Olive secoiridoids and semisynthetic bioisostere analogues for the control of metastatic breast cancer

(?)-Oleocanthal (1) and ligstroside aglycone (2) are common bioactive olive oil secoiridoids. Secoiridoid 1 has been previously reported as a c-MET inhibitor. Chemically, (?)-oleocanthal is the elenolic acid ester of the common olive phenolic alcohol tyrosol. Therefore, several analogues (4–13) were synthesized by esterification and carbamoylation of tyrosol using diverse phenolic naturally occurring in olive and heterocyclic acids as elenolic acid bioisosteres to assess the effect of replacing the acid moiety of (?)-oleocanthal. Their c-MET inhibitory activity as well as their antiproliferative, antimigratory, and anti-invasive activities against the highly metastatic human breast cancer cell line MDA-MB231 has been assessed. Ligstroside aglycone (2) showed the best antimigratory activity. Generally, tyrosol esters showed better activities versus carbamate analogues. Tyrosol sinapate (5) showed the best c-MET phosphorylation inhibitory activity in Z′-LYTE? kinase assay. Both 1 and 5 competitively inhibited the ATP binding into its pocket in the c-MET catalytic domain. Compound 5 showed selective activities against tumor cells without toxicity to the non-tumorigenic human breast MCF10A epithelial cell line. Tyrosol esters with a phenolic acid containing hydrogen bond donor and/or acceptor groups at the para-position have better anticancer and c-MET inhibitory activities. Olive oil secoiridoids are excellent scaffolds for the design of novel c-MET inhibitors.     

Identification and Molecular Characterization of the Homogentisate Pathway Responsible for Pyomelanin Production,the Major Melanin Constituents in Aeromonas media WS

The pigmentation of many Aeromonas species has been thought to be due to the production of a L-DOPA (L-3,4-dihydroxyphenylalanine) based melanin. However, in this study we found that although L-DOPA synthesis occurs in the high-melanin-yielding Aeromonas media strain WS, it plays a minor, if any, role in pigmentation. Instead, the pigmentation of A. media strain WS is due to the production of pyomelanin through HGA (homogentisate). Gene products of phhA (encodes phenylalanine hydroxylase), tyrB and aspC (both encode aromatic amino acid aminotransferase), and hppD (encodes 4-hydroxyphenylpyruvate dioxygenase) constitute a linear pathway of converting phenylalanine to HGA and disruption of any one of these genes impairs or blocks pigmentation of A. media strain WS. This HGA biosynthesis pathway is widely distributed in Aeromonas, but HGA is only detectable in the cultures of pigmented Aeromonas species. Heterologous expression of HppD from both pigmented and non-pigmented Aeromonas species in E. coli leads to the production of pyomelanin and thus pigmentation, suggesting that most Aeromonas species have the critical enzymes to produce pyomelanin through HGA. Taken together, we have identified a widely conserved biosynthesis pathway of HGA based pyomelanin in Aeromonas that may be responsible for pigmentation of many Aeromonas species.     

Biodegradation of polycyclic aromatic hydrocarbons by a halophilic microbial consortium

In this study we investigated the phenanthrene degradation by a halophilic consortium obtained from a saline soil sample. This consortium, named Qphe, could efficiently utilize phenanthrene in a wide range of NaCl concentrations, from 1% to 17% (w/v). Since none of the purified isolates could degrade phenanthrene, serial dilutions were performed and resulted in a simple polycyclic aromatic hydrocarbon (PAH)-degrading culture named Qphe-SubIV which was shown to contain one culturable Halomonas strain and one unculturable strain belonging to the genus Marinobacter. Qphe-SubIV was shown to grow on phenanthrene at salinities as high as 15% NaCl (w/v) and similarly to Qphe, at the optimal NaCl concentration of 5% (w/v), could degrade more than 90% of the amended phenanthrene in 6 days. The comparison of the substrate range of the two consortiums showed that the simplified culture had lost the ability to degrade chrysene but still could grow on other polyaromatic substrates utilized by Qphe. Metabolite analysis by HPLC and GC–MS showed that 2-hydroxy 1-naphthoic acid and 2-naphthol were among the major metabolites accumulated in the Qphe-SubIV culture media, indicating that an initial dioxygenation step might proceed at C1 and C2 positions. By investigating the growth ability on various substrates along with the detection of catechol dioxygenase gene, it was postulated that the uncultured Marinobacter strain had the central role in phenanthrene degradation and the Halomonas strain played an auxiliary role in the culture by utilizing phenanthrene metabolites whose accumulation in the media could be toxic.     

群体感应动态调控促进大肠杆菌合成酪醇

酪醇是一种多酚类天然产物,广泛应用于化工、医药和食品等领域。目前大肠杆菌(Escherichia coli)从头合成酪醇存在发酵菌体密度低和产量低等问题。为此,本研究将前期获得苯丙酮酸脱羧酶突变体ARO10F138L/D218G与不同来源的醇脱氢酶融合表达,最优组合ARO10F138L/D218G-L-YahK酪醇产量达到1.09 g/L。为进一步提高酪醇产量,敲除了4-羟基苯乙酸竞争途径关键基因feaB,使酪醇产量提高了21.15%,达到1.26g/L。针对酪醇发酵菌体密度低的问题,通过群体感应系统动态调控酪醇合成途径,减轻酪醇对底盘细胞的毒性作用,缓解生长抑制,使其产量提高了33.82%,达到1.74 g/L。在2 L发酵罐中,群体感应动态调控工程菌TRFQ5的酪醇产量达到4.22g/L,OD600值达到42.88,分别较静态诱导表达工程菌TRF5提高了38.58%和43.62%。本研究应用基因敲除技术,阻断了酪醇合成竞争途径;同时结合群体感应动态调控策略,减轻了酪醇毒性对底盘细胞的生长抑制,从而有效地提高了酪醇产量。本研究对其他高毒性化学品的生物合成具有良好的借鉴和应用价值。     

Multi-modular engineering of Saccharomyces cerevisiae for high-titre production of tyrosol and salidroside

Tyrosol and its glycosylated product salidroside are important ingredients in pharmaceuticals, nutraceuticals and cosmetics. Despite the ability of Saccharomyces cerevisiae to naturally synthesize tyrosol, high yield from de novo synthesis remains a challenge. Here, we used metabolic engineering strategies to construct S. cerevisiae strains for high-level production of tyrosol and salidroside from glucose. First, tyrosol production was unlocked from feedback inhibition. Then, transketolase and ribose-5-phosphate ketol-isomerase were overexpressed to balance the supply of precursors. Next, chorismate synthase and chorismate mutase were overexpressed to maximize the aromatic amino acid flux towards tyrosol synthesis. Finally, the competing pathway was knocked out to further direct the carbon flux into tyrosol synthesis. Through a combination of these interventions, tyrosol titres reached 702.30 ± 0.41 mg l⁻¹ in shake flasks, which were approximately 26-fold greater than that of the WT strain. RrU8GT33 from Rhodiola rosea was also applied to cells and maximized salidroside production from tyrosol in S. cerevisiae. Salidroside titres of 1575.45 ± 19.35 mg l⁻¹ were accomplished in shake flasks. Furthermore, titres of 9.90 ± 0.06 g l⁻¹ of tyrosol and 26.55 ± 0.43 g l⁻¹ of salidroside were achieved in 5 l bioreactors, both are the highest titres reported to date. The synergistic engineering strategies presented in this study could be further applied to increase the production of high value-added aromatic compounds derived from the aromatic amino acid biosynthesis pathway in S. cerevisiae.     

Homogentisic acid is the primary precursor of melanin synthesis in Vibrio cholerae, a Hyphomonas strain, and Shewanella colwelliana.

The enzyme p-hydroxyphenylpyruvate hydroxylase (HPPH) is involved in pigmentation (pyomelanin) via homogentisic acid (HGA). Pyomelanin formation is correlated with HGA production and expression of HPPH in three disparate marine species: Vibrio cholerae, a Hyphomonas strain, and Shewanella colwelliana. Induction of pigmentation in V. cholerae 569B by nutrient limitation also correlated with production of HGA.     

New insights and advances on pyomelanin production: from microbial synthesis to applications

Pyomelanin is a brown-black phenolic polymer and results from the oxidation of homogentisic acid (HGA) in the L-tyrosine pathway. As part of the research for natural and active ingredients issued from realistic bioprocesses, this work re-evaluates the HGA pigment and makes an updated inventory of its syntheses, microbial pathways, and properties, with tracks and recent advances for its large-scale production. The mechanism of the HGA polymerization is also well documented. In alkaptonuria, pyomelanin formation leads to connective tissue damage and arthritis, most probably due to the ROS issued from HGA oxidation. While UV radiation on human melanin may generate degradation products, pyomelanin is not photodegradable, is hyperthermostable, and has other properties better than L-Dopa melanin. This review aims to raise awareness about the potential of this pigment for various applications, not only for skin coloring and protection but also for other cells, materials, and as a promising (semi)conductor for bioelectronics and energy.     

o‐Phthalate derived from plastics’ plasticizers and a bacterium's solution to its anaerobic degradation

Phthalic acid esters (phthalates) are anthropogenic compounds that are used as plasticizers. Unfortunately, because phthalates are non‐covalently intercalated into plastic polymers they leach into the environment, accumulating in anoxic sediments. This has negative consequences for animal and human health. Denitrifying Betaproteobacteria, such as Aromatoleum aromaticum, can use ortho‐phthalate, derived by ester hydrolysis, as a carbon and energy source. Mergelsberg et al. ( 2018 ) deconstruct the pathway whereby ortho‐phthalic acid is converted, via the highly unstable phthaloyl‐CoA, to the central intermediate of anaerobic aromatic degradation, benzoyl‐CoA. The latter reaction is catalysed by UbiD‐like phthaloyl‐CoA decarboxylase (PCD). Succinyl‐CoA:o‐phthalate CoA‐transferase (SPT) generates phthaloyl‐CoA, which accumulates at only sub‐micromolar concentrations, while the K_m of PCD for phthaloyl‐CoA is two‐orders of magnitude higher. This seemingly insurmountable kinetic barrier is overcome because A. aromatoleum massively over‐produces PCD and because the decarboxylation reaction is irreversible. These features of the pathway facilitate capture of phthaloyl‐CoA as it is released from SPT without the need for direct substrate‐channelling. The authors provide strong evidence from both in vivo and in vitro studies to support their conclusions. This work reveals how these anaerobic bacteria have rapidly evolved a stop‐gap measure to allow them to completely degrade an otherwise recalcitrant aromatic xenobiotic.     

Rewiring central carbon metabolism for tyrosol and salidroside production in Saccharomyces cerevisiae

Metabolic engineering of Saccharomyces cerevisiae for high-level production of aromatic chemicals has received increasing attention in recent years. Tyrosol production from glucose by S. cerevisiae is considered an environmentally sustainable and safe approach. However, the production of tyrosol and salidroside by engineered S. cerevisiae has been reported to be lower than 2 g/L to date. In this study, S. cerevisiae was engineered with a push-pull-restrain strategy to efficiently produce tyrosol and salidroside from glucose. The biosynthetic pathways of ethanol, phenylalanine, and tryptophan were restrained by disrupting PDC1, PHA2, and TRP3. Subsequently, tyrosol biosynthesis was enhanced with a metabolic pull strategy of introducing PcAAS and EcTyrA^M53I/A354V. Moreover, a metabolic push strategy was implemented with the heterologous expression of phosphoketolase (Xfpk), and then erythrose 4-phosphate was synthesized simultaneously by two pathways, the Xfpk-based pathway and the pentose phosphate pathway, in S. cerevisiae. Furthermore, the heterologous expression of Xfpk alone in S. cerevisiae efficiently improved tyrosol production compared with the coexpression of Xfpk and phosphotransacetylase. Finally, the tyrosol yield increased by approximately 135-folds, compared with that of parent strain. The total amount of tyrosol and salidroside with glucose fed-batch fermentation was over 10 g/L and reached levels suitable for large-scale production.     

Halomonas sinaiensis sp. nov., a novel halophilic bacterium isolated from a salt lake inside Ras Muhammad Park, Egypt

An alkalitolerant and halotolerant bacterium, designated strain Sharm was isolated from a salt lake inside Ras Muhammad. The morphological, physiological and genetic characteristics were compared with those of related species of the genus Halomonas. The isolate grew optimally at pH 7.0, 5–15% NaCl at 35°C. The cells were Gram-negative rods, facultative anaerobes. They accumulated glycine-betaine, as a major osmolyte, and ectoine and glutamate as minor components. The strain Sharm^T biosynthetised α-glucosidase. The polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and a novel phosphoglycolipid as major components. Ubiquinone with nine repetitive unities (Q9) was the only quinone found and, nC16:0 and C19:0 with cyclopropane were the main cellular fatty acids, accounting for 87.3% of total fatty acids. The G + C content of the genomic DNA was 64.7 mol %. The 16S rRNA sequence analysis indicated that strain Sharm was a member of the genus Halomonas. The closest relatives of the strain Sharm were Halomonas elongata and Halomonas eurihalina. However, DNA–DNA hybridisation results clearly indicated that strain Sham was a distinct species of Halomonas. On the basis of the evidence, we propose to assign strain Sharm as a new species of the genus Halomonas, H. sinaiensis sp. nov, with strain Sharm^T as the type strain (DSM 18067^T; ATCC BAA-1308^T). The EMBL accession number for the 16S rRNA sequence of Halomonas sinaiensis strain Sharm^T is AM238662.     

Simultaneous Degradation of Atrazine and Phenol by Pseudomonas sp. Strain ADP: Effects of Toxicity and Adaptation

The strain Pseudomonas sp. strain ADP is able to degrade atrazine as a sole nitrogen source and therefore needs a single source for both carbon and energy for growth. In addition to the typical C source for Pseudomonas, Na₂-succinate, the strain can also grow with phenol as a carbon source. Phenol is oxidized to catechol by a multicomponent phenol hydroxylase. Catechol is degraded via the ortho pathway using catechol 1,2-dioxygenase. It was possible to stimulate the strain in order to degrade very high concentrations of phenol (1,000 mg/liter) and atrazine (150 mg/liter) simultaneously. With cyanuric acid, the major intermediate of atrazine degradation, as an N source, both the growth rate and the phenol degradation rate were similar to those measured with ammonia as an N source. With atrazine as an N source, the growth rate and the phenol degradation rate were reduced to ~35% of those obtained for cyanuric acid. This presents clear evidence that although the first three enzymes of the atrazine degradation pathway are constitutively present, either these enzymes or the uptake of atrazine is the bottleneck that diminishes the growth rate of Pseudomonas sp. strain ADP with atrazine as an N source. Whereas atrazine and cyanuric acid showed no significant toxic effect on the cells, phenol reduces growth and activates or induces typical membrane-adaptive responses known for the genus Pseudomonas. Therefore Pseudomonas sp. strain ADP is an ideal bacterium for the investigation of the regulatory interactions among several catabolic genes and stress response mechanisms during the simultaneous degradation of toxic phenolic compounds and a xenobiotic N source such as atrazine.     

Novel metabolic routes during the oxidation of hydroxylated aromatic acids by the yeast Arxula adeninivorans

Aim: To complete our study on tannin degradation via gallic acid by the biotechnologically interesting yeast Arxula adeninivorans as well as to characterize new degradation pathways of hydroxylated aromatic acids. Methods and Results: With glucose‐grown cells of A. adeninivorans, transformation experiments with hydroxylated derivatives of benzoic acid were carried out. The 12 metabolites were analysed and identified by high performance liquid chromatography and GC/MS. The yeast is able to transform the derivatives by oxidative and nonoxidative decarboxylation as well as by methoxylation. The products of nonoxidative decarboxylation of protocatechuate and gallic acid are substrates for further ring fission. Conclusion: Whereas other organisms use only one route of transformation, A. adeninivorans is able to carry out three different pathways (oxidative, nonoxidative decarboxylation and methoxylation) on one hydroxylated aromatic acid. The determination of the K_M‐values for protocatechuate and gallic acid in crude extracts of cells of A. adeninivorans cultivated with protocatechuate and gallic acid, respectively, suggests that the decarboxylation of protocatechuate and gallic acid may be catalysed by the same enzyme. Significance and Impact of the Study: This transformation pathway of protocatechuate and gallic acid via nonoxidative decarboxylation up to ring fission is novel and has not been described so far. This is also the first report of nonoxidative decarboxylation of gallic acid by a eukaryotic micro‐organism.     

Bioconversion of tyrosol into hydroxytyrosol and 3,4-dihydroxyphenylacetic acid under hypersaline conditions by the new Halomonas sp. strain HTB24

This paper reports the characterization of a Halomonas sp. strain (named HTB24) isolated from olive-mill wastewater and capable of transforming tyrosol into hydroxytyrosol (HT) and 3,4-dihydroxyphenylacetic acid (DHPA) in hypersaline conditions. This is the first time that a halophile has been shown to perform such reactions. The potent natural antioxidant HT was obtained through a C3 hydroxylation on the ring cycle, whereas DHPA was synthesized via the 4-hydroxyphenylacetic acid (HPA) pathway, which has been well described from other bacterial sources. HT was produced first, and then DHPA was detected in the medium accompanied by traces of HPA. HPA involved another pathway resulting from the activity of an aryl-dehydrogenase, which is suggested to be responsible for both tyrosol and hydroxytyrosol oxidation. Maximal HT content (2.30 mM) and maximal DHPA (5.15+/-0.42 mM) were obtained from a culture inoculated in the presence of 20 mM tyrosol and 0.5 g L(-1) yeast extract. Following this, DHPA was quickly degraded into 5-carboxymethyl-2-hydroxymuconic semialdehyde by a 2,3-dioxygenase, finally resulting in succinate and pyruvate. Phylogenetic analysis of the 16S rRNA gene revealed that this isolate was a member of the genus Halomonas. Strain HTB24, with a G+C content of 55.3 mol%, is closely related to Halomonas neptunia DSM 15720(T), 'Halomonas alkaliantarctica' DSM 15686(T) and Halomonas boliviensis DSM 15516(T).     

Tyrosol and tryptophol produced by <Emphasis Type="Italic">Ceratocystis adiposa</Emphasis>

Extracellular tyrosol and tryptophol were produced and isolated from the phytopathogen fungus Ceratocystis adiposa cultivated in a rich broth. These compounds have not been previously reported in this Ceratocystis species, as its identification was established through spectroscopic methods (¹H NMR, ¹³C NMR and MS). Productivity of tyrosol and tryptophol was 65.3 and 44 μg l⁻¹ day⁻¹, respectively, which were values higher than those reported to date. Tyrosol is a compound of pharmaceutical interest showing antioxidating activity, a property used in the treatment of atherosclerosis. Tryptophol is reported to have antibiotic and phytotoxic activity.     

深渊沉积物中盐单胞菌(Halomonas sp.)NyZ771菌株的分离培养及其降解芳香酸的研究

【背景】海洋是地球上最大的碳库,也是地球生物最大的栖息地。在这个庞大的生态系统中拥有多种多样的微生物,它们在全球碳循环中扮演了重要的角色。海斗深渊(海平面6 000 m以下的海域)由于高静水压和表层沉积汇集了大量有机质,形成了包含丰富生物资源的特殊生境。【目的】从马里亚纳海沟海斗深渊沉积物样品中分离培养能够以芳香酸为唯一碳源和能源生长的微生物,并研究其降解特性。【方法】通过模拟原位高压环境富集培养和常压条件下芳香酸选择性分离培养获得深渊来源的纯培养细菌,并根据形态学观察和16S rRNA基因序列系统发育分析进行种属鉴定,利用不同芳香酸进行培养和生物转化,通过HPLC和LC/MS鉴定芳香酸代谢中间产物。【结果】从马里亚纳海沟6 300 m沉积物样本中分离获得了一株盐单胞菌(Halomonas sp.)NyZ771。该菌株能够利用苯甲酸和4-羟基苯甲酸作为唯一碳源生长。其代谢4-羟基苯甲酸的中间产物鉴定为原儿茶酸。【结论】从深渊沉积物样本分离得到一株能降解苯甲酸和4-羟基苯甲酸的盐单胞菌NyZ771,丰富了深渊来源的微生物资源,为今后研究深渊中微生物的芳香酸降解及海洋微生物驱动的碳循环提供了一定的理论基础。     

Enantioseparation and enantioselective behavior of trichlorfon enantiomers in sediments

Trichlorfon (TF), an organophosphorus insecticide, has been widely used in seawater aquaculture; it is easily degraded to the highly toxic insecticide, dichlorvos (DDVP). In this study, the enantioseparation of TF enantiomers, as well as their degradation behavior and product (DDVP) formation in mariculture pond sediments, was investigated. The results show that both TF enantiomers degrade into DDVP, which is the main degradation product. Furthermore, S ‐(+)‐TF is preferentially degraded under natural conditions, suggesting that TF enantiomers degrade enantioselectively. Nevertheless, the degradation behavior of TF enantiomers is not enantiospecific under sterile conditions. The formation of DDVP and the enantiospecific degradation of TF enantiomers are attributed to the activities of microbes present in the sediments.     

Effect of olive oil minor components on oxidative stress and arachidonic acid mobilization and metabolism by macrophages RAW 264.7

Minor components of virgin olive oil may explain the healthy effects of the Mediterranean diet on the cardiovascular system and cancer development. The uncontrolled production of reactive oxygen species (ROS) and arachidonic acid (AA) metabolites contributes to the pathogenesis of cardiovascular disease and cancer, and inflammatory cells infiltrated in the atheroma plaque or tumor are a major source of ROS and eicosanoids. We aimed to determine the effects of squalene, beta-sitosterol, and tyrosol, which are representative of the hydrocarbons, sterols, and polyphenols of olive oil, respectively, on superoxide anion (O2(-)), hydrogen peroxide (H2O2), and nitric oxide (*NO) levels. We also studied AA release and eicosanoid production by phorbol esters (PMA)-stimulated macrophages RAW 264.7. beta-Sitosterol and tyrosol decreased the O2(-) and H2O2 production induced by PMA, and tyrosol scavenged the O2(-) released by a ROS generating system. These effects were correlated with the impairment of [3H]AA release, cyclooxygenase-2 (COX-2) expression, and prostaglandin E(2)/leukotriene B(4) synthesis in RAW 264.7 cultures stimulated by PMA. beta-Sitosterol exerted its effects after 3-6 h of preincubation. Tyrosol inhibited the [3H]AA release induced by exogenous ROS. beta-Sitosterol and tyrosol also reduced the *NO release induced by PMA, which was correlated with the impairment of inducible nitric oxide synthase (iNOS) levels. This may be correlated with the modulation of NF-kappaB activation. Further studies are required to gain more insight into the potential healthy effects of minor components of extra virgin olive oil.     

Isolation and characterization of marine-derived Mucor sp. for the fermentative production of tyrosol

An investigation of marine red algae surface-associated fungal communities led to the isolation of a previously undescribed Mucor strain. Many characteristic features of the genus Mucor, including sporangiophores, sporangium, sporangiospores and columellae, were apparent in scanning electron microscope (SEM) images of the isolated strain. The sequence of the internal transcribed spacer (ITS) rDNA revealed that the strain exhibits 97% homology to the genus Mucor. In a search for active compounds isolated by an ethyl acetate extract of the strain, tyrosol (2-(4-hydroxyphenyl) ethanol) was identified as a major secondary metabolite present in the culture broth. Remarkably, the tyrosol production level was considerably higher than that of reported tyrosol producing microorganisms. The optimal conditions for the fermentative production of tyrosol by the new strain were identified, including culture media, incubation period, temperature and pH. These findings clearly demonstrate that this novel strain has the potential for development as a natural source of tyrosol for industrial purposes.     

The influence of microbial communities for triadimefon enantiomerization in soils with different pH values

Enantiomers of chiral molecules can undergo interconversion leading to markedly different toxicities, which can introduce significant uncertainty when evaluating biological and environmental fates. However, enantiomerization (the reversible conversion of one enantiomer into the other) related to soil microorganism is rarely understood. For better understanding, S‐triadimefon and R‐triadimefon enantiopure were incubated in different soils with different pH value. Both high‐performance liquid chromatography and high‐throughput sequencing technology were used to explore target analytes quantitatively and microbial taxa related to the conversion process. Results revealed a significant enantiomerization among the soils. The alkaline soil from Beijing had a faster conversion than neutral soil from Changchun, while acidic soil from Wuhan had no conversion. At the same results, analysis of bacteria community showed higher abundance of Arthrobacter and Halomonas genus in alkaline soil than neutral soil after treatments, but the acidic soil was lower. Moreover, Arthrobacter and Halomonas were responsible for converting S‐triadimefon to R‐triadimefon and R‐triadimefon to S‐triadimefon in alkaline and neutral soil, respectively. Thus, these genera may be one of the reasons to explain the enantiomerization in different soils observed in this study. Thus, research at microbial level is necessary for efficient ecological risk assessment of chiral fungicide.     

Exploration of interaction mechanism of tyrosol as a potent anti-inflammatory agent

Abstract

Drug discovery for a vigorous and feasible lead candidate is a challenging scientific mission as it requires expertise, experience, and huge investment. Natural products and their derivatives having structural diversity are renowned source of therapeutic agents since many years. Tyrosol (a natural phenylethanoid) has been extracted from olive oil, and its structure was confirmed by elemental analysis, FT-IR, FT-NMR, and single crystal X-ray crystallography. The conformational analysis for tyrosol geometry was performed by Gaussian 09 in terms of density functional theory. Validation of bond lengths and bond angles obtained experimentally as well as theoretically were performed with the help of curve fitting analysis, and values of correlation coefficient (R) obtained as 0.988 and 0.984, respectively. The charge transfer within the tyrosol molecule was confirmed by analysis of HOMO→LUMO molecular orbitals. In molecular docking with COX-2 (PDB ID: 5F1A), tyrosol was found to possess satisfactory binding affinity as compared to other NSAIDs (Aspirin, Ibuprofen, and Naproxen) and a COX-2 selective drug (Celecoxib). ADMET prediction, drug-likeness and bioactivity score altogether confirm the lead/drug like potential of tyrosol. Further investigation of simulation quality plot, RMSD and RMSF plots, ligands behavior plot as well as post simulation analysis manifest the consistency of 5F1A-tyrosol complex throughout the 20?ns molecular simulation process that signifies its compactness and stability within the receptor pocket. Abbreviations ADMET Absorption, Distribution, Metabolism, Excretion and Toxicity

Å Angstrom

COX-2 Cyclooxygenase-2

DFT Density Functional Theory

DMF Dimethylformamide

FMO Frontier Molecular Orbital

FT-IR Fourier-transform Infrared Spectroscopy

FT-NMR Nuclear Magnetic Resonance Spectroscopy

HOMO Highest Occupied Molecular Orbital

LUMO Lowest Unoccupied Molecular Orbital

MD Molecular Dynamics

NS Nanosecond

NSAIDs Non-steroidal anti-inflammatory drugs

OPE Osiris Property Explorer

RMSD Root-Mean-Square Deviation

RMSF Root Sean Square Fluctuation

Communicated by Ramaswamy H. Sarma