Bixin

Ethanol-Based Extraction of Annatto (Bixa Orellana L.) and Characterization of the Bixin and Norbixin.[Pubmed:38680327]

ACS Omega. 2024 Apr 13;9(16):18273-18277.

Bixa orellana dye was extracted from the seeds by the method of Soxhlet extraction using ethanol where a percentage yield of 31.4 was obtained. Recrystallization of the dye was performed using chloroform and ethanol to increase the degree of purity. Thin layer chromatography was used to separate the dye into Bixin and NorBixin with R(f) values of 0.56 and 0.42, respectively. The extracted dye was pH sensitive, insoluble in water but soluble in some organic solvents like ethanol and acetic acid. Characterization by UV-vis spectroscopy showed maximum absorption for Bixin and NorBixin at 457 and 453 nm, respectively. Fourier transform infrared spectra of Bixin and NorBixin showed bands confirming similar functional groups for both Bixin and NorBixin except for -CO- stretching and methylene vibration which was absent in NorBixin. With the yield, purity level, and other properties of the extracted dye, the comparative advantages of ethanol-based Soxhlet extraction of the dye were well established.

Study on synergistic effects of curcumin and bixin against foodborne pathogens.[Pubmed:38628195]

Food Sci Nutr. 2024 Feb 22;12(4):2426-2435.

Various studies have shown that natural colorants, in addition to their coloring attributes, have valuable biological effects such as antioxidant, anti-inflammation, and anticarcinogenic properties. Moreover, their use as a food colorant can restrict the potential disadvantages of synthetic additives and turn foods into functional products. In this study, in vitro antimicrobial activities of two natural colorants of Bixin and curcumin against some important foodborne pathogens: Staphylococcus aureus (S. aureus), Listeria innocua (L. innocua), and Escherichia coli (E. coli) were investigated by disk diffusion method. Minimum inhibitory concentration and minimum bactericidal concentration values were determined by agar dilution and broth microdilution methods. The synergistic activity of the colorants against selected microorganisms was assayed by the checkerboard microdilution method. The results showed that the inhibitory effects of Bixin against S. aureus were more pronounced than E. coli and L. innocua. The lowest concentration of curcumin (0.6 mg/mL) in the disk diffusion method was not inhibited by any tested bacteria. However, it was effective at the higher concentrations against three microorganisms, but its diameter of inhibition zones was lower than gentamicin in all concentrations. Synergetic effects were observed by curcumin and Bixin combination against S. aureus (FICI

Sustainable appraisal of lipstick tree seeds (Bixa orellana)-based bixin natural orange colorant for green mordanted silk fabrics and wool yarns.[Pubmed:38614468]

Sci Prog. 2024 Apr-Jun;107(2):368504241242282.

This research aims to optimize the silk and wool dyeing process using natural dyes from Bixa orellana (annatto) through response surface methodology. Central composite design experiments highlight the significant enhancement of color outcomes achieved through microwave treatment. For silk, the optimal conditions (80 degrees C for 40 min) with annatto extract yield a color strength (K/S) of 17.8588, while wool achieves a K/S of 7.5329. Introducing eco-friendly bio-mordants, such as pomegranate peel and red sumac tannins, enhances color strength. Pre-dyeing treatments with 2% red sumac, 1.5% pomegranate peel, and weld flower extracts for silk produce high color strength, with K/S values of 16.4063, 16.3784, and 12.1658, respectively. Post-dyeing, the K/S values increase to 40.1178, 17.4779, and 21.6494. Wool yarn exhibits similar improvements, with pre-dyeing K/S values of 13.1353, 13.5060, and 16.3232, escalating to 10.5892, 15.3141, and 23.4850 post-dyeing. Furthermore, this research underscores improved colorfastness properties, including notable enhancements in light, wash, and rubbing fastness for both silk fabric and wool yarn. These findings underscore the efficacy of the proposed sustainable dyeing methods, offering valuable insights for eco-friendly textile production.

Gene structure and potential regulation of the lycopene cyclase genes in Bixa orellana L.[Pubmed:38076759]

Physiol Mol Biol Plants. 2023 Oct;29(10):1423-1435.

Lycopene cyclases (LCYs) are a key branching point in regulating the carotenoid biosynthesis pathway in plants. Bixa orellana L. is characterized by the presence in its seed of Bixin, an apocarotenoid of significant importance in the food, pharmaceutical, and cosmetic industries. Gene analysis provides the opportunity to investigate the LCY gene structure in plant species and its relationship with the synthesis of carotenoids. Coding sequences of the LCY genes were retrieved from a B. orellana genome DNA. Bobeta-LCY1 and Bobeta-LCY2 genes exhibit 100% of identity to their respective cDNA accessions, and exhibit a single coding region of 1512 bp (504 aa) and 1495 bp (498 aa), respectively. In contrast, Boepsilon-LCY gene shows a coding region of 1581 bp (527 aa) with 10 introns of diverse lengths. Putative Transcription Factors (TFs) binding sites were upstream (3000 bp) identified for each LCY gene. TFs cover two groups, one with the categories of photosynthesis, reproduction, and oxidative processes that are frequent. The second one with the categories of defense, cell cycle, signaling, and carbohydrate metabolism, which are poorly represented. Besides, repetitive DNA elements showed motifs and proteins related to LTR from the Ty3/Gypsy family, were associated with the TFs regions. In general, TFs vary in the different BoLCY genes, being more abundant in the Boepsilon-LCY gene. LCY expression analyzed from a transcriptome database, and validated by RT-qPCR, shows an upregulation of the three LCYs, mainly oriented to the synthesis of essential carotenoids in photosynthetic tissues (leaves), as well as an upregulation of the Bobeta-LCY2 gene in the non-photosynthetic tissues (firsts seed development stages) related to the Bixin accumulation. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-023-01384-8.

Comprehensive Update on Carotenoid Colorants from Plants and Microalgae: Challenges and Advances from Research Laboratories to Industry.[Pubmed:38002140]

Foods. 2023 Nov 10;12(22):4080.

The substitution of synthetic food dyes with natural colorants continues to be assiduously pursued. The current list of natural carotenoid colorants consists of plant-derived annatto (Bixin and norBixin), paprika (capsanthin and capsorubin), saffron (crocin), tomato and gac fruit lycopene, marigold lutein, and red palm oil (alpha- and beta-carotene), along with microalgal Dunaliella beta-carotene and Haematococcus astaxanthin and fungal Blakeslea trispora beta-carotene and lycopene. Potential microalgal sources are being sought, especially in relation to lutein, for which commercial plant sources are lacking. Research efforts, manifested in numerous reviews and research papers published in the last decade, have been directed to green extraction, microencapsulation/nanoencapsulation, and valorization of processing by-products. Extraction is shifting from conventional extraction with organic solvents to supercritical CO(2) extraction and different types of assisted extraction. Initially intended for the stabilization of the highly degradable carotenoids, additional benefits of encapsulation have been demonstrated, especially the improvement of carotenoid solubility and bioavailability. Instead of searching for new higher plant sources, enormous effort has been directed to the utilization of by-products of the fruit and vegetable processing industry, with the application of biorefinery and circular economy concepts. Amidst enormous research activities, however, the gap between research and industrial implementation remains wide.

Plant age-dependent dynamics of annatto pigment (bixin) biosynthesis in Bixa orellana.[Pubmed:37975812]

J Exp Bot. 2024 Feb 28;75(5):1390-1406.

Age affects the production of secondary metabolites, but how developmental cues regulate secondary metabolism remains poorly understood. The achiote tree (Bixa orellana L.) is a source of Bixin, an apocarotenoid used in diverse industries worldwide. Understanding how age-dependent mechanisms control Bixin biosynthesis is of great interest for plant biology and for economic reasons. Here we overexpressed miRNA156 (miR156) in B. orellana to comprehensively study the effects of the miR156-SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) module on age-dependent Bixin biosynthesis in leaves. Overexpression of miR156 in annatto plants (miR156ox) reduced BoSPL transcript levels, impacted leaf ontogeny, lessened Bixin production, and increased abscisic acid levels. Modulation of expression of BoCCD4-4 and BoCCD1, key genes in carotenoid biosynthesis, was associated with diverting the carbon flux from Bixin to abscisic acid in miR156ox leaves. Proteomic analyses revealed an overall low accumulation of most secondary metabolite-related enzymes in miR156ox leaves, suggesting that miR156-targeted BoSPLs may be required to activate several secondary metabolic pathways. Our findings suggest that the conserved BomiR156-BoSPL module is deployed to regulate leaf dynamics of Bixin biosynthesis, and may create novel opportunities to fine-tune Bixin output in B. orellana breeding programs.

Validation, Measurement Uncertainty, and Determination of Bixin and Norbixin in Processed Foods of Animal Resources Distributed in Korea.[Pubmed:37969328]

Food Sci Anim Resour. 2023 Nov;43(6):949-960.

This research aimed to validate a high-performance liquid chromatography method for the quantitative determination of Bixin and norBixin in various foods. The Diode Array Detector (495 nm) technique was used. Method was validated for specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy, and the measurement uncertainty was assessed. The calibration curve showed excellent linearity (r(2)>/=0.9999) over the tested concentration range of 0.2-25 mg/L. The LOD and LOQ were 0.03-0.11 and 0.02-0.05 mg/L for Bixin and norBixin, respectively. The intra- and inter-day accuracies and precisions were 88.0+/-1.3-97.0+/-0.5% and 0.2%-2.6% relative SD (RSD) for Bixin and 88.2+/-0.8-105.8+/-0.8% and 0.3%-2.7% RSD for norBixin, respectively. Inter-laboratory validation for accuracy and precision was conducted in three laboratories, and these results all met the AOAC guidelines. In addition, the relative expanded uncertainty (<22%) satisfied the CODEX recommendation. Furthermore, products distributed in Korea were monitored for annatto extracts using the proposed method to demonstrate its application. The developed analytical method is reliable for quantifying Bixin and norBixin in various foods.

Exploring bixin from Bixa orellana L. seeds: quantification and in silico insights into its anti-cancer potential.[Pubmed:37837422]

J Biomol Struct Dyn. 2023 Oct 14:1-15.

Bixin, the key pigment of Bixa orellana L., is an apo-carotenoid found in the seed arils. The present study aimed to quantitatively determine the Bixin content of seeds and explore its anti-cancer activity through in silico studies. The Bixin content from the seeds of the local genotype, TNMTP8, quantified by RP-HPLC was 4.58 mg per gram. The prediction of pharmacological activity suggested that Bixin may serve as a BRAF, MMP9, TNF expression inhibitors, and TP53 expression enhancer. According to molecular docking analysis, Bixin interacted with eight different skin cancer targets and had the lowest binding energy compared to the standard drug, 5-fluorouracil. The binding score between Bixin and the targets ranged from -4.7 to -8.7 kcal/mol. The targets BRAF and SIRT3 interacted well with Bixin, with binding energies as low as -8.3 and -8.7 kcal/mol, respectively. Hence, the dynamic behavior of these two docked complexes throughout a 500 ns trajectory run was investigated further. The Root Mean Square Deviation (RMSD), Root Mean Square Fluctuation (RMSF) values, and total contacts as a function of time recorded during scrutiny suggest that both complexes were stable. This was validated by post-molecular dynamics analysis using Molecular Mechanics Generalized Born Surface Area (MM-GBSA). Principal component analysis (PCA) was used to analyze the significant differences in motion exhibited by BRAF-Bixin and SIRT3-Bixin. The results showed that Bixin is a promising source for potential treatment interventions in skin cancer therapies.Communicated by Ramaswamy H. Sarma.

Therapeutic potential of bixin on inflammation: a mini review.[Pubmed:37781110]

Front Nutr. 2023 Sep 13;10:1209248.

Chronic inflammation is the underlying mechanism for many diseases. Thus, inflammatory signaling pathways are valuable targets for new treatment modalities. Natural products have gained interest as a potential source of bioactive compounds which provide health benefits in combating inflammatory-related diseases. Recent reports have linked the medicinal values of Bixa orellana L. with its anti-inflammatory activities. Therefore, this review aims to examine the therapeutic potential of Bixin, a major bioactive constituent found in the seeds of B. orellana, on inflammatory-related diseases based on existing in vitro and in vivo evidence. Additionally, the anti-inflammatory mechanism of Bixin via signaling pathways is explored and possible toxic effects are addressed. The findings suggest that Bixin may ameliorate inflammation via inhibition of toll-like receptor 4/nuclear factor-kappa B (TLR4/NF-kappaB), phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) and thioredoxin-interacting protein/NOD-like receptor protein 3 (TXNIP/NLRP3) inflammasome mechanisms. More well-planned clinical studies should be performed to verify its effectiveness and safety profile.

Dietary carotenoids to improve hypertension.[Pubmed:37662767]

Heliyon. 2023 Aug 25;9(9):e19399.

Hypertension is one of the major risk factors for cardiovascular diseases and the main reason for premature death in older adults. Although antihypertensive medications have been used frequently, hypertension prevalence has increased in the last decade. Lifestyle improvement is a cornerstone of hypertension prevention and control. High dietary consumptions of fruits and vegetables are linked to reduced risks of high blood pressure. Carotenoids are natural tetraterpene pigments produced by bacteria, fungi, algae, some animals, and various plants. Because of their high pharmacological potential and safety, they have been mentioned as unique therapeutic agents for a diverse range of diseases. Carotenoids modulate high blood pressure. They also have several additional benefits for the cardiovascular system, including antioxidative, anti-inflammatory, anti-atherogenic, and antiplatelet effects. They improve endothelial function and metabolic profile, as well. In the present article, we reviewed the literature data regarding carotenoids' influence on hypertension in both preclinical and clinical studies. Furthermore, we reviewed the underlying mechanisms associated with antihypertensive properties derived from in vitro and in vivo studies. Suppressing reactive oxygen species (ROS) production, Inhibiting angiotensin-II, endothelin-1, and oxidized low-density lipoprotein; and also nitric oxide enhancement are some of the mechanisms by which they lower blood pressure. The present article indicated that astaxanthine, beta-carotene, Bixin, capsanthin, lutein, crocin, and lycopene have antihypertensive properties. Having significant antioxidant properties, they can decrease high blood pressure and concomitant comorbidities.

The role of bixin as antioxidant, anti-inflammatory, anticancer, and skin protecting natural product extracted from Bixa orellana L.[Pubmed:37454777]

Fitoterapia. 2023 Sep;169:105612.

Since long, medicinal plants or herbs are being used in different traditional treatment systems as therapeutic agents to treat a variety of illnesses. Bixa orellana L., an medicinal plant (family: Bixaceae), is an Ayurvedic herb used to treat dyslipidemia, diarrhoea, and hepatitis since ancient times. B. orellana L., seeds contain an orange-red coloured component known as Bixin (C(25)H(30)O(4)), which constitutes 80% of the extract.Chemically, Bixin is a natural apocarotenoid, biosynthesized through the oxidative degradation of C40 carotenoids. Bixin helps to regulate the Nrf2/MyD88/TLR4 and TGF-1/PPAR-/Smad3 pathways, which further give it antifibrosis, antioxidant, and anti-inflammatory properties. This current review article presents a comprehensive review of Bixin as an anti-inflammatory, antioxidant, anticancer,and skin protecting natural product. In addition, the biosynthesis and molecular target of Bixin, along with Bixin extraction techniques, are also presented.

Bixin and Fuxoxanthin Alone and in Combination with Cisplatin Regulate ABCC1 and ABCC2 Transcription in A549 Lung Cancer Cells.[Pubmed:37313537]

J Pharm Bioallied Sci. 2023 Jan-Mar;15(1):15-20.

BACKGROUND: The ATP-Binding Cassette (ABC) transporter has long been studied to confer drug resistance in human tumors and play important role in metabolic processes and cellular signaling. The overexpression of ABCB1, ABCC1, ABCC2, ABCC3, and ABCG2 leads to decreased sensitivity of lung cancer to cisplatin. At the transcription level, the expression of ABC transporters is highly regulated and requires the complex interplay of factors involved in differentiation and development, cell survival and apoptosis upon intrinsic and environmental stress. The p53 regulation of drug-resistance genes is also complex yet not well understood. Previously, we demonstrated the synergistic interaction between Bixin or fucoxanthin with cisplatin in A549 lung cancer cells. OBJECTIVES: Current study aims to identify whether carotenoids enhancing therapeutic effect of Cisplatin due to the ability to reverse drug resistance associated proteins, such as ABC transporter and regulating the tumor suppressor corresponding gene, p53. METHODS: Real-Time Quantitative-Polymerase Chain Reaction (RT-qPCR) was performed to estimate the expression level of ABCC1 and ABCC2, and p53 of A549 cell lines in response to carotenoids alone and in combination with cisplatin. RESULTS AND CONCLUSION: The administration of Bixin or fucoxanthin decreases the expression of ABCC1 and ABCC2. Both carotenoids, either alone or in combination with cisplatin, upregulated p53 gene expression indicating the mechanism of proliferation inhibition and apoptosis occurs via the p53 caspase-independent pathway.

Aqueous extract from urucum (Bixa orellana L.): antimicrobial, antioxidant, and healing activity.[Pubmed:37213245]

Porto Biomed J. 2023 Feb 7;8(1):e183.

Background: Annatto was obtained from seed B orellana (urucum) and is commonly used in food and cosmetic industries. The objective of this study was to identify the antimicrobial and antioxidant activity of the aqueous extract from urucum seeds and its skin healing potential in exposed cutaneous lesions in rats treated with the gel containing the extract. Methods: Three types of extracts from seeds were made using chloroform, sodium hydroxide, water, and estimated Bixin and norBixin. In the presence of antioxidants, antibacterial was observed and then evaluated the skin healing in rats using aqueous extract. Results: Annatto dyes have been evaluated in all three extracts. When the seeds were extracted with chloroform, Bixin was detected. If extraction was performed by sodium hydroxide or water, norBixin was detected. For healing use, 10% of aqueous extract was mixed in a gel base. The finding obtained from the antioxidant assay revealed that the activities of the water extract could be used as a source of polyphenolic compounds. In chloroform extract, the antioxidant was not effective because it has weak radical scavengers. With respect to antimicrobial activity, it has been observed that aqueous extract has more effect. For skin healing assay, a total of 3 study groups were tested: negative control group (gel base), positive control group (fibrinase), and test group (gel with urucum aqueous extract). After 7 days of treatment, animals treated with fibrinase had an improvement of 4.7% in total wound area when compared with the negative control while those treated with urucum aqueous extract presented an improvement of 51.55% in comparison. After 14 days, the total wound area of animals within the test group had a decrease of 94.97% when compared with the negative control (gel base) results while the control group presented an improvement of 56.58% in total wound area. These results indicate that wounds treated with urucum aqueous extract were 38.39% more efficient than fibrinase, a cream used for skin healing. Conclusions: It is possible to conclude that gel with aqueous extract is effective in skin healing in rats, being used as a phytotherapic, besides possessing antioxidant and antimicrobial activity.

Ultra-small magnetic Candida antarctica lipase B nanoreactors for enzyme synthesis of bixin-maltitol ester.[Pubmed:37094396]

Food Chem. 2023 Sep 30;421:136132.

Bixin has desirable bioactivities but poor water solubility, which limits its practical applications. Enzymatic transesterification of methyl to alditol groups in Bixin by Candida antarctica lipase B (CALB) improves Bixin water solubility. Herein, magnetic CALB nanoreactors with diameter of 11.7 nm and CALB layer thickness of 3.5 nm were developed by covalently linking CALB onto silicon covered Fe(3)O(4) nanoparticles. The CALB loading capacity in nanoreactors achieved 30%. The Michaelis constant (Km) and maximum reaction rate of magnetic CALB nanoreactors were 56.1 mmol/L and 0.2 mmol/(L.min). Magnetic CALB nanoreactors could circularly catalyze Bixin-maltitol ester synthesis and keep catalytic efficiency of 62.6% after eight repetitive enzymatic reactions. Additionally, the optimal Bixin-maltitol ester synthesis procedure was heating Bixin-maltitol mixture at molar ratio of 1:7 in anhydrous 2-methyl-2-butanol-dimethylsulfoxide (8:2, v/v) at 50 degrees C for 24 h. Bixin-maltitol ester showed improved water solubility at pH 5.5 and 7.0.