Combination Strategy of Genetic Dereplication and Manipulation of Epigenetic Regulators Reveals a Novel Compound from Plant Endophytic Fungus

The strategies of genetic dereplication and manipulation of epigenetic regulators to activate the cryptic gene clusters are effective to discover natural products with novel structure in filamentous fungi. In this study, a combination of genetic dereplication (deletion of pesthetic acid biosynthetic gene, PfptaA) and manipulation of epigenetic regulators (deletion of histone methyltransferase gene PfcclA and histone deacetylase gene PfhdaA) was developed in plant endophytic fungus Pestalotiopsis fici. The deletion of PfptaA with PfcclA and/or PfhdaA led to isolation of 1 novel compound, pestaloficiol X (1), as well as another 11 known compounds with obvious yield changes. The proposed biosynthesis pathway of pestaloficiol X was speculated using comparative analysis of homologous biosynthetic gene clusters. Moreover, phenotypic effects on the conidial development and response to oxidative stressors in the mutants were explored. Our results revealed that the new strain with deletion of PfcclA or PfhdaA in ΔPfptaA background host can neutralise the hyperformation of conidia in the PfptaA mutant, and that the ΔPfptaA ΔPfhdaA mutant was generally not sensitive to oxidative stressors as much as the ΔPfptaA ΔcclA mutant in comparison with the single mutant ΔPfptaA or the parental strains. This combinatorial approach can be applied to discover new natural products in filamentous fungi.     

Biosynthesis of Colabomycin E,a New Manumycin‐Family Metabolite,Involves an Unusual Chain‐Length Factor

Colabomycin E is a new member of the manumycin‐type metabolites produced by the strain Streptomyces aureus SOK1/5‐04 and identified by genetic screening from a library of streptomycete strains. The structures of colabomycin E and accompanying congeners were resolved. The entire biosynthetic gene cluster was cloned and expressed in Streptomyces lividans. Bioinformatic analysis and mutagenic studies identified components of the biosynthetic pathway that are involved in the formation of both polyketide chains. Recombinant polyketide synthases (PKSs) assembled from the components of colabomycin E and asukamycin biosynthetic routes catalyzing the biosynthesis of “lower” carbon chains were constructed and expressed in S. aureus SOK1/5‐04 ΔcolC11–14 deletion mutant. Analysis of the metabolites produced by recombinant strains provided evidence that in both biosynthetic pathways the length of the lower carbon chain is controlled by an unusual chain‐length factor supporting biosynthesis either of a triketide in asukamycin or of a tetraketide in colabomycin E. Biological activity assays indicated that colabomycin E significantly inhibited IL‐1β release from THP‐1 cells and might thus potentially act as an anti‐inflammatory agent.     

A Synthetic,Species‐Specific Activator of Secondary Metabolism and Sporulation in Streptomyces coelicolor

The secondary metabolites produced by bacterial species serve many clinically useful purposes, and Streptomyces have been an abundant source of such compounds. However, a poor understanding of their regulatory cascades leads to an inability to isolate all of the secondary metabolites this genus is capable of producing. This study focuses on comparing synthetic small molecules that were found to alter the production of secondary metabolites in Streptomyces coelicolor. A survey of these molecules suggests that each has a distinct mechanism of action, and hence, could be used as a unique probe of secondary metabolism. A comparative analysis of two of these molecules, ARC2 and ARC6, confirmed that they modulate secondary metabolites in different ways. In a separate study, ARC2 was shown to give rise to a different phenotype through the inhibition of a target in fatty acid biosynthesis. The results of this study suggest that ARC6 does not have the same target, although it might target the same metabolic system. Furthermore, the results demonstrate that ARC2 and ARC6 act through distinct mechanisms and further suggest that chemical probes can be important tools in enhancing our understanding of secondary metabolism and the streptomycete life cycle.     

源自微生物次生代谢产物的3,4-二氢异香豆素

结合自己的工作,对分离自真菌、细菌等微生物次生代谢产物的3,4-二氢异香豆素的来源、结构特征、生物活性等方面的研究进展进行了综述,引用文献28篇,涉及26种真菌或细菌的32个3,4-二氢异香豆素类代谢产物。     

Aromatics from Lignocellulosic Biomass: Economic Analysis of the Production of p‐Xylene from 5‐Hydroxymethylfurfural

A novel production of biobased p‐xylene from hydroxymethylfurfual has been investigated and presented. This is an important part of a process for the production of aromatics from lignocellulosic biomass. The detailed flow sheets are designed based on laboratory experiments by Leshkov et al. and William et al. and economic analysis has been performed. The minimum biobased p‐xylene cost is estimated as $3,962/metric ton, of which the main contribution comes from the HMF cost. Sensitivity analysis has been used to assess the impact of uncertainties of economic parameters and also to determine the most significant reaction factor in the technological development, i.e., selectivity and conversion. It has been shown that high selectivity is favored at the same yield. © 2013 American Institute of Chemical Engineers AIChE J, 59: 2079–2087, 2013     

Chemistry of the Lichen <Emphasis Type="Italic">Hypogymnia physodes</Emphasis> Transplanted to an Industrial Region

Lichens produce a great number of secondary metabolites that participate in ecological interactions and respond to environmental changes. We examined the influence of heavy metal accumulations on lichen secondary metabolism. Thalli of Hypogymnia physodes were transplanted for 6 months to the Cracow–Silesia industrial region. Based on heavy metal accumulations in lichen, two of the investigated sites were classified as highly polluted. The highest concentrations of Cd, Pb, and Zn were found in lichens transplanted in the vicinity of a Zn–Pb smelter. Significant accumulations of Cr and Ni were detected in Hypogymnia transplanted near a chemical industry. Physodic, physodalic, hydroxyphysodic acids, and atranorin were identified and analyzed in extracts obtained from specimen samples. The most detrimental changes were observed in lichen transplanted into the vicinity of a chemical industry producing chromium, phosphor, and sulfur compounds that contained 340-fold higher Cr levels than control thalli. Decreases in the levels of physodic acid, hydroxyphysodic acid, and atranorin were detected, and one additional polar compound (probably product of degradation of lichen acids) appeared in the extract. The content of physodalic acid increased in every thalli sample transplanted, suggesting a possible role of this compound in defense against stress caused by accumulated pollutants. The levels of physodic acid decreased in thalli from both of the most polluted sites compared to those of the controls—but were not changed in thalli transplanted to less polluted sites. Our results illustrate that lichen compounds are sensitive to heavy metal accumulation and could be used as biomarkers in environmental studies.     

Variation in Secondary Metabolite Concentrations in Yellow and Grey Morphs of the Red Sea Soft Coral Parerythropodium fulvum fulvum: Possible Ecological Implications

Secondary metabolites in yellow and grey morphs of the soft coral Parerythropodium fulvum fulvum were compared between colonies collected from different depths and reef sites along the Red Sea. The concentrations of fulfulvene, the major metabolite in the yellow morph, varied considerably among samples, with significant differences between shallow and deep colonies. The concentrations of 5-hydroxy-8-methoxy-calamenene and 5-hydroxy-8-methoxy-calamenene-6-al, the major metabolites in the grey morph, also exhibited significant differences between shallow and deep colonies. The ecological implications of these variations in secondary metabolites are discussed.     

Development of Tolerance to the Dietary Plant Secondary Metabolite 1,8-cineole by the Brushtail Possum (<Emphasis Type="Italic">Trichosurus vulpecula</Emphasis>)

The common brushtail possum (Trichosurus vulpecula) is a generalist herbivore whose diet includes Eucalyptus leaves that are well defended by plant secondary metabolites (PSM) such as the terpene 1,8-cineole (cineole). We accustomed possums to a terpene-free diet, then challenged them with the addition of 2% cineole to the diet. Initially, there was a 50% reduction in total overnight food consumption associated with a marked decrease in the mass of the major feeding bout. After nine nights, however, cineole tolerance had developed as total food consumption had returned to the control amount. Compared to the control diet, the cineole diet was eaten in a larger number of smaller bouts, which were also eaten at a slower rate. The experiment was repeated with animals that had been accustomed to day-time feeding to take blood samples during feeding sessions. Feeding variables and blood concentration data for cineole were compared on the first and seventh day of the cineole diet. Although the total food consumed increased 2.5-fold after 7 days of the cineole diet, there was no increase in average blood cineole concentration, measured as the area under the concentration–time curve. This indicates that induction of liver enzymes resulted in greater pre-systemic metabolism of cineole and reduced bioavailability. The maximum tolerated blood concentration of cineole also increased, suggesting some adaptation of the central nervous system to the cineole aversive effects. This appears to be the first report in a vertebrate herbivore that consumption of a dietary PSM leads to metabolism induction and that this contributes to development of tolerance to the PSM. Overall, herbivores adapt to newly encountered dietary PSMs by immediate changes in feeding behavior followed by development of increased metabolism of PSM and probably diminished cellular responsiveness to effects.     

Supercritical CO2 extraction of accumulated capsidiol from biotic elicitor‐activated Capsicum annuum L fruit tissues

This work investigates the supercritical CO₂ extraction of capsidiol from pepper fruit tissues activated with Alternaria alternate (Fr) Keissler suspension culture as a biotic elicitor. Capsidiol production in the fruit tissue was markedly increased by the treatment with a biotic elicitor and reached its maximum level after 4 days of elicitation. The effects of separation parameters such as temperature, pressure, supercritical solvent flow rate, particle diameter and also initial capsidiol concentration were investigated on solubility, initial extraction rate and extraction yield. The optimal extraction conditions were obtained at the temperature of 40 °C, the pressure of 400 bar, the supercritical CO₂ flow rate of 2 cm³ min^?1, and the average particle diameter of 116 µm. The results showed that the ratio of the supercritical CO₂ extraction yield to the organic solvent extraction yield was changed from 84 to 97 wt‐% depending on the initial capsidiol concentration. Copyright © 2004 Society of Chemical Industry     

Rapid Absorption of Dietary 1,8-Cineole Results in Critical Blood Concentration of Cineole and Immediate Cessation of Eating in the Common Brushtail Possum (<Emphasis Type="Italic">Trichosurus vulpecula</Emphasis>)

The blood concentration of 1,8-cineole and its metabolites was measured in six male brushtail possums while they voluntarily fed on diets laced with varying concentrations of cineole for 3 d. On the third day, blood samples were collected during and after each bout of feeding for 3 hr. Blood cineole was measured by using headspace solid-phase microextraction (SPME), while cineole metabolites were measured by liquid–liquid extraction followed by gas chromatography-mass spectroscopy. Feeding patterns were measured by continual recording of residual food weight and time. Cineole absorption was rapid, resulting in a peak blood concentration at the end of each feeding bout. The blood concentration of cineole did not exceed a critical value (51.8 ± 14.1 μmol/l) regardless of the concentration in the diet. Food and, therefore, cineole intake was regulated. The amount of food ingested in the first feeding bout decreased from 236 ± 52 g on the control diet to 36 ± 20 g on the 4% cineole diet. The amount of cineole ingested in the first bout (1.18 ± 1.10 g) was the same regardless of the dietary concentration and was controlled by the size of the meal. Total food eaten during the 7-hr feeding session decreased by 64% from 368 ± 94 g (control diet) to 131 ± 52 g (4% diet). Total cineole intake increased from 2.47 ± 0.60 g (1% diet) to 5.05 ± 2.41 g (4% diet). Cineole metabolites accumulated throughout the sampling period and were generally still rising at the end of blood sampling period. Blood levels of metabolites were at least 10-fold higher than cineole levels. The immediate control of feeding seems to be regulated by blood levels of cineole, whereas metabolites are likely to be more important in regulating the chronic ingestion of cineole.     

Synthesis and properties of polyamides from 2,5‐furandicarboxylic acid

The aim of this study is to investigate the synthesis of 2,5‐furandicarboxylic acid (FDCA)‐based copolyamides. Indeed, FDCA monomer may be a potential bio‐based alternative to phthalic acids. A series of polyamides and copolyamides, PA 6‐I(x)/6‐F(y), are synthesized in a pilot scale reactor by melt polycondensation of salts based on FDCA, isophthalic acid (IPA), and 1,6‐hexamethylenediamine. The chemical structure and composition of the resulting copolymers are extensively characterized by NMR (¹H, ¹³C, and 2D), MALDI‐TOF as well as size exclusion chromatography and solution viscosimetry. Their thermal properties are studied by differential scanning calorimetry and TGA and also by a molecular modeling technique. It is pointed out that, during the synthesis, FDCA‐rich polymers overcome a massive decarboxylation of FDCA, preventing the production of high molar mass polymers. It is also found that all polymers are amorphous and that the glass transition temperatures decrease as the amount of FDCA in the polymer increases. This is confirmed by the results from molecular dynamics. In addition, it is observed that the copolymers become more hygroscopic when the amount of FDCA in the copolyamides increases. However, the water uptake of PA 6‐I(x)/6‐F(y) with 90 > x > 50 is lower than those of PA6 and PA 66 polyamides. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2018 , 135, 45901.     

Volatile Compounds on the Leaf Surface of Intact and Regrowth Tarbush (<Emphasis Type="Italic">Flourensia cernua</Emphasis> DC) Canopies

Shrub expansion into desert grasslands is a serious problem resulting in loss of forage and rangeland productivity. Flourensia cernua DC (tarbush) is one such shrub contributing to the decline of Chihuahuan Desert grasslands. Our previous research has shown tarbush consumption by sheep and goats to be negatively related to leaf surface concentration of individual terpenes and epicuticular wax. Concentrations of compounds such as terpenes often change with plant age and phenology. Our objective was to examine the effect of altering the vegetative state of tarbush on volatile chemicals. Ninety tarbush plants were randomly selected, and all biomass within 10 cm of the soil surface was removed from 45 plants during winter dormancy. Leaves were collected the following summer during active growth from the canopy of intact controls and resprouts. Leaf surface volatiles were analyzed by gas chromatography-mass spectroscopy and subjected to univariate analysis of variance and stepwise discriminate analysis. Of the 87 compounds present on tarbush leaves, 35 were greater in canopy samples and 16 were greater in regrowth samples based on univariate analysis (P < 0.05). Mean concentration of total volatiles on canopy leaves tended to be less (P = 0.062) than that of regrowth (3,642 vs. 4,684 μg/g DM). Nine compounds in the discriminant analysis (α-muurolene, iso-borneol, unknown#6, p-cymen-8-ol, unknown#7, sabinene, β-caryophyllene, δ-cadinene, and α-copaene) explained 95% of the variation between canopy and regrowth samples. Lower cumulative concentration of volatile compounds in canopy than regrowth samples suggests repsrouts may be less vulnerable to herbivory than intact tarbush.     

The Role of Secondary Interactions in the Asymmetric Palladium‐Catalysed Hydrosilylation of Olefins with Monophosphane Ligands

A structure‐activity survey of monophosphane ligands used in the asymmetric palladium‐catalysed hydrosilylation reaction of olefins is presented and used to assess the role of secondary interactions on catalyst activity and selectivity.     

Salinity Stress Alters the Secondary Metabolic Profile of M. sativa,M. arborea and Their Hybrid (Alborea)

Increased soil salinity, and therefore accumulation of ions, is one of the major abiotic stresses of cultivated plants that negatively affect their growth and yield. Among Medicago species, only Medicago truncatula, which is a model plant, has been extensively studied, while research regarding salinity responses of two important forage legumes of Medicago sativa (M. sativa) and Medicago arborea (M. arborea) has been limited. In the present work, differences between M. arborea, M. sativa and their hybrid Alborea were studied regarding growth parameters and metabolomic responses. The entries were subjected to three different treatments: (1) no NaCl application (control plants), (2) continuous application of 100 mM NaCl (acute stress) and (3) gradual application of NaCl at concentrations of 50-75-150 mM by increasing NaCl concentration every 10 days. According to the results, M. arborea maintained steady growth in all three treatments and appeared to be more resistant to salinity. Furthermore, results clearly demonstrated that M. arborea presented a different metabolic profile from that of M. sativa and their hybrid. In general, it was found that under acute and gradual stress, M. sativa overexpressed saponins in the shoots while M. arborea overexpressed saponins in the roots, which is the part of the plant where most of the saponins are produced and overexpressed. Alborea did not perform well, as more metabolites were downregulated than upregulated when subjected to salinity stress. Finally, saponins and hydroxycinnamic acids were key players of increased salinity tolerance.     

Structure and Biosynthesis of Fimsbactins A–F,Siderophores from Acinetobacter baumannii and Acinetobacter baylyi

Novel chatechol/hydroxamate siderophores (named “fimsbactins”) were identified in Acinetobacter baumannii ATCC 17978 and Acinetobacter baylyi ADP1. The major compound, fimsbactin A, was isolated from low‐iron cultures of A. baylyi ADP1, and its chemical structure was elucidated by mass spectrometry, and detailed ¹H, ¹³C and ¹⁵N NMR spectroscopy. From inverse feeding experiments following HPLC‐MS analysis, the structures of five additional derivatives were elucidated. The gene cluster encoding the fimsbactin synthetase (fbs) was identified in both genomes, and mutants in fbs genes in A. baylyi were analyzed, thus allowing prediction of the fimsbactin biosynthesis pathway.     

First Application of Secondary Phosphines as Supporting Ligands for the Palladium‐Catalyzed Heck Reaction: Efficient Activation of Aryl Chlorides

Secondary dialkylphosphines were successfully used for the first time as efficient supporting ligands for the palladium‐catalyzed Heck reaction of electron‐rich and electron‐poor aryl chlorides with olefins such as acrylate, ethylene, styrene, and n‐butyl vinyl ether. The yields with HP(t‐butyl)₂ and HP(adamantyl)₂ were comparable or better than those obtained with known systems of tertiary phosphines such as P(cyclohexyl)₃ and P(t‐butyl)₃, especially at a catalyst loading of <1 mol %. In comparison with tertiary phosphines, the secondary phosphines have the advantage of being readily available at low cost on a technical scale, and are comparable with respect to handling and oxygen sensitivity.     

Fermentative Production of L‐Lysine‐L‐lactate with Fractionated Press Juices from the Green Biorefinery

Lactic acid (LA) is a platform chemical for a future bio‐based economy. Aminium lactates act as intermediates for the production of lactic acid sequence products like lactide. It could be demonstrated that, after separation of proteins from alfalfa press juice, the supernatant can be used as fermentation media for the production of the aminium lactate, L‐lysine‐L‐lactate.     

Enrichment of Secondary Wastewater Sludge for Production of Hydrogen from Crude Glycerol and Comparative Evaluation of Mono-, Co- and Mixed-Culture Systems

Anaerobic digestion using mixed-culture with broader choice of pretreatments for hydrogen (H₂) production was investigated. Pretreatment of wastewater sludge by five methods, such as heat, acid, base, microwave and chloroform was conducted using crude glycerol (CG) as substrate. Results for heat treatment (100 °C for 15 min) showed the highest H₂ production across the pretreatment methods with 15.18 ± 0.26 mmol/L of medium at 30 °C in absence of complex media and nutrient solution. The heat-pretreated inoculum eliminated H₂ consuming bacteria and produced twice as much as H₂ as compared to other pretreatment methods. The fermentation conditions, such as CG concentration (1.23 to 24 g/L), percentage of inoculum size (InS) (1.23% to 24% v/v) along with initial pH (2.98 to 8.02) was tested using central composite design (CCD) with H₂ production as response parameter. The maximum H₂ production of 29.43 ± 0.71 mmol/L obtained at optimum conditions of 20 g/L CG, 20% InS and pH 7. Symbiotic correlation of pH over CG and InS had a significant (p-value: 0.0011) contribution to H₂ production. The mixed-culture possessed better natural acclimatization activity for degrading CG, at substrate inhibition concentration and provided efficient inoculum conditions in comparison to mono- and co-culture systems. The heat pretreatment step used across mixed-culture system is simple, cheap and industrially applicable in comparison to mono-/co-culture systems for H₂ production.     

Production of activated carbon from agricultural by‐products

The production of activated carbon from agricultural by‐products is a research field of increasing interest as it deals with the problem of the disposal of agro‐residues, at the same time producing an added‐value product that can be used in a number of environmental applications. The paper presents an overview of the latest developments in processes for the production of activated carbon from agricultural by‐products, with emphasis on the methodology applied, the effect of critical process parameters such as retention time, temperature, chemical to material ratio, as well as the adsorbing capacity of the activated carbons produced in removing select compounds from synthetic and real wastewaters. Agricultural by‐products can be a source for activated carbon production with high surface areas and high adsorption capacity. Copyright © 2008 Society of Chemical Industry