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OBJECTIVE: Primary tumor treatment through surgical resection and adjuvant therapy has been extensively studied, but there is a lack of effective strategies and drugs for the treatment of tumor metastases. Here, we describe a functional product based on a combination of compounds, which can be used as an adjuvant therapy and has well-known mechanisms for inhibiting cancer metastases, improving anti-cancer treatment, and enhancing immunity and antioxidant capacity. Our designed combination, named MVBL, consists of four inexpensive compounds: L-selenium-methylselenocysteine (MSC), D-‍α‍-tocopheryl succinic acid (VES), β‍-carotene (β‍-Ca), and L-lysine (Lys). METHODS: The effects of MVBL on cell viability, cell cycle, cell apoptosis, cell migration, cell invasion, reactive oxygen species (ROS), and paclitaxel (PTX)-combined treatment were studied in vitro. The inhibition of tumor metastasis, antioxidation, and immune enhancement capacity of MVBL were determined in vivo. RESULTS: MVBL exhibited higher toxicity to tumor cells than to normal cells. It did not significantly affect the cell cycle of cancer cells, but increased their apoptosis. Wound healing, adhesion, and transwell assays showed that MVBL significantly inhibited tumor cell migration, adhesion, and invasion. MVBL sensitized MDA-MB-231 breast cancer cells to PTX, indicating that it can be used as an adjuvant to enhance the therapeutic effect of chemotherapy drugs. In mice, experimental data showed that MVBL inhibited tumor metastasis, prolonged their survival time, and enhanced their antioxidant capacity and immune function. CONCLUSIONS: This study revealed the roles of MVBL in improving immunity and antioxidation, preventing tumor growth, and inhibiting metastasis in vitro and in vivo. MVBL may be used as an adjuvant drug in cancer therapy for improving the survival and quality of life of cancer patients.

Bitter melon (Momordica charantia L.) is a widely cultivated food and medicinal plant native to the world's subtropics and tropics. Constraints affecting cultivation of Bitter melon affect productivity of β-carotene. Knowing the mechanism that controls the transcription of the β-carotene biosynthesis genes in Bitter melon will be of great value in improving the yield of this important metabolite.The expressions of β-carotene biosynthetic genes such as Phytoene Desaturase (PDS) and Phytoene Synthase (PSY) were evaluated in Bitter melon accessions 'GBK027049', 'NS1026', 'Mahy-ventura', '453B' and 'Sibuka532'. Transcript expression level analysis of PSY and PDS, and amount of β-carotene in leaf, stem, and fruit, were determined using quantitative polymerase chain reaction and high-performance liquid chromatography (HPLC). Root transcript expression was used as a negative control for determining relative fold change in other tissues. Expression of PSY in fruit (6 to 27-fold compared to the control) was higher than in the other organs for all accessions. This was also the case of PDS expression (10 to 29-fold compared to the control). Leaves had the highest β-carotene concentration (17.92-45.35 µg∙gBitter melon leaves, especially those of 'GBK027049' and '453B' accessions, are an alternative to alleviate the β-carotene deficiencies in the world and especially in Africa.

Oxidative stress is one of the common factors leading to age-related eye diseases in older adults. Factors such as high oxygen consumption, high concentration of polyunsaturated fatty acids, and cumulative exposure to high-energy visible light in the eyes, lead to excessive generation of reactive oxygen species, hence triggering apoptosis of ocular cells and giving rise to ophthalmic diseases. Dietary supplements such as carotenoids, anthocyanins, and vitamins have antioxidant properties which may be of benefit in retaining better vision or reversing vision impairment; thus, studies have been conducted to understand the role of dietary supplements in the treatment or prevention of ophthalmic diseases. While high concentration of carotenoids such as lutein and zeaxanthin decrease the risk of developing age-related macular disease, anthocyanins and vitamins play a role in the treatment and prevention of other ophthalmic diseases: saffron extract reduced intraocular pressure in glaucoma patients; bilberry extract prevented impairments in lenses and retina, as well as alleviate symptoms of dry eye disease; high concentration of beta-carotene may reduce the risk of developing cataract. Further studies with clinical measurements are required to investigate the effectiveness of antioxidants on visual function and ophthalmic diseases.

For the greater utilization of β-carotene in antioxidant material, β-carotene-loaded emulsion stabilized by alkali lignin (AL) was successfully electrospinning with poly (vinyl alcohol) (PVA) (PVA/AL/β-carotene nanofiber). Transmission electron microscopy demonstrated the core-shell structure of nanofiber with the average diameter being 356.31 nm, and 85.7 % of β-carotene was effectively encapsulated into the core section. Fourier transform infrared spectra and differential scanning calorimetry revealed the good compatibility and decreased crystallinity of β-carotene, favoring its stability and solubility, respectively. As expected, the PVA/AL/β-carotene nanofiber exhibited higher antioxidant activity than free β-carotene due to the protection of AL matrix and the special structure of nanofiber, as the DPPH free radical scavenging rate being 90.7 % at 7th day. The sustained release behavior of β-carotene and AL from fiber followed Fickian diffusion model, contributing to the greater protection for fish oil than that of emulsion. Thus, this study provides an approach to develop hydrophobic compounds-loaded emulsion electrospun antioxidant material with controlled release property and enhanced activity.

Pumpkin seed and sunflower oil are rich in bioactive compounds, but are prone to oxidation during storage. Their fatty acids, carotenoid and volatile compounds and their Fourier-transform infrared (FTIR) profiles were studied during 8 months storage in order to assess the overall quality, but also to assess the impact of the oleogelation as conditioning process.The fatty acids methyl esters were analyzed by gas chromatography-mass spectrometry (GC-MS). The linoleic acid was the most abundant in the oils (604.6 g kgDuring 8 months storage, the decrease of polyunsaturated fatty acid total amount was 5.72% for the pumpkin seed oil and 3.55% for the oleogel, while in the sunflower oil samples of 2.93% and 3.28% for the oleogel. It was concluded that oleogelation might protect specific carotenoid compounds, since the oleogels displayed higher content of β-carotene at each storage time. Hexanal and heptanal were detected during storage, regardless of the oil or oleogel type. FTIR analysis depicts the differences in the constituent fatty acids resulting due to thermal oxidation or due to storage. © 2022 Society of Chemical Industry.

The aim of the current review was to explore the association between various dietary antioxidants and the risk of developing Parkinson's disease (PD). PubMed, Scopus, Web of Science, and Google Scholar were searched up to March 2021. Prospective, observational cohort studies, nested case-control, and case-control designs that investigated the association between antioxidants and PD risk were included. A random-effects model was used to pool the RRs. The certainty of the evidence was rated using the GRADE (Grading of Recommendations Assessment, Development, and Evaluations) scoring system. In addition, a dose-response relation was examined between antioxidant intake and PD risk. Six prospective cohort studies and 2 nested case-control (total n = 448,737 with 4654 cases), as well as 6 case-control (1948 controls, 1273 cases) studies were eligible. The pooled RR was significantly lower for the highest compared with the lowest intake categories of vitamin E (n = 7; 0.84; 95% CI: 0.71, 0.99) and anthocyanins (n = 2; 0.76; 95% CI: 0.61, 0.96) in cohort studies. Conversely, a significantly higher risk of PD was observed for higher lutein intake (n = 3; 1.86; 95% CI: 1.20, 2.88) among case-control studies. Dose-response meta-analyses indicated a significant association between a 50-mg/d increase in vitamin C (n = 6; RR: 0.94; 95% CI: 0.88, 0.99), a 5-mg/d increment in vitamin E (n = 7; RR: 0.84; 95% CI: 0.70, 0.99), a 2-mg/d increment in β-carotene (n = 6; RR: 0.94, 95% CI: 0.89, 0.99), and a 1-mg/d increment in zinc (n = 1; OR: 0.65; 95% CI: 0.49, 0.86) and a reduced risk of PD. Overall, higher intake of antioxidant-rich foods may be associated with a lower risk of PD. Future well-designed prospective studies are needed to validate the present findings. The protocol was registered in the International Prospective Register of Systematic Reviews (PROSPERO) database (https://www.crd.york.ac.uk/PROSPERO, CRD42021242511).

Observational studies reported conflicting results regarding the association between circulating concentrations of nutrition-related factors and atrial fibrillation (AF). The aim of this study was to evaluate the potential causal effect of 8 circulating nutrition-related factors (vitamin B12, vitamin E, folate, retinol, β-carotene, iron, zinc, and copper) on AF risk using mendelian randomization (MR). Summary-level data for the nutrition-related factors and AF were obtained from genome-wide association studies conducted among individuals of European ancestry. The genome-wide association study on AF included 60,620 cases and 970,216 controls. A 2-sample MR design was applied for evaluating the causal association. In the primary MR analyses, the inverse variance-weighted method did not identify any causal effect of circulating concentrations of vitamin B12 [β = 0.000, standard error (SE) = 0.021, P = 0.994], vitamin E (β = 0.080, SE = 0.152, P = 0.600), retinol (β = 0.098, SE = 0.397, P = 0.806), folate (β = -0.006, SE = 0.052, P = 0.901), β-carotene (β = 0.014, SE = 0.025, P = 0.560), iron (β = -0.009, SE = 0.072, P = 0.905), zinc (β = 0.038, SE = 0.032, P = 0.239), and copper (β = -0.012, SE = 0.023, P = 0.589) on AF. The MR-Egger and MR pleiotropy residual sum and outlier (MR-PRESSO) analyses did not suggest the presence of pleiotropy. In addition, the lack of association remained in the leave-one-out analysis. This MR study indicates no causal association of circulating concentrations of vitamin B12, vitamin E, folate, retinol, β-carotene, iron, zinc, and copper with AF.

To apply starch nanoparticles (SNP) as host materials for β-carotene encapsulation, aqueous SNP dispersions (10, 25, 50, and 100 mg/10 mL) and β-carotene in acetone (10, 50, 100, 150, and 200 μg/mL) were mixed. The acetone in the mixture was evaporated to prepare SNP and β-carotene composites, which were homogeneously dispersed in aqueous media with over 90 % solubility. When SNP content was higher than 50 mg, over 80 % of β-carotene was encapsulated in the composite matrix. X-ray diffraction, nuclear magnetic resonance spectroscopy, and transmission electron microscopic analyses confirmed the micellar-shaped composite particles with diameters120 nm and an amorphous structure. High SNP content in the composites enhanced β-carotene stability under extremely hot and acidic conditions as well as against ultraviolet rays and oxidation reactions. The encapsulated β-carotene was not readily released in simulated gastric fluid, but was gradually released in simulated intestinal fluid via SNP digestion in the composites.

In these studies, Fluorescence spectroscopy has been utilized for the characterization of pure and commercially available corn oil. The best excitation wavelength of 380 nm has been investigated, where maximum spectral information can be assessed. The emission spectra from pure and commercial corn oil samples disclosed that pure corn oil contained oleic acid, beta-carotenes, chlorophylls, isomers of vitamin E and traces of oxidized products which exhibit fluorescence at 406, 525, 675, 440 and 435/475 nm respectively. Whereas, commercial corn oils lack these valuable ingredients and only contain fats along with their primary and secondary oxidized products that emit a broad emission band centred at 440 nm. The study has also depicted that Fluorescence spectroscopy can even be used to select best quality corn oil among pure corn oil samples with different varieties and seed origins. In addition, the effect of temperature on the composition of pure and commercial corn oil samples have also been investigated by heating them at 100, 120,140, 160, 180 and 200 °C each sample for 30 min. This was done because corn oil is being used for cooking where it is generally heated up to 120 °C and for deep frying up to 180 °C. On heating, in pure corn oil, deterioration of Vitamin-E and beta-carotenes occurred with an increase in the oxidation products, whereas, in commercial oil samples, only the concentration of oxidation products increased. However, it was found that up to 140 °C, pure corn oil can be used safely for cooking purpose where it does not lose much of its valuable ingredients while in commercial corn oils, fat composition does not alter much up to 180 °C and after that oxidized products start to increase rapidly.

Raman spectroscopy is an emerging dermatological technique with the potential to discriminate biochemically between cell types in a label-free and non-invasive manner. Here, we use live single-cell Raman spectroscopy and principal component analysis (PCA) to fingerprint mouse melanoblasts, melanocytes, keratinocytes and melanoma cells. We show the differences in their spectra are attributable to biomarkers in the melanin biosynthesis pathway and that melanoma cells are a heterogeneous population that sit on a trajectory between undifferentiated melanoblasts and differentiated melanocytes. We demonstrate the utility of Raman spectroscopy as a highly sensitive tool to probe the melanin biosynthesis pathway and its immediate response to ultraviolet (UV) irradiation revealing previously undescribed opposing responses to UVA and UVB irradiation in melanocytes. Finally, we identify melanocyte-specific accumulation of β-carotene correlated with a stabilisation of the UVR response in lipids and proteins consistent with a β-carotene-mediated photoprotective mechanism. In summary, our data show that Raman spectroscopy can be used to determine the differentiation status of cells of the melanocyte lineage and describe the immediate and temporal biochemical changes associated with UV exposure which differ depending on cell type, differentiation status and competence to synthesise melanin. Our work uniquely applies Raman spectroscopy to discriminate between cell types by biological function and differentiation status while they are growing in culture. In doing so, we demonstrate for the first time its utility as a tool with which to probe the melanin biosynthesis pathway.

Genome mutagenesis drives the evolution of organisms. Here, we developed a

Epidermal growth factor (EGF) has many important biological functions. It plays an important role in regulating the growth, survival, migration, apoptosis, proliferation, and differentiation of intestinal tissues and cells. However, until now, the effect of inflammation on the biological activity of EGF in intestinal cells or tissues is still unclear. For this reason, in the current research, we have conducted a detailed study on this issue. Using the rat small intestinal crypt epithelial cell line (IEC6) was used as an in vitro model, and Confocal laser scanning microscope (CLSM), Flow cytometry (FCM), Indirect immunofluorescence assay (IFA), Western-blotting (WB), and Quantitative real-time RT-PCR (QRT-PCR) methods were used to explore the effects of inflammation on EGF/EGFR biological activity and signal transduction profiles. We found that the EGF/EGFR nuclear signal almost disappeared in the inflammatory state, and the phosphorylation levels of EGFR, AKT, and STAT3 were all significantly down-regulated. In addition, we also studied the effect of β-carotene on the biological activity of EGF, and found that when cells were pretreated with β-carotene, the cellular behavior, biological activity, and nuclear signal of EGF/EGFR under inflammation stimulation were partially restored. In summary, the current study shows that inflammation can disrupt EGF/EGFR-mediated signaling in IEC6 cells, suggesting that inflammation negatively regulates the biological activity of EGF/EGFR. Furthermore, we found that β-carotene not only attenuated lipopolysaccharide (LPS)-induced inflammation but also partially restored the biological activity of EGF in IEC6 cells, laying a solid foundation for studying the biological functions of EGF and β-carotene.

There has been increasing demands worldwide for bioactive compounds of natural origins, especially for the nutraceutical and food-supplement sectors. In this context, microalgae are viewed as sustainable sources of molecules with an array of health benefits. For instance, astaxanthin is a xanthophyll pigment with powerful antioxidant capacity produced by microalgae such as the chlorophyte Haematococcus sp., which is regarded as the most suitable organism for the mass production of this pigment. In this study, three Haematococcus sp. strains were cultivated using a batch mode under favourable conditions to promote vegetative growth. Their environment was altered in a second phase using a higher and constant illumination regime combined with either exposure to blue LED light, an osmotic shock (with NaCl addition) or supplementation with a phytohormone (gibberellic acid, GA3), a plant extract (ginger), an herbicide (molinate) or an oxidant reagent (hydrogen peroxide). The effects of these stressors were evaluated in terms of antioxidant response and astaxanthin and β-carotene accumulation. Overall, strain CCAP 34/7 returned the highest Trolox Equivalent Antioxidant Capacity (TEAC) response (14.1-49.1 µmoL Trolox eq. g

In this study the potential of liposomes as a vitamin E (VE) and β-carotene (βC) delivery system was examined. The co-encapsulated liposomes of βC and VE (L-VE-βC) were prepared and characterized. Their antioxidant properties were evaluated by free radical scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), hydroxyl radical and lipid peroxidation assay. The in vitro sustained release behaviour was then investigated and discussed.VE and βC were co-encapsulated in liposomes with high encapsulation efficiency, up to 92.49% and 86.16% for βC and VE, respectively. The antioxidant activities of L-VE-βC samples were significantly higher than that of single loaded liposome. Among different ratios of VE/βC, L-VE-βC at 5:3 exhibited the highest radical scavenging rates, with 66.80%, 56.58% and 34.39% for DPPH, ABTS and OH radical, respectively. L-VE-βC samples also had a good ability to inhibit lipid peroxidation, especially the sample with ratios of VE/βC at 5:3 and 3:1. In simulated gastrointestinal release, L-VE-βC exhibited an excellent sustained release behaviour in SGF with the accumulated rate at about 20%, while the release rate in SIF increased to over 80%, where they should be absorbed. The release kinetics analysis indicated that βC was released in the Higuchi model in stomach, and the Korsmeyr-Peppas model in intestine.Compared to single loaded liposomes, the combined-loaded liposomes exhibited higher antioxidant activity and bioavailability, suggesting the potential applications in functional foods. © 2022 Society of Chemical Industry.

Background: Anchusa azurea Mill. (AA) is a medicinal plant largely used traditionally in folk medicine in Algeria; it is locally named hamham. It is effective in the treatment of various diseases. Objectives: The aim of the present study is to determine the antioxidant, anti-inflammatory, and anti- hemolytic effects of phenolic fractions from Anchusa azurea Mill. Methods: In this study, various extracts from Anchusa azurea Mill. (AA) using solvents with increasing polarity were prepared. The quantification of polyphenols and flavonoids was determined. The anti-radical activity of the different extracts was evaluated using DPPH and by measuring the inhibition of the oxidative degradation of β-carotene. The In-vitro antihemolytic effect of the plant extracts is determined (CrE, ChE, AcE, and AqE). For each extract, four concentrations were tested: 10.59, 21.18, 42.37, 84.74 μg/ml. Vitamin C is used as a standard. The free-radical attack was measured by measuring the HT50 (Half-Hemolysis Time). The anti-inflammatory effect using PMA on mice of the methanolic extract (CrE) was evaluated. Results: The quantification of polyphenols and flavonoids showed that ethyl acetate extract (AcE) contains a higher amount of polyphenols. However, chloroform extract (ChE) presents a higher amount of flavonoids. AcE showed an important scavenging activity using the DPPH radical (IC50= 68.35 μg/ml). The results showed that AcE also exhibited asignificant inhibition effect on the oxidation of β-carotene/linoleic acid (84.33 %). All extracts increased the HT50 values (Half-Hemolysis Time) in a dose-dependent manner. The three highest concentrations (21.18, 42.37, and 84.74 μg / ml) of ChE caused a very significant delay (p ≤ 0.001) of hemolysis compared to the negative control and the positive control “VIT C”. The anti-inflammatory effect of using PMA on mice showed that the methanolic extract (CrE) of AA reduced the weight of the ear edema. Conclusions: This plant has a strong pharmacological power, which supports its traditional medicinal use.

Astaxanthin is a kind of ketone carotenoid belonging to tetraterpenoids with an excellent antioxidant activity and it is widely used in nutrition and health-care industries. This study aimed to explore the effect of different abiotic stresses on carotenoid production in Schizochytrium sp. Firstly, the characteristics of carotenoid accumulation were studied in Schizochytrium sp. by monitoring the change of carotenoid yields and gene expressions. Then, different abiotic stresses were systematically studied to regulate the carotenoid accumulation. Results showed that low temperature could advance the astaxanthin accumulation, while ferric ion could stimulate the conversion from carotene to astaxanthin. The glucose and monosodium glutamate ratio of 100:5 was helpful for the accumulation of β-carotene. In addition, micro-oxygen supply conditions could increase the yield of β-carotene and astaxanthin by 25.47% and 14.92%, respectively. This study provided the potential regulation strategies for carotenoid production which might be used in different carotenoid-producing strains.

Research on the morphophysiological behavior of forest seeds during germination with respect to climate change is scarce. To date, there have been no studies on the biochemical or morphological aspects of Ormosia spp. In this study, we subjected Ormosia coarctata seeds to various temperature conditions to investigate temperature-dependent impacts on morphology, reactive oxygen species (ROS) generation, antioxidant systems, and storage systems. Analyses were performed on seeds exposed to 25, 35, and 40 °C for 48, 96, and 144 h. The morphology was evaluated by radiation using a Faxitron MX-20 device. ROS production (superoxide anion and hydrogen peroxide), malonaldehyde (MDA), carbonylated proteins, antioxidant enzyme activity (superoxide dismutase [SOD], ascorbate peroxidase [APX], catalase [CAT], and peroxidase [POX]), β-carotene, lycopene, glucose, and reserve enzyme activity (α- and β-amylase, lipase, and protease) were analyzed by spectrophotometry. Heat stress (40 °C) decreased germination by 76.2% and 78.1% (compared to 25 and 35 °C, respectively), caused damage to the external morphology of the seed, increased the content of ROS, MDA, and carbonylated proteins, and reduced APX, CAT, and POX activity. Furthermore, heat stress decreased glucose content and α-amylase activity. These results suggest that an increase of 5 °C in temperature negatively affects germination, promotes oxidative stress, and induces deterioration in O. coarctata seeds.

High blood levels of β-carotene and increased intake in the diets are inversely proportional to incidence of many cancer types. Antioxidant activity of β-carotene was proposed to be related with its antitumor effect. Despite this plant derivative substance being sought in many cancer types, the effectiveness of β-carotene against malignant mesothelioma remained unclear. Therefore, the present study aims to explore the impact of β-carotene on cell viability, apoptosis, and oxidative stress in mesothelioma cells. Human mesothelioma cell SPC212 were treated with β-carotene (3.125-200 μM) for 24, 48, 72, and 96 h. Cytotoxicity was measured with the MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide). Annexin-V/propidium iodide (PI) and caspase 3/7 biomarkers were used to identify apoptotic cells. Finally, the oxidative stress was evaluated with flow cytometry. The results of the measurements indicated a significant decline in viable mesothelioma cancer cell numbers upon β-carotene treatment in time- and concentration-dependent manner when compared to control cells. Furthermore, β-carotene treatment led to apoptosis induction according to both annexin V/PI and caspase 3/7 assays. Furthermore, β-carotene increased oxidative stress in SPC212 cells. These results show how β-carotene affects proliferative, apoptotic, and oxidative properties in SPC212 malignant pleural mesothelioma cells and provide useful insights into future studies.

Insufficient vitamin content in the diet is a risk factor for the development of various diseases and their progression, and drug therapy can also contribute to the development of a deficiency of these micronutrients. Data on vitamin sufficiency of patients with type 2 diabetes mellitus (T2DM) and diabetic nephropathy (DN) are needed to develop measures to improve it.

In this study, we investigated the spray-drying microencapsulation of β-carotene in oil co-stabilized by soy protein isolate-epigallocatechin-3-gallate conjugate (SPE) and small molecule surfactants [sodium dodecyl sulfate (SDS), hexadecyl trimethyl ammonium bromide (CTAB), and tea saponin (TS)] of different concentrations [0.1, 0.5, and 1.0% (w/v)], as a prospective approach to stabilize β-carotene. The results show that different surfactant types and concentrations significantly affect the encapsulation efficiency, water dispersibility, microstructure, and digestion of the microcapsules. Interactions between the surfactants and the SPE at the interface were found to include both synergistic and competitive effects, and they depended on the surfactant type and concentration. Moreover, the addition of SDS and TS before spray drying significantly improved the microencapsulation performance of the microcapsules and the water dispersion behavior of the corresponding spray-dried powders. The highest encapsulation efficiency was achieved for the SPE-0.1TS-encapsulated β-carotene microcapsules. In contrast, the addition of CTAB was not conducive to microcapsule formation, resulting in poor encapsulation efficiency, water dispersibility, thermal stability, β-carotene retention rate, and oxidation stability.

The antioxidant capacity of β-carotene has been studied in terms of H-atom abstraction reactions using quantum chemical methods. These oxidation reactions are studied for the all-trans as well as 15,15'-cis isomers (15Z) of β-carotene, as the latter is only ~ 10 kJ/mol less stable than the all-trans isomer in the gas phase and about 9 kJ/mol less stable in aqueous solution. Hydrogen abstraction from the rotamers obtained through C-C single and double bond rotations has been shown to play an important role in determining the antioxidant capacity of β-carotene. Hydrogen abstraction from the C4 and C5-CH3 positions of the β-ionone rings and the C7 and C9 positions along the polyene chain of β-carotene by hydroxyl radical have been studied. In the all-trans form the most favorable H-atom abstraction reaction occurs at the C4 position of the terminal regions of the polyene -system of β-carotene, closely followed by hydrogen abstraction from the C5 methyl position. The H-atom abstraction reactions are more exothermic in water than in the gas phase due to solvation energies for the water product.

This study improved the bioaccessibility of puerarin, a bioactive flavonoid, through incorporating co-encapsulated puerarin and β-carotene into a food-grade high internal-phase Pickering emulsion (HIPPE) system stabilised with meat proteins.

Background/Aim: Fucoxanthinol (FxOH), a marine carotenoid, induces apoptosis and anoikis in human colorectal cancer (CRC) DLD-1 cells via the down-regulation of chloride intracellular channel 4 (CLIC4) expression, a key molecule for apoptosis. However, whether FxOH is susceptible to CLIC4 expression and its regulatory mechanisms in human CRC cells remains unknown. We investigated the inhibitory effects of FxOH on six types of human CRC cells with CLIC4 regulation. Materials and Methods: The association between FxOH and CLIC4 was investigated using gene knockdown, overexpression, and transcriptome analyses. Results: CLIC4 expression in CRC cells was a significant factor associated with sensitivity to FxOH. CLIC4 regulates many cancer-related signals and participates in growth inhibition in FxOH-treated DLD-1 cells. Both CLIC4 knockdown and overexpression attenuated the inhibitory effects of FxOH on DLD-1 cells. Conclusion: Our findings suggest that the protein expression of CLIC4 and its regulating mechanisms play significant roles regarding cell death in human CRC cells by FxOH treatment. Further investigation by in vitro and in vivo models is needed to determine the effect of CLIC4.

Background: The study aimed to identify 2 beta-carotene 15,15′-monooxygenase (BCMO1) mutations, namely R267S and A379V, and determine their association with vitamin A status among Filipinos 6 to 19 years old respondents of the 2013 Philippine National Nutrition Survey living in the National Capital Region. Materials and Methods: This study followed cross-sectional design. Whole blood specimen was collected in the morning and was used as source of genomic DNA and serum for retinol concentration determination. Fisher exact test was performed to determine whether genotype frequencies were associated to retinol concentrations/vitamin A deficiency status. A level of P < .05 was identified as significant. Results: A total of 693 Filipino children and adolescents were included. Of the 693, there were at least 7.6% who bear the combined mutations for R267S + A379V. Association analysis showed that an inverse relationship exists between the A379V TT variant and vitamin A status, although the exact role of these identified polymorphisms on retinol/carotenoid metabolism need to be confirmed in dedicated functional studies. Conclusion: This study has identified for the first time the presence of 2 nonsynonymous genetic variants/mutations in the coding region of BCMO1 gene. Interestingly, one of these 2 variants, the A379V T, was found to be associated with vitamin A status. It is, therefore, warranted to investigate the role of BCMO1 variants for the success of supplementation programs and fortification efforts among vulnerable populations in this region. Genetic variability should be considered for future provitamin A supplementation recommendations among children and adolescents in the Philippines.

Background In this study, flowers of nine mullein species (Verbascum erianthum, V. songaricum, V. speciosum, V. szovitsianum, V. stachydiforme, V. sinuatum, V. haussknechtianum, V. cheirantifolium, and V. saccatum) were collected from Iran. Total flavonoid content (TFC), antioxidant activity, total carotenoid (TCC) and β-carotene content and the amount of phenolic compounds were determined in all of the collected mullein species. Quantitative analysis of phenolic compounds was performed by using HPLC-DAD. Results V. saccatum and V. songaricum, in particular, were found rich in total flavonoid so exhibit the highest antioxidant capacity. The HPLC-DAD analysis revealed that V. erianthum possesses the highest amount of caffeic acid (0.022 g kg-1 DW), chlorogenic acid (2.649 g kg-1 DW), p-coumaric acid (0.253 g kg-1 DW), and apigenin (0.066 g kg-1 DW) and the highest amount of gallic acid (0.134 g kg-1 DW), rutin (5.254 g kg-1 DW), quercetin (1.303 g kg-1 DW), and cinnamic acid (1.031 g kg-1 DW) content were observed in V. saccatum. Our results demonstrate the considerable variations in the TFC, antioxidant activity, TCC and β-carotene content among the mullein species. Conclusions The findings of this study provide remarkable information for schematization breeding strategies, and for choosing the best species with high phenolic compounds content to produce natural antioxidant compounds as a medicinal plant for the medical uses and pharmaceutical industries. This article is protected by copyright. All rights reserved.

Background According to the International Agency for Research on Cancer, some hair dye chemicals are considered mutagenic and carcinogenic in humans. One hospital-based study reported a positive association between hair dye use and prostate cancer risk, but no prospective analyses have been conducted. Methods This study investigated the association between hair dye use and prostate cancer risk in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study cohort, a large, well-characterized cohort of 29,133 male Finnish smokers. Participants completed questionnaires regarding lifestyle, medical, and risk factor information, including the use of hair dye, which was available for 98.8% of the cohort (28,795 men). Prostate cancer cases were identified through linkage with the Finnish Cancer Registry and the Finnish Mortality Register. Hazard ratios (HRs) and confidence intervals (CIs) were estimated via multivariable Cox proportional hazards regression. Results During a 28-year period of observation, 2703 incident prostate cancer cases were diagnosed. As reported at the baseline, 75 men used hair dye, and 13 of these men were subsequently diagnosed with prostate cancer. After adjustments for potential confounders, men who used hair dyes experienced substantially higher prostate cancer risk than men who did not (HR, 1.77; 95% CI, 1.03-3.05). Conclusions This first prospective investigation of hair dye use and prostate cancer suggests that personal hair dye use may be related to increased risk. The findings warrant re-examination in other prospective cohorts along with studies evaluating specific hair dyes and possible underlying biological mechanisms.

Author(s): Yu, Shu; Li, Michelle; Dubcovsky, Jorge; Tian, Li | Abstract: Grains of tetraploid wheat (Triticum turgidum L.) mainly accumulate the non-provitamin A carotenoid lutein-with low natural variation in provitamin A β-carotene in wheat accessions necessitating alternative strategies for provitamin A biofortification. Lycopene e-cyclase (LCYe) and β-carotene hydroxylase (HYD) function in diverting carbons from β-carotene to lutein biosynthesis and catalyzing the turnover of β-carotene to xanthophylls, respectively. However, the contribution of LCYe and HYD gene homoeologs to carotenoid metabolism and how they can be manipulated to increase β-carotene in tetraploid wheat endosperm (flour) is currently unclear. We isolated loss-of-function Targeting Induced Local Lesions in Genomes (TILLING) mutants of LCYe and HYD2 homoeologs and generated higher order mutant combinations of lcye-A, lcye-B, hyd-A2, and hyd-B2. Hyd-A2 hyd-B2, lcye-A hyd-A2 hyd-B2, lcye-B hyd-A2 hyd-B2, and lcye-A lcye-B hyd-A2 hyd-B2 achieved significantly increased β-carotene in endosperm, with lcye-A hyd-A2 hyd-B2 exhibiting comparable photosynthetic performance and light response to control plants. Comparative analysis of carotenoid profiles suggests that eliminating HYD2 homoeologs is sufficient to prevent β-carotene conversion to xanthophylls in the endosperm without compromising xanthophyll production in leaves, and that β-carotene and its derived xanthophylls are likely subject to differential catalysis mechanisms in vegetative tissues and grains. Carotenoid and gene expression analyses also suggest that the very low LCYe-B expression in endosperm is adequate for lutein production in the absence of LCYe-A. These results demonstrate the success of provitamin A biofortification using TILLING mutants while also providing a roadmap for guiding a gene editing-based approach in hexaploid wheat.

Blakeslea trispora has great potential uses in industrial production because of the excellent capability of producing a large quantity of carotenoids. However, the mechanism of light induced carotenoid biosynthesis even the structural and regulatory genes in pathways remain unclear. In this paper, we reported the first transcriptome study in B. trispora in which we have carried out global survey of expression changes of genes participated in blue light response. We verified that the yield of β-carotene reaching to 3-fold when transferred from darkness to blue light for 24 h and the enhancement of transcription levels of carRA and carB presented a positive correlation with the increase in carotenoid production. RNA-seq analysis revealed that 1124 genes were upregulated and 740 genes were downregulated respectively after blue light exposure. Annotation through GO, KEGG, Swissprot and COG databases showed 11119 unigenes compared well with known gene sequences, 5514 unigenes were classified into Gene Ontology, and 4675 unigenes were involved in distinct pathways. Among the blue light responsive genes, 4 genes (carG1, carG3, carRA and carB) identified to function in carotenoid metabolic pathways were dominantly upregulated. We also discovered that 142 TF genes belonging to 45 different superfamilies showed significant differential expression (p≤ 0.05), 62 of which were obviously repressed by blue light. The detailed profile of transcription data will not only allow us to conduct further functional genomics study in B. trispora, but also enhance our understanding of potential metabolic pathway and regulatory network involved in light regulated carotenoid synthesis.

Rice bran protein is an emerging protein source from rice milling that possesses health benefits and emulsifying capacity suitable for hypoallergenic encapsulation applications, especially for lipophilic compounds such as β-carotene. The purpose of this study is to develop and characterize β-carotene encapsulates with maltodextrin and rice bran protein. Rice bran protein was prepared using conventional alkali extraction. β-carotene was added to the composite wall materials (50:50 of 4%, 8%, 12%, and 16% solids content) and spray-dried. Encapsulation efficiency (85–98%) and radical scavenging activity (11–43%) varied proportionally with rice bran protein. Across increasing maltodextrin and rice bran protein content of the feed, carbohydrate content of the microcapsules varied proportionally (50–66%) but protein content was uniform (10–13%). Scanning electron microscopy, differential scanning calorimetry, and Fourier transform infrared spectroscopy data suggested successful encapsulation. Release profiles showed decreasing trend with increasing rice bran protein content; co-digestion with rice mitigated negative impacts of rice bran protein. Microcapsules with nutritive potential and health-promoting properties were developed as potential carotenoid delivery systems.

We aimed to explore the relationship between intakes of food-derived antioxidants (zinc, vitamin E, β-carotene, and selenium) and semen parameters in an infertile male population. This is a cross-sectional study among infertile men (> 18 years) referred to Isfahan Fertility and Infertility Center in Iran from March 2019 to October 2019. The main outcome measures were semen volume, sperm count, concentration, total motility, and morphology. Micronutrients considered in this analysis were zinc, selenium, β-carotene, and vitamin E. A total of 350 infertile men with a mean (SE) age of 34.77 (0.42) were included in this study. Selenium intake tended to be positively associated with higher semen volume (β = 0.79; 95% CI: - 0.01, 1.59) and sperm total motility after adjustment for main confounders (β = 2.64; 95% CI: - 3.61, 8.89). Intake of β-carotene was only associated with higher sperm total motility (β = 5.46; 95% CI: - 0.84, 11.77). No significant association was detected between intakes of zinc and vitamin E and semen parameters.In a population of infertile adult men, selenium intake was associated with higher sperm total motility and semen volume, and in the case of β-carotene, better sperm total motility. Further, prospective studies are warranted.

Carotenoids are important natural pigments that give bright colors to plants. The difference in the accumulation of carotenoids is one of the key factors in the formation of various colors in carrot taproots. Carotenoid cleavage dioxygenases (CCDs), including CCD and nine-cis epoxycarotenoid dioxygenase (NCED), are the main enzymes involved in the cleavage of carotenoids in plants. Seven CCD genes have been annotated from the carrot genome. In this study, through expression analysis, we found that the expression level of DcCCD4 was significantly higher in the taproot of white carrot (low carotenoid content) than orange carrot (high carotenoid content). The overexpression of DcCCD4 in orange carrots caused the taproot color to be pale yellow, and the contents of α-carotene and s-carotene decreased sharply. Mutant carrot with loss of DcCCD4 function exhibited yellow color (the taproot of the control carrot was white). The accumulation of s-carotene was also detected in taproot. Functional analysis of the DcCCD4 enzyme in vitro showed that it was able to cleave α-carotene and s-carotene at the 9, 10 (9', 10') double bonds. In addition, the number of colored chromoplasts in the taproot cells of transgenic carrots overexpressing DcCCD4 was significantly reduced compared with that in normal orange carrots. Results demonstrated that DcCCD4 affects the accumulation of carotenoids through cleavage of α-carotene and s-carotene in carrot taproot.

Vitamin A is present in multivitamin products mainly in the form of retinol esters: retinyl acetate, retinyl palmitate, and beta carotene—retinol precursor (dimer) found in plants, which is capable of converting into retinol in liver cells. Retinol is determined in medicinal products primarily by high performance liquid chromatography (HPLC), with preliminary purification and vitamin isolation by liquid-liquid extraction. However, scientific literature also describes other methods of sample preparation and analysis of such compounds. An important issue is differentiation of vitamin A from other fat-soluble vitamins often included as components in multivitamin products. The aim of the study was to analyse and summarise data on current methods used for determination of vitamin A and its derivatives in medicinal products. The authors analysed the range of vitamin A products authorised in the Russian Federation, and the test methods described in their product specification files. The study demonstrated that the test method most often used for determination of retinol esters was HPLC with isocratic elution mode using octadecylsilyl packing in the reverse-phase mode, and, less frequently, aminopropylsilyl packing in the normal phase mode. Determination of beta carotene in medicinal products is most often performed using spectrophotometry.

Background and purpose An effect of dietary carotenes on risk of cardiovascular disease (CVD) is uncertain. We aimed to investigate whether the association between dietary carotenes intake and risk of CVD mortality will persist after controlling for the intakes of potential cardioprotective dietary factors that correlate with dietary alpha- and/or beta-carotenes. Methods and results We followed up a total of 58,646 Japanese between 1988 and 1990 and 2009. We used a food frequency questionnaire (FFQ) to determine the dietary intakes of carotenes, and estimated the hazard ratios (HRs) and 95% confidence intervals (CIs) of CVD mortality in relation to carotene intake by the proportional hazard regression developed by David Cox. During 965,970 person-years of follow-up (median 19.3 years), we identified 3388 total CVD deaths. After adjusting for demographic and lifestyle factors, dietary intakes of alpha-carotene were significantly associated with the reduced risk of mortality from coronary heart disease (CHD); adjusted HR (95% CI) in the highest versus lowest quintiles of intake was 0.75 (0.58–0.96; P-trend = 0.02) and dietary intakes of beta-carotene were significantly associated with the reduced risk of mortality from CVD, CHD, and other CVD; adjusted HRs (95% CIs) were 0.88 (0.79–0.98; P-trend = 0.04), 0.78 (0.61–0.99; P-trend = 0.01), and 0.81 (0.67–0.98; P-trend = 0.04), respectively. However, after further adjusting for the dietary intakes of potassium, calcium, vitamins C, E, or K, these associations disappeared. Conclusions —Dietary alpha- and beta-carotene intakes were not associated with risk of CVD mortality after controlling for intakes of other potential cardioprotective nutrients.

The scientific interest in the medicinal properties of Kombucha beverages, a carbonated drink with live microorganisms, has increased recently. Hence, the aim of this study was to determine the chemical profile and to examine the antioxidant, antidiabetic and antineurodegenerative potential of unfermented and also Kombucha fermented

Patients with chronic kidney disease (CKD) occasionally need to restrict their consumption of vegetables and fruits. However, recent evidence suggests that plant-based diets have beneficial effects in patients with CKD. We aimed to determine the sufficiency of β carotene and dietary fiber intake in patients with CKD. We conducted a cross-sectional study among 4476 patients registered in the Fukuoka Kidney Disease Registry (FKR) study, a Japanese prospective cohort study of patients with CKD. Data from 3545 patients were analyzed after excluding cases with insufficient information. We evaluated the relationship between CKD stages and the intake of vegetables and fruits. The intake of β carotene and dietary fiber in CKD stages was evaluated using analysis of covariance. As the CKD stage advanced, the intake of vegetables, green leafy vegetables, and fruits significantly decreased (P-value for all trends

β-Carotene is a dietary source of vitamin A, and its physiological functions, such as anti-inflammatory activity, immune regulation, and improvement of intestinal flora, are attracting increasing attention. Recent studies have shown that the development of food allergy is closely related to intestinal dysfunction. Therefore, the present study investigated the potential anti-food allergy activity of β-carotene and its regulatory intestinal homeostasis pathway. The results obtained using an ovalbumin (OVA)-induced food allergy mouse model indicated that the clinical allergic symptoms were alleviated, and the levels of anaphylactic mediators (such as immunoglobulin (Ig) E, IgG, and histamine) were reduced after β-carotene supplementation at 5.00 mg per kg per day. In addition, the expression of tight junction (TJ) proteins (claudin-1, occludin, and ZO-1) increased by 38.58%, 24.39%, and 26.23%, respectively. Additionally, the secretion of secretory IgA (sIgA) and the regeneration of islet-derived protein (Reg) IIIγ were promoted in the intestinal mucous after β-carotene administration. Furthermore, the alpha and beta diversity analysis showed that the composition and diversity of the intestinal flora in the β-carotene group tended to be normalized compared to the model group. Higher levels of beneficial bacteria, such as

Background Abnormal metabolism and perturbations in metabolic pathways play significant roles in the development and progression of prostate cancer; however, comprehensive metabolomic analyses of human data are lacking and needed to elucidate the interrelationships. Methods We examined the serum metabolome in relation to prostate cancer survival in a cohort of 1812 cases in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. Using an ultrahigh-performance LC-MS/MS platform, we identified 961 known metabolites in prospectively collected serum. Median survival time from diagnosis to prostate cancer-specific death (N=472) was 6.6 years (interquartile range=2.9–11.1 years). Cox proportional hazards regression models estimated hazard ratios and 95% confidence intervals of the associations between the serum metabolites (in quartiles) and prostate cancer death, adjusted for age at baseline and diagnosis, disease stage, and Gleason sum. In order to calculate risk scores, we first randomly divided the metabolomic data into a discovery set (70%) and validated in a replication set (30%). Results Overall, 49 metabolites were associated with prostate cancer survival after Bonferroni correction. Notably, higher levels of the phospholipid choline, amino acid glutamate, long-chain polyunsaturated fatty acid (n6) arachidonate (20:4n6), and glutamyl amino acids gamma-glutamylglutamate, gamma-glutamylglycine, and gamma-glutamylleucine were associated with increased risk of prostate cancer-specific mortality (fourth versus first quartile HRs=2.07–2.14; P-values −5). By contrast, the ascorbate/aldarate metabolite oxalate, xenobiotics S-carboxymethyl-L-cysteine, fibrinogen cleavage peptides ADpSGEGDFXAEGGGVR and fibrinopeptide B (1-12) were related to reduced disease-specific mortality (fourth versus first quartile HRs=0.82–0.84; P-value −5). Further adjustment for years from blood collection to cancer diagnosis, body mass index, smoking intensity and duration, and serum total and high-density lipoprotein cholesterol did not alter the results. Participants with a higher metabolic score based on the discovery set had an elevated risk of prostate cancer-specific mortality in the replication set (fourth versus first quartile, HR=3.9, P-value for trend Conclusions The metabolic traits identified in this study, including for choline, glutamate, arachidonate, gamma-glutamyl amino acids, fibrinopeptides, and endocannabinoid and redox pathways and their composite risk score, corroborate our previous analysis of fatal prostate cancer and provide novel insights and potential leads regarding the molecular basis of prostate cancer progression and mortality.

The beneficial effects of dietary β-carotene and vitamin A on Parkinson disease (PD) have been confirmed, but some studies have yielded questionable results. Therefore, this meta-analysis investigated the effect of dietary β-carotene and vitamin A on the risk of PD.The following databases were searched for relevant paper: PubMed, Embase, Medline, Scopus, Cochrane Library, CNKI, Wanfang Med online, and Weipu databases for the relevant paper from 1990 to March 28, 2022. The studies included were as follows: β-carotene and vitamin A intake was measured using scientifically recognized approaches, such as food frequency questionnaire (FFQ); evaluation of odds ratios using OR, RR, or HR; β-carotene and vitamin A intake for three or more quantitative categories; and PD diagnosed by a neurologist or hospital records.This study included 11 studies (four cohort studies, six case-control studies, and one cross-sectional study). The high β-carotene intake was associated with a significantly lower chance of developing PD than low β-carotene intake (pooled OR = 0.83, 95%CI = 0.74-0.94). Whereas the risk of advancement of PD was not significantly distinctive among the highest and lowest vitamin A intake (pooled OR = 1.08, 95%CI = 0.91-1.29).Dietary β-carotene intake may have a protective effect against PD, whereas dietary vitamin A does not appear to have the same effect. More relevant studies are needed to include into meta-analysis in the further, as the recall bias and selection bias in retrospective and cross-sectional studies cause misclassifications in the assessment of nutrient intake.

A novel approach to producing high-purity fucoxanthinol (FXOH) was exploited as a sustainable method to maximize fucoxanthin (FX) utilization. Through fusing the genes of cholesterol esterase and SpyTag and then expressing them in Escherichia coli, the fusion chimera was self-assembled into insoluble active aggregates by SpyTag, which could be regarded as carrier-free immobilization. The immobilization yield of the active cholesterol esterase aggregates could reach 60%. They have expressed good activity retention at 92.48% and 60.13% after 3 and 12 cycles, respectively, which is an exciting finding. The conversion ratio of FX to FXOH is 95.02%, which is remarkably higher than those realized via the conventional chemical reduction method (55.86%) and the enzymatic hydrolysis method by free cholesterol esterases (84.51%). The purity of FXOH obtained by this method is as high as 98%, which is much higher than those obtained by other methods. Thus, a promising method for simultaneously purifying and immobilizing active cholesterol esterase aggregates is demonstrated in this study by SpyTag tailoring. In addition, this study provides an eco-friendly method for producing high-purity FXOH from FX in a highly efficient manner.

Recent studies indicated a strong relationship between carotenoids and gut microflora. However, their structure-activity relationship remains unclear. This study evaluated the interaction between four typical carotenoids (β-carotene, lutein, lycopene, and astaxanthin) and gut microflora using an

Epidemiological studies reported that resilience, generally regarded as the ability to manage stress in the face of adversity, correlates with mental health in middle-aged and older adults. Currently, there is limited information on eating habits that affect resilience. Therefore, this cross-sectional study investigated the relationship between vitamin intake and resilience based on sex in community-dwelling middle-aged and older individuals in Shika town, Ishikawa Prefecture, Japan. A total of 221 participants (106 men and 115 women) aged 40 years or older were included in the analysis. We assessed vitamin intake and resilience using a brief-type self-administered diet history questionnaire (BDHQ) and the resilience scale (RS), respectively. A two-way analysis of covariance (ANCOVA) revealed that higher intakes of β-carotene and vitamin K were associated with higher RS in women, but not in men. Furthermore, a multiple logistic regression analysis stratified by sex showed that β-carotene and vitamin K were significant independent variables for RS only in women. The present study suggests that higher intakes of β-carotene and vitamin K were associated with higher resilience among middle-aged and older women. The results obtained demonstrate that β-carotene and vitamin K intakes may enhance resilience by strengthening stress tolerance.

In this study, we utilized different types of particles to stabilize β-carotene-loaded Pickering emulsions: spherical hydrophobic zein colloidal particles (ZCPs) (517.3 nm) and rod-shaped hydrophilic cellulose nanocrystals (CNCs) (115.2 nm). Either of the particles was incapable of stabilizing Pickering emulsions owing to their inappropriate wettability. When the mass ratio of ZCPs and CNCs was 1:4, the Pickering emulsion showed the best physical and photothermal stability. Compared to the ZCP-stabilized Pickering emulsion (9.29%), the retention rate of β-carotene in the Pickering emulsion costabilized by ZCPs and CNCs was increased to 60.23% after 28 days of storage at 55 °C. Confocal microscopy and cryoscanning electron microscopy confirmed that different types of particles could form a multilayered structure or induce the formation of an interparticle network. Furthermore, the complexation of ZCPs and CNCs delayed the lipolysis of the emulsion during in vitro digestion. The free fatty acid (FFA) release rate of Pickering emulsions in the small intestinal phase was reduced from 19.46 to 8.73%. Accordingly, the bioaccessibility of β-carotene in Pickering emulsions ranged from 9.14 to 27.25% through adjusting the mass ratio and addition sequence of distinct particles at the interface. The Pickering emulsion with the novel particle-particle complex interface was designed in foods and pharmaceuticals for purpose of enhanced stability, delayed lipolysis, or sustained nutrient release.

Microalgae are a potential source of a wide range of food and novel value-added products. The versatility of microalgae to produce different kind of pigments is gaining interest as a sustainable source of natural carotenoids. Currently, commercial production of carotenoids from selected microalgae requires special culture conditions which are difficult to maintain. The present study has been undertaken to optimize culture conditions for growth and carotenoid production by a new isolate Scenedesmus quadricauda PUMCC 4.1.40. The results revealed that test organism produced 1.54 mg dry biomass/ml with a content of 40 μg carotenoids/mg dry biomass during stationary phase. The growth and carotenoid production was increased by 2.4-fold under combined optimized culture conditions. The optimized conditions were growth medium, Chu-10; pH 8.5; temperature, 30°C; nitrogen, 20 mM nitrate; phosphate, 0.22 mM; NaCl, 0.42 mM and blue light. Separation and identification of four important carotenoids through high-performance thin-layer chromatography (HPTLC) followed by purification using flash chromatography and quantification by HPLC revealed 23.8, 19.0, 6.5, and 4.0 μg astaxanthin, β-carotene, lutein, and canthaxanthin /mg dry biomass, respectively. The amount of total carotenoids (98 μg/mg dry biomass) containing 40% valuable astaxanthin and β-carotene produced under optimized conditions was significantly higher than control cultures. This justifies that S. quadricauda is a promising candidate for scale-up production of carotenoid.

The market-expanding lutein is currently mainly supplied by plant extraction, with microbial fermentation using engineered cell factory emerging as a promising substitution. During construction of lutein-producing yeast, α-carotene formation through asymmetric ε- and β-cyclization of lycopene was found as the main limiting step, attributed to intra-pathway competition of the cyclases for lycopene, forming β-carotene instead. To solve this problem, temperature-responsive expression of β-cyclase was coupled to constitutive expression of ε-cyclase for flux redirection to α-carotene by allowing ε-cyclization to occur first. Meanwhile, the ε-cyclase was engineered and re-localized to the plasma membrane for further flux reinforcement towards α-carotene. Finally, pathway extension with proper combination of carotenoid hydroxylases enabled lutein (438 μg/g dry cells) biosynthesis in S. cerevisiae. The success of heterologous lutein biosynthesis in yeast suggested temporospatial pathway control as a potential strategy in solving intra-pathway competitions, and may also be applicable for promoting the biosynthesis of other natural products.

In this work, the effects of arginine (Arg) on biochemical responses and antioxidant enzyme activity in the green microalga Dunaliella salina grown at different salt concentrations were investigated. Suspensions adapted with the concentrations of 1, 2, and 3 M NaCl were treated at the exponential growth phase with a concentration of 5 mM Arg. Salt stress was associated with a large decrease in the number of cells and non-reducing sugar levels but accumulated higher amounts of chlorophyll, β-carotene, reducing sugar, starch, total protein, free amino acid, and glycerol. Increased levels of protein carbonylation, lipid peroxidation, proteolysis, hydrogen peroxide, and antioxidant enzyme activity also occurred during salinity. Arg treatment changed the pattern of biochemical responses in the cells grown at high salinity by directing carbon flow to the biosynthesis of non-reducing sugars instead of starch, lowering levels of hydrogen peroxide, and downregulating antioxidant enzyme activity, but the levels of lipid peroxidation, glycerol, and β-carotene remained nearly unchanged. These results suggest that Arg treatment alleviates salinity-induced oxidative stress in D. salina cells by modifying carbon partitioning and inducing signaling molecules rather than antioxidant enzymes.

CRISPR Cas12a is an RNA-programmable endonuclease particularly suitable for gene regulation. This is due to its preference for T-rich PAMs that allows it to more easily target AT-rich promoter sequences, and built-in RNase activity which can process a single CRISPR RNA array encoding multiple spacers into individual guide RNAs (gRNAs), thereby simplifying multiplexed gene regulation. Here, we develop a flexible dCas12a-based CRISPRi system for Saccharomyces cerevisiae and systematically evaluate its design features. This includes the role of the NLS position, use of repression domains, and the position of the gRNA target. Our optimal system is comprised of dCas12a E925A with a single C-terminal NLS and a Mxi1 or a MIG1 repression domain, which enables up to 97% downregulation of a reporter gene. We also extend this system to allow for inducible regulation via an RNAP II-controlled promoter, demonstrate position-dependent effects in crRNA arrays, and use multiplexed regulation to stringently control a heterologous β-carotene pathway. Together these findings offer valuable insights into the design constraints of dCas12a-based CRISPRi and enable new avenues for flexible and efficient gene regulation in S. cerevisiae.

Engineered cyanobacterium Synechococcus elongatus can use light and CO2 to produce sucrose, making it a promising candidate for use in co-cultures with heterotrophic workhorses. However, this process is challenged by the mutual stresses generated from the multispecies microbial culture. Here we demonstrate an ecosystem where S. elongatus is freely grown in a photo-bioreactor (PBR) containing an engineered heterotrophic workhorse (either β-carotene-producing Yarrowia lipolytica or indigoidine-producing Pseudomonas putida) encapsulated in calcium-alginate hydrogel beads. The encapsulation prevents growth interference, allowing the cyanobacterial culture to produce high sucrose concentrations enabling the production of indigoidine and β-carotene in the heterotroph. Our experimental PBRs yielded an indigoidine titer of 7.5 g/L hydrogel and a β-carotene titer of 1.3 g/L hydrogel, amounts 15–22-fold higher than in a comparable co-culture without encapsulation. Moreover, 13C-metabolite analysis and protein overexpression tests indicated that the hydrogel beads provided a favorable microenvironment where the cell metabolism inside the hydrogel was comparable to that in a free culture. Finally, the heterotroph-containing hydrogels were easily harvested and dissolved by EDTA for product recovery, while the cyanobacterial culture itself could be reused for the next batch of immobilized heterotrophs. This co-cultivation and hydrogel encapsulation system is a successful demonstration of bioprocess optimization under photobioreactor conditions.

Carotenoids are bioactive molecules known to promote human health. Many extreme halophilic archaea synthesize carotenoids, mainly represented by C

In this study, the effect of media composition, N/P ratio and cultivation strategy on the formation of carotenoids in a

The apple fruit is one of the most widely cultivated temperate fruits and a rich source of bioactive compounds. Since a growing number of consumers are increasingly seeking safe and healthy food production, organic apple production provides this possibility. In this study, the content of bioactive compounds in organic apples depending on temperature and storage time were determined. 'Chopin' apples were stored for 7, 14 and 21 days at 5 °C and 20 °C. All analyses were performed using the HPLC method. The control samples of fresh apples contained the highest content of vitamin C (21.87 mg/100 g FW), L-ascorbic (11.50 mg/100 g FW), DHA (10.37 mg/100 g FW), total carotenoids (2.82 mg/100 g FW), β-carotene (0.21 mg/100 g FW) and lutein (2.41 mg/100 g FW). Samples stored at 20 °C had the highest content of total carotenoids and lutein, but samples stored at 5 °C contained the most β-carotene and zeaxanthin. Apples stored at 20 °C contained significantly more chlorophyll a (0.05 g/100 g FW). The samples stored at 5 °C contained the most total polyphenols. Samples stored for 14 days were characterized by the highest content of total flavonoids. Samples stored for 21 days were characterized by the highest content of epigallocatechin and quercetin-3-O-rutinoside, but the highest content of quercetin and kaempferol was found in control samples. The highest content of total flavonoids was found in apples stored at 5 °C.