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1.
Int J Mol Sci ; 25(13)2024 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-39000258

RESUMO

Currently, there is a dearth of in-depth analysis and research on the impact of canthaxanthin on the production performance, egg quality, physical characteristics, and offspring health of laying hens. Furthermore, the metabolic mechanism of cantharidin in the body remains unclear. Therefore, to solve the above issues in detail, our study was conducted with a control group (C group), a low-dose canthaxanthin group (L group), and a high-dose canthaxanthin group (H group), each fed for a period of 40 days. Production performance was monitored during the experiment, in which L and H groups showed a significant increase in ADFI. Eggs were collected for quality analysis, revealing no significant differences in qualities except for yolk color (YC). The YC of the C group almost did not change, ranging from 6.08 to 6.20; however, the trend in YC change in other groups showed an initial intense increase, followed by a decrease, and eventually reached dynamic equilibrium. By detecting the content of canthaxanthin in the yolk, the YC change trend was found to be correlated with canthaxanthin levels in the yolk. The content of unsaturated fatty acid increased slightly in L and H groups. Following the incubation period, the physical characteristics and blood biochemical indices of chicks were evaluated. It was observed that the shank color of chicks in the L and H groups was significantly higher than that in the C group at birth. However, by the 35th day, there were no significant differences in shank color among the three groups. Further investigation into the metabolic mechanism involving canthaxanthin revealed that the substance underwent incomplete metabolism upon entering the body, resulting in its accumulation as well as metabolic by-product accumulation in the yolk. In summary, this study highlighted the importance of understanding canthaxanthin's role in production performance, egg quality, and offspring health, providing valuable insights for breeders to optimize feeding strategies.


Assuntos
Cantaxantina , Galinhas , Gema de Ovo , Animais , Cantaxantina/metabolismo , Feminino , Gema de Ovo/metabolismo , Gema de Ovo/química , Ovos/análise , Ração Animal/análise , Dieta/veterinária , Suplementos Nutricionais
2.
Photosynth Res ; 156(1): 3-17, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36063303

RESUMO

Our analysis of the X-ray crystal structure of canthaxanthin (CAN) showed that its ketolated ß-ionone rings can adopt two energetically equal, but structurally distinct puckers. Quantum chemistry calculations revealed that the potential energy surface of the ß-ionone ring rotation over the plane of the conjugated π-system in carotenoids depends on the pucker state of the ß-ring. Considering different pucker states and ß-ionone ring rotation, we found six separate local minima on the potential energy surface defining the geometry of the keto-ß-ionone ring-two cis and one trans orientation for each of two pucker states. We observed a small difference in energy and no difference in relative orientation for the cis-minima, but a pronounced difference for the position of trans-minimum in alternative pucker configurations. An energetic advantage of ß-ionone ring rotation from a specific pucker type can reach up to 8 kJ/mol ([Formula: see text]). In addition, we performed the simulation of linear absorption of CAN in hexane and in a unit cell of the CAN crystal. The electronic energies of [Formula: see text] transition were estimated both for the CAN monomer and in the CAN crystal. The difference between them reached [Formula: see text], which roughly corresponds to the energy gap between A and B pucker states predicted by theoretical estimations. Finally, we have discussed the importance of such effects for biological systems whose local environment determines conformational mobility, and optical/functional characteristics of carotenoid.


Assuntos
Carotenoides , Norisoprenoides , Carotenoides/química , Norisoprenoides/química , Conformação Molecular , Cantaxantina
3.
Phys Chem Chem Phys ; 25(48): 33000-33012, 2023 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-38032096

RESUMO

The orange carotenoid protein (OCP) functions as a sensor of the ambient light intensity and as a quencher of bilin excitons when it binds to the core of the cyanobacterial phycobilisome. We show herein that the photoactivation mechanism that converts the resting, orange-colored state, OCPO, to the active red-colored state, OCPR, requires a sequence of two reactions, each requiring absorption of a single photon by an intrinsic ketocarotenoid chromophore. Global analysis of absorption spectra recorded during continuous illumination of OCPO preparations from Synechocystis sp. PCC 6803 detects the reversible formation of a metastable intermediate, OCPI, in which the ketocarotenoid canthaxanthin exhibits an absorption spectrum with a partial red shift and a broadened vibronic structure compared to that of the OCPO state. While the dark recovery from OCPR to OCPI is a first-order, unimolecular reaction, the subsequent conversion of OCPI to the resting OCPO state is bimolecular, involving association of two OCPO monomers to form the dark-stable OCPO dimer aggregate. These results indicate that photodissociation of the OCPO dimer to form the monomeric OCPO intermediate is the first step in the photoactivation mechanism. Formation of the OCPO monomer from the dimer increases the mean value and broadens the distribution of the solvent-accessible surface area of the canthaxanthin chromophore measured in molecular dynamics trajectories at 300 K. The second step in the photoactivation mechanism is initiated by absorption of a second photon, by canthaxanthin in the OCPO monomer, which obtains the fully red-shifted and broadened absorption spectrum detected in the OCPR product state owing to displacement of the C-terminal domain and the translocation of canthaxanthin more than 12 Å into the N-terminal domain. Both steps in the photoactivation reaction of OCP are likely to involve changes in the structure of the C-terminal domain elicited by excited-state conformational motions of the ketocarotenoid.


Assuntos
Proteínas de Bactérias , Synechocystis , Proteínas de Bactérias/química , Cantaxantina , Luz , Synechocystis/metabolismo , Carotenoides/química
4.
J Fish Biol ; 103(1): 13-21, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37060354

RESUMO

Colouring has a great influence on the commercialization of ornamental fish. The aim of this study was to evaluate different concentrations of canthaxanthin in the diet of the blood swordtail Xiphophorus helleri in an effort to obtain a more intense red colour. Six concentrations of canthaxanthin (0, 50, 100, 250, 400 and 600 mg kg-1 diet) were used. The experiment lasted 60 days. Fish were evaluated for increased red pigmentation through the use of photographs (performed by smartphone) and imaging applications considering the Hunter method and the CMYK and productive performance. No significant differences were observed for productive performance. The use of photographs by means of a smartphone and the use of imaging applications proved to be adequate to assess differences in colour in the species. Swordtail red pigmentation did not show significant increase regardless of canthaxanthin dosages. Varieties of ornamental fish bred for red coloration may have limits for increased colour due to the storage capacity of carotenoids by chromatophores.


Assuntos
Cantaxantina , Ciprinodontiformes , Animais , Cor , Carotenoides , Dieta/veterinária
5.
Molecules ; 28(7)2023 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-37050034

RESUMO

Green alga Caulerpa racemosa is an underexploited species of macroalgae, even though it is characterized by a green color that indicates an abundance of bioactive pigments, such as chlorophyll and possibly xanthophyll. Unlike chlorophyll, which has been well explored, the composition of the carotenoids of C. racemosa and its biological activities have not been reported. Therefore, this study aims to look at the carotenoid profile and composition of C. racemose and determine their biological activities, which include antidiabetic, anti-obesity, anti-oxidative, anti-inflammatory, and cytotoxicity in vitro. The detected carotenoids were all xanthophylls, which included fucoxanthin, lutein, astaxanthin, canthaxanthin, zeaxanthin, ß-carotene, and ß-cryptoxanthin based on orbitrap-mass spectrometry (MS) and a rapid ultra-high performance liquid chromatography (UHPLC) diode array detector. Of the seven carotenoids observed, it should be highlighted that ß-carotene and canthaxanthin were the two most dominant carotenoids present in C. racemosa. Interestingly, the carotenoid extract of C. racemosa has good biological activity in inhibiting α-glucosidase, α-amylase, DPPH and ABTS, and the TNF-α and mTOR, as well as upregulating the AMPK, which makes it a drug candidate or functional antidiabetic food, a very promising anti-obesity and anti-inflammatory. More interestingly, the cytotoxicity value of the carotenoid extract of C. racemosa shows a level of safety in normal cells, which makes it a potential for the further development of nutraceuticals and pharmaceuticals.


Assuntos
Caulerpa , Clorófitas , Carotenoides/química , Antioxidantes/química , beta Caroteno/química , Cantaxantina , Hipoglicemiantes/farmacologia , Luteína/química , Zeaxantinas , Anti-Inflamatórios/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/química
6.
J Anim Physiol Anim Nutr (Berl) ; 107(2): 650-658, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-35534942

RESUMO

This study examined the impacts of dietary spirulina (SP) and canthaxanthin (CX) supplementation to laying hens on reproductive performance and cock's semen quality evaluation. Three hundred and sixty of indigenous Sinai chickens (300 hens + 60 cocks), 48-weeks-old, were randomly divided into five treatments; a control and four supplemented (0.6 g SP mg/kg, 0.8 g SP mg/kg, 6 mg CX/kg diet, and 8 mg CX/kg diet) with six replications of 10 females and 1 male per treatment for laying trial and 1 male for semen evaluation. The animals were fed with experimental diets for 12 weeks. Supplementing 6 mg CX/kg diet led to an increase in the final body weight and egg production traits, while 8 mg CX/kg diet resulted in an improving feed conversion ratio. Supplementation of 0.6 g SP mg/kg or 6.0 mg CX/kg diet resulted in the highest egg production. The heaviest egg weight and egg mass were induced by 0.8 g SP mg/kg. Eggs laid from hens fed 0.6 g SP/kg diet had a higher concentration of cholesterol than those fed 0.8 g SP/kg and those fed 6.0 and 8.0 mg CX/kg. Cocks fed 6 mg CX/kg and those fed 0.8 g SP/kg diet had the greatest sperm concentration. The supplementation of 0.8 g SP mg/kg diet or both 6 and 8 mg CX/kg diet enhanced hatchability. So, dietary spirulina (0.8 mg/kg) and canthaxanthin (6 mg/kg) supplementation to Sinai laying hens and cocks could be used to improve productive and reproductive and performance.


Assuntos
Gema de Ovo , Spirulina , Animais , Masculino , Feminino , Galinhas , Cantaxantina/farmacologia , Análise do Sêmen/veterinária , Sêmen , Óvulo , Suplementos Nutricionais/análise , Dieta/veterinária , Colesterol , Ração Animal/análise , Ovos/análise
7.
BMC Plant Biol ; 22(1): 530, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36380271

RESUMO

BACKGROUND: The rich yellow-orange to vividly deep red bark of willow (Salix spp.) branches have high ornamental and economic value. However, the mechanism underlying the regulation of willow branch color remains unknown. Therefore, we performed metabolomics and transcriptomics analyses of purple, green, and red willow barks to elucidating the mechanisms regulating color development. RESULTS: Seven anthocyanins were isolated; pelargonidin, petunidin 3-O-rutinoside, and cyanin chloride were the most abundant in red bark, whereas pelargonin chloride was most abundant in purple bark. The green bark contained the highest level of malvidin; however, the malvidin level was not significantly higher than in the red bark. The purple bark contained the largest amount of canthaxanthin, a carotenoid pigment. The integrated pathways of flavonoid biosynthesis, carotenoid biosynthesis, and porphyrin and chlorophyll metabolism were constructed for the willow barks. Among the three barks, the expression of the structural genes ANS, ANR, and BZ1, which are involved in anthocyanin synthesis, was the highest in red bark, likely causing anthocyanin accumulation. The expression of CrtZ, which participates in the carotenoid pathway, was the highest in purple bark, likely leading to canthaxanthin accumulation. The high expression of DVR, POR, and CRD1 may be associated with green pigment synthesis in the chlorophyll biosynthesis pathway. CONCLUSIONS: Purple bark color is co-regulated by anthocyanins and carotenoids, whereas red bark is characterized by anthocyanin accumulation and chlorophyll degradation. The green pigment is regulated by maintaining chlorophyll synthesis. BZ1 and CrtZ are candidate genes regulating anthocyanin and canthaxanthin accumulation in red and purple barks respectively. Collectively, our results may facilitate the genetic breeding and cultivation of colorful willows with improved color and luster.


Assuntos
Antocianinas , Regulação da Expressão Gênica de Plantas , Transcriptoma , Cantaxantina , Cloretos , Cor , Melhoramento Vegetal , Carotenoides , Clorofila
8.
Microb Cell Fact ; 21(1): 71, 2022 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-35468798

RESUMO

BACKGROUND: The bifunctional enzyme ß-carotene hydroxylase (CrtZ) catalyzes the hydroxylation of carotenoid ß-ionone rings at the 3, 3' position regardless of the presence of keto group at 4, 4' position, which is an important step in the synthesis of astaxanthin. The level and substrate preference of CrtZ may have great effect on the amount of astaxanthin and the accumulation of intermediates. RESULTS: In this study, the substrate preference of PCcrtZ from Paracoccus sp. PC1 and PAcrtZ from Pantoea Agglomerans were certified and were combined utilization for increase astaxanthin production. Firstly, PCcrtZ from Paracoccus sp. PC1 and PAcrtZ from P. Agglomerans were expressed in platform strains CAR032 (ß-carotene producing strain) and Can004 (canthaxanthin producing strain) separately to identify their substrate preference for carotenoids with keto groups at 4,4' position or not. The results showed that PCcrtZ led to a lower zeaxanthin yield in CAR032 compared to that of PAcrtZ. On the contrary, higher astaxanthin production was obtained in Can004 by PCcrtZ than that of PAcrtZ. This demonstrated that PCCrtZ has higher canthaxanthin to astaxanthin conversion ability than PACrtZ, while PACrtZ prefer using ß-carotene as substrate. Finally, Ast010, which has two copies of PAcrtZ and one copy of PCcrtZ produced 1.82 g/L of astaxanthin after 70 h of fed-batch fermentation. CONCLUSIONS: Combined utilization of crtZ genes, which have ß-carotene and canthaxanthin substrate preference respectively, can greatly enhance the production of astaxanthin and increase the ratio of astaxanthin among total carotenoids.


Assuntos
Escherichia coli , Paracoccus , Cantaxantina , Carotenoides/química , Escherichia coli/genética , Oxigenases/genética , Xantofilas , beta Caroteno
9.
Biotechnol Appl Biochem ; 69(6): 2422-2436, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34841569

RESUMO

Purple nonsulfur bacteria (PNSB) were investigated for their carotenoid production and anti-vibrio activity against acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus. To test carotenoid production, selected strains were cultivated in basic isolation medium (BIM), glutamate acetate medium, G5 medium and artificial acetic acid wastewater (AAW) medium. From 144 PNSB, Rhodopseudomonas palustris KTSSG46 was selected to produce carotenoids under microaerobic light conditions in BIM. When the culture medium was optimized, strain KTSSG46 grown in BIM modified with l-glutamate at 1 g/L more effectively inhibited AHPND-causing V. parahaemolyticus strains than standard BIM with 1 g/L (NH4 )2 SO4 . BIM was further modified with 1.23 g/L MgSO4 ·7H2 O and carotenoid production increased 40.22%. Carotenoid production at day 2 by strain KTSSG46 grown in BIM modified with l-glutamate at 1 and 1.23 g/L MgSO4 ·7H2 O was the same as production in BIM modified with monosodium glutamate (MSG). Culture supernatants from all BIM formulations showed similar activity against the resistant AHPND strain SR2. Based on high-performance liquid chromatography, carotenoids of strain KTSSG46 might be canthaxanthin. Grown in BIM modified with MSG, strain KTSSG46 could produce inexpensive carotenoids and release anti-vibrio compounds that, applied as shrimp feed additive, would prevent AHPND strains.


Assuntos
Penaeidae , Vibrio parahaemolyticus , Animais , Glutamato de Sódio/farmacologia , Penaeidae/microbiologia , Doença Aguda , Cantaxantina/farmacologia , Necrose
10.
Mar Drugs ; 20(7)2022 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-35877709

RESUMO

The carotenogenic thraustochytrid Thraustochytrium sp. RT2316-16 was grown in batch and repeated-batch cultures using different feeds containing glucose, or glycerol, and yeast extract, for the production of lipids, phospholipids and carotenoids. RT2316-16 produced canthaxanthin, astaxanthin and ß-carotene. The effects of biotin, ascorbic acid, light and temperature were evaluated in some of the experiments. In 2-day-old batch cultures, the combined mass percentage of eicosapentaenoic acid and docosahexaenoic acid in total lipids was between 16.5% (glycerol-based medium in the dark; biomass concentration = 4.2 ± 1.1 g L-1) and 42.6% (glucose-based medium under light; biomass concentration = 3.3 ± 0.1 g L-1), decreasing to 3.8% and 6.1%, respectively, after day 4. In repeated-batch cultures, the total lipids in the biomass increased after glucose or glycerol was fed alone, whereas the total carotenoids (168 ± 7 µg g-1 dry weight (DW)) and phospholipids in the biomass increased after feeding with yeast extract. The biomass with the highest content of phospholipids (28.7 ± 4.3 mg g-1 DW) was obtained using a feed medium formulated with glycerol, yeast extract and ascorbic acid. Glycerol was the best carbon source for the production of a biomass enriched with total lipids (467 ± 45 mg g-1 DW). The composition of carotenoids depended strongly on the composition of the feed. Repeated-batch cultures fed with yeast extract contained canthaxanthin as the main carotenoid, whereas in the cultures fed only with glucose, the biomass contained mainly ß-carotene.


Assuntos
Técnicas de Cultura Celular por Lotes , Estramenópilas , Ácido Ascórbico , Biomassa , Cantaxantina , Carotenoides , Glucose , Glicerol , Fosfolipídeos , beta Caroteno
11.
Andrologia ; 54(1): e14304, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34773278

RESUMO

Antioxidants are used to minimize oxidative stress during liquid semen storage. The main aim of current study was to elucidate effect of supplementing melatonin and canthaxanthin in Tris-based extender could enhance seminal quality of ram at 4°C up to 72 h. A total of 48 ejaculates were collected from breeding Magra rams (n = 8) and were preliminarily subjected for various macroscopic and microscopic semen evaluation tests. These ejaculates were pooled and divided into three equal aliquots. Two aliquots were diluted (1:10) using extender encompassing final concentration of 1mM melatonin and 25 µM canthaxanthin and stored at 4°C. Third aliquot with extender only was kept as control. Structural and functional seminal changes were observed at different time points of preservation. Results revealed that mean values for progressive sperm motility, viability and total antioxidant capacity were significantly higher (p < 0.05) in melatonin group while hypo-osmotic swelling test was significantly (p < 0.05) higher in canthaxanthin group. Total sperm abnormalities and malondialdehyde levels were significantly (p < 0.05) lower in both treatment groups indicating their antioxidant efficacy in protection of spermatozoa from oxidative stress. Results of study indicated that supplementation of these antioxidants to ram semen could be used to enhance storage life of liquid semen at 4°C up to 72 h.


Assuntos
Melatonina , Preservação do Sêmen , Animais , Cantaxantina , Criopreservação , Masculino , Melatonina/farmacologia , Estresse Oxidativo , Ovinos , Motilidade dos Espermatozoides , Espermatozoides
12.
World J Microbiol Biotechnol ; 38(8): 129, 2022 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-35689122

RESUMO

The green algae Tetraspora sp. CU2551 was previously identified as a strain with high potential for biohydrogen production; however, its algal biomass characteristics changed from green to reddish orange within 43 days of biohydrogen production. The crude pigments were extracted, partially purified, and characterized by chemical determination. The present study focused on elucidating the carotenoid composition of the selected green alga Tetraspora sp. CU2551. The pigment extract was partially purified and fractionated using thin layer chromatography, and yielded two major and two minor carotenoid bands. The fractions were confirmed by high-performance liquid chromatography with a diode array detector (HPLC-DAD) before being identified and confirmed using Liquid Chromatograph-Quadrupole Time of Flight-Mass Spectrometry (LC-QTOF-MS). The spectral data of these fractions revealed four sub-fractions of interest that were lutein, canthaxanthin, neochrome, and ß-carotene, which had percentages in the crude extracts of 30.57%, 25.47%, 7.89%, and 0.71%, respectively. Lutein and canthaxanthin were found to be the major carotenoid pigments present. Our findings in this present study are the first reporting of Tetraspora sp. CU2551 as a potential alternate source for carotenoid pigment production.


Assuntos
Carotenoides , Clorófitas , Cantaxantina/análise , Carotenoides/análise , Clorófitas/química , Cromatografia Líquida de Alta Pressão/métodos , Luteína/análise , Xantofilas , beta Caroteno
13.
J Zoo Wildl Med ; 53(3): 561-572, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36214241

RESUMO

The greater flamingo (Phoenicopterus roseus) is often maintained in managed care. Although values for plasma concentrations of minerals and vitamins have been published for this species, limited studies investigate the effects of diet changes on these values. Plasma concentrations of selected vitamins (A [and the provitamin A carotenoid ß-carotene] and E), minerals (Ca, Cu, Fe, K, Mg, Mn, Na, inorganic P, Se, and Zn), and total protein (TP) were measured in a zoo collection of greater flamingos in November 2018 (n = 48) and in November 2019 (n = 52), 30 d after a change from the summer diet, which was the same in both years, to different winter diets. Chloride, cholesterol, and triglycerides were only measured once in 2019. The nutrient profiles of the two complete diets (labeled and analyzed) differed considerably, especially for the canthaxanthin concentration. The plasma concentrations of ß-carotene (P < 0.0001), Cu (P < 0.0001), Fe (P < 0.0001), Mg (P < 0.0001), and Se (P < 0.0001) were significantly higher and vitamin E (P < 0.0001), Ca (P = 0.0014), Mn (P < 0.0001), Na (P = 0.0005), and TP (P < 0.0001) were significantly lower in 2018 than in 2019. Four minerals (Cu, Fe, Mg, and Zn) were measured using both spectrophotometry and inductively coupled plasma mass spectrometry, and all of these minerals showed significant (P < 0.05) differences between the two methods. Based on the results of the study, both the influence of diet and the detection method used for circulating nutrients must be considered when interpreting the plasma vitamin and mineral concentrations in greater flamingos.


Assuntos
Vitaminas , beta Caroteno , Animais , Aves/metabolismo , Cantaxantina , Cloretos , Dieta/veterinária , Minerais , Provitaminas/análise , Sódio , Triglicerídeos , Vitamina A , Vitamina E
14.
Environ Microbiol ; 23(2): 1009-1019, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33048442

RESUMO

Membrane vesicles (MVs) released from bacteria act as extracellular vehicles carrying various functional cargoes between cells. MVs with different cargoes play multiple roles in stress adaptation, nutrient acquisition and microbial interactions. However, previous studies have primarily focused on MVs from Gram-negative bacteria, while the characteristics of cargoes in MVs from Gram-positive bacteria and their involvement in microbial interactions remain to be elucidated. Here, we used a Gram-positive strain, Dietzia sp. DQ12-45-1b from Corynebacteriales, to analyse the characteristics and functions of MVs. We identified the 'antioxidant' canthaxanthin is stored within MVs by LC-MS/MS. In addition, nearly the entire genomic content of strain DQ12-45-1b are evenly distributed in MVs, suggesting that MVs from DQ12-45-1b might involve in horizontal gene transfer. Finally, the mycobactin-type siderophores were detected in MVs. The iron-loaded MVs effectively mediate iron binding and delivery to homologous bacteria from the order Corynebacteriales, but not to more distantly related species from the orders Pseudomonadales, Bacillales and Enterobacterales. These results revealed that the iron-loaded MVs are shared between homologous species. Together, we report the Gram-positive bacterium Dietzia sp. DQ12-45-1b released MVs that contain canthaxanthin, DNA and siderophores and prove that MVs act as public goods between closely related species.


Assuntos
Actinomycetales/metabolismo , Vesículas Extracelulares/metabolismo , Ferro/metabolismo , Actinomycetales/citologia , Actinomycetales/genética , Cantaxantina/metabolismo , DNA Bacteriano/metabolismo , Transferência Genética Horizontal , Interações Microbianas , Sideróforos/metabolismo , Especificidade da Espécie
15.
Appl Environ Microbiol ; 87(6)2021 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-33452022

RESUMO

Yarrowia lipolytica has been extensively used to produce essential chemicals and enzymes. As in most other eukaryotes, nonhomologous end joining (NHEJ) is the major repair pathway for DNA double-strand breaks in Y. lipolytica Although numerous studies have attempted to achieve targeted genome integration through homologous recombination (HR), this process requires the construction of homologous arms, which is time-consuming. This study aimed to develop a homology-independent and CRISPR/Cas9-mediated targeted genome integration tool in Y. lipolytica Through optimization of the cleavage efficiency of Cas9, targeted integration of a hyg fragment was achieved with 12.9% efficiency, which was further improved by manipulation of the fidelity of NHEJ repair, the cell cycle, and the integration sites. Thus, the targeted integration rate reached 55% through G1 phase synchronization. This tool was successfully applied for the rapid verification of intronic promoters and iterative integration of four genes in the pathway for canthaxanthin biosynthesis. This homology-independent integration tool does not require homologous templates and selection markers and achieves one-step targeted genome integration of the 8,417-bp DNA fragment, potentially replacing current HR-dependent genome-editing methods for Y. lipolyticaIMPORTANCE This study describes the development and optimization of a homology-independent targeted genome integration tool mediated by CRISPR/Cas9 in Yarrowia lipolytica This tool does not require the construction of homologous templates and can be used to rapidly verify genetic elements and to iteratively integrate multiple-gene pathways in Y. lipolytica This tool may serve as a potential supplement to current HR-dependent genome-editing methods for eukaryotes.


Assuntos
Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Yarrowia/genética , Cantaxantina/metabolismo , Reparo do DNA por Junção de Extremidades , Edição de Genes , Genoma Fúngico , Yarrowia/metabolismo , beta Caroteno/metabolismo
16.
FASEB J ; 34(10): 13430-13444, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32812278

RESUMO

Transmembrane 16A (TMEM16A), also known as anoctamin 1, is the molecular basis of the calcium-activated chloride channels. TMEM16A is present in interstitial cells of Cajal, which are the pacemaker cells that control smooth muscle contraction. TMEM16A is implicated in gastrointestinal disorders. Activation of TMEM16A is believed to promote the gastrointestinal muscle contraction. Here, we report a highly efficient, nontoxic, and selective activator of TMEM16A, canthaxanthin (CX). The study using molecular docking and site-directed mutation revealed that CX-specific binging site in TMEM16A is K769. CX was also found to promote the contraction of smooth muscle cells in gastrointestinal tract through activation of TMEM16A channels, which provides an excellent basis for development of CX as a chemical tool and potential therapeutic for gastrointestinal dysfunction.


Assuntos
Anoctamina-1/fisiologia , Cantaxantina/farmacologia , Íleo/efeitos dos fármacos , Contração Muscular/efeitos dos fármacos , Animais , Gastroenteropatias/metabolismo , Cobaias , Células HEK293 , Humanos , Masculino , Ligação Proteica
17.
Biogerontology ; 22(1): 19-34, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-32926226

RESUMO

It has been earlier reported that partially saturated canthaxanthin (PSC) from Aspergillus carbonarius mutant is non-toxic, has anti-lipid peroxidation activity and can induce apoptosis in prostate cancer cell lines. In the present study, the antiaging effect of PSC was explored in D-galactose administered male wistar rats. 8-10 weeks old, male wistar rats were randomly divided into (i) Vehicle Control Group (VCG), (ii) Aged Control Group (ACG), (iii) Aged + α Lipoic Acid Group (ALG) and (iv) Aged + Partially saturated canthaxanthin Group (APG). Rats received D-galactose (300 mg /kg bwt/day; i.p.) alone (ACG) or together with PSC (APG) (20 mg/kg bwt/day; oral) and α Lipoic Acid (ALG) (80 mg/kg bwt/day; oral) for 10 weeks. Rats in VCG were injected with the same volume of physiological saline (i.p.) and fed with olive oil (vehicle). In vitro protein oxidation and DNA oxidation inhibition, in vivo malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), acetylcholinesterase (AChE) and monoamine oxidase (MAO) activities were determined. In addition, brain neurotransmitters, dopamine and serotonin were estimated by NMR. PSC treatment showed inhibition against protein and DNA oxidation. PSC effectively improved D-galactose induced aging rats by inducing a protective effect through up-regulation of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and brain neurotransmitters and downregulated malondialdehyde (MDA) and monoamineoxidase (MAO) levels. Thus, PSC appears to be a functional compound having antioxidant and antiaging properties.


Assuntos
Cantaxantina , Galactose , Envelhecimento , Animais , Antioxidantes/farmacologia , Aspergillus , Catalase/metabolismo , Glutationa Peroxidase/metabolismo , Peroxidação de Lipídeos , Masculino , Malondialdeído , Estresse Oxidativo , Ratos , Ratos Wistar , Superóxido Dismutase/metabolismo
18.
J Appl Microbiol ; 130(5): 1730-1744, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33078530

RESUMO

AIM: Nano-biotechnologically synthesizing silver nanoparticles via canthaxanthin pigment extracted from Dietzia maris AURCCBT01 and assessing their cytotoxic therapeutic potential against human keratinocyte cell line (HaCaT) were the key objectives of this study. METHODS AND RESULTS: The pigment extracted from D. maris AURCCBT01 was identified as canthaxanthin using UV-VIS spectroscopy, FTIR, NMR (1 H NMR and 13 C NMR) and MS. Canthaxanthin, treated with silver nitrate solution, produced canthaxanthin-mediated silver nanoparticles and they were characterized by UV-VIS spectroscopy, FTIR, XRD, FESEM-EDX and TEM-SAED techniques. UV-VIS spectroscopy pointed out an absorption band at 420 nm, relating to the surface plasmon resonance of silver nanoparticles. FTIR findings suggested that the diverse functional groups of canthaxanthin bio-molecules played a significant task in capping the silver nanoparticles. XRD analysis exhibited 40·20 nm for the crystal size of nanoparticles. FESEM and TEM exhibited that the biosynthesized silver nanoparticles were spherical in shape with crystalline nature and the particle size was 40-50 nm. Moreover, the cytotoxicity assessment of the synthesized nanoparticles in HaCaT revealed significant cytotoxicity in the cultured cells with an IC50 value of 43 µg ml-1 . CONCLUSION: Stable silver nanoparticles synthesized using canthaxanthin from D. maris AURCCBT01 were found effective for application in wound healing activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Biosynthesized silver nanoparticles via canthaxanthin bacterial pigment exhibited their cytotoxicity effect in HaCaT and testified their eventual therapeutic potential in the wound healing activity with no side effects in a cost effective and eco-friendly process.


Assuntos
Actinobacteria/metabolismo , Cantaxantina/isolamento & purificação , Queratinócitos/efeitos dos fármacos , Nanopartículas Metálicas/química , Nanopartículas Metálicas/toxicidade , Prata , Linhagem Celular , Química Verde , Células HaCaT , Humanos , Concentração Inibidora 50 , Nanotecnologia , Tamanho da Partícula , Espectroscopia de Infravermelho com Transformada de Fourier , Cicatrização
19.
Molecules ; 26(9)2021 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-33922133

RESUMO

The increased interest in sequencing cyanobacterial genomes has allowed the identification of new homologs to both the N-terminal domain (NTD) and C-terminal domain (CTD) of the Orange Carotenoid Protein (OCP). The N-terminal domain homologs are known as Helical Carotenoid Proteins (HCPs). Although some of these paralogs have been reported to act as singlet oxygen quenchers, their distinct functional roles remain unclear. One of these paralogs (HCP2) exclusively binds canthaxanthin (CAN) and its crystal structure has been recently characterized. Its absorption spectrum is significantly red-shifted, in comparison to the protein in solution, due to a dimerization where the two carotenoids are closely placed, favoring an electronic coupling interaction. Both the crystal and solution spectra are red-shifted by more than 50 nm when compared to canthaxanthin in solution. Using molecular dynamics (MD) and quantum mechanical/molecular mechanical (QM/MM) studies of HCP2, we aim to simulate these shifts as well as obtain insight into the environmental and coupling effects of carotenoid-protein interactions.


Assuntos
Proteínas de Bactérias/química , Cantaxantina/química , Cianobactérias , Simulação de Dinâmica Molecular , Conformação Proteica , Domínios Proteicos , Teoria Quântica
20.
Arch Microbiol ; 202(6): 1285-1293, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32128608

RESUMO

Mycelial morphogenesis and the production of fungal secretory proteins are still largely unknown. A mutant strain of Aspergillus carbonarius UV-10046 produced abundant polygalacturonase (PG) along with partially saturated canthaxanthin (PSC) at low pH conditions. In the present study, the relationship between PG secretion and PSC biosynthesis was studied using carotenogenic inhibitors and SDS-PAGE electrophoresis. Also the correlation between morphogenesis and PG secretion was investigated by analysing through microscopic studies. From the results, it was observed that secretion of PG was positively influenced by the PSC biosynthesis. The results also showed that the mutant with hairy mycelial structure resulted in higher PG activity when compared to the wild type that lacks hyper branching. From the results, it was confirmed that a mutation might have occurred in the isoprenoid pathway that has helped mutant for survival at acidic conditions. Further, an alteration in the morphogenesis and hyper branching development caused over secretion of PG enzyme in the mutant.


Assuntos
Aspergillus/enzimologia , Aspergillus/genética , Poligalacturonase/metabolismo , Aspergillus/citologia , Cantaxantina/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Morfogênese , Mutação/genética
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