Comparative metabolic and transcriptomic analysis of macrozoids and palmelloid cells of Haematococcus pluvialis related to astaxanthin accumulation
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ABSTRACT: In this work, we propose a metabolomic and transcriptomic comparative study between a vegetative state and intermediate reddish palmelloid state growing in continuous culture conditions, giving rise to a profound understanding of the first steps of the adaptation of Haematococcus pluvialis to nitrogen limitation and the correlation with astaxanthin biosynthesis. In addition, other related metabolic pathways as fatty acid biosynthesis, starch accumulation, citric acid cycle, and amino acid biosynthesis have been analyzed. Understanding the molecular basis of astaxanthin accumulation will be useful for the optimization of astaxanthin production.
Project description:Haematococcus pluvialis transcriptome analysis The genetic transcriptome details of astaxanthin biosynthesis were analysed by exposing the algal cells to 25 mg/L of SA and JA for 1, 6 and 24 hours, plus to the control (no stress). SRP062153 PRJNA292140
Project description:Astaxanthin alleviates hepatic lipid accumulation and peroxidation, inflammation, and fibrosis in mice with high-cholesterol, high-cholate, and high-fat (CL) diet-induced nonalcoholic steatohepatitis (NASH). It has been proposed as a potential new treatment to inhibit the progression of NASH in humans. Therefore, we compared hepatic gene expression profiles after treatment with astaxanthin or the antioxidant vitamin E in mice with CL diet-induced NASH. Comprehensive gene expression analyses of the livers of mice fed a standard, CL, or CL diet containing astaxanthin or vitamin E for 12 weeks were performed using a DNA microarray. Both astaxanthin and vitamin E effectively improved gene expression associated with eukaryotic initiation factor-2 (EIF2) signaling, which is suppressed in NASH by endoplasmic reticulum (ER) stress in the liver. Astaxanthin but not vitamin E was predicted to suppress the actions of ligand-dependent nuclear receptors peroxisome proliferator-activated receptors (PPAR) α and PPARδ and to affect related molecules. Establishing a new therapy using astaxanthin will require elucidation of astaxanthin’s molecular action on the functions of PPARα and related molecules in the livers of mice with diet-induced NASH.
Project description:The differential expression of the high-yield astaxanthin Phaffia rhodozyma BPAX-A1 with the wild-type Phaffia rhodozyma CBS6938 was obtained and analysed to understand metabolic changes leading to an improved astaxanthin production.
Project description:Phaffia rhodozyma is an ideal microbial astaxanthin resource. However, the low productivity of astaxanthin in this yeast obstructs the process of industrial production. Although P. rhodozyma is isolated from plant material and the phytohormone has proved to be an effective stimulator for microbial production, the effects and mechanisms of phytohormones on astaxanthin synthesis in P. rhodozyma have been rarely reported. In this study, a concentration of down to 0.5 mg/L exogenous salicylic acid (SA) could promote biomass, astaxanthin content and yield by 20.8%, 95.8% and 135.3% in P. rhodozyma, respectively. Further transcriptomic analysis showed that SA could discriminate well the transcriptomic profile of P. rhodozyma cells. The differently expressed genes (DEGs) in P. rhodozyma cells between the SA-treatment and the SA-free groups were identified, and involved in astaxanthin and its competitive metabolite synthesis, material supply, biomolecules metabolism and transportation, anti-stress and signal transductions at a global-cell level. A regulation mechanism of astaxanthin synthesis induced by SA, including perception and transduction of the SA signal, expression regulations of the downstream genes by transcription factors, and cellular stress response to the SA, is put forward in this study. The polyamine transporter gene (PT), as an up-regulated DEG, was overexpressed in the P. rhodozyma to obtain the transformant Prh-PT-006. It was found that biomass, astaxanthin content and yield of the Prh-PT-006 under 0.5 mg/L SA treatment could reach 6.6 g/L, 0.35 mg/g DCW and 2.3 mg/L, 24.5%, 143.1% and 199.0% higher than the wild strain at the SA-free condition, respectively. The result will provide the engineering targets for constructing strains with high-yield of astaxanthin, and help to accelerate the industrialization process of microbial astaxanthin.
Project description:The hyperglycemic state in diabetes mellitus (DM) induces oxidative stress and inflammation, thus contributing to diabetic tissue damage and associated complications. Astaxanthin, a potent antioxidant carotenoid, has been investigated for its potential in preventing and managing diabetes across various species, but its effect on client-owned dogs is not well-studied. This study explored the impact of astaxanthin supplementation on canine DM using a proteomic approach. A total of 18 client-owned dogs were enrolled: 6 dogs with DM and 12 clinically healthy dogs. The diabetic dogs received their standard treatment regimen alongside daily oral supplementation of 12 mg of astaxanthin (1.5–2.4 mg/kg) for 90 days. Plasma samples were collected at the beginning and end of the study period for proteomic analysis. After astaxanthin supplementation, there were significant alterations in protein expression associated with the complement system, coagulation cascade, JAK-STAT signaling, and protein kinase C signaling, which all contribute to inflammation and oxidative stress. Astaxanthin demonstrated a protective effect against diabetes-associated complications, including insulin resistance, vascular dysfunction, nephropathy, and cardiac complications. These findings highlight the potential of astaxanthin as a complementary therapeutic agent for controlling DM in canines.
Project description:Doxorubicin (DOX) and other anthracyclines are effective chemotherapeutic agents, however, their use is influenced by the risk of cardiotoxicity. We still have an incomplete understanding of the cardiomyocyte protective pathways activated after anthracycline-induced cardiotoxicity (AIC).Danshen injection (DSI), astaxanthin (AXT) and diosmetin (DMT) are effective in the treatment of cardiovascular diseases, but the mechanism of protection against adriamycin-induced cardiotoxicity is unclear. Here, we performed RNA-seq screening in H9c2 cardiomyocytes to determine the potential protective mechanisms of Danshen injection, astaxanthin and diosmetin against AIC.
Project description:Astaxanthin is a dark red keto-carotenoid found in aquatic animals such as salmon and shrimp, and algae (Haematococcus pluvialis). Astaxanthin has a unique molecular structure that may facilitate anti-oxidative, immunomodulatory, and anti-inflammatory effects during physiological stress. The primary objective of this study was to examine the efficacy of 4-weeks ingestion of astaxanthin in moderating exercise-induced inflammation and immune dysfunction using a multi-omics approach.
Project description:A koji mold Aspergillus kawachii is used for making a Japanese distilled spirit, shochu. During making shochu, A. kawachii is grown in a solid-state culture comprised of steamed grains such as rice or barley, termed koji, to convert the starch to glucose and also to produce citric acid. During this process, cultivation temperature of A. kawachii is generally controlled by raising to 40M-BM-0C and then lowering to 30M-BM-0C. The cultivation at 40M-BM-0C and the following at lower temperature of 30M-BM-0C has important roles in the elevated activity of amylase and starting an accumulation of a large amount of citric acid, respectively. In this study, we investigated the effect of temperature on the gene expression of A. kawachii when barley was used as ingredient for making koji. The results of DNA microarray and gene ontology analysis showed that the expression of genes involved in the metabolic processes of glycerol, trehalose, and pentose phosphate that branch from glycolysis was down-regulated by shifting the cultivation temperature from 40M-BM-0C from 30M-BM-0C. In addition, reduction in the expression of the genes related with heat shock responses and increasing in the expression of the genes related with amino acid transport after the temperature lowering were found to be significant change. These results suggested that the cultivation at 40M-BM-0C is stressful event for the A. kawachii and the heat adaption lead to the depression of citric acid accumulation through activation of the branching pathways from glycolysis. The gene expression profile obtained in this study will help to understand the gene regulation during koji-making process to optimize A. kawachii as industrial microorganism. Gene expression of Aspergillus kawachii in barley koji was measured at 25, 26.5, and 44 hours at two different temperature control. Three independent experiments were performed at each time (25, 26.5, or 44 hours) using different barley koji for each expreriment.
Project description:Purpose of study was to investigate whole genome expression changes of a strain with deletion of the two-component system TctD-TctE and determine genes dysregulate relative to the parental wildtype to gain insight into possible regulatory targets of TctD-TctE. TctD-TctE is a two-component system in Pseudomonas aeruginosa that responds to and regulates uptake of tricarboxylic acids such as citric acid. It accomplishes this through derepression of the porin encoding the gene opdH, thereby regulating influx of citrate metabolites from the surrounding environment. Deletion of the tctED operon (ΔtctED) resulted in a reduced growth phenotype when citric acid is present in media. In the ΔtctED strain the presence of citric acid was found to have an inhibitory effect on growth. When the alternative carbon source arginine was present, wildtype levels of growth could not be restored. Static cultures of ΔtctED had low cell density in the presence of citric acid but maintained the same levels of biofilm formation compared to conditions when no citric acid was present. This suggests a dysregulation of biofilm formation in the presence of citric acid. In the ΔtctED strain there was also greater accumulation of tobramycin within the biofilm compared to the PA14 wildtype strain. Additionally, analysis of whole-genome expression found that multiple metabolic genes were dysregulated in ΔtctED. Here it is concluded that TctD-TctE is involved in biofilm tolerance to tobramycin in the presence of citrate metabolites.
Project description:Sirtuin is considered to play a significant role in the growth phase-dependent gene expression. In this study, we characterized sirtuin in the white koji fungus, Aspergillus kawachii, to examine their role on regulation of amylolytic enzymes and citric acid productions during the solid-state culture (koji). Characterization of rice-koji made using five putative sirtuin gene disruptants indicated that they are involved in amylolytic activity and acidity of rice koji; especially the sirD disruptant showed lower levels of α-amylase activity and citric acid production per mycelial weight in the koji compared the control strain. In addition, the sirD disruptant also showed a change in mycelial pigmentation, higher sensitivity to cell wall biogenesis inhibitor such as calcofluor white and Congo red, and reduced conidia formation, indicating that SirD is required for secondary metabolism, cell wall integrity, and conidial development. The cap analysis gene expression (CAGE) indicated that the transcriptional changes related to the characteristic phenotype of the sirD disruptant (e.g., a reduced transcript level of acid-stable α-amylase gene and a citric acid exporter) in rice koji. These results indicated that SirD has a significant role on the global transcriptional regulation including productions of α-amylase and citric acid in A. kawachii during the solid-state fermentation process.