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1.
Molecules ; 26(6)2021 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-33804230

RESUMEN

The fruit of Lycium barbarum L. (goji berry) is used as traditional Chinese medicine, and has the functions of immune regulation, anti-tumor, neuroprotection, anti-diabetes, and anti-fatigue. One of the main bioactive components is L. barbarum polysaccharide (LBP). Nowadays, LBP is widely used in the health market, and it is extracted from the fruit of L. barbarum. The planting of L. barbarum needs large amounts of fields, and it takes one year to harvest the goji berry. The efficiency of natural LBP production is low, and the LBP quality is not the same at different places. Goji berry-derived LBP cannot satisfy the growing market demands. Engineered Saccharomyces cerevisiae has been used for the biosynthesis of some plant natural products. Recovery of LBP biosynthetic pathway in L. barbarum and expression of them in engineered S. cerevisiae might lead to the yeast LBP production. However, information on LBP biosynthetic pathways and the related key enzymes of L. barbarum is still limited. In this review, we summarized current studies about LBP biosynthetic pathway and proposed the strategies to recover key enzymes for LBP biosynthesis. Moreover, the potential application of synthetic biology strategies to produce LBP using engineered S. cerevisiae was discussed.


Asunto(s)
Medicamentos Herbarios Chinos/metabolismo , Lycium/metabolismo , Saccharomyces cerevisiae/metabolismo , Animales , Vías Biosintéticas/fisiología , Fitoterapia/métodos , Biología Sintética/métodos
2.
Int J Mol Sci ; 22(6)2021 Mar 16.
Artículo en Inglés | MEDLINE | ID: mdl-33809693

RESUMEN

The N-terminal of Myc-like basic helix-loop-helix transcription factors (bHLH TFs) contains an interaction domain, namely the MYB-interacting region (MIR), which interacts with the R2R3-MYB proteins to regulate genes involved in the anthocyanin biosynthetic pathway. However, the functions of MIR-domain bHLHs in this pathway are not fully understood. In this study, PbbHLH2 containing the MIR-domain was identified and its function investigated. The overexpression of PbbHLH2 in "Zaosu" pear peel increased the anthocyanin content and the expression levels of late biosynthetic genes. Bimolecular fluorescence complementation showed that PbbHLH2 interacted with R2R3-MYB TFs PbMYB9, 10, and 10b in onion epidermal cells and confirmed that MIR-domain plays important roles in the interaction between the MIR-domain bHLH and R2R3-MYB TFs. Moreover, PbbHLH2 bound and activated the dihydroflavonol reductase promoter in yeast one-hybrid (Y1H) and dual-luciferase assays. Taken together these results suggested that the MIR domain of PbbHLH2 regulated anthocyanin biosynthesis in pear fruit peel.


Asunto(s)
Antocianinas/biosíntesis , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/química , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Vías Biosintéticas , Frutas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Pyrus/metabolismo , Secuencia de Aminoácidos , Frutas/genética , Regulación de la Expresión Génica de las Plantas , Cebollas/citología , Filogenia , Epidermis de la Planta/citología , Proteínas de Plantas/genética , Regiones Promotoras Genéticas , Unión Proteica , Dominios Proteicos , Pyrus/genética , Relación Estructura-Actividad
3.
Nat Commun ; 12(1): 2056, 2021 04 06.
Artículo en Inglés | MEDLINE | ID: mdl-33824341

RESUMEN

Biotin is an essential micro-nutrient across the three domains of life. The paradigm earlier step of biotin synthesis denotes "BioC-BioH" pathway in Escherichia coli. Here we report that BioZ bypasses the canonical route to begin biotin synthesis. In addition to its origin of Rhizobiales, protein phylogeny infers that BioZ is domesticated to gain an atypical role of ß-ketoacyl-ACP synthase III. Genetic and biochemical characterization demonstrates that BioZ catalyzes the condensation of glutaryl-CoA (or ACP) with malonyl-ACP to give 5'-keto-pimeloyl ACP. This intermediate proceeds via type II fatty acid synthesis (FAS II) pathway, to initiate the formation of pimeloyl-ACP, a precursor of biotin synthesis. To further explore molecular basis of BioZ activity, we determine the crystal structure of Agrobacterium tumefaciens BioZ at 1.99 Å, of which the catalytic triad and the substrate-loading tunnel are functionally defined. In particular, we localize that three residues (S84, R147, and S287) at the distant bottom of the tunnel might neutralize the charge of free C-carboxyl group of the primer glutaryl-CoA. Taken together, this study provides molecular insights into the BioZ biotin synthesis pathway.


Asunto(s)
Vías Biosintéticas , Biotina/biosíntesis , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Proteína Transportadora de Acilo/metabolismo , Acilcoenzima A/metabolismo , Agrobacterium/crecimiento & desarrollo , Secuencia de Aminoácidos , Biocatálisis , Cristalografía por Rayos X , Simulación del Acoplamiento Molecular , Filogenia , Multimerización de Proteína , Homología Estructural de Proteína , Especificidad por Sustrato
4.
Science ; 372(6541): 460-461, 2021 04 30.
Artículo en Inglés | MEDLINE | ID: mdl-33926938
5.
Int J Mol Sci ; 22(6)2021 Mar 23.
Artículo en Inglés | MEDLINE | ID: mdl-33806795

RESUMEN

Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, is important for bacterial viability in general and host-pathogen interactions in particular. Negative charges at its core oligosaccharide (core-OS) contribute to membrane integrity through bridging interactions with divalent cations. The molecular structure and synthesis of the core-OS have been resolved in various bacteria including the mammalian pathogen Pseudomonas aeruginosa. A few core-OS structures of plant-associated Pseudomonas strains have been solved to date, but the genetic components of the underlying biosynthesis remained unclear. We conducted a comparative genome analysis of the core-OS gene cluster in Pseudomonas syringae pv. tomato (Pst) DC3000, a widely used model pathogen in plant-microbe interactions, within the P. syringae species complex and to other plant-associated Pseudomonas strains. Our results suggest a genetic and structural conservation of the inner core-OS but variation in outer core-OS composition within the P. syringae species complex. Structural analysis of the core-OS of Pst DC3000 shows an uncommonly high phosphorylation and presence of an O-acetylated sugar. Finally, we combined the results of our genomic survey with available structure information to estimate the core-OS composition of other Pseudomonas species.


Asunto(s)
Lipopolisacáridos/biosíntesis , Lipopolisacáridos/química , Oligosacáridos/química , Pseudomonas syringae/metabolismo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Vías Biosintéticas , Regulación Bacteriana de la Expresión Génica , Orden Génico , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Estructura Molecular , Pseudomonas syringae/genética
6.
Nat Commun ; 12(1): 2330, 2021 04 20.
Artículo en Inglés | MEDLINE | ID: mdl-33879780

RESUMEN

The hypothalamus plays a central role in monitoring and regulating systemic glucose metabolism. The brain is enriched with phospholipids containing poly-unsaturated fatty acids, which are biologically active in physiological regulation. Here, we show that intraperitoneal glucose injection induces changes in hypothalamic distribution and amounts of phospholipids, especially arachidonic-acid-containing phospholipids, that are then metabolized to produce prostaglandins. Knockdown of cytosolic phospholipase A2 (cPLA2), a key enzyme for generating arachidonic acid from phospholipids, in the hypothalamic ventromedial nucleus (VMH), lowers insulin sensitivity in muscles during regular chow diet (RCD) feeding. Conversely, the down-regulation of glucose metabolism by high fat diet (HFD) feeding is improved by knockdown of cPLA2 in the VMH through changing hepatic insulin sensitivity and hypothalamic inflammation. Our data suggest that cPLA2-mediated hypothalamic phospholipid metabolism is critical for controlling systemic glucose metabolism during RCD, while continuous activation of the same pathway to produce prostaglandins during HFD deteriorates glucose metabolism.


Asunto(s)
Glucosa/metabolismo , Fosfolipasas A2 Citosólicas/metabolismo , Prostaglandinas/biosíntesis , Núcleo Hipotalámico Ventromedial/metabolismo , Animales , Ácido Araquidónico/metabolismo , Vías Biosintéticas , Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Técnicas de Silenciamiento del Gen , Fosfolipasas A2 Grupo IV/antagonistas & inhibidores , Fosfolipasas A2 Grupo IV/genética , Fosfolipasas A2 Grupo IV/metabolismo , Hiperglucemia/metabolismo , Resistencia a la Insulina , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Fosfolipasas A2 Citosólicas/antagonistas & inhibidores , Fosfolipasas A2 Citosólicas/genética , Fosfolípidos/metabolismo
7.
Plant Sci ; 306: 110863, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33775368

RESUMEN

Unlike animals, plants possess diverse L-serine (Ser) biosynthetic pathways. One of them, the Phosphorylated Pathway of Serine Biosynthesis (PPSB) has been recently described as essential for embryo, pollen and root development, and required for ammonium and sulfur assimilation. The first and rate limiting step of PPSB is the reaction catalyzed by the enzyme phosphoglycerate dehydrogenase (PGDH). In Arabidopsis, the PGDH family consists of three genes, PGDH1, PGDH2 and PGDH3. PGDH1 is characterized as being the essential gene of the family. However, the biological significance of PGDH2 and PGDH3 remains unknown. In this manuscript, we have functionally characterized PGDH2 and PGDH3. Phenotypic, metabolomic and gene expression analysis in PGDH single, double and triple mutants indicate that both PGDH2 and PGDH3 are functional, affecting plant metabolism and development. PGDH2 has a stronger effect on plant growth than PGDH3, having a partial redundant role with PGDH1. PGDH3, however, could have additional functions in photosynthetic cells unrelated to Ser biosynthesis.


Asunto(s)
Arabidopsis/crecimiento & desarrollo , Arabidopsis/genética , Arabidopsis/metabolismo , Fosfoglicerato-Deshidrogenasa/genética , Fosfoglicerato-Deshidrogenasa/metabolismo , Serina/biosíntesis , Serina/genética , Vías Biosintéticas , Regulación Enzimológica de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas
8.
Nat Commun ; 12(1): 1760, 2021 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-33741955

RESUMEN

Plant natural products (PNPs) and their derivatives are important but underexplored sources of pharmaceutical molecules. To access this untapped potential, the reconstitution of heterologous PNP biosynthesis pathways in engineered microbes provides a valuable starting point to explore and produce novel PNP derivatives. Here, we introduce a computational workflow to systematically screen the biochemical vicinity of a biosynthetic pathway for pharmaceutical compounds that could be produced by derivatizing pathway intermediates. We apply our workflow to the biosynthetic pathway of noscapine, a benzylisoquinoline alkaloid (BIA) with a long history of medicinal use. Our workflow identifies pathways and enzyme candidates for the production of (S)-tetrahydropalmatine, a known analgesic and anxiolytic, and three additional derivatives. We then construct pathways for these compounds in yeast, resulting in platforms for de novo biosynthesis of BIA derivatives and demonstrating the value of cheminformatic tools to predict reactions, pathways, and enzymes in synthetic biology and metabolic engineering.


Asunto(s)
Productos Biológicos/metabolismo , Vías Biosintéticas/genética , Biología Computacional/métodos , Ingeniería Metabólica/métodos , Noscapina/metabolismo , Saccharomyces cerevisiae/metabolismo , Alcaloides/biosíntesis , Bencilisoquinolinas/metabolismo , Noscapina/química , Plantas/genética , Plantas/metabolismo , Saccharomyces cerevisiae/genética , Programas Informáticos
9.
Molecules ; 26(5)2021 Feb 27.
Artículo en Inglés | MEDLINE | ID: mdl-33673610

RESUMEN

Generally, bacteriochlorophyllides were responsible for the photosynthesis in bacteria. Seven types of bacteriochlorophyllides have been disclosed. Bacteriochlorophyllides a/b/g could be synthesized from divinyl chlorophyllide a. The other bacteriochlorophyllides c/d/e/f could be synthesized from chlorophyllide a. The chemical structure and synthetic route of bacteriochlorophyllides were summarized in this review. Furthermore, the potential applications of bacteriochlorophyllides in photosensitizers, immunosensors, influence on bacteriochlorophyll aggregation, dye-sensitized solar cell, heme synthesis and for light energy harvesting simulation were discussed.


Asunto(s)
Bacterias/metabolismo , Clorofílidas/biosíntesis , Clorofílidas/química , Complejos de Coordinación/química , Técnicas Biosensibles , Vías Biosintéticas , Hemo/química , Hemo/metabolismo , Fármacos Fotosensibilizantes/química , Fotosíntesis , Energía Solar
10.
Int J Mol Sci ; 22(4)2021 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-33669861

RESUMEN

Microsorum species produce a high amount of phytoecdysteroids (PEs), which are widely used in traditional medicine in the Pacific islands. The PEs in two different Microsorum species, M. punctatum (MP) and M. scolopendria (MS), were examined using high-performance liquid chromatography (HPLC). In particular, MS produces a high amount of 20-hydroxyecdysone, which is the main active compound in PEs. To identify genes for PE biosynthesis, we generated reference transcriptomes from sterile frond tissues using the NovaSeq 6000 system. De novo transcriptome assembly after deleting contaminants resulted in 57,252 and 54,618 clean transcripts for MP and MS, respectively. The clean Microsorum transcripts for each species were annotated according to gene ontology terms, UniProt pathways, and the clusters of the orthologous group protein database using the MEGAN6 and Sma3s programs. In total, 1852 and 1980 transcription factors were identified for MP and MS, respectively. We obtained transcripts encoding for 38 and 32 enzymes for MP and MS, respectively, potentially involved in mevalonate and sterol biosynthetic pathways, which produce precursors for PE biosynthesis. Phylogenetic analyses revealed many redundant and unique enzymes between the two species. Overall, this study provides two Microsorum reference transcriptomes that might be useful for further studies regarding PE biosynthesis in Microsorum species.


Asunto(s)
Ecdisteroides/metabolismo , Helechos/enzimología , Helechos/genética , Transcriptoma/genética , Vías Biosintéticas/genética , Regulación de la Expresión Génica de las Plantas , Biblioteca de Genes , Ontología de Genes , Ácido Mevalónico/metabolismo , Anotación de Secuencia Molecular , Fenotipo , Filogenia , Hojas de la Planta/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo , Factores de Transcripción/metabolismo
11.
Molecules ; 26(4)2021 Feb 18.
Artículo en Inglés | MEDLINE | ID: mdl-33670502

RESUMEN

Promising research over the past decades has shown that some types of pentacyclic triterpenes (PTs) are associated with the prevention of type 2 diabetes (T2D), especially those found in foods. The most abundant edible sources of PTs are those belonging to the ursane and oleanane scaffold. The principal finding is that Cecropia telenitida contains abundant oleanane and ursane PT types with similar oxygenation patterns to those found in food matrices. We studied the compositional profile of a rich PT fraction (DE16-R) and carried out a viability test over different cell lines. The biosynthetic pathway connected to the isolated PTs in C. telenitida offers a specific medicinal benefit related to the modulation of T2D. This current study suggests that this plant can assemble isobaric, positional isomers or epimeric PT. Ursane or oleanane scaffolds with the same oxygenation pattern are always shared by the PTs in C. telenitida, as demonstrated by its biosynthetic pathway. Local communities have long used this plant in traditional medicine, and humans have consumed ursane and oleanane PTs in fruits since ancient times, two key points we believe useful in considering the medicinal benefits of C. telenitida and explaining how a group of molecules sharing a closely related scaffold can express effectiveness.


Asunto(s)
Vías Biosintéticas , Cecropia (Planta)/química , Suplementos Dietéticos , Triterpenos Pentacíclicos/metabolismo , Animales , Muerte Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Fraccionamiento Químico , Cromatografía Líquida de Alta Presión , Humanos , Espectroscopía de Resonancia Magnética , Ratones , Triterpenos Pentacíclicos/química , Triterpenos Pentacíclicos/farmacología
12.
Molecules ; 26(4)2021 Feb 18.
Artículo en Inglés | MEDLINE | ID: mdl-33670642

RESUMEN

We investigated the relationship between the blue-light photoreceptor cryptochrome (CRY) and melatonin biosynthesis by generating RNA interference (RNAi) transgenic rice plants that suppress the cryptochrome 1b gene (CRY1b). The resulting CRY1b RNAi rice lines expressed less CRY1b mRNA, but not CRY1a or CRY2 mRNA, suggesting that the suppression is specific to CRY1b. The growth of CRY1b RNAi rice seedlings was enhanced under blue light compared to wild-type growth, providing phenotypic evidence for impaired CRY function. When these CRY1b RNAi rice plants were challenged with cadmium to induce melatonin, wild-type plants produced 100 ng/g fresh weight (FW) melatonin, whereas CRY1b RNAi lines produced 60 ng/g FW melatonin on average, indicating that melatonin biosynthesis requires the CRY photoreceptor. Due to possible feedback regulation, the expression of melatonin biosynthesis genes such as T5H, SNAT1, SNAT2, and COMT was elevated in the CRY1b RNAi lines compared to the wild-type plants. In addition, laminar angles decreased in the CRY1b RNAi lines via the suppression of brassinosteroid (BR) biosynthesis genes such as DWARF. The main cause of the BR decrease in the CRY1b RNAi lines seems to be the suppression of CRY rather than decreased melatonin because the melatonin decrease suppressed DWARF4 rather than DWARF.


Asunto(s)
Vías Biosintéticas/genética , Brasinoesteroides/biosíntesis , Criptocromos/genética , Genes de Plantas , Melatonina/biosíntesis , Oryza/genética , Tolerancia a la Sal/genética , Vías Biosintéticas/efectos de los fármacos , Criptocromos/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Oryza/efectos de los fármacos , Fenotipo , Plantas Modificadas Genéticamente , Interferencia de ARN , ARN Mensajero/genética , ARN Mensajero/metabolismo , Tolerancia a la Sal/efectos de los fármacos , Plantones/efectos de los fármacos , Plantones/genética , Serotonina/metabolismo , Cloruro de Sodio/farmacología
13.
Plant Mol Biol ; 106(1-2): 157-172, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33704646

RESUMEN

KEY MESSAGE: Our results provide insights into the flavonol biosynthesis regulation of M. truncatula. The R2R3-MYB transcription factor MtMYB134 emerged as tool to improve the flavonol biosynthesis. Flavonols are plant specialized metabolites with vital roles in plant development and defense and are known as diet compound beneficial to human health. In leguminous plants, the regulatory proteins involved in flavonol biosynthesis are not well characterized. Using a homology-based approach, three R2R3-MYB transcription factor encoding genes have been identified in the Medicago truncatula reference genome sequence. The gene encoding a protein with highest similarity to known flavonol regulators, MtMYB134, was chosen for further experiments and was characterized as a functional flavonol regulator from M. truncatula. MtMYB134 expression levels are correlated with the expression of MtFLS2, encoding a key enzyme of flavonol biosynthesis, and with flavonol metabolite content. MtMYB134 was shown to activate the promoters of the A. thaliana flavonol biosynthesis genes AtCHS and AtFLS1 in Arabidopsis protoplasts in a transactivation assay and to interact with the Medicago promoters of MtCHS2 and MtFLS2 in yeast 1-hybrid assays. To ascertain the functional aspect of the identified transcription factor, we developed a sextuple mutant, which is defective in anthocyanin and flavonol biosynthesis. Ectopic expression of MtMYB134 in a multiple myb A. thaliana mutant restored flavonol biosynthesis. Furthermore, overexpression of MtMYB134 in hairy roots of M. truncatula enhanced the biosynthesis of various flavonol derivatives. Taken together, our results provide insight into the understanding of flavonol biosynthesis regulation in M. truncatula and provides MtMYB134 as tool for genetic manipulation to improve flavonol synthesis.


Asunto(s)
Vías Biosintéticas , Flavonoles/biosíntesis , Medicago truncatula/metabolismo , Proteínas de Plantas/metabolismo , Factores de Transcripción/metabolismo , Secuencia de Aminoácidos , Arabidopsis/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Medicago truncatula/enzimología , Medicago truncatula/genética , Mutación/genética , Sistemas de Lectura Abierta/genética , Especificidad de Órganos/genética , Fenotipo , Proteínas de Plantas/química , Raíces de Plantas/genética , Regiones Promotoras Genéticas , Unión Proteica , Factores de Transcripción/química
14.
Plant Mol Biol ; 106(1-2): 173-192, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33738678

RESUMEN

KEY MESSAGE: A transcriptome analysis reveals the transcripts and alleles differentially expressed in sugarcane genotypes with contrasting lignin composition. Sugarcane bagasse is a highly abundant resource that may be used as a feedstock for the production of biofuels and bioproducts in order to meet increasing demands for renewable replacements for fossil carbon. However, lignin imparts rigidity to the cell wall that impedes the efficient breakdown of the biomass into fermentable sugars. Altering the ratio of the lignin units, syringyl (S) and guaiacyl (G), which comprise the native lignin polymer in sugarcane, may facilitate the processing of bagasse. This study aimed to identify genes and markers associated with S/G ratio in order to accelerate the development of sugarcane bioenergy varieties with modified lignin composition. The transcriptome sequences of 12 sugarcane genotypes that contrasted for S/G ratio were compared and there were 2019 transcripts identified as differentially expressed (DE) between the high and low S/G ratio groups. These included transcripts encoding possible monolignol biosynthetic pathway enzymes, transporters, dirigent proteins and transcriptional and post-translational regulators. Furthermore, the frequencies of single nucleotide polymorphisms (SNPs) were compared between the low and high S/G ratio groups to identify specific alleles expressed with the phenotype. There were 2063 SNP loci across 787 unique transcripts that showed group-specific expression. Overall, the DE transcripts and SNP alleles identified in this study may be valuable for breeding sugarcane varieties with altered S/G ratio that may provide desirable bioenergy traits.


Asunto(s)
Regulación de la Expresión Génica de las Plantas , Lignina/metabolismo , Saccharum/genética , Saccharum/metabolismo , Alelos , Transporte Biológico , Vías Biosintéticas/genética , Pared Celular/metabolismo , Perfilación de la Expresión Génica , Ontología de Genes , Genes de Plantas , Genotipo , Lignina/biosíntesis , Lignina/química , Anotación de Secuencia Molecular , Polimerizacion , ARN Mensajero/genética , ARN Mensajero/metabolismo
15.
Sheng Wu Gong Cheng Xue Bao ; 37(2): 404-417, 2021 Feb 25.
Artículo en Chino | MEDLINE | ID: mdl-33645144

RESUMEN

Curcuminoids are rare diketone compounds in plants and can be found in the rhizome of Curcuma longa as well as other Zingiberaceae and Araceae. Curcuminoids have been widely used in food and medical area owing to the yellow colors, as well as the antioxidant and many other pharmacological activities. Curcuminoids are a mixture of compounds containing curcumin, demethoxycurcumin and bisdemethoxycurcumin, which have distinct benzene ring substituents. Currently, curcuminoids are exclusively produced through plant extraction, which do not satisfy the meeting of the market demand. Empowered with new synthetic biology tools and metabolic engineering strategies, there is renewed interest in production of curcuminoids using microorganisms. Heterologous production of curcuminoids has been achieved using Escherichia coli, Yarrowia lipolytica, Pseudomonas putida and Aspergillus oryzae via engineering of curcuminoids biosynthesis pathway. In this review, we first describe the biological activities and various applications of curcuminoids. Next, we summarize the biosynthetic pathway of curcuminoids in Curcuma longa and discuss the catalytic mechanisms of curcumin synthases. Then, we thoroughly explore recent advances in the use of distinct microorganisms for the production of curcuminoids with a special focus on metabolic engineering strategies. Finally, we prospect the microbial production of curcuminoids by highlighting some promising techniques and approaches.


Asunto(s)
Curcumina , Diarilheptanoides , Antioxidantes , Vías Biosintéticas/genética , Ingeniería Metabólica , Extractos Vegetales
16.
Nat Commun ; 12(1): 1513, 2021 03 08.
Artículo en Inglés | MEDLINE | ID: mdl-33686068

RESUMEN

3-Hydroxypropionic acid (3HP), an important three carbon (C3) chemical, is designated as one of the top platform chemicals with an urgent need for improved industrial production. Halomonas bluephagenesis shows the potential as a chassis for competitive bioproduction of various chemicals due to its ability to grow under an open, unsterile and continuous process. Here, we report the strategy for producing 3HP and its copolymer poly(3-hydroxybutyrate-co-3-hydroxypropionate) (P3HB3HP) by the development of H. bluephagenesis. The transcriptome analysis reveals its 3HP degradation and synthesis pathways involving endogenous synthetic enzymes from 1,3-propanediol. Combing the optimized expression of aldehyde dehydrogenase (AldDHb), an engineered H. bluephagenesis strain of whose 3HP degradation pathway is deleted and that overexpresses alcohol dehydrogenases (AdhP) on its genome under a balanced redox state, is constructed with an enhanced 1.3-propanediol-dependent 3HP biosynthetic pathway to produce 154 g L-1 of 3HP with a yield and productivity of 0.93 g g-1 1,3-propanediol and 2.4 g L-1 h-1, respectively. Moreover, the strain could also accumulate 60% poly(3-hydroxybutyrate-co-32-45% 3-hydroxypropionate) in the dry cell mass, demonstrating to be a suitable chassis for hyperproduction of 3HP and P3HB3HP.


Asunto(s)
Vías Biosintéticas , Halomonas/genética , Halomonas/metabolismo , Ácido Láctico/análogos & derivados , Ácido Láctico/biosíntesis , Ingeniería Metabólica , Proteínas Bacterianas/metabolismo , Biopolímeros/metabolismo , Vías Biosintéticas/genética , Edición Génica , Regulación Bacteriana de la Expresión Génica , Halomonas/enzimología , Hidroxibutiratos/metabolismo , Poliésteres/metabolismo , Glicoles de Propileno/metabolismo
17.
Sci Total Environ ; 771: 145336, 2021 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-33736184

RESUMEN

Symbiotic corals receive energy not only by ingesting food (e.g. plankton, inorganic/organic matter, i.e. heterotrophy), but also by endosymbiosis, which supplies photosynthates (dissolved inorganic carbon, i.e. autotrophy). These two sources of energy have distinct fatty acid (FA) profiles, which can be used to differentiate corals by their primary feeding mode. FA profiles have been applied as biomarkers to evaluate the quality of nutrition in the midst of environmental change. However, species-specific responses of coral FA profiles and biosynthetic pathway under cultural eutrophication are still unknown. We collected two coral species (Acropora samoensis, Platygyra carnosa) from sites with different levels of eutrophication to test for variations in FA profiles. Gas Chromatography-Mass Spectrometry (GC-MS) was performed to identify FA profiles and quantify their concentration. Our main findings are threefold: 1) chronic eutrophication inhibits corals' ability to synthesize essential FA; 2) PUFA:SFA ratio and certain FA biomarkers or their pathway can be successfully utilized to determine the relative degree of autotrophy and heterotrophy in corals; 3) under eutrophication, different FA profiles of coral host tissue are attributed to different feeding strategies. Thus, our research provides significant new insights into the roles of FA as a risk assessment tool in coral reef ecosystems under the pressure of eutrophication.


Asunto(s)
Antozoos , Animales , Vías Biosintéticas , Arrecifes de Coral , Ecosistema , Eutrofización , Ácidos Grasos , Simbiosis
18.
Nat Commun ; 12(1): 1696, 2021 03 16.
Artículo en Inglés | MEDLINE | ID: mdl-33727542

RESUMEN

Cystobactamids are myxobacteria-derived topoisomerase inhibitors with potent anti-Gram-negative activity. They are formed by a non-ribosomal peptide synthetase (NRPS) and consist of tailored para-aminobenzoic acids, connected by a unique α-methoxy-L-isoasparagine or a ß-methoxy-L-asparagine linker moiety. We describe the heterologous expression of the cystobactamid biosynthetic gene cluster (BGC) in Myxococcus xanthus. Targeted gene deletions produce several unnatural cystobactamids. Using in vitro experiments, we reconstitute the key biosynthetic steps of linker formation and shuttling via CysB to the NRPS. The biosynthetic logic involves a previously uncharacterized bifunctional domain found in the stand-alone NRPS module CysH, albicidin biosynthesis and numerous BGCs of unknown natural products. This domain performs either an aminomutase (AM) or an amide dehydratase (DH) type of reaction, depending on the activity of CysJ which hydroxylates CysH-bound L-asparagine. Furthermore, CysQ O-methylates hydroxyl-L-(iso)asparagine only in the presence of the AMDH domain. Taken together, these findings provide direct evidence for unique steps in cystobactamid biosynthesis.


Asunto(s)
Amidas/metabolismo , Antibacterianos/biosíntesis , Amidas/química , Asparagina/metabolismo , Vías Biosintéticas , Hidroxilación , Modelos Biológicos , Peso Molecular , Myxococcus xanthus/metabolismo , Especificidad por Sustrato
19.
Zhongguo Zhong Yao Za Zhi ; 46(5): 1084-1093, 2021 Mar.
Artículo en Chino | MEDLINE | ID: mdl-33787101

RESUMEN

In order to enrich the transcriptome data of Fagopyrum dibotrys plants, analyze the genes encoding key enzyme involved in flavonoid biosynthesis pathway, and mine their functional genes, in this study, we performed RNA sequencing analysis for the rhizomes, roots, flowers, leaves and stems of F. dibotrys on the BGISEQ-500 sequencing platform. After de novo assembly of transcripts, a total of 205 619 unigenes were generated and 132 372 unigenes were obtained and annotated into seven public databases, of which, 81 327 unigenes were mapped to the GO database and most of the unigenes were annotated in cellular process, biological regulation, binding and catalytic activity. Besides, 86 922 unigenes were enriched in 136 pathways using KEGG database' and we identified 82 unigenes that encodes key enzymes involved in flavonoid biosynthesis. Comparing rhizome with root, flower, leaf or stem in F. dibotrys, 27 962 co-expressed differentially expressed genes(DEGs) were obtained. Among them, 23 515 DEGs of rhizome tissue-specific were enriched into 132 pathways and 13 unigenes were significantly enriched in biosynthesis of flavone and flavonol. In addition, we also identified 3 427 unigenes encoding 60 transcription factor(TFs) families as well as four unigenes encoding bHLH TFs were enriched in flavonoid biosynthesis. Our results greatly enriched the transcriptome database of plants, provided a reference for the analysis of key enzymes involved in flavonoid biosynthesis in plants, and will facilitate the study of the functions and regulatory mechanisms of key enzymes involved in flavonoid biosynthesis in F. dibotrys at the genetic level.


Asunto(s)
Fagopyrum , Vías Biosintéticas/genética , Flavonoides , Flores , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Humanos , Transcriptoma/genética
20.
Nat Commun ; 12(1): 1389, 2021 03 02.
Artículo en Inglés | MEDLINE | ID: mdl-33654100

RESUMEN

Sulfated glycosaminoglycans (GAGs) are a class of important biologics that are currently manufactured by extraction from animal tissues. Although such methods are unsustainable and prone to contamination, animal-free production methods have not emerged as competitive alternatives due to complexities in scale-up, requirement for multiple stages and cost of co-factors and purification. Here, we demonstrate the development of single microbial cell factories capable of complete, one-step biosynthesis of chondroitin sulfate (CS), a type of GAG. We engineer E. coli to produce all three required components for CS production-chondroitin, sulfate donor and sulfotransferase. In this way, we achieve intracellular CS production of ~27 µg/g dry-cell-weight with about 96% of the disaccharides sulfated. We further explore four different factors that can affect the sulfation levels of this microbial product. Overall, this is a demonstration of simple, one-step microbial production of a sulfated GAG and marks an important step in the animal-free production of these molecules.


Asunto(s)
Vías Biosintéticas , Sulfatos de Condroitina/biosíntesis , Escherichia coli/metabolismo , Transporte Biológico , Escherichia coli/enzimología , Fermentación , Oxidorreductasas/metabolismo , Sulfotransferasas/metabolismo
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