β-carotene production from soap stock by loofa-immobilized Rhodotorula rubra in an airlift photobioreactor

In this study, the soap stock as a sole carbon source was used for growing a carotenoid producing yeast (Rhodotorula rubra). The application of soap stock resulted in increase of carotenoids yield up to 5.36 folds when compared with the grown cultures on glucose. On the best Monod equation fitted on the specific growth rate (μ) data, the maximum specific growth rate (μ_m) and half-saturation concentration (K_S) were respectively determined at 0.064 h⁻¹ and 3.26 g L⁻¹ for total fatty acids presented in soap stock. Further tests on the carotenogenesis process were carried out in a cell-immobilized airlift photobioreactor where the natural loofa sponge was used for immobilization of the cells. The performance of the bioreactor was statistically studied by the response surface methodology (RSM) where aeration rate of 0.11 vvm and light irradiation intensity of 2517 Lx provided an optimum condition for producing β-carotene with a specific production rate of 22.65 mg g_cell⁻¹ day⁻¹.     

Glycerol as substrate for biomass and β-carotene production byRhodotorula lactosa

Glycerol was used as the sole carbon and energy source for growingRhodotorula lactosa. The maximum biomass yield (0.53 g/g substrate) was obtained after 20 h with 21.5 g glycerol/l; growth was inhibited with 28.0 g glycerol/l and cell morphology was changed. At this time, the cells were not pigmented. After 48 h of cultivation, -carotene was at 1.8 mg/g dry cells, yielding 22.0 mg/l. When cells were grown for 20 h, washed, suspended in distilled water and aerated for 24 hours, more -carotene (2.66 mg/g dry cells or 28.0 mg/l of the original culture) was produced. Cell protein content after 48 h was 36 to 38% (w/w) before extraction and 45 to 47% (w/w) for acetone-extracted cells.     

Combinatorial expression of different β-carotene hydroxylases and ketolases in Escherichia coli for increased astaxanthin production

Journal of Industrial Microbiology & Biotechnology - In natural produced bacteria, β-carotene hydroxylase (CrtZ) and β-carotene ketolase (CrtW) convert β-carotene into...     

Exploitation of Dunaliella for β-carotene production

Halotolerant microalga Dunaliella, which is exploited for the production of dried biomass or cell extract, is used as a medicinal food. With the advancement in this field in recent years, the production of bio-organic compounds such as β-carotene is established in many countries. Large-scale production of β-carotene is controlled by numerous stress factors like high light intensity, high salinity, temperature and availability of nutrients. The state-of-the-art strategies in industries in closed systems under new set of inductive factors will additionally promote the ease of commercial production of β-carotene. This review mainly focuses on the different methodologies employed recently for the optimum production of β-carotene from Dunaliella species.     

Improved production of lycopene and β-carotene by Blakeslea trispora with oxygen-vectors

Lycopene and β-carotene production were increased when oxygen-vectors, n-hexane and n-dodecane, were added to cultures of Blakeslea trispora because of the enhanced dissolved oxygen concentrations. With 1% (v/v) n-hexane or n-dodecane added in the medium, lycopene production was 51% or 78% higher and β-carotene production was 44% or 65% higher than that of the control, respectively. The highest lycopene and β-carotene production, 533 mg l⁻¹and 596 mg l⁻¹, were obtained when 1% (v/v) n-dodecane and 0.1% (w/v) Span 20 were added together, which were 2.1-fold and 1.8-fold of the control, respectively.     

Astaxanthin production in transgenicArabidopsis with chyB gene encoding β;-carotene Hydroxylase

Oxycarotenoids, produced through the oxidation of carotenoids, play critical roles in plants. This reaction is mediated by a specific enzyme, β;-carotene hydroxylase, which adds hydroxyl groups to the β;-rings of carotenes. To investigate the effect of the β;-carotene hydroxylase gene (Chyb) on oxycarotenoid biosynthesis, we generated transgenicArabidopsis plants that over-expressedChyb under the control of a 35S promoter. Their levels of zeaxanthin and neoxanthin were two- to three-fold greater relative to the WT, while that of violaxanthin, a final product in the xanlthophyll pathway, was 1.3-fold higher than the control. In contrast, the amount of β;-carotene declined as much as 2.4-fold, depending on the particular transgenic line. Interestingly, astaxanthin was produced in the transgenics, but not in the WT. These data suggest that, with the aid of unknown factors in the host, carotenoids could be converted into metabolites in the astaxanthin biosynthetic pathway. Microarray analysis was used lo identify several genes that were consistently up-or down-regulated in transgenic chyB leaves compared with the controls. Here, we also discuss possible modifications in leaf carotenoids, and the importance of these data from a nutritional standpoint. These authors contributed equally to this work.     

β-carotene ameliorates CUS-induced circadian alternations of locomotor activity and melatonin patterns in rats

Circadian and stress systems have a crucial role in adaptation of organisms to environmental challenges. This study investigates the ability of Oscillatoria brevis (O. brevis) β-carotene extract (βC) in modulating the circadian alternations of locomotor activity (LA) and serum melatonin (M) rhythms under chronic unpredictable stress (CUS) in rats. Twenty rats (5 rats/group) were used in monitoring LA using running wheels. Eighty rats (20 rats/group) were used in observing circadian serum M profile. Rats were randomly divided into four groups, viz. control, βC-treated, CUS-exposed, and “βC-treated&CUS-exposed” groups. CUS-exposure was applied for 21 days. One hour before exposure, βC was daily administered (10 mg/kg), intraperitoneally (IP). Blood was sampled at 6-h intervals for 5 rats/time point at Zeitgeber (ZT) 3, 9, 15, and 21. Results demonstrated that unstressed rats exhibited circadian M pattern and nocturnal LA rhythm with acrophase around ZT 21 and ZT 15, respectively. CUS-exposure revealed a disturbance in these patterns. Phase shifting of M and LA profiles was recorded. A decrease M acrophase and a significant decrease in LA (p < 0.05) were recorded at ZT 9. Daily βC administration in stressed rats modulates the CRs alternation induced by CUS. It may be concluded that βC ameliorated the induced alternations in circadian rhythms.     

Strain selection for β-carotene production by Dunaliella

Four strains of Dunaliella were grown at 25°C and pH 8±0.5, with continous illumination at 200 W/m². Their maximum specific growth rates ranged from 0.093 day^-1 to 0.234 day^-1, nitrate yields from 3.0 to 7.8 g cells/g NaNO₃ and lipid contents from 3% to 6% of the dry wt, with carotenes 50 to 80% of the lipids. Of the carotenes, -carotene made up 7 to 19%; all-trans--carotene 32 to 52% and 9-cis--carotene 29 to 55%. There are, therefore, considerable intra-specific differences between strains of Dunaliella.     

TGFβ2-mediated production of hyaluronan is important for the induction of epicardial cell differentiation and invasion

In the developing heart, the epicardium is a major source of progenitor cells that contribute to the formation of the coronary vessel system. These epicardial progenitors give rise to the different cellular components of the coronary vasculature by undergoing a number of morphological and physiological changes collectively known as epithelial to mesenchymal transformation (EMT). However, the specific signaling mechanisms that regulate epicardial EMT are yet to be delineated. In this study we investigated the role of TGFβ2 and hyaluronan (HA) during epicardial EMT and how signals from these two molecules are integrated during this important process. Here we show that TGFβ2 induces MEKK3 activation, which in turn promotes ERK1/2 and ERK5 phosphorylation. TGFβ2 also increases Has2 expression and subsequent HA production. Nevertheless, inhibition of MEKK3 kinase activity, silencing of ERK5 or pharmacological disruption of ERK1/2 activation significantly abrogates this response. Thus, TGFβ2 promotes Has2 expression and HA production through a MEKK3/ERK1/2/5-dependent cascade. Furthermore, TGFβ2 is able to induce epicardial cell invasion and differentiation but not proliferation. However, inhibition of MEKK3-dependent pathways, degradation of HA by hyaluronidases or blockade of CD44, significantly impairs the biological response to TGFβ2. Taken together, these findings demonstrate that TGFβ2 activation of MEKK3/ERK1/2/5 signaling modulates Has2 expression and HA production leading to the induction of EMT events. This is an important and novel mechanism showing how TGFβ2 and HA signals are integrated to regulate changes in epicardial cell behavior.     

MicroRNA-98 inhibits TGF-β1-induced differentiation and collagen production of cardiac fibroblasts by targeting TGFBR1

To investigate the effects of miR-98 on TGF-β1-induced cardiac fibrosis in human cardiac fibroblasts (HCFs), and to establish the mechanism underlying these effects, HCFs were transfected with miR-98 inhibitor or mimic, and then treated with or without TGF-β1. The level of miR-98 was determined by qRT-PCR in TGF-β1-induced HCFs. Cell differentiation and collagen accumulation of HCFs were detected by qRT-PCR and Western blot assays, respectively. The mRNA and protein expressions of TGFBR1 were determined by qRT-PCR and Western blotting. In this study, the outcomes showed that TGF-β1 could dramatically decrease the level of miR-98 in a time- and concentration-dependent manner. Upregulation of miR-98 dramatically improved TGF-β1-induced increases in cell differentiation and collagen accumulation of HCFs. Moreover, bioinformatics analysis predicted that the TGFBR1 was a potential target gene of miR-98. Luciferase reporter assay demonstrated that miR-98 could directly target TGFBR1. Inhibition of TGFBR1 had the similar effect as miR-98 overexpression. Downregulation of TGFBR1 in HCFs transfected with miR-98 inhibitor partially reversed the protective effect of miR-98 overexpression on TGF-β1-induced cardiac fibrosis in HCFs. Upregulation of miR-98 ameliorates TGF-β1-induced differentiation and collagen accumulation of HCFs by downregulation of TGFBR1. These results provide further evidence for protective effect of miR-98 overexpression on TGF-β1-induced cardiac fibrosis.     

Biosynthesis of plastoquinone and β-carotene in isolated chloroplasts

Intact, isolated spinach chloroplasts incorporated ¹⁴C from ¹⁴CO₂ into plastoquinone and β-carotene under photosynthetic conditions. Addition of unlabelled l-tyrosine, p-hydroxyphenylpyruvate, or homogentisate increased the incorporation of ¹⁴C into plastoquinone, but decreased that into β-carotene.     

β-Amyloid-aluminum complex alters cytoskeletal stability and increases ROS production in cortical neurons

Several lines of evidence have supported the potential involvement of metal ions in the etiology of Alzheimer’s Disease (AD). However, the molecular mechanisms underlying this interaction are still partially unknown. Previous work from our laboratory has shown that β-amyloid peptide (Aβ) aggregation was strongly influenced by the conjugation of the peptide with few metal ions (aluminum, copper, zinc, and iron) that are found in high concentrations in the senile plaque core. The binding of aluminum (Al) to Aβ specifically stabilized the peptide in an oligomeric conformation. Here, we show that the aggregation of Aβ-Al was boosted by sodium dodecyl sulfate, a detergent that mimics some characteristics of biological membrane, suggesting a potential role for membrane components in the Aβ aggregation process. Notably, we also found that Aβ-Al caused mitochondrial dysfunction and reactive oxygen species production in primary cortical neurons. Aβ-Al strongly promoted also alterations in cytoskeleton network as shown by the increased F-actin expression and the occurrence of neuritic beading. Interestingly, the neurotoxic effect of this metal complex was associated with a decreased mRNA expression of ubiquitin thiolesterase, an ubiquitin-dependent protein involved in catabolic process, and by the increased expression of glutaminyl cyclase, responsible for pathological post-translational modification of Aβ. These results suggest that, in neuronal cells, Aβ-Al can induce relevant detrimental changes that resemble pathological hallmarks of AD.     

Improved β-carotene production of Rhodotorula glutinis in fermented radish brine by continuous cultivation

Fermentation kinetics of growth and β-carotene production by Rhodotorula glutinis DM28 in batch and continuous cultures using fermented radish brine, a waste generated from fermented vegetable industry, as a cultivation medium were investigated. The suitable brine concentration for β-carotene production by R. glutinis DM28 was 30 g l^?1. Its growth and β-carotene production obtained by batch culture in shake flasks were 2.2 g l^?1 and 87 μg l^?1, respectively, while, in a bioreactor were 2.6 g l^?1 and 186 μg l^?1, respectively. Furthermore, its maximum growth rate and β-carotene productivity in continuous culture obtained at the dilution rate of 0.24 h^?1 were 0.3 g l^?1 h^?1 and 19 μg l^?1 h^?1, respectively, which were significantly higher than those in the batch. Therefore, improved growth rate and β-carotene productivity of R. glutinis in fermented radish brine could be accomplished by continuous cultivation.     

β-carotene improves fecal dysbiosis and intestinal dysfunctions in a mouse model of vitamin A deficiency

Vitamin A deficiency (VAD) results in intestinal inflammation, increased redox stress and reactive oxygen species (ROS) levels, imbalanced inflammatory and immunomodulatory cytokines, compromised barrier function, and perturbations of the gut microbiome. To combat VAD dietary interventions with β-carotene, the most abundant precursor of vitamin A, are recommended. However, the impact of β-carotene on intestinal health during VAD has not been fully clarified, especially regarding the VAD-associated intestinal dysbiosis. Here we addressed this question by using Lrat^?/-Rbp^?/? (vitamin A deficient) mice deprived of dietary preformed vitamin A and supplemented with β-carotene as the sole source of the vitamin, alongside with WT (vitamin A sufficient) mice. We found that dietary β-carotene impacted intestinal vitamin A status, barrier integrity and inflammation in both WT and Lrat^?/-Rbp^?/? (vitamin A deficient) mice on the vitamin A-free diet. However, it did so to a greater extent under overt VAD. Dietary β-carotene also modified the taxonomic profile of the fecal microbiome, but only under VAD. Given the similarity of the VAD-associated intestinal phenotypes with those of several other disorders of the gut, collectively known as Inflammatory Bowel Disease (IBD) Syndrome, these findings are broadly relevant to the effort of developing diet-based intervention strategies to ameliorate intestinal pathological conditions.     

Inhibitory effects of α- and β-carotene on croton oil-induced or enzymatic lipid peroxidation and hydroperoxide production in mouse skin epidermis

• 1.1. The effects of carotenes (α- and β-) on edema, MDA contents and peroxidizability ofcroton oil-treated mouse skin epidermis, hydroperoxide production and enzymatic lipid peroxidation of epidermal homogenates were studied. Edema was determined as ear punch weight and the intensity of lipid peroxidation was measured using malondialdehyde formation.
• 2.2. Carotenes (α- and β-) significantly suppressed edema formation, hydroperoxide production, lipid peroxidation caused by croton oil, Fe + 3-ADP/NADPH or paraquat/NADPH in vivo as well as in vitro.
• 3.3. These results indicate that both α- and β-carotene have chemopreventive effects on croton oil-induced tumor promotion in skin tumorigenesis by scavenging oxygen free radicals, indirectly determined as carotene inhibition of lipid peroxidation and hydroperoxide formation.

     

Improved β-carotene production by oxidative stress in Blakeslea trispora induced by liquid paraffin

When 3 % (v/v) liquid paraffin was added to the medium, β-carotene production increased from 397 to 715 mg l^?1 in mated cultures of Blakeslea trispora. Liquid paraffin also enhanced the oxygen concentration and induce high oxidative stress, as observed by the increase in activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD). After 84 h of cultivation in the presence of liquid paraffin, the activities of SOD, CAT and POD in B. trispora increased 77, 52.5 and 76.6 %, respectively.