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1.
Nature ; 578(7796): 577-581, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-32076270

RESUMEN

Hydrogen peroxide (H2O2) is a major reactive oxygen species in unicellular and multicellular organisms, and is produced extracellularly in response to external stresses and internal cues1-4. H2O2 enters cells through aquaporin membrane proteins and covalently modifies cytoplasmic proteins to regulate signalling and cellular processes. However, whether sensors for H2O2 also exist on the cell surface remains unknown. In plant cells, H2O2 triggers an influx of Ca2+ ions, which is thought to be involved in H2O2 sensing and signalling. Here, by using forward genetic screens based on Ca2+ imaging, we isolated hydrogen-peroxide-induced Ca2+ increases (hpca) mutants in Arabidopsis, and identified HPCA1 as a leucine-rich-repeat receptor kinase belonging to a previously uncharacterized subfamily that features two extra pairs of cysteine residues in the extracellular domain. HPCA1 is localized to the plasma membrane and is activated by H2O2 via covalent modification of extracellular cysteine residues, which leads to autophosphorylation of HPCA1. HPCA1 mediates H2O2-induced activation of Ca2+ channels in guard cells and is required for stomatal closure. Our findings help to identify how the perception of extracellular H2O2 is integrated with responses to various external stresses and internal cues in plants, and have implications for the design of crops with enhanced fitness.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Peróxido de Hidrógeno/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Calcio/metabolismo , Canales de Calcio/metabolismo , Señalización del Calcio , Cisteína/química , Cisteína/metabolismo , Activación Enzimática , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Mutación , Oxidación-Reducción , Células Vegetales/metabolismo , Dominios Proteicos , Proteínas Serina-Treonina Quinasas/química , Proteínas Serina-Treonina Quinasas/genética
2.
Nature ; 572(7769): 341-346, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-31367039

RESUMEN

Salinity is detrimental to plant growth, crop production and food security worldwide. Excess salt triggers increases in cytosolic Ca2+ concentration, which activate Ca2+-binding proteins and upregulate the Na+/H+ antiporter in order to remove Na+. Salt-induced increases in Ca2+ have long been thought to be involved in the detection of salt stress, but the molecular components of the sensing machinery remain unknown. Here, using Ca2+-imaging-based forward genetic screens, we isolated the Arabidopsis thaliana mutant monocation-induced [Ca2+]i increases 1 (moca1), and identified MOCA1 as a glucuronosyltransferase for glycosyl inositol phosphorylceramide (GIPC) sphingolipids in the plasma membrane. MOCA1 is required for salt-induced depolarization of the cell-surface potential, Ca2+ spikes and waves, Na+/H+ antiporter activation, and regulation of growth. Na+ binds to GIPCs to gate Ca2+ influx channels. This salt-sensing mechanism might imply that plasma-membrane lipids are involved in adaption to various environmental salt levels, and could be used to improve salt resistance in crops.


Asunto(s)
Arabidopsis/citología , Arabidopsis/metabolismo , Señalización del Calcio , Calcio/metabolismo , Glicoesfingolípidos/metabolismo , Células Vegetales/metabolismo , Cloruro de Sodio/metabolismo , Arabidopsis/genética , Glucuronosiltransferasa/genética , Glucuronosiltransferasa/metabolismo , Potenciales de la Membrana/efectos de los fármacos , Mutación , Estrés Salino/genética , Estrés Salino/fisiología , Cloruro de Sodio/farmacología , Intercambiadores de Sodio-Hidrógeno/metabolismo
3.
Nat Prod Rep ; 41(3): 331-346, 2024 Mar 20.
Artículo en Inglés | MEDLINE | ID: mdl-37743806

RESUMEN

Covering: 2018 to Jun of 2023The efficiency of traditional antibiotics has been undermined by the proliferation of antibiotic-resistant pathogenic microorganisms, necessitating the pursuit of innovative therapeutic agents. Antimicrobial peptides (AMPs), which are part of host defence peptides found ubiquitously in nature, exhibiting a wide range of activity towards bacteria, fungi, and viruses, offer a highly promising candidate solution. The efficacy of AMPs can frequently be augmented via alterations to their amino acid sequences or structural adjustments. Given the vast reservoir of marine life forms and their distinctive ecosystems, marine AMPs stand as a burgeoning focal point in the quest for alternative peptide templates extracted from natural sources. Advances in identification and characterization techniques have accelerated the discoveries of marine AMPs, thereby stimulating AMP customization, optimization, and synthesis research endeavours. This review presents an overview of recent discoveries related to the intriguing qualities of marine AMPs. Emphasis will be placed upon post-translational modifications (PTMs) of marine AMPs and how they may impact functionality and potency. Additionally, this review considers ways in which marine PTM might support larger-scale, heterologous AMP manufacturing initiatives, providing insights into translational applications of these important biomolecules.


Asunto(s)
Antiinfecciosos , Antiinfecciosos/farmacología , Antiinfecciosos/química , Péptidos Antimicrobianos , Organismos Acuáticos/química , Ecosistema , Péptidos Catiónicos Antimicrobianos/química , Antibacterianos
4.
Nucleic Acids Res ; 50(16): 9339-9354, 2022 09 09.
Artículo en Inglés | MEDLINE | ID: mdl-35971620

RESUMEN

Chemical modifications are important for RNA function and metabolism. N4-acetylcytidine (ac4C) is critical for the translation and stability of mRNA. Although ac4C is found in RNA viruses, the detailed mechanisms through which ac4C affects viral replication are unclear. Here, we reported that the 5' untranslated region of the enterovirus 71 (EV71) genome was ac4C modified by the host acetyltransferase NAT10. Inhibition of NAT10 and mutation of the ac4C sites within the internal ribosomal entry site (IRES) suppressed EV71 replication. ac4C enhanced viral RNA translation via selective recruitment of PCBP2 to the IRES and boosted RNA stability. Additionally, ac4C increased the binding of RNA-dependent RNA polymerase (3D) to viral RNA. Notably, ac4C-deficient mutant EV71 showed reduced pathogenicity in vivo. Our findings highlighted the essential role of ac4C in EV71 infection and provided insights into potential antiviral treatments.


Asunto(s)
Enterovirus Humano A , Enterovirus , Enterovirus Humano A/genética , ARN Viral/genética , Enterovirus/genética , Virulencia/genética , Sitios Internos de Entrada al Ribosoma/genética , Regiones no Traducidas 5' , Replicación Viral/genética
5.
Ecotoxicol Environ Saf ; 281: 116609, 2024 Jun 20.
Artículo en Inglés | MEDLINE | ID: mdl-38905937

RESUMEN

Copper (Cu) is a necessary mineral nutrient for plant growth and development and is involved in several morphological, physiological, and biochemical processes; however, high concentrations of Cu can negatively impact these processes. The role of stomata in responding to various biotic and abiotic stimuli has not been studied in Bruguiera gymnorhiza, particularly in terms of their coordinated interactions at the molecular, physiological, and biochemical levels. Moreover, numerous plants employ strategies such as the presence of thick waxy cuticles on their leaf epidermis and the closing of stomata to reduce water loss. Thus, this study investigates the accumulation of Cu in B. gymnorhiza and its effect on leaf morphology and the molecular response under different Cu treatments (0, 200, and 400 mg L⁻¹, Cu0, Cu200, and Cu400, respectively) during a two years stress period. The results show that Cu stress affected accumulation and transport, increased the activities of peroxidase and ascorbate peroxidase, concentrations of soluble sugar, proline, and H2O2, and decreased the activity of catalase and content of malondialdehyde. Also, Cu-induced stress decreased the uptake of phosphorus and nitrogen and inhibited plant photosynthesis, which consequently led to reduced plant growth. Scanning electron microscopy combined with gas chromatography-mass spectrometry showed that B. gymnorhiza leaves had higher wax crystals and compositions under increased Cu stress, which forced the leaf's stomata to be closed. Also, the contents of alkanes, alcohols, primary alcohol levels (C26:0, C28:0, C30:0, and C32:0), n-Alkanes (C29 and C30), and other wax loads were significantly higher, while fatty acid (C12, C16, and C18) was lower in Cu200 and Cu400 compared to Cu0. Furthermore, the transcriptomic analyses revealed 1240 (771 up- and 469 downregulated), 1000 (723 up- and 277 down-regulated), and 1476 (808 up- and 668 downregulated) differentially expressed genes in Cu0 vs Cu200, Cu0 vs Cu400, and Cu200 vs Cu400, respectively. RNA-seq analyses showed that Cu mainly affected eight pathways, including photosynthesis, cutin, suberin, and wax biosynthesis. This study provides a reference for understanding mangrove response to heavy metal stress and developing novel management practices.

6.
J Biol Chem ; 298(6): 102006, 2022 06.
Artículo en Inglés | MEDLINE | ID: mdl-35504354

RESUMEN

The discovery of reduced flavin mononucleotide and fatty aldehydes as essential factors of light emission facilitated study of bacterial luminescence. Although the molecular mechanisms underlying bacterial luminescence have been studied for more than 60 years, the structure of the bacterial fatty acid reductase complex remains unclear. Here, we report the cryo-EM structure of the Photobacterium phosphoreum fatty acid reductase complex LuxC-LuxE to a resolution of 2.79 Å. We show that the active site Lys238/Arg355 pair of LuxE is >30 Å from the active site Cys296 of LuxC, implying that catalysis relies on a large conformational change. Furthermore, mutagenesis and biochemical experiments support that the L-shaped cleft inside LuxC plays an important role in substrate binding and reaction. We obtained a series of mutants with significantly improved activity as measured by in vitro bioluminescence assays and demonstrated that the double mutant W111A/F483K displayed the highest activity (370% of the WT). Our results indicated that the activity of LuxC significantly affects the bacterial bioluminescence reaction. Finally, we expressed this mutated lux operon in Escherichia coli but observed that the in vivo concentrations of ATP and NADPH limited the enzyme activity; thus, we conclude that the luminous intensity mainly depends on the level of metabolic energy.


Asunto(s)
Aldehído Oxidorreductasas , Proteínas Bacterianas , Oxidorreductasas , Photobacterium , Aldehído Oxidorreductasas/química , Aldehído Oxidorreductasas/genética , Aldehído Oxidorreductasas/metabolismo , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Microscopía por Crioelectrón , Escherichia coli/genética , Luminiscencia , Complejos Multiproteicos/química , Complejos Multiproteicos/metabolismo , Operón , Oxidorreductasas/química , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Photobacterium/genética
7.
Plant Mol Biol ; 113(6): 415-430, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37566350

RESUMEN

Cadmium (Cd) is a non-essential heavy metal, assimilated in plant tissue with other nutrients, disturbing the ions' homeostasis in plants. The plant develops different mechanisms to tolerate the hazardous environmental effects of Cd. Recently studies found different miRNAs that are involved in Cd stress. In the current study, miR397 mutant lines were constructed to explore the molecular mechanisms of miR397 underlying Cd tolerance. Compared with the genetically modified line of overexpressed miR397 (artificial miR397, amiR397), the lines of downregulated miR397 (Short Tandem Target Mimic miR397, STTM miR397) showed more substantial Cd tolerance with higher chlorophyll a & b, carotenoid and lignin content. ICP-OES revealed higher cell wall Cd and low total Cd levels in STTM miR397 than in the wild-type and amiR397 plants.Further, the STTM plants produced fewer reactive oxygen species (ROS) and lower activity of antioxidants enzymes (e.g., catalase [CAT], malondialdehyde [MDA]) compared with amiR397 and wild-type plants after stress, indicating that silencing the expression of miR397 can reduce oxidative damage. In addition, the different family transporters' gene expression was much higher in the amiR397 plants than in the wild type and STTM miRNA397. Our results suggest that miR397 plays a role in Cd tolerance in Arabidopsis thaliana. Overexpression of miR397 could decrease Cd tolerance in plants by regulating the expression of LAC 2/4/17, changing the lignin content, which may play an important role in inducing different stress-tolerant mechanisms and protecting the cell from a hazardous condition. This study provides a basis to elucidate the functions of miR397 and the Cd stress tolerance mechanism in Arabidopsis thaliana.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Cadmio/metabolismo , Lignina/metabolismo , Clorofila A/metabolismo , Antioxidantes/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Regulación de la Expresión Génica de las Plantas
8.
Crit Rev Biotechnol ; : 1-23, 2023 May 09.
Artículo en Inglés | MEDLINE | ID: mdl-37158096

RESUMEN

Thraustochytrids are eukaryotes and obligate marine protists. They are increasingly considered to be a promising feed additive because of their superior and sustainable application in the production of health-benefiting bioactive compounds, such as fatty acids, carotenoids, and sterols. Moreover, the increasing demand makes it critical to rationally design the targeted products by engineering industrial strains. In this review, bioactive compounds accumulated in thraustochytrids were comprehensively evaluated according to their chemical structure, properties, and physiological function. Metabolic networks and biosynthetic pathways of fatty acids, carotenoids, and sterols were methodically summarized. Further, stress-based strategies used in thraustochytrids were reviewed to explore the potential methodologies for enhancing specific product yields. There are internal relationships between the biosynthesis of fatty acids, carotenoids, and sterols in thraustochytrids since they share some branches of the synthetic routes with some intermediate substrates in common. Although there are classic synthesis pathways presented in the previous research, the metabolic flow of how these compounds are being synthesized in thraustochytrids still remains uncovered. Further, combined with omics technologies to deeply understand the mechanism and effects of different stresses is necessary, which could provide guidance for genetic engineering. While gene-editing technology has allowed targeted gene knock-in and knock-outs in thraustochytrids, efficient gene editing is still required. This critical review will provide comprehensive information to benefit boosting the commercial productivity of specific bioactive substances by thraustochytrids.

9.
Fish Shellfish Immunol ; 136: 108713, 2023 May.
Artículo en Inglés | MEDLINE | ID: mdl-36990258

RESUMEN

In shellfish aquaculture, antibiotics are commonly used to address Vibrio infections. However, antibiotic abuse has increased the risk of environment pollution, which has also raised food safety concerns. Antimicrobial peptides (AMPs) are considered safe and sustainable alternatives to antibiotics. Hence, in this study, we aimed to develop a transgenic Tetraselmis subcordiformis line harboring AMP-PisL9K22WK for reducing the use of antibiotics in mussel aquaculture. Toward this, pisL9K22WK was assembled into nuclear expression vectors of T. subcordiformis. Post particle bombardment, several stable transgenic lines were selected after 6 months of herbicide resistance culture. Subsequently, Vibrio-infected mussels (Mytilus sp.) were orally fed transgenic T. subcordiformis to test the efficacy of this drug delivery system. The results showed that the transgenic line as an oral antimicrobial agent significantly improved the resistance of mussels to Vibrio. The growth rate of the mussels fed transgenic T. subcordiformis was considerably higher than that of mussels fed wild-type algae (10.35% versus 2.44%). In addition, the possibility of using the lyophilized powder of the transgenic line as drug delivery system was also evaluated; however, compared to that observed after feeding with live cells, the lyophilized powder did not improve the low growth rate caused by Vibrio infection, suggesting that fresh microalgae are more beneficial for the delivery of the PisL9K22WK to mussels than the lyophilized powder. In summary, this is a promising step toward the development of safe and environment-friendly antimicrobial baits.


Asunto(s)
Microalgas , Mytilus , Vibriosis , Vibrio , Animales , Péptidos Antimicrobianos , Polvos , Animales Modificados Genéticamente , Antibacterianos/farmacología
10.
Mar Drugs ; 22(1)2023 Dec 28.
Artículo en Inglés | MEDLINE | ID: mdl-38248646

RESUMEN

Crocin is one of the most valuable components of the Chinese medicinal plant Crocus sativus and is widely used in the food, cosmetics, and pharmaceutical industries. Traditional planting of C. sativus is unable to fulfill the increasing demand for crocin in the global market, however, such that researchers have turned their attention to the heterologous production of crocin in a variety of hosts. At present, there are reports of successful heterologous production of crocin in Escherichia coli, Saccharomyces cerevisiae, microalgae, and plants that do not naturally produce crocin. Of these, the microalga Dunaliella salina, which produces high levels of ß-carotene, the substrate for crocin biosynthesis, is worthy of attention. This article describes the biosynthesis of crocin, compares the features of each heterologous host, and clarifies the requirements for efficient production of crocin in microalgae.


Asunto(s)
Chlorophyceae , Microalgas , Carotenoides , beta Caroteno , Industria Farmacéutica , Escherichia coli , Saccharomyces cerevisiae
11.
Int J Mol Sci ; 24(19)2023 Sep 27.
Artículo en Inglés | MEDLINE | ID: mdl-37834043

RESUMEN

Chlamydomonas reinhardtii (C. reinhardtii) is a single-cell green alga that can be easily genetically manipulated. With its favorable characteristics of rapid growth, low cost, non-toxicity, and the ability for post-translational protein modification, C. reinhardtii has emerged as an attractive option for the biosynthesis of various valuable products. To enhance the expression level of exogenous genes and overcome the silencing of foreign genes by C. reinhardtii, synthetic promoters such as the chimeric promoter AR have been constructed and evaluated. In this study, a synthetic promoter GA was constructed by hybridizing core fragments from the natural promoters of the acyl carrier protein gene (ACP2) and the glutamate dehydrogenase gene (GDH2). The GA promoter exhibited a significant increase (7 times) in expressing GUS, over the AR promoter as positive control. The GA promoter also displayed a strong responsiveness to blue light (BL), where the GUS expression was doubled compared to the white light (WL) condition. The ability of the GA promoter was further tested in the expression of another exogenous cadA gene, responsible for catalyzing the decarboxylation of lysine to produce cadaverine. The cadaverine yield driven by the GA promoter was increased by 1-2 times under WL and 2-3 times under BL as compared to the AR promoter. This study obtained, for the first time, a blue light-responsive GDH2 minimal fragment in C. reinhardtii, which delivered a doubling effect under BL when used alone or in hybrid. Together with the strong GA synthetic promoter, this study offered useful tools of synthetic biology to the algal biotechnology field.


Asunto(s)
Chlamydomonas reinhardtii , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Cadaverina/metabolismo , Regiones Promotoras Genéticas , Biotecnología , Luz
12.
Int J Mol Sci ; 24(8)2023 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-37108242

RESUMEN

One important feature of tumour development is the regulatory role of metabolic plasticity in maintaining the balance of mitochondrial oxidative phosphorylation and glycolysis in cancer cells. In recent years, the transition and/or function of metabolic phenotypes between mitochondrial oxidative phosphorylation and glycolysis in tumour cells have been extensively studied. In this review, we aimed to elucidate the characteristics of metabolic plasticity (emphasizing their effects, such as immune escape, angiogenesis migration, invasiveness, heterogeneity, adhesion, and phenotypic properties of cancers, among others) on tumour progression, including the initiation and progression phases. Thus, this article provides an overall understanding of the influence of abnormal metabolic remodeling on malignant proliferation and pathophysiological changes in carcinoma.


Asunto(s)
Carcinoma , Fosforilación Oxidativa , Humanos , Glucólisis , Mitocondrias/metabolismo , Transformación Celular Neoplásica/metabolismo , Carcinoma/metabolismo
13.
Int J Mol Sci ; 24(24)2023 Dec 09.
Artículo en Inglés | MEDLINE | ID: mdl-38139139

RESUMEN

Copper-containing amine oxidases (CuAOs) are known to have significant involvement in the process of polyamine catabolism, as well as serving crucial functions in plant development and response to abiotic stress. A genome-wide investigation of the CuAO protein family was previously carried out in sweet orange (Citrus sinensis) and sweet cherry (Prunus avium L.). Six CuAO (KoCuAO1-KoCuAO6) genes were discovered for the first time in the Kandelia obovata (Ko) genome through a genome-wide analysis conducted to better understand the key roles of the CuAO gene family in Kandelia obovata. This study encompassed an investigation into various aspects of gene analysis, including gene characterization and identification, subcellular localization, chromosomal distributions, phylogenetic tree analysis, gene structure analysis, motif analysis, duplication analysis, cis-regulatory element identification, domain and 3D structural variation analysis, as well as expression profiling in leaves under five different treatments of copper (CuCl2). Phylogenetic analysis suggests that these KoCuAOs, like sweet cherry, may be subdivided into three subgroups. Examining the chromosomal location revealed an unequal distribution of the KoCuAO genes across four out of the 18 chromosomes in Kandelia obovata. Six KoCuAO genes have coding regions with 106 and 159 amino acids and exons with 4 and 12 amino acids. Additionally, we discovered that the 2.5 kb upstream promoter region of the KoCuAOs predicted many cis elements linked to phytohormones and stress responses. According to the expression investigations, CuCl2 treatments caused up- and downregulation of all six genes. In conclusion, our work provides a comprehensive overview of the expression pattern and functional variety of the Kandelia obovata CuAO gene family, which will facilitate future functional characterization of each KoCuAO gene.


Asunto(s)
Amina Oxidasa (conteniendo Cobre) , Rhizophoraceae , Rhizophoraceae/genética , Amina Oxidasa (conteniendo Cobre)/metabolismo , Filogenia , Cobre/metabolismo , Aminoácidos/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
14.
Int J Mol Sci ; 24(13)2023 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-37446180

RESUMEN

Ginsenosides are major bioactive compounds found in Panax ginseng that exhibit various pharmaceutical properties. Dammarenediol-II, the nucleus of dammarane-type ginsenosides, is a promising candidate for pharmacologically active triterpenes. Dammarenediol-II synthase (DDS) cyclizes 2,3-oxidosqualene to produce dammarenediol-II. Based on the native terpenoids synthetic pathway, a dammarane-type ginsenosides synthetic pathway was established in Chlamydomonas reinhardtii by introducing P. ginseng PgDDS, CYP450 enzyme (PgCYP716A47), or/and Arabidopsis thaliana NADPH-cytochrome P450 reductase gene (AtCPR), which is responsible for producing dammarane-type ginsenosides. To enhance productivity, strategies such as "gene loading" and "culture optimizing" were employed. Multiple copies of transgene expression cassettes were introduced into the genome to increase the expression of the key rate-limiting enzyme gene, PgDDS, significantly improving the titer of dammarenediol-II to approximately 0.2 mg/L. Following the culture optimization in an opt2 medium supplemented with 1.5 mM methyl jasmonate under a light:dark regimen, the titer of dammarenediol-II increased more than 13-fold to approximately 2.6 mg/L. The C. reinhardtii strains engineered in this study constitute a good platform for the further production of ginsenosides in microalgae.


Asunto(s)
Chlamydomonas reinhardtii , Ginsenósidos , Panax , Triterpenos , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Triterpenos/metabolismo , Panax/genética , Damaranos
15.
Fish Shellfish Immunol ; 121: 239-244, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-34990807

RESUMEN

Mytilin is one of the most important CS-αß peptides involved in innate immune response in Mytilidae. In this study, we successfully identified four mytilin-like antimicrobial peptides (pernalins) from Asian green mussel Perna viridis by aligning the P. viridis transcriptome with 186 mytilins and myticins related sequences collected from the transcriptome data of six Mytilus species. Analysis on gene structure showed that pernalin genes had high conservation with mytilin B of Mediterranean mussel Mytilus galloprovincialis. Interestingly, all pernalin genes have a similar tissue expression feature, evidenced by the highest transcription level observed in the hemocytes and followed by the mantle. The lowest transcription level was observed in the foot and gills. qRT-PCR analysis showed that all pernalin genes were significantly down-regulated at each time points from 3 h to 48 h after Vibrio parahaemolyticus infection, suggesting their timely immune responses after bacterial infection.


Asunto(s)
Péptidos Antimicrobianos/genética , Mytilus , Perna , Animales , Péptidos Catiónicos Antimicrobianos , Clonación Molecular , Mytilus/genética , Perna/genética
16.
Mar Drugs ; 21(1)2022 Dec 22.
Artículo en Inglés | MEDLINE | ID: mdl-36662180

RESUMEN

Six new sesquiterpene aminoglycosides, trichaspside F (2) and cyclonerosides A-E (5-9), two new diterpene aminoglycosides, harzianosides A and B (10, 11), and three known sesquiterpenes, trichodermoside (1), cycloneran-3,7,10,11-tetraol (3), and cyclonerodiol (4), have been isolated from the n-butanol extract of Trichoderma sp. SCSIOW21 (Hypocreaceae), a deep-sea-sediment-derived fungus. The structures and relative configurations of the new compounds were determined using spectroscopic techniques and comparisons with those reported in the literature. The absolute configurations of the aglycone part of cyclonerosides A-E (5-9) were tentatively proposed based on optical rotation and biogenic considerations. Cyclonerosides A-E (5-9) represent the first glycosides of cyclonelane-type sesquiterpenes generated from Trichoderma. The NO-production-inhibitory activities were evaluated using macrophage RAW264.7 cells. Among the isolated compounds, trichaspside F (2) and cyclonerosides B-E (6-9) exhibited the strongest NO-production-inhibitory activities with IC50 values of 54.8, 50.7, 57.1, 42.0, and 48.0 µM, respectively, compared to the IC50 value of 30.8 µM for the positive control (quercetin). When tested for anti-fungal activities against several pathogenic fungi, none of the compounds exhibited significant activities at a concentration of 100 µM.


Asunto(s)
Diterpenos , Sesquiterpenos , Trichoderma , Aminoglicósidos/farmacología , Trichoderma/química , Sesquiterpenos/química , Antibacterianos/química , Estructura Molecular
17.
Mar Drugs ; 20(9)2022 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-36135753

RESUMEN

Alginate is an acidic polysaccharide mainly extracted from kelp or sargassum, which comprises 40% of the dry weight of algae. It is a linear polymer consisting of ß-D-mannuronic acid (M) and α-L-guluronic acid (G) with 1,4-glycosidic linkages, possessing various applications in the food and nutraceutical industries due to its unique physicochemical properties and health benefits. Additionally, alginate is able to form a gel matrix in the presence of Ca2+ ions. Alginate properties also affect its gelation, including its structure and experimental conditions such as pH, temperature, crosslinker concentration, residence time and ionic strength. These features of this polysaccharide have been widely used in the food industry, including in food gels, controlled-release systems and film packaging. This review comprehensively covers the analysis of alginate and discussed the potential applications of alginate in the food industry and nutraceuticals.


Asunto(s)
Alginatos , Suplementos Dietéticos , Alginatos/química , Preparaciones de Acción Retardada , Geles , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Iones , Polímeros
18.
Mar Drugs ; 20(5)2022 Apr 21.
Artículo en Inglés | MEDLINE | ID: mdl-35621927

RESUMEN

The alga Chlamydomonas reinhardtii is a potential platform for recombinant protein expression in the future due to various advantages. Dozens of C. reinhardtii strains producing genetically engineered recombinant therapeutic protein have been reported. However, owing to extremely low protein expression efficiency, none have been applied for industrial purposes. Improving protein expression efficiency at the molecular level is, therefore, a priority. The 3'-end poly(A) tail of mRNAs is strongly correlated with mRNA transcription and protein translation efficiency. In this study, we identified a canonical C. reinhardtii poly(A) polymerase (CrePAPS), verified its polyadenylate activity, generated a series of overexpressing transformants, and performed proteomic analysis. Proteomic results demonstrated that overexpressing CrePAPS promoted ribosomal assembly and enhanced protein accumulation. The accelerated translation was further verified by increased crude and dissolved protein content detected by Kjeldahl and bicinchoninic acid (BCA) assay approaches. The findings provide a novel direction in which to exploit photosynthetic green algae as a recombinant protein expression platform.


Asunto(s)
Chlamydomonas reinhardtii , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Biosíntesis de Proteínas , Proteómica , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/metabolismo
19.
Mar Drugs ; 20(9)2022 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-36135766

RESUMEN

Eukaryotic green microalgae show considerable promise for the sustainable light-driven biosynthesis of high-value fine chemicals, especially terpenoids because of their fast and inexpensive phototrophic growth. Here, the novel isopentenol utilization pathway (IUP) was introduced into Chlamydomonas reinhardtii to enhance the hemiterpene (isopentenyl pyrophosphate, IPP) titers. Then, diphosphate isomerase (IDI) and limonene synthase (MsLS) were further inserted for limonene production. Transgenic algae showed 8.6-fold increase in IPP compared with the wild type, and 23-fold increase in limonene production compared with a single MsLS expressing strain. Following the culture optimization, the highest limonene production reached 117 µg/L, when the strain was cultured in a opt2 medium supplemented with 10 mM isoprenol under a light: dark regimen. This demonstrates that transgenic algae expressing the IUP represent an ideal chassis for the high-value terpenoid production. The IUP will facilitate further the metabolic and enzyme engineering to enhance the terpenoid titers by significantly reducing the number of enzyme steps required for an optimal biosynthesis.


Asunto(s)
Chlamydomonas reinhardtii , Ingeniería Metabólica , Chlamydomonas reinhardtii/metabolismo , Difosfatos/metabolismo , Hemiterpenos/metabolismo , Isomerasas/metabolismo , Limoneno/metabolismo , Pentanoles , Terpenos/metabolismo
20.
Ecotoxicol Environ Saf ; 242: 113903, 2022 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-35870349

RESUMEN

Selenium incorporates into selenocysteine (Sec) which is a key component of selenoproteins implicated in antioxidant defense and redox homeostasis. Methionine sulfoxide reductases (Msr) play crucial roles in cellular defense against environmental stress. Whereas mammals have the MsrB selenoprotein form, unicellular organisms have MsrA. The Sec residue at the conserved catalytic sites of selenoprotein MsrA confers a metabolic advantage over the non-selenoprotein type MsrA. In the present study, the novel selenoprotein HpMsrA from Haematococcus pluvialis was cloned by the rapid amplification of cDNA ends and transformed into the model green alga Chlamydomonas reinhardtii. Alignment of homologs revealed the presence of the conserved catalytic domain GUFW and showed that the HpMsrA protein comprises Sec (U) at the N-terminus but no recycled Cys at the C-terminus. We studied the response of HpMsrA expression to selenite, high light intensity, hydrogen peroxide, cadmium nitrate, and glyphosate exposure via real-time quantitative PCR and enzyme activity analysis. The results demonstrated that HpMsrA protects cellular proteins against oxidative and environmental stressors. Compared with wild type C. reinhardtii, the transformant exhibited a superior antioxidant ability. The discoveries made herein shed light on the antioxidant physiology and environmental stress resistance mechanisms of the selenoproteins in microalgae. This information may aid in conducting environmental risk assessments of aquatic ecosystems involving microalgae known to respond rapidly and quantitatively to abiotic stress factors promoting excessive reactive oxygen species generation.


Asunto(s)
Antioxidantes , Metionina Sulfóxido Reductasas , Animales , Cadmio/toxicidad , Ecosistema , Glicina/análogos & derivados , Peróxido de Hidrógeno , Mamíferos/metabolismo , Metionina Sulfóxido Reductasas/genética , Metionina Sulfóxido Reductasas/metabolismo , Selenocisteína/metabolismo , Selenoproteínas/genética , Selenoproteínas/metabolismo , Glifosato
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