mutagenized using ultraviolet-diethyl sulfate (UV-DES), and cultivation conditions were simultaneously optimized to enhance the carotenoid synthesis ability of the mutant strain (M5). The carotenoid content was significantly increased following UV-DES compound mutagenesis (CK = 249.34 μg/g, M5 = 535.28 μg/g). The carotenoid content of M5 was increased to 763.85 μg/g under optimal culture conditions. In addition, mutagenesis altered the carotenoid components, and torulene and torularhodin levels were also increased following mutation, leading to changes in antioxidant capacity. Real-time quantitative PCR results showed that expression of six genes (CAR1, CAR2, CrtE, CrtI, ME1, HMG1) related to carotenoid accumulation were significantly upregulated by UV-DES composite mutagenesis. In conclusion, combined UV-DES mutagenesis and optimal culture conditions promoted the biosynthesis of carotenoids in R. toruloides.

Genome-Wide Association Study of Sweet Potato Storage Root Traits Using GWASpoly, a Gene Dosage-Sensitive Model.

Sweet potato (Ipomoea batatas) is an important food crop that plays a pivotal role in preserving worldwide food security. Due to its polyploid genome, high heterogeneity, and phenotypic plasticity, sweet potato genetic characterization and breeding is challenging. Genome-wide association studies (GWASs) can provide important resources for breeders to improve breeding efficiency and effectiveness. GWASpoly was used to identify 28 single nucleotide polymorphisms (SNPs), comprising 21 unique genetic loci, associated with sweet potato storage root traits including dry matter (4 loci), subjective flesh color (5 loci), flesh hue angle (3 loci), and subjective skin color and skin hue angle (9 loci), in 384 accessions from the USDA sweet potato germplasm collection. The I. batatas 'Beauregard' and I. trifida reference genomes were utilized to identify candidate genes located within 100 kb from the SNPs that may affect the storage traits of dry matter, flesh color, and skin color. These candidate genes include transcription factors (especially Myb, bHLH, and WRKY family members), metabolite transporters, and metabolic enzymes and associated proteins involved in starch, carotenoid, and anthocyanin synthesis. A greater understanding of the genetic loci underlying sweet potato storage root traits will enable marker-assisted breeding of new varieties with desired traits. This study not only reinforces previous research findings on genes associated with dry matter and β-carotene content but also introduces novel genetic loci linked to these traits as well as other root characteristics.

Gene editing technology combined with response surface optimization to improve the synthesis ability of lycopene in Pantoea dispersa MSC14.

The aim of this study is to engineer Pantoea dispersa MSC14 into a strain capable of producing lycopene and to enhance its lycopene content. Our laboratory isolated the strain P. dispersa MSC14 from petroleum-contaminated soil in a mining area. Whole-genome sequencing confirmed the existence of a carotenoid synthesis pathway in this strain. This study employed an optimized CRISPR/Cas9 system to perform a traceless gene knockout of the lycopene cyclase gene crtY and to overexpress the octahydrolycopene dehydrogenase gene crtI in the P. dispersa MSC14. This strategic genetic modification successfully constructed the lycopene-producing strain MSC14-LY, which exhibited a notable lycopene content with a biomass productivity of 553μg of lycopene per gram dry cell weight(DCW). Additionally, the components of the lycopene fermentation medium were optimized using Plackett-Burman (PB) design and Response Surface Methodology (RSM). The average lycopene content was increased to 5.13mg g -1DCW in the optimized LY fermentation medium. Through genetic engineering, P. dispersa MSC14 was transformed into a strain capable of producing lycopene, achieving a yield of 5.13mg g-1 DCW after medium optimization. Genetic engineering successfully transformed P. dispersa MSC14 into a strain capable of producing lycopene, achieving a yield of 5.13mg g-1 DCW after medium optimization.

Impact of emerging pollutants mixtures on marine and brackish phytoplankton: diatom Phaeodactylum tricornutum and cyanobacterium Microcystis aeruginosa

Pharmaceuticals and ionic liquids (ILs) are emerging as significant micropollutants with environmental presence and potential ecological impacts. The possible simultaneous occurrence of these two groups of pollutants in aquatic environments raises complex challenges due to their diverse chemical properties and potential for interactive effects. Given the documented widespread presence of pharmaceuticals and the emerging concerns about ILs, the study aims to evaluate the adverse effects of binary mixtures of imidazolium ionic liquid IM1-8C(CN)3 and two representatives of pharmaceuticals: antibiotic oxytetracycline (OXTC) and metabolite carbamazepine 10,11 epoxide (CBZ-E) on the brackish cyanobacterium Microcystis aeruginosa and the marine diatom Phaeodactylum tricornutum during chronic exposure experiments. A comprehensive approach was employed, incorporating various endpoints including oxidative stress, chlorophyll a fluorescence, detailed photoprotective and photosynthetic pigment profiles of target microorganisms to assess modes of action and identify the mixture effects of the selected substances.The observed alterations in pigment production affecting carotenoids synthesis in both selected species may be attributed to the differential impacts of these substances on the photosynthetic pathways and metabolic processes in the cyanobacterial and diatom cells. Changes in chlorophyll a fluorescence-specific parameters suggest impairment of the photosynthetic activity, particularly affecting the efficiency of photosystem II.The application of Concentration Addition (CA) and Independent Action (IA) mathematical models, complemented by the evaluation of Model Deviation Ratios (MDR), revealed predominantly antagonistic interactions within the studied mixtures. The findings of this study provide important insights into the effects of mixtures of organic micropollutants and their potential impact on environment including brackish and marine waters.

Mining and expression analysis of color related genes in Bougainvillea glabra bracts based on transcriptome sequencing.

Bract coloration is one of the key ornamental traits in Bougainvillea, yet research has predominantly focused on phenotypic color traits and pigment composition, with limited understanding of the molecular mechanisms underlying color formation. This gap hinders the improvement and innovation in bract coloration. To elucidate the regulatory mechanisms of bract coloration in Bougainvillea and to enhance the utilization of its germplasm resources, this study employed the Illumina Novaseq 6000 sequencing platform to conduct transcriptomic sequencing on 21 samples of bracts exhibiting seven distinct phenotypes. Comparative analysis against Nr, Pfam, EggNOG, GO, and KEGG databases annotated 90,279 unigenes. The highest annotation rates were achieved with the Nr (40.13%), GO (30.44%), and EggNOG (25.64%) databases. Among the species annotated, Beta vulgaris (20.08%) and Chenopodium quinoa (14.58%) shared the highest homology with Bougainvillea bract transcriptomes. WGCNA analysis identified 12 positively correlated tissue-specific modules, of which 2 are related to bract color formation. By comparing transcriptome data and genes within these specific modules against the KEGG database, a total of 321 unigenes associated with bract color formation in Bougainvillea were discovered. Among these, 220 unigenes are involved in anthocyanin synthesis, 43 unigenes are involved in betalain synthesis, 23 unigenes are annotated as Chlorophyll a-b binding protein genes, and 35 unigenes participate in carotenoid synthesis. Quantitative real-time PCR (qRT-PCR) validation of 16 differentially expressed genes (DEGs) including PAL2, CHS1, ANS, BZ1, 6GT, CDOPA5GT, ANR, CHS2, and DOPA, revealed significant expression differences among magenta, yellow, white, and cherry-colored bracts, suggesting their potential as candidate genes for bract color development. This study not only enriches the transcriptomic data of Bougainvillea but also identifies genes associated with bract coloration, providing a valuable theoretical basis for future gene cloning, genetic engineering, and breeding efforts in Bougainvillea.

A low frequency magnetic field regulated the synthesis of carotenoids in Rhodotorula mucilaginosa by influencing iron metabolism

The purpose of this study is to investigate the effect of low frequency magnetic fields (LFMF) on carotenoid synthesis and iron metabolism in Rhodotorula mucilaginosa and to explore potential connections between the two processes. Various LFMF intensities were employed to assess carotenoid production, intracellular iron content, and the expression levels of genes related to carotenoid synthesis and iron metabolism in R. mucilaginosa. The results show that a LFMF could promote carotenoid synthesis, resulting in varying increases in intracellular iron content. Specifically, the expression of the iron metabolism-related gene transferrin receptor (TFR) was upregulated, while the expression of ferroportin 1 (FPN1) was downregulated. Meanwhile, the expressions of the carotenoid synthesis-related genes geranylgeranyl diphosphate synthases (GGPPS) and phytoene synthase (PSY) displayed varying degrees of upregulation at different time points. Further, the TFR and FPN1 gene knockout strains of R. mucilaginosa were subjected to treatment with a 3.5-mT LFMF, the results of which showed that an LFMF is capable of influencing the iron metabolism levels and subsequently regulating carotenoid synthesis in R. mucilaginosa.

The role of SlCHRC in carotenoid biosynthesis and plastid development in tomato fruit

Chromoplasts are specialized plastids in plants involved in carotenoid synthesis, accumulation, and stress resistance. In tomatoes (Solanum lycopersicum), the Chromoplast-associated carotenoid binding protein (CHRC) regulates chromoplast development and carotenoid accumulation, although its precise mechanisms are not yet fully understood. To investigate the role of SlCHRC in carotenoid biosynthesis, we generated transgenic tomatoes using overexpression (oe-SlCHRC) and CRISPR/Cas9-mediated gene editing (cr-SlCHRC) techniques. The results demonstrated inhibited fruit ripening and delayed onset of color break in both transgenic lines. The oe-SlCHRC lines exhibited increased carotenoid accumulation, particularly (E/Z)-phytoene, lycopene, and γ-carotene, with abundant plastoglobules and carotenoid crystals observed via TEM. In contrast, cr-SlCHRC mutants showed a greener phenotype, reduced carotenoid content, and fewer plastoglobules at the BK + 10 stage. Transcriptome analysis indicated that SlCHRC influences key genes in carotenoid biosynthesis, such as SlNCED2, as well as genes related to chloroplast development, photosynthesis, and plastoglobule formation. Additionally, SlCHRC enhances heat stress tolerance in tomato fruits by upregulating heat shock proteins (HSPs), antioxidants, and proline accumulation. These findings indicate that SlCHRC plays a crucial role in improving tomato fruit quality under heat stress conditions.

Enhancing carotenoid accumulation in Dunaliella bardawil by combined treatments with fulvic acid and optimized culture conditions

Natural carotenoids from microalgae have received more attention as an alternative source. In this study, fulvic acid (FA), a plant growth regulator, was used to enhance carotenoid accumulation in microalgae Dunaliella bardawil rich in lutein. However, the addition of FA promoted pigment synthesis but also exhibited an inhibitory effect on biomass. Therefore, the optimization of culture conditions was performed to further enhance carotenoid accumulation, including high light stress (10,000 lx) and the two-stage cultivation comprising 1-aminocyclopropane-1-carboxylic acid (ACC) and FA. Under both culture conditions, the growth inhibition caused by FA was alleviated, leading to a further increase in the contents of chlorophylls and carotenoids. HPLC analysis revealed that the production of lutein, α-carotene and β-carotene increased by 0.44-, 0.37- and 0.54-fold under the treatment of 400 mg/L FA with high light intensity and 0.91-, 1.15- 0.29-fold under the two-stage cultivation comprising 11 mM ACC and 500 mg/L FA. Furthermore, algal cells under FA treatment and the two-stage cultivation stained with Bodipy505/515 emitted stronger fluorescence under a laser confocal microscope, suggesting that lipid accumulation was increased. Additionally, the transcription levels of carotenogenic genes were also found to be up-regulated by qRT-PCR. These results indicated an enhancement in both the storage capacity and synthesis of carotenoids in D. bardawil. This study revealed the potential application of plant growth regulators in promoting carotenoid accumulation in D. bardawil which could be further improved by optimizing the culture conditions, providing a reference for efficient carotenoid production in microalgae.

Transcriptome Analysis Revealed the Anabolic Regulation of Chlorophyll and Carotenoids in Curcuma alismatifolia Bracts.

Curcuma alismatifolia is an attractive ornamental plant in the ginger family. Its bracts come in a variety of colors and are commonly used as cut flowers, potted plants, and landscaping. To investigate the regulation of bract pigmentation in C. alismatifolia, we examined the pigment levels of chlorophyll and carotenoids in the pure color part (PC) and variegated part (VA) of three C. alismatifolia varieties, i.e., "Siam TM Sitrone," "Chiang Mai Pink," and "Snow White." To mine the color mechanisms of the pure color and variegated parts of the bract, we conducted RNA-seq analysis on C. alismatifolia. We identified a total of 89,975 unigenes, and there were 3584 differentially expressed genes identified post-screening. Furthermore, 1858 DEGs were annotated in the GO database and 681 in the KEGG database. We pinpointed key genes responsible for the diverse bract colors in C. alismatifolia, including ZEP for carotenoid synthesis and GAGA2 in the chlorophyll synthesis pathway. This study provides valuable insights into understanding the pigmentation mechanism of bracts in C. alismatifolia and the breeding process.

Extraction and Synthesis of Typical Carotenoids: Lycopene, β-Carotene, and Astaxanthin.

Carotenoids are tetraterpene compounds acting as precursors to vitamin A, with functions that include protecting eyesight, enhancing immunity, promoting cell growth and differentiation, and providing antioxidative benefits. Lycopene, β-carotene, and astaxanthin are particularly critical for health and have diverse applications in food, health products, and medicine. However, natural carotenoids are encased within cell structures, necessitating mechanical methods to disrupt the cell wall for their extraction and purification-a process often influenced by environmental conditions. Thus, improving the efficiency of carotenoid extraction from natural resources is of great interest. This review delves into the research progress made on the extraction processes, structures, and biological functions of carotenoids, focusing on lycopene, β-carotene, and astaxanthin. Traditional extraction methods primarily involve organic solvent-assisted mechanical crushing. With deeper research and technological advancements, more environmentally friendly solvents, advanced machinery, and suitable methods are being employed to enhance the extraction and purification of carotenoids. These improvements have significantly increased extraction efficiency, reduced preparation time, and lowered production costs, laying the groundwork for new carotenoid product developments.

The transcription factor RppA regulates chlorophyll and carotenoid biosynthesis to improve photoprotection in cyanobacteria.

Chlorophyll is an essential photosynthetic pigment but also a strong photosensitizer. Excessive free chlorophyll and its precursors can cause oxidative damage to photosynthetic organisms. Cyanobacteria are the oldest oxygenic photosynthetic organisms and the ancestors of the chloroplast. Owing to their complex habitats, cyanobacteria require precise regulation of chlorophyll synthesis to respond to environmental factors, especially changes in light. Chlorophyll synthase, encoded by chlG, is the enzyme catalyzing the final step of chlorophyll biosynthesis, which is closely related to photosynthesis biogenesis. However, the transcriptional regulation on chlG remains unclear. Here, the transcription factor, regulator of photosynthesis and photopigment-related gene expression A (RppA) was identified to bind to the chlG promoter by screening a yeast one-hybrid library in the cyanobacterium Synechocystis sp. PCC 6803. The rppA knock-out mutant showed a phenotype of slow growth and severe oxidative damage under dark-light transition conditions. The up-regulated transcriptional expression of chlG was significantly higher and more chlorophyll and its precursors accumulated in the rppA knock-out mutant than those in the wild-type strain during the transition from darkness to light, indicating RppA represses the expression of chlG in Synechocystis. Meanwhile, RppA could synchronously promote the transcription of carotenoids biosynthesis-related genes to enhance carotenoids synthesis during the dark-light transition. These results reveal synergistic regulation of chlorophyll and carotenoids biosynthesis in cyanobacteria in response to frequent dark-light transitions, which slows down chlorophyll biosynthesis while promoting carotenoids biosynthesis to avoid oxidative damage caused by excessive reactive oxygen species accumulation.

The Plasma Membrane H+-ATPase Promoter Driving the Expression of FADX Enables Highly Efficient Production of Punicic Acid in Rhodotorula toruloides Cultivated on Glucose and Crude Glycerol.

Punicic acid (PuA) is a conjugated fatty acid with a wide range of nutraceutical properties naturally present in pomegranate seed oil. To meet the rising demand for pomegranate seed oil, a single-cell oil enriched in PuA provides a sustainable biomass-derived alternative. This study describes the production of a PuA-enriched single-cell oil through the engineering of the red yeast Rhodotorula toruloides grown in glucose and a low-cost substrate, crude glycerol. The gene for Punica granatum fatty acid conjugase, PgFADX, was randomly integrated into the genome of R. toruloides without disrupting the carotenoid synthesis. In shake flask studies, the effects of three promoters (PPGI1, PNAR1, and PPMA1) on PuA production were evaluated. PuA titers of 105.77 mg/L and 72.81 mg/L were obtained from engineered cells expressing PgFADX from the PPMA1 promoter cultivated for 72 h in glucose and for 168 h in crude glycerol, respectively. Furthermore, the detailed lipid analysis revealed a high enrichment PuA in the triacylglycerol lipid structures, even without substantial modifications to the metabolic pathways. This report demonstrates the high potential of R. toruloides in the upcycling of a low-cost substrate, crude glycerol, into a value-added product such as PuA. The findings support the feasibility of using engineered R. toruloides for sustainable production of PuA-enriched single-cell oil.

Unraveling the Genetic Control of Pigment Accumulation in Physalis Fruits.

Physalis pubescens and Physalis alkekengi, members of the Physalis genus, are valued for their delicious and medicinal fruits as well as their different ripened fruit colors-golden for P. pubescens and scarlet for P. alkekengi. This study aimed to elucidate the pigment composition and genetic mechanisms during fruit maturation in these species. Fruit samples were collected at four development stages, analyzed using spectrophotometry and high-performance liquid chromatography (HPLC), and complemented with transcriptome sequencing to assess gene expression related to pigment biosynthesis. β-carotene was identified as the dominant pigment in P. pubescens, contrasting with P. alkekengi, which contained both lycopene and β-carotene. The carotenoid biosynthesis pathway was central to fruit pigmentation in both species. Key genes pf02G043370 and pf06G178980 in P. pubescens, and TRINITY_DN20150_c1_g3, TRINITY_DN10183_c0_g1, and TRINITY_DN23805_c0_g3 in P. alkekengi were associated with carotenoid production. Notably, the MYB-related and bHLH transcription factors (TFs) regulated zeta-carotene isomerase and β-hydroxylase activities in P. pubescens with the MYB-related TF showing dual regulatory roles. In P. alkekengi, six TF families-bHLH, HSF, WRKY, M-type MADS, AP2, and NAC-were implicated in controlling carotenoid synthesis enzymes. Our findings highlight the intricate regulatory network governing pigmentation and provide insights into Physalis germplasm's genetic improvement and conservation.

Benzophenone-4 inhibition in marine diatoms: Physiological and molecular perspectives

Benzophenone-4 (BP-4), a widely utilized organic ultraviolet (UV) filter, is recognized as a pseudo-persistent contaminant in aquatic environments. To elucidate the effects and mechanisms of BP-4 on marine diatoms, an investigation was conducted on the growth rate, photosynthetic pigment content, photosynthetic parameters, antioxidant enzyme activity, malondialdehyde (MDA) levels, cellular structure, and transcriptome profile of the model species, Phaeodactylum tricornutum. The results showed a pronounced inhibition of algal growth upon exposure to BP-4, with a 144 h-EC50 value of 201 mg·L−1. In addition, BP-4 exposure resulted in a significant reduction in biomass, disruption of cell membrane integrity, and increased MDA accumulation, with levels escalating 3.57-fold at 125 mg·L−1 of BP-4. In the BP-4-treated samples, 1556 differentially expressed genes (DEGs) were identified, of which 985 were upregulated and 571 were downregulated. Gene ontology and KEGG pathway enrichment analysis revealed that the carbon fixation and carbon metabolism processes in P. tricornatum were disrupted in response to BP-4 exposure, along with excessive reactive oxygen species (ROS) production. The upregulation of genes associated with photosynthetic pigment (chlorophyll and carotenoids) synthesis, phospholipid synthesis, ribosome biogenesis, and translation-related pathways may be regarded as a component of P. tricornatum's tolerance mechanism towards BP-4. These results provide preliminary insights into the toxicity and tolerance mechanisms of BP-4 on P. tricornatum. They will contribute to a better understanding of the ecotoxicological impacts of BP-4 on the marine ecosystem and provide valuable information for elimination of BP-4 in aquatic environment by bioremediation.

Optimizing tomato waste hydrolysate for enhanced fucoxanthin biosynthesis in mixotrophic cultivation of Isochrysis galbana

Vegetable waste, rich in bioactive compounds, offers a promising resource for producing value-added products. This study explored the use of tomato waste, containing glucose (40 mg/g), lycopene (95.12 μg/g), and β-carotene (24.31 μg/g), for cultivating fucoxanthin-rich Isochrysis galbana. Water-soluble lycopene (2.0 μg/mL) and β-carotene (0.4 μg/mL) effectively upregulated key carotenoid synthesis genes and boosted cell growth and fucoxanthin production (3.64 and 3.60 pg/cell, respectively) within 10 days in a mixotrophic culture. Optimized tomato waste hydrolysate achieved a high cell density of 1.21 × 107 cells/mL, 2.13 g/L biomass, and 21.02 mg/g fucoxanthin. This study highlights the potential of combining tomato waste with microalgae for a novel and innovative approach towards waste management and resource utilization.