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Background: The high-value aryl tetralin lignan (+)-pinoresinol is the main precursor of many plant lignans including (-)-podophyllotoxin, which is used for the synthesis of chemotherapeutics. As (-)-podophyllotoxin is traditionally isolated from endangered and therefore limited natural sources, there is a particular need for biotechnological production. Recently, we developed a reconstituted biosynthetic pathway from (+)-pinoresinol to (-)-deoxypodophyllotoxin, the direct precursor of (-)-podophyllotoxin, in the recombinant host Escherichia coli. However, the use of the expensive substrate (+)-pinoresinol limits its application from the economic viewpoint. In addition, the simultaneous expression of multiple heterologous genes from different plasmids for a multi-enzyme cascade can be challenging and limits large-scale use. Results: In this study, recombinant plasmid-free E. coli strains for the multi-step synthesis of pinoresinol from ferulic acid were constructed. To this end, a simple and versatile plasmid toolbox for CRISPR/Cas9-assisted chromosomal integration has been developed, which allows the easy transfer of genes from the pET vector series into the E. coli chromosome. Two versions of the developed toolbox enable the efficient integration of either one or two genes into intergenic high expression loci in both E. coli K-12 and B strains. After evaluation of this toolbox using the fluorescent reporter mCherry, genes from Petroselinum crispum and Zea mays for the synthesis of the monolignol coniferyl alcohol were integrated into different E. coli strains. The product titers achieved with plasmid-free E. coli W3110(T7) were comparable to those of the plasmid-based expression system. For the subsequent oxidative coupling of coniferyl alcohol to pinoresinol, a laccase from Corynebacterium glutamicum was selected. Testing of different culture media as well as optimization of gene copy number and copper availability for laccase activity resulted in the synthesis of 100 mg/L pinoresinol using growing E. coli cells. Conclusions: For efficient and simple transfer of genes from pET vectors into the E. coli chromosome, an easy-to-handle molecular toolbox was developed and successfully tested on several E. coli strains. By combining heterologous and endogenous enzymes of the host, a plasmid-free recombinant E. coli growing cell system has been established that enables the synthesis of the key lignan pinoresinol. [ABSTRACT FROM AUTHOR]

Abstract Background The high-value aryl tetralin lignan (+)-pinoresinol is the main precursor of many plant lignans including (-)-podophyllotoxin, which is used for the synthesis of chemotherapeutics. As (-)-podophyllotoxin is traditionally isolated from endangered and therefore limited natural sources, there is a particular need for biotechnological production. Recently, we developed a reconstituted biosynthetic pathway from (+)-pinoresinol to (-)-deoxypodophyllotoxin, the direct precursor of (-)-podophyllotoxin, in the recombinant host Escherichia coli. However, the use of the expensive substrate (+)-pinoresinol limits its application from the economic viewpoint. In addition, the simultaneous expression of multiple heterologous genes from different plasmids for a multi-enzyme cascade can be challenging and limits large-scale use. Results In this study, recombinant plasmid-free E. coli strains for the multi-step synthesis of pinoresinol from ferulic acid were constructed. To this end, a simple and versatile plasmid toolbox for CRISPR/Cas9-assisted chromosomal integration has been developed, which allows the easy transfer of genes from the pET vector series into the E. coli chromosome. Two versions of the developed toolbox enable the efficient integration of either one or two genes into intergenic high expression loci in both E. coli K-12 and B strains. After evaluation of this toolbox using the fluorescent reporter mCherry, genes from Petroselinum crispum and Zea mays for the synthesis of the monolignol coniferyl alcohol were integrated into different E. coli strains. The product titers achieved with plasmid-free E. coli W3110(T7) were comparable to those of the plasmid-based expression system. For the subsequent oxidative coupling of coniferyl alcohol to pinoresinol, a laccase from Corynebacterium glutamicum was selected. Testing of different culture media as well as optimization of gene copy number and copper availability for laccase activity resulted in the synthesis of 100 mg/L pinoresinol using growing E. coli cells. Conclusions For efficient and simple transfer of genes from pET vectors into the E. coli chromosome, an easy-to-handle molecular toolbox was developed and successfully tested on several E. coli strains. By combining heterologous and endogenous enzymes of the host, a plasmid-free recombinant E. coli growing cell system has been established that enables the synthesis of the key lignan pinoresinol.

Eucommiae Cortex (EC) is frequently used alone or in combination with other active ingredients to treat a range of illnesses. An efficient technical instrument for changing cheap or plentiful organic chemicals into rare or costly counterparts is biotransformation. It combines EC with biotransformation techniques with the aim of producing some novel active ingredients, using different strains of bacteria that were introduced to biotransform EC in an aseptic environment. The high-quality strains were screened for identification after the fermentation broth was found using HPLC, and the primary unidentified chemicals were separated and purified in order to be structurally identified. Strain 1 was identified as Aspergillus niger and strain 2 as Actinomucor elegans; the main transformation product A was identified as pinoresinol (Pin) and B as dehydrodiconiferyl alcohol (DA). The biotransformation of EC utilizing Aspergillus niger and Actinomucor elegans is reported for the first time in this study’s conclusion, resulting in the production of Pin and DA. [ABSTRACT FROM AUTHOR]

Mallotus paxii is a Euphorbiaceae plant belonging to the genus Mallotus Lour. This genus mainly distributes in tropical and subtropical Asia, and there are about 140 species in the world. There are 36 species in China, including 11 varieties, which mainly grow in Guangxi, Guangdong and Hainan. To study the compounds of Mallotus paxii, twelve lignans were isolated from the leaves of M. paxii by means of various column chromatographic techniques, including silica gel, ODS, Sephadex LH-20 and preparative RP-HPLC methods. The structures of the isolated lignans were determined on the basis of analyses of spectroscopic methods (1H- and 13C-NMR spectroscopy), high-resolution electrospray ionization mass spectrometry (HR-ESI-MS), and comparison of their spectroscopic data with previously reported data. The lignans were identified as 7″,8″-threo-buddlenol D (1), buddlenol D (2), 7″,8″-threo-buddlenol C (3), buddlenol C (4), (+)-syringaresinol (5), epipinoresinol (6), pinoresinol (7), 1-acetoxyl-2e,6e-dipiperonyl-3,7-dioxabicyclo-［3,3,0］-octane (8), ciwujiatone (9), lariciresinol-4′-O-β-D-glucopyranoside (10), rel-(2α,3β)-7-O-methylcedrusin (11), and dihydrodehydrodiconiferyl-alcohol 4′-O-β-D-glucoside (12). All compounds were isolated from M. paxii for the first time. Some lignans have good pharmacological activities such as antibacterial, anti-inflammatory, anti-tumor and anti-virus. The results of this study show that the leaves of M. paxii are rich in lignans with good activity, indicating that its good medicinal value may be related to these lignans.

Mallotus paxii is a Euphorbiacea plant belonging to the genus Mallotus Lour. This genus mainly distributes in tropical and subtropical Asia, and there are about 140 species in the world. There are 36 species in China, including 11 varieties, which mainly grow in the provinces of Guangxi, Guangdong and Hainan. The plants of this genus have antibacterial, anti-inflammatory, anti-tumor and anti-viral effects. To study the compounds of Mallotus paxii Pamp, twelve lignans were isolated from the leaves of Mallotus paxii Pamp by means of various column chromatographic techniques, including silica gel, ODS, Sephadex LH-20 and preparative RP-HPLC methods. The structures of the isolated lignans were determined on the basis of analyses of spectroscopic methods(¹H-and 13C-NMR spectroscopy), LH-20 and preparative RP-HPLC methods. The structures of the isolated lignans were determined on the basis of analyses of spectroscopic methods high-resolution electrospray ionization mass spectrometry(HR-ESI-MS), and comparison of their spectroscopic data with previously reported data. The lignans were identified as 7″,8″-threo-buddlenol D(1), buddlenol D(2), 7″,8″-threo-buddlenol C(3), buddlenol C(4),(+)-syringaresinol(5), epipinoresinol(6), pinoresinol(7), 1-acetoxyl-2e,6e-dipiperonyl-3,7-dioxabicyclo-[3,3,0]-octane(8), ciwujiatone(9), lariciresinol-4'-O-β-D-glucopyranoside(10), rel-(2α,3β)-7-O-methylcedrusin(11), and dihydrodehydrodiconiferyl-alcohol 4'-O-β-D-glucoside(12). All compounds were isolated from M. paxii for the first time. Some lignans have good pharmacological activities such as antibacterial, anti-inflammatory, anti-tumor and anti-virus. The results of this study show that the leaves of M. paxii are rich in lignans with good activity, indicating that its good medicinal value may be related to these lignans. [ABSTRACT FROM AUTHOR]

The coronavirus disease 2019 (COVID-19) was first found in Wuhan, China, in December 2019. Because the virus spreads quickly, it quickly became a global worry. Coronaviridae is the family that contains both SARS-CoV-2 and the viruses that came before (i.e., MERS-CoV and SARS-CoV). Recent sources portray that the COVID-19 virus has affected 344,710,576 people worldwide and killed about 5,598,511 people in the last 2 years. The B.1.1.529 strain, later called "Omicron," was named a Variant of Concern on November 24, 2021. The SARS-CoV-2 virus has gone through a never-ending chain of changes that have never happened before. As a result, it has many different traits. Most of these changes have occurred in the spike protein, where antibodies bind. Because of these changes, the Omicron type is very contagious and easy to pass on. There have been a lot of studies done to try to figure out this new challenge in the COVID-19 strains race, but there is still a lot that needs to be explained. This study focuses on virtual screening, docking, and molecular dynamic analysis; we aimed to identify therapeutic candidates for the SARS-CoV-2 variant Omicron based on their ability to inhibit non-structural proteins. We investigate the prediction of the properties of a substantial database of drug molecules obtained from the OliveNet™ database. Compounds that did not exhibit adequate gastrointestinal absorption and failed the Lipinski test are not considered for further research. The filtered compounds were coupled with our primary target, SARS-CoV-2 Omicron spike protein. We focused on SARS-CoV-2 Omicron spike protein and filtering potent olive compounds. Pinoresinol, the most likely candidate, is bound best (− 8.5 kcal/mol). Pinoresinol's strong interaction with the active site made the complex's dynamic structure more resilient. MD simulations explain the protein–ligand complex's stability and function. Pinoresinol may be a promising SARS-CoV-2 Omicron spike protein receptor lead drug, and additional research may assist the scientific community. [ABSTRACT FROM AUTHOR]

The chemistry of the phenolic compounds found in virgin olive oil (VOO) is very complex due, not only to the different classes of polyphenols that can be found in it, but, above all, due to the existence of a very specific phenol class found only in oleaceae plants: the secoiridoids. Searching in the Scopus data base the keywords flavonoid, phenolic acid, lignin and secoiridoid, we can find a number of 148174, 79435, 11326 and 1392 research articles respectively, showing how little is devote to the latter class of compounds. Moreover, in contrast with other classes, that include only phenolic compounds, secoiridoids may include phenolic and non-phenolic compounds, being the articles concerning phenolic secoiridoids much less than the half of the abovementioned articles. Therefore, it is important to clarify the structures of these compounds and their chemistry, as this knowledge will help understand their bioactivity and metabolism studies, usually performed by researchers with a more health science's related background. In this review, all the structures found in many research articles concerning VOO phenolic compounds chemistry and metabolism was gathered, with a special attention devoted to the secoiridoids, the main phenolic compound class found in olives, VOO and olive leaf. [ABSTRACT FROM AUTHOR]

Trewia nudiflora Linn. is a valuable forest resource due to its economic, ethnomedicinal, and ecological properties; however, its allelopathic potential is undocumented. Therefore, this research was designed to investigate the allelopathic impacts of T. nudiflora leaf extracts on alfalfa (Medicago sativa L.) and barnyard grass (Echinochloa crus-galli (L.) P. Beauv.) growth, as well as to isolate and to identify the active allelopathic substances responsible for these effects. A bioassay experiment with eight different treatments (0.1, 0.3, 1, 3, 10, 30, 100, and 300 mg dry weight (DW) equivalents of T. nudiflora extracts in mL−1) was carried out. The results showed that the growth of both plants decreased with increasing contents of T. nudiflora extracts, with the effect depending on the plant species and reaching its peak at a concentration of a 300 mg DW equivalent of T. nudiflora extract in mL−1. Active substances were isolated and identified using an HPLC system, which revealed the presences of methyl gallate and pinoresinol in aqueous methanol extracts of T. nudiflora. The shoot and root lengths of the alfalfa and the barnyard grass decreased significantly when they were treated with the methyl gallate and the pinoresinol. The allelopathic inhibition increased with increasing compound concentrations, with the root growth being more sensitive to the pinoresinol than to the methyl gallate, and it was significantly higher at the concentration of 10 mM. These results indicated that the T. nudiflora leaf extracts limited the growth of the treated plants, and the methyl gallate and pinoresinol in the extracts may have caused the inhibition of the T. nudiflora extracts. Thus, the leaf extracts of T. nudiflora and the substances methyl gallate and pinoresinol could be incorporated into sustainable agricultural practices or used to develop bioherbicides that would promote sustainable weed management practices. [ABSTRACT FROM AUTHOR]

Dirigent proteins (DIRs) are a new class of proteins that were identified during the 8–8′ lignan biosynthetic pathway and involves the formation of (+) or (−)-pinoresinol through stereoselective coupling from E-coniferyl alcohol. These proteins are known to play a vital role in the development and stress response in plants. Various studies have reported the functional and structural characterization of dirigent gene family in different plants using in silico approaches. Here, we have summarized the importance of dirigent proteins in plants and their role in plant stress tolerance by analyzing the genome-wide analysis including gene structure, mapping of chromosomes, phylogenetic evolution, conserved motifs, gene structure, and gene duplications in important plants. Overall, this review would help to compare and clarify the molecular and evolutionary characteristics of dirigent gene family in different plants. [ABSTRACT FROM AUTHOR]

Trewia nudiflora Linn. is a valuable forest resource due to its economic, ethnomedicinal, and ecological properties; however, its allelopathic potential is undocumented. Therefore, this research was designed to investigate the allelopathic impacts of T. nudiflora leaf extracts on alfalfa (Medicago sativa L.) and barnyard grass (Echinochloa crus-galli (L.) P. Beauv.) growth, as well as to isolate and to identify the active allelopathic substances responsible for these effects. A bioassay experiment with eight different treatments (0.1, 0.3, 1, 3, 10, 30, 100, and 300 mg dry weight (DW) equivalents of T. nudiflora extracts in mL−1) was carried out. The results showed that the growth of both plants decreased with increasing contents of T. nudiflora extracts, with the effect depending on the plant species and reaching its peak at a concentration of a 300 mg DW equivalent of T. nudiflora extract in mL−1. Active substances were isolated and identified using an HPLC system, which revealed the presences of methyl gallate and pinoresinol in aqueous methanol extracts of T. nudiflora. The shoot and root lengths of the alfalfa and the barnyard grass decreased significantly when they were treated with the methyl gallate and the pinoresinol. The allelopathic inhibition increased with increasing compound concentrations, with the root growth being more sensitive to the pinoresinol than to the methyl gallate, and it was significantly higher at the concentration of 10 mM. These results indicated that the T. nudiflora leaf extracts limited the growth of the treated plants, and the methyl gallate and pinoresinol in the extracts may have caused the inhibition of the T. nudiflora extracts. Thus, the leaf extracts of T. nudiflora and the substances methyl gallate and pinoresinol could be incorporated into sustainable agricultural practices or used to develop bioherbicides that would promote sustainable weed management practices.

Growth and maintenance of skeletal muscle is essential for athletic performance and a healthy life. Stimulating the proliferation and differentiation of muscle cells may help prevent loss of muscle mass. To discover effective natural substances enabling to mitigate muscle loss without side effects, we evaluated muscle growth with several compounds extracted from Catalpa bignonioides Walt. Among these compounds, pinoresinol and vanillic acid increased C2C12, a mouse myoblast cell line, proliferation being the most without cytotoxicity. These substances activated the Akt/mammalian target of the rapamycin (mTOR) pathway, which positively regulates the proliferation of muscle cells. In addition, the results of in silico molecular docking study showed that they may bind to the active site of insulin-like growth factor 1 receptor (IGF-1R), which is an upstream of the Akt/mTOR pathway, indicating that both pinoresinol and vanillic acid stimulate myoblast proliferation through direct interaction with IGF-1R. These results suggest that pinoresinol and vanillic acid may be a natural supplement to improve the proliferation of skeletal muscle via IGF-1R/Akt/mTOR signaling and thus strengthen muscles. [ABSTRACT FROM AUTHOR]

α-EG is a unique substance that was first found in the leaves and fruits of Morinda citrifolia (Mc) growing in Thailand using GC-MS at 52.33% and 54.12%. It was then concentrated and its abundance quantified, along with that of pinoresinol, via GC, compared to the standards in leaves, ufp, rfp, rawfs, and seeds. α-EG and pinoresinol, which have collagen stimulating, skin whitening, and an inhibitory effect on wrinkle formation, were found in different concentrations and amounts. Three different concentrations of the five Mc part extracts were tested on NHDF for gene expression related to the aforementioned activities, COL1A1, COL1A2, and COL3A1, FGF1 and FGF7 by qRT-PCR. The results showed various expression levels, both stimulatory and inhibitory, with different concentrations of plant parts and genes. Similar results were revealed when the experiments were performed with Morus alba (Ma), which was found to contain 20.48 g protein p/100 g leaves at concentrations of 3.11 mg/mL. The studied Mc parts seem to have advantages based on the stated objectives, gene type and level of activity of each plant part. Rawfs and leaves supplemented with Ma samples were selected for toxicity tests with PBMCs. The lack of both cell and DNA toxicity from the rawfs indicated that they can be used safely. [ABSTRACT FROM AUTHOR]

We describe a novel nature-derived epoxy resin monomer (ERM) derived from the plant lignan pinoresinol. Epoxy resins are thermosetting materials in global usage owing to their excellent technical properties such as flexibility and durability. However, their adverse health effects are often not considered and affect users of epoxy resins worldwide. Components of epoxy resin systems are strong skin sensitizers and cause allergic contact dermatitis. The reported prevalence attributable to epoxy chemicals is between 11.7 and 12.5% of all cases of occupational allergic contact dermatitis. We are committed to developing epoxy resins with reduced allergenic effect, while maintaining their excellent properties. The novel ERM, pinoresinol diglycidyl ether (PinoDGE), was synthesized in one step from pinoresinol and epichlorohydrin in 88% yield. It was not classified as a skin sensitizer in the in vivo local lymph node assay, at concentrations up to 0.17 m, as it did not cause a stimulation index >3 compared to control. Pinoresinol diglycidyl ether reacted with the model peptide AcPHCKRM in a reactivity assay and was predicted to be a skin sensitizer in the KeratinoSens assay. Preliminary cross-linking studies indicate that it has promising properties compared to commercially used ERMs. Pinoresinol diglycidyl ether could be seen as a lead compound for further development of alternative ERMs with a better safety profile based on natural and renewable sources for construction of epoxy resin polymers. [ABSTRACT FROM AUTHOR]

Pinoresinol (PINL) and pinoresinol diglucoside (PDG), two natural lignans found in Eucommia ulmoides Oliv. (Duzhong), have several pharmacological activities. However, there is no report available on their absorption, distribution, metabolism, and elimination (ADME) properties. Given the possible wide spectrum of their application in therapeutic areas, this area should be investigated. This work studied the in vitro ADME properties of PDG and PINL, including their kinetic solubility, permeability across monolayer cells (PAMPA), protein binding, and metabolic stabilities in liver microsomes. The in vivo pharmacokinetic study and in vitro vasorelaxant effects on isolated phenylephrine-induced aortic rings of PINL and PDG were also investigated. It was found that both of their kinetic solubility in PBS (pH 7.4) was greater than 100 μM, indicating that they are both soluble compounds. The permeability investigations (Peff) by PAMPA indicated that PINL had higher permeability than PDG (p < 0.05). Both components represented moderate plasma protein binding activities (average binding rate in human plasma: PINL 89.03%, PDG 45.21%) and low metabolic rate (t1/2 in human liver microsome: PINL 1509.5 min, PDG 1004.8 min). Furthermore, the results of pharmacokinetic studies indicated that PINL might be eliminated less quickly than PDG from the rat plasma, and its cumulative urinary excretion was much lower than that of PDG. The phenylephrine-induced aortic rings demonstrated concentration-dependent vasorelaxation in PDG, PINL, or their combination group. The vasorelaxant effects of PINL were more obvious than those of PDG, whereas the vasorelaxant effect of the combinations was significantly better than that of the single component (p < 0.05). The similarity or difference between PINL and its diglucoside in these pharmaceutical aspects may offer valuable insights into the further exploration of lignans and might contribute to relevant studies involving natural products with similar molecular structure and their glucosides.

Pinoresinol (PINL) and pinoresinol diglucoside (PDG), two natural lignans found in Eucommia ulmoides Oliv. (Duzhong), have several pharmacological activities. However, there is no report available on their absorption, distribution, metabolism, and elimination (ADME) properties. Given the possible wide spectrum of their application in therapeutic areas, this area should be investigated. This work studied the in vitro ADME properties of PDG and PINL, including their kinetic solubility, permeability across monolayer cells (PAMPA), protein binding, and metabolic stabilities in liver microsomes. The in vivo pharmacokinetic study and in vitro vasorelaxant effects on isolated phenylephrine-induced aortic rings of PINL and PDG were also investigated. It was found that both of their kinetic solubility in PBS (pH 7.4) was greater than 100 μM, indicating that they are both soluble compounds. The permeability investigations (P eff ) by PAMPA indicated that PINL had higher permeability than PDG (p < 0.05). Both components represented moderate plasma protein binding activities (average binding rate in human plasma: PINL 89.03%, PDG 45.21%) and low metabolic rate (t 1/2 in human liver microsome: PINL 1509.5 min, PDG 1004.8 min). Furthermore, the results of pharmacokinetic studies indicated that PINL might be eliminated less quickly than PDG from the rat plasma, and its cumulative urinary excretion was much lower than that of PDG. The phenylephrine-induced aortic rings demonstrated concentration-dependent vasorelaxation in PDG, PINL, or their combination group. The vasorelaxant effects of PINL were more obvious than those of PDG, whereas the vasorelaxant effect of the combinations was significantly better than that of the single component (p < 0.05). The similarity or difference between PINL and its diglucoside in these pharmaceutical aspects may offer valuable insights into the further exploration of lignans and might contribute to relevant studies involving natural products with similar molecular structure and their glucosides. [ABSTRACT FROM AUTHOR]

Growth and maintenance of skeletal muscle is essential for athletic performance and a healthy life. Stimulating the proliferation and differentiation of muscle cells may help prevent loss of muscle mass. To discover effective natural substances enabling to mitigate muscle loss without side effects, we evaluated muscle growth with several compounds extracted from Catalpa bignonioides Walt. Among these compounds, pinoresinol and vanillic acid increased C2C12, a mouse myoblast cell line, proliferation being the most without cytotoxicity. These substances activated the Akt/mammalian target of the rapamycin (mTOR) pathway, which positively regulates the proliferation of muscle cells. In addition, the results of in silico molecular docking study showed that they may bind to the active site of insulin-like growth factor 1 receptor (IGF-1R), which is an upstream of the Akt/mTOR pathway, indicating that both pinoresinol and vanillic acid stimulate myoblast proliferation through direct interaction with IGF-1R. These results suggest that pinoresinol and vanillic acid may be a natural supplement to improve the proliferation of skeletal muscle via IGF-1R/Akt/mTOR signaling and thus strengthen muscles.

α-EG is a unique substance that was first found in the leaves and fruits of Morinda citrifolia (Mc) growing in Thailand using GC-MS at 52.33% and 54.12%. It was then concentrated and its abundance quantified, along with that of pinoresinol, via GC, compared to the standards in leaves, ufp, rfp, rawfs, and seeds. α-EG and pinoresinol, which have collagen stimulating, skin whitening, and an inhibitory effect on wrinkle formation, were found in different concentrations and amounts. Three different concentrations of the five Mc part extracts were tested on NHDF for gene expression related to the aforementioned activities, COL1A1, COL1A2, and COL3A1, FGF1 and FGF7 by qRT-PCR. The results showed various expression levels, both stimulatory and inhibitory, with different concentrations of plant parts and genes. Similar results were revealed when the experiments were performed with Morus alba (Ma), which was found to contain 20.48 g protein p/100 g leaves at concentrations of 3.11 mg/mL. The studied Mc parts seem to have advantages based on the stated objectives, gene type and level of activity of each plant part. Rawfs and leaves supplemented with Ma samples were selected for toxicity tests with PBMCs. The lack of both cell and DNA toxicity from the rawfs indicated that they can be used safely.

A new sesquiterpene lactone 1a along with other four known compounds 1b, 2, 3 and 4 were isolated from fresh leaves of Cynara scolymus L. using ordinary chromatographic techniques. The structures of the isolated compounds were determined via spectroscopic analysis. [ABSTRACT FROM AUTHOR]

Eucommiae Cortex (EC) is frequently used alone or in combination with other active ingredients to treat a range of illnesses. An efficient technical instrument for changing cheap or plentiful organic chemicals into rare or costly counterparts is biotransformation. It combines EC with biotransformation techniques with the aim of producing some novel active ingredients, using different strains of bacteria that were introduced to biotransform EC in an aseptic environment. The high-quality strains were screened for identification after the fermentation broth was found using HPLC, and the primary unidentified chemicals were separated and purified in order to be structurally identified. Strain 1 was identified as Aspergillus niger and strain 2 as Actinomucor elegans ; the main transformation product A was identified as pinoresinol (Pin) and B as dehydrodiconiferyl alcohol (DA). The biotransformation of EC utilizing Aspergillus niger and Actinomucor elegans is reported for the first time in this study's conclusion, resulting in the production of Pin and DA.

(+)-Pinoresinol (Pin), a biologically active lignan and widely found in many dietary plants, was reported to have antifungal, anti-inflammatory, antioxidation, and hypoglycemic activities. In the present study, the capability of Pin to inhibit human hepatocellular carcinoma HepG2 cells was evaluated and the mechanism was analyzed. Pin exhibited significant inhibition of HepG2 proliferation and revealed a pro-apoptotic effect. Pin led to activation of Fas, FasL, caspase-8, and Bid, which provoked death receptor apoptotic signals. Significant loss of mitochondrial membrane potential was also observed, as well as the altered expression of mitochondrial apoptotic proteins, including increased Cyto-c, Bax/Bcl-2 ratio, and caspase 3. Pin also exhibited inhibitory effect on the migration and invasion of HepG2 in a concentration-dependent manner by increasing E-cadherin and decreasing VCAM-1, ICAM-1 and MMP-9 expression. All the results indicated Pin could inhibit HepG2 via the induction of apoptosis through the death receptor pathway and/or mitochondrial pathway and the inhibition of invasion via disturbing the cytoskeleton.

Abstract Dementia is a category of brain diseases that cause a decrease in cognitive functions. Alzheimer's disease (AD) is the most frequently mentioned neurodegenerative disease showing dementia. Although many useful drugs for dementia were developed, we still need better and safer drugs. Here, we tested pinoresinol, a lignan found in sesame seed and olive oil, whether it could be a candidate for this purpose. Pinoresinol (25 mg/kg, p.o.) ameliorated memory impairment in dementia model induced by cholinergic blockade in the passive avoidance test in a dose-dependent manner. Moreover, pinoresinol (50 μM) facilitated induction of hippocampal long-term potentiation, a cellular model of learning and memory. Pinoresinol blocked acetylcholinesterase (AchE), an acetylcholine-degrading enzyme, activity in a concentration-dependent manner. Moreover, pinoresinol (50 μM) facilitated calcium influx into neuro2a cell. These results suggest that pinoresinol improves memory impairment and facilitates hippocampal LTP induction and these results might be related to the effect of pinoresinol on AChE and calcium influx. Highlights • Pinoresinol ameliorated memory impairment in dementia model induced by cholinergic blockade. • Pinoresinol facilitated induction of hippocampal LTP. • Pinoresinol blocked AChE. • Pinoresinol facilitated calcium influx into neuro2a cell. [ABSTRACT FROM AUTHOR]

No clear picture has yet been elaborated concerning the mechanism of lignin growth, and thus this topic is the focus of the present paper. Namely, the enzymatic dehydrogenative polymerization (DHP formation) of coniferyl alcohol (CA, as a monolignol) and three dilignols and their reaction kinetics were investigated. The dilignols [guaiacylglycerol-β-coniferyl ether (IβO4), dehydrodiconiferyl alcohol (IIβ5), and pinoresinol (IIIββ)] and CA as a monolignol [(3-OCD3)-coniferyl alcohol (CAOCD3)] were synthesized and subjected to enzymatic DHP formation. The dilignol derived from CAOCD3 could be identified by its higher molecular weight in comparison with the starting dilignols (IβO4, IIβ5, and IIIββ). Based on the observed consumption rate of the CA and its dilignols, it was proposed that a radical transfer system exists between the dilignols, which is generated from the CA and the starting substrates. [ABSTRACT FROM AUTHOR]

Highlights • Pinoresinol was detected for the first time in cold-pressed grape seed oils. • Total phenolic content ranged between 0.83 mg/kg to 15.16 mg/kg. • Good correlation between total phenolic content and inhibitory power of PTP-1B. • Pinoresinol, p -coumaric acid and quercetin give the major contribute to inhibition. • Ethyl esters of gallic and caffeic acid were detected for the first time. Abstract This paper investigates the phenolic composition of 17 monocultivar commercial cold-pressed grape seed oils. Chromatographic profiles showed the presence of more than 28 molecules, 11 of which were successfully identified by HPLC-DAD-MS-TOF and HPLC-FLD analysis. Pinoresinol, ethyl caffeate and ethyl gallate were detected for the first time in these oils. The total phenolic content ranged between 0.83 mg/kg for Viognier sample to 15.16 mg/kg for Merlot org sample. The detected ethyl esters can be suggested as markers to evaluate the intensity of fermentation in grape seeds before oil extraction, and to control the sensorial quality of the produced oils. In addition, the inhibitory power of these phenolic extracts against Protein Tyrosine Phosphatase 1B enzyme (PTP-1B), overexpressed in type-two diabetes, was investigated for the first time. Data highlighted a good correlation between total phenolic content and inhibitory power, with pinoresinol, p -coumaric acid and quercetin making the greater contributions. [ABSTRACT FROM AUTHOR]

Graphical abstract Schematic representation of PLR down-regulation effect on phenylpropanoids. Abstract Lignans are diphenolic compounds produced in plants via coupling of two coniferyl alcohol molecules with the aid of a dirigent protein to form pinoresinol (PINO). The latter is reduced via lariciresinol (LARI) to secoisolariciresinol by the bifunctional pinoresinol–lariciresinol reductase (PLR). In this study, we clarified the consequences of altered lignan biosynthesis on amino acids, phenolics compounds and lignin in the hairy roots of Linum album with an ihpRNAi construct to silence PLR gene expression. Down-regulation of PLR-La1 resulted in up to an 8.3 and 3.3-time increased PINO and LARI content respectively, and reduced levels of podophyllotoxin (PTOX) and 6-methoxy podophyllotoxin (6-MPTOX). By Suppression of PLR expression, the metabolites belonging to shikimate and phenylpropanoid pathways are conducted to phenolic compounds and lignin accumulations. Although PINO and LARI were induced in response to fungal elicitor, the accumulation of PTOX and 6-MPTOX did not occur in PLR down-regulated roots. Our result also demonstrated variation in amino acids, phenolic compounds and lignin levels in presence of the fungal elicitation in PLR down regulated-roots. This data assert the accumulation of aryltetralin lignans in interactions with plant pathogens by PLR activity and the importance this enzyme for defense against pathogens in L. album. [ABSTRACT FROM AUTHOR]

The aim of our study was to investigate the presence and quantitative contents of lignans in the tissues of Taxus ×media. The presence of the lignans: pinoresinol, matairesinol and secoisolariciresinol was assessed in needles, shoots cultures and suspension culture. Pinoresinol was the only lignan found in the tissue of T. ×media. The total pinoresinol content in the needles and in the shoots was 1.24 mg/g dry weight (dw) and 0.69 mg/g dw, respectively. Most of the pinoresinol identified was appeared glycosidically bound. In needles, the amount of glycosidically bound pinoresinol (0.81 mg/g dw) was about twice as high as that of free pinoresinol (0.43 mg/g dw). The content of free and glycosidically bound pinoresinol showed the level of 0.18 mg/g dw and 0.51 mg/g dw, respectively in the in vitro shoot cultures. In the cell culture, no pinoresinol was found.

(+)-Pinoresinol (Pin), a biologically active lignan and widely found in many dietary plants, was reported to have antifungal, anti-inflammatory, antioxidation, and hypoglycemic activities. In the present study, the capability of Pin to inhibit human hepatocellular carcinoma HepG2 cells was evaluated and the mechanism was analyzed. Pin exhibited significant inhibition of HepG2 proliferation and revealed a pro-apoptotic effect. Pin led to activation of Fas, FasL, caspase-8, and Bid, which provoked death receptor apoptotic signals. Significant loss of mitochondrial membrane potential was also observed, as well as the altered expression of mitochondrial apoptotic proteins, including increased Cyto-c, Bax/Bcl-2 ratio, and caspase 3. Pin also exhibited inhibitory effect on the migration and invasion of HepG2 in a concentration-dependent manner by increasing E-cadherin and decreasing VCAM-1, ICAM-1 and MMP-9 expression. All the results indicated Pin could inhibit HepG2 via the induction of apoptosis through the death receptor pathway and/or mitochondrial pathway and the inhibition of invasion via disturbing the cytoskeleton. [ABSTRACT FROM AUTHOR]

This work deals with the combined effect of agronomical and technological variables on the content of virgin olive oils in phenolic compounds with special attention to flavonoids (luteolin and apigenin) and lignans (pinoresinol and 1-acetoxypinoresinol). Olives from irrigated or rainfed trees at different fruit ripening were studied using Response Surface Methodology. Relevant variables included malaxation temperature (20–60 °C); time (20–60 min); and talc dosage (5–25 g/kg g olives). According to the models, the highest concentrations were 7.83, 289.4, 38.0 and 4.70 mg/kg of hydroxytyrosol, oleocanthal, luteolin and pinoresinol, respectively. Results show that the content of phenolic compounds is mostly affected by the malaxation temperature, increasing when the temperature rises; however, the time exerts an irregular influence, while the dosage of adjuvant has little influence. The flavonoid content, both luteolin and apigenin, decreased with increasing temperature and malaxation time. The pinoresinol content increased with temperature and malaxation time. Non-irrigated olive oils have a higher content in phenolic compounds. The flavonoid contents presented a maximum around maturity index 4. Moreover, the contents usually were higher for olives coming from rainfed. Pinoresinol content decreased with the increase of the index of maturity, the oils from fruits grown in rainfed containing more pinoresinol. [ABSTRACT FROM AUTHOR]

Pinoresinol is a dimer of two β-β=-linked coniferyl alcohol molecules. It is both a plant defense molecule synthesized through the shikimic acid pathway and a representative of several β-β-linked dimers produced during the microbial degradation of lignin in dead plant material. Until now, little has been known about the bacterial catabolism of such dimers. Here we report the isolation of the efficient (β)- pinoresinol-mineralizing Pseudomonas sp. strain SG-MS2 and its catabolic pathway. Degradation of pinoresinol in this strain is inducible and proceeds via a novel oxidative route, which is in contrast to the previously reported reductive transformation by other bacteria. Based on enzyme assays and bacterial growth, cell suspension, and resting cell studies, we provide conclusive evidence that pinoresinol degradation in strain SG-MS2 is initiated by benzylic hydroxylation, generating a hemiketal via a quinone methide intermediate, which is then hydrated at the benzylic carbon by water. The hemiketal, which stays in equilibrium with the corresponding keto alcohol, undergoes an aryl-alkyl cleavage to generate a lactone and 2-methoxyhydroquinone. While the fate of 2-methoxyhydroquinone is not investigated further, it is assumed to be assimilated by ring cleavage. The lactone is further metabolized via two routes, namely, lactone ring cleavage and benzylic hydroxylation via a quinone methide intermediate, as described above. The resulting hemiketal again exists in equilibrium with a keto alcohol. Our evidence suggests that both routes of lactone metabolism lead to vanillin and vanillic acid, which we show can then be mineralized by strain SG-MS2. [ABSTRACT FROM AUTHOR]

The stability and chemical reactions of lignans during alkaline cooking is not yet fully clarified although their degradation products may have a considerable effect on the pulp and its bleaching, such as spotting and color contamination. To fill this gap, a phenolic lignan (pinoresinol) and four non-phenolic furofuran lignans (gmelinol, paulownin, eudesmin and sesamin) were submitted to alkaline cooking at various conditions. The effect of cooking duration shows that non-phenolic lignans are more stable during a 3-h reaction time than pinoresinol. After a 24-h reaction, most of the lignans were decomposed or polymerized. The effect of cooking temperature shows that pinoresinol, eudesmin and sesamin are more susceptible to degradation with increasing temperature. The concentration of the alkaline liquors only affect significantly to pinoresinol because of its acidity. It was also observed that pinoresinol, gmelinol and eudesmin give rise to small amounts of vanillin and acetovanillone, while paulownin and sesamin were mainly polymerized or degraded. To confirm the reaction mechanism of the occurrence of acetovanillone in the reaction, 13C-labeled pinoresinol was also submitted to alkaline cooking. It was revealed that the cleavage at β-β′ position occurred during alkaline reaction. The stability of these compounds during alkaline cooking are not affected by the hydroxyl group in the C-β′ position. [ABSTRACT FROM AUTHOR]

Pinoresinol from the leaves of Cinnamomum camphora used as a model natural compound was extracted and isolated. The chemical characterization of the compound was confirmed using UV, IR, MS, H NMR and C HMR. The purity of the compound (86%) was determined using HPLC technique and it was used for further experiments. Its antibacterial activities against five food-related bacteria ( Escherichia coli , Pseudomonas aeruginosa , Staphylococcus aureus , Bacillus subtilis , Salmonella enterica ) were detected using both broth dilution method and disk diffusion method. Results indicated that 62.5 μg/mL pinoresinol completely inhibit the growth of five bacteria. Much effort was focused on elucidating the mechanism of antibacterial action of pinoresinol against P . aeruginosa and B . subtilis by determination of cell permeability and observing the changes of cell microstructure using scanning electron microscope (SEM). The leakage of the soluble saccharides and proteins were detected and up to 13.89 and 21.76 μg/mL, respectively for P . aeruginosa and 11.08 and 19.05 μg/mL, respectively for B . subtilis . Measurements of the release of the soluble saccharides and proteins confirmed the disruptive action of pinoresinol on cytoplasmic membrane. The damages of the pinoresinol on cell wall were confirmed using Gram staining and scanning electron microscopy (SEM). We concluded that the antibacterial action mechanism of the pinoresinol is attributed to the physiological and morphological changes in bacterial cells. [ABSTRACT FROM AUTHOR]

ABSTRACT Pinoresinol is a natural lignan with a high market value that has potential pharmacological and food supplement applications. Pinoresinol is currently isolated from plants, which suffers from low efficiency and yield. To produce pinoresinol from inexpensive and industrially available eugenol, an in vivo enzymatic cascade composed of vanillyl alcohol oxidase and peroxidase was designed, which scavenges H2O2 automatically and eliminates protein purification and cofactor addition. Two peroxidases were screened and identified from Escherichia coli BL21 (DE3), and tested in the enzymatic cascade. To balance the flux, different genetic architectures were constructed by using ePathBrick and fusion gene approaches. Scavenging H2O2 alleviated by-product toxicity and enzyme inhibition, and led to efficient pinoresinol production. Optimization of the reaction conditions achieved a titer of 11.29 g/L pinoresinol. The molar yield and productivity were 52.77% and 1.03 g/(L × h), respectively. The elegant strategy developed herein utilizes the harmful by-product to drive the biosynthetic reaction forward and simultaneously detoxify cells, thereby preventing enzyme inhibition. Biotechnol. Bioeng. 2017;114: 2066-2074. © 2017 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

High amount of the valuable lignan pinoresinol (PR) was determined in Carduus nutans fruit (7.8 mg/g) for the first time. A preparative separation method using two consecutive, identical steps of centrifugal partition chromatography (CPC) was developed in order (i) to isolate PR and (ii) to subsequently isolate PR and its 7′ epimer epipinoresinol (EPR) simultaneously after an optimized acid treatment which resulted in PR epimerization forming equal amounts of PR and EPR, from C. nutans fruit. As optimal conditions, a two-phase solvent system consisting of methyl tert -butyl ether:acetone:water (4:3:3, v/v/v) for CPC separation, and an acid treatment performed at 50 °C for 30 min for the epimerization were applied. Thus, 33.7 mg and 32.8 mg PR and EPR, in as high as 93.7% and 92.3% purity, were isolated from 10.0 g C. nutans fruit, representing 86.4% and 84.1% efficiency, respectively. Conversion characteristic of PR and EPR in acidic medium, determined as a function of time and temperature of acid treatment provides their unambiguous identification by on-line high performance liquid chromatography (HPLC). Antiproliferative assay of isolated PR and EPR in two different types of colon cancer cell lines (HCT116 and SW480) confirmed that both epimers caused a more significant decrease of viability in HCT116 cells than in SW480 cells, suggesting their similar mechanism of antiproliferative action. [ABSTRACT FROM AUTHOR]

Background: Dietary intakes of plant lignans have been hypothesized to be inversely associated with the risk of developing cardiovascular disease and cancer. Earlier studies were based on a Finnish lignan database (Fineli®) with two lignan precursors, secoisolariciresinol (SECO) and matairesinol (MAT). More recently, a Dutch database, including SECO and MAT and the newly recognized lignan precursors lariciresinol (LARI) and pinoresinol (PINO), was compiled. The objective was to re-estimate and re-evaluate plant lignan intakes and to identify the main sources of plant lignans in five European countries using the Finnish and Dutch lignan databases, respectively. Methods: Forty-two food groups known to contribute to the total lignan intake were selected and attributed a value for SECO and MAT from the Finnish lignan database (Fineli®) or for SECO, MAT, LARI, and PINO from the Dutch database. Total intake of lignans was estimated from food consumption data for adult men and women (19–79 years) from Denmark, Finland, Italy, Sweden, United Kingdom, and the contribution of aggregated food groups calculated using the Dutch lignin database. Results: Mean dietary lignan intakes estimated using the Dutch database ranged from 1 to 2 mg/day, which was approximately four-fold higher than the intakes estimated from the Fineli® database. When LARI and PINO were included in the estimation of the total lignan intakes, cereals, grain products, vegetables, fruit and berries were the most important dietary sources of lignans. Conclusion: Total lignin intake was approximately four-fold higher in the Dutch lignin database, which includes the lignin precursors LARI and PINO, compared to estimates based on the Finnish database based only on SECO and MAT. The main sources of lignans according to the Dutch database in the five countries studied were cereals and grain products, vegetables, fruit, berries, and beverages.

Insect gut microbiota have been widely reported to help the insects to overcome host tree defense. Streltzoviella insularis (Lepidoptera: Cossidae) is one of the most common wood borers in China, attacking various hosts, including ash trees ( Fraxinus sp.), but little is known about its gut microbial associates and their involvement in host tree defense. We isolated gut bacteria of S. insularis larvae, analyzed their ability to degrade pinoresinol (a defense compound of ash trees) and cellulose, and identified pinoresinol degradation products. Larval mortality increased with increasing pinoresinol concentration (reflecting natural variation observed in the host trees). All the five detected gut bacteria isolates were able to degrade pinoresinol, two of which were also capable of cellulose degradation. Furthermore, gut bacteria were also shown to degrade pinoresinol via the gluconeogenesis pathway. These results suggest that S. insularis -associated microorganisms help to overcome host pinoresinol defense and possibly contribute to insects or gut microbial nutrition via carbohydrate synthesis.