Characterization of terpenoids and norisoprenoids from base and retail Qingke Baijiu by GC×GC-TOFMS and multivariate statistical analysis

Qingke(highland barley)Baijiu is a special Chinese Baijiu which is mainly produced from Qinghai-Tibet Plateau.Since the pine board is used as the bottom of the fermentation pit,we deduced that the terpenoids and norisoprenoids in pine board might be introduced into Qingke Baijiu.Thus,the terpenoids and norisoprenoids in Qingke Baijiu were investigated by headspace solid phase microextraction(HS-SPME)combined with comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry(GC×GC-TOFMS).The results showed thatγ-terpinene(0.70-530.72μg/L),α-phellandrene(0.34-256.66μg/L),longicyclene(0-38.55μg/L),α-pinene(1.21-35.54μg/L)and limonene(0.93-23.69μg/L)were the top 5 terpenoids/norisoprenoids in Qingke Baijiu.Pulegone was detected and reported for the first time in Baijiu,and the concentrations in Qingke Baijiu were 0.78μg/L(fresh)and 3.90μg/L(7 years old).According to the principal component analysis(PCA)plot,young,aged,and retail Qingke Baijiu could be differentiated clearly.Fold change(FC)and t-tests analysis indicated thatβ-pinene,γ-terpinene,andα-selinene were the most different terpenoids/norisoprenoids between young and aged Qingke Baijiu,and longifolene was the most different terpenoids/norisoprenoids between base and retail Qingke Baijiu.The terpenoids and norisoprenoids,such as(E)-β-ionone,isoborneol andβ-cyclocitral,could be potential markers indicating the ageing process of Qingke Baijiu.

Remodeling Isoprene Pyrophosphate Metabolism for Promoting Terpenoids Bioproduction

Terpenoids are the largest family of natural products.They are made from the building block isoprene pyrophosphate(IPP),and their bioproduction using engineered cell factories has received a great deal of attention.To date,the insufficient metabolic supply of IPP remains a great challenge for the efficient synthesis of terpenoids.In this work,we discover that the imbalanced metabolic flux distribution between the central metabolism and the IPP supply hinders IPP accumulation in Bacillus subtilis(B.subtilis).Therefore,we remodel the IPP metabolism using a series of genetically encoded two-input-multioutput(TIMO)circuits that are responsive to pyruvate or/and malonyl-CoA,resulting in an IPP pool that is significantly increased by up to four-fold.As a proof-of-concept validation,we design an IPP metabolism remodeling strategy to improve the production of three valuable terpenoids,including menaquinone-7(MK-7,4.1-fold),lycopene(9-fold),andβ-carotene(0.9-fold).In particular,the titer of MK-7 in a 50-L bioreactor reached 1549.6 mg·L^(-1),representing the highest titer reported so far.Thus,we propose a TIMO genetic circuits-assisted IPP metabolism remodeling framework that can be generally used for the synergistic fine-tuning of complicated metabolic modules to achieve the efficient bioproduction of terpenoids.

Terpenoids as potential chemopreventive and therapeutic agents in liver cancer

Despite significant advances in medicine, liver cancer, predominantly hepatocellular carcinoma remains a major cause of death in the United States as well as the rest of the world. As limited treatment options are currently available to patients with liver cancer, novel preventive control and effective therapeutic approaches are considered to be reasonable and decisive measures to combat this disease. Several naturally occurring dietary and non-dietary phytochemicals have shown enormous potential in the prevention and treatment of several cancers, especially those of the gastrointestinal tract. Terpenoids, the largest group of phytochemicals, traditionally used for medicinal purposes in India and China, are currently being explored as anticancer agents in clinical trials. Terpenoids (also called "isoprenoids") are secondary metabolites occurring in most organisms, particularly plants. More than 40 000 individual terpenoids are known to exist in nature with new compounds being discovered every year. A large number of terpenoids exhibit cytotoxicity against a variety of tumor cells and cancer preventive as well as anticancer efficacy in preclinical animal models. This review critically examines the potential role of naturally occurring terpenoids, from diverse origins, in the chemoprevention and treatment of liver tumors. Both in vitro and in vivo effects of these agents and related cellular and molecular mechanisms are highlighted. Potential challenges and future directions involved in the advancement of these promising natural compounds in the chemoprevention and therapy of human liver cancer are also discussed.

Study on effect and mechanism of Celastrus Orbiculatus total terpenoids in regulating lipid metabolism

Objective To investigate the effect and mechanism of Celastrus Orbiculatus total terpenoids on lipid metabolism in hyperlipidemia mice.Methods ICR mice were selected as investigated subject.The hyperlipemia mice models were made with feeding high-fat forage and were randomly divided into six groups:the normal group,the model group,the positive control group(treated with simvastatin)and the three groups treated with Celastrus Orbiculatus total terpenoids with low,medium and high dosage,respectively.Each group included eight mice.The control group was fed normal forage,but other groups were fed high fat forage.All groups were allowed to drink water freely.Since the first day when the models were made,intragastric administration had been adopted.The normal group was fed normal forage without intragastric administration;the model group was received physiological saline 20 mL·kg-1·d-1 with intragastric administration;the positive control group received simvastatin 2.6 mg·kg-1·d-1 with intragastric administration;the three treated groups received Celastrus Orbiculatus total terpenoids 60 mg·kg-1·d-1,120 mg·kg-1·d-1,200 mg·kg-1·d-1 with intragastric administration respectively.Each group was weighed once a week.On the basis of establishing hyperlipidemia mice model,blood lipids,lipid metabolic enzyme,antioxidative capacity were investigated after 21 days feeding of high-fat forage.Results Compared with model group,TC and TG in mice treated with Celastrus Orbiculatus total terpenoids all reduced and HDL-C raised obviously(P<0.01).Celastrus orbiculatus total terpenoids was shown to decreased MDA content in both serum and liver,increased serum SOD activity and inhibited the activity of the cholesteryl ester transfer protein(CETP).Conclusions Celastrus Orbiculatus total terpenoids could remarkably modulate the lipid metabolic disorder in hyperlipidemia mice,and has a certain regulating function on lipoprotein,inferring that it could reduce the occur of atherosclerosis.The mechanism of regulating lipid metabolism might be related with decreasing the activity of CETP and increasing antioxidative capacity.

Study of the mechanism underlying inhibitory effect of terpenoids from Daphne plants on liver cancer cells using a network pharmacology-based approach

Daphne plants,belonging to Thymelaeaceae,have been used as traditional Chinese medicine for thousands of years.Previous studies suggested that they exerted anti-tumor activities and reported a number of compounds with anti-tumor activities,isolated from the genus Daphne.However,the targets of these compounds have not been determined yet.In this study,network pharmacology was used to analyze 128 terpenoid compounds from the genus Daphne and 49 target proteins of liver cancer.The results suggested that the target with the most number of directed edges was 5yz0.The compound which docked with 5yz0 best is AR.DT.25(acutilobin D),and the compounds with total docking score greater than 10 were almost all Daphnane diterpenoids.The network pharmacology results predicted that the 5yz0 protein was one of the potential targets of the terpenoids from the genus Daphne and then suggested several new targets and compounds for further liver cancer research.

Impact of oxygen supply on production of terpenoids by microorganisms: State of the art

Terpenoids are a class of high value-added natural products with a variety of biological functions.Genetically engineered microorganisms,such as those of Escherichia coli and Saccharomyces cerevisiae,have merits in producing plant or fungus-derived terpenoids,due to their mature genetic manipulation,simple nutrient demand and fast growth.Oxygen,as a key environmental factor,is particularly important to microbial metabolism and growth,and suitable oxygen supply is viewed as a prerequisite for realizing highly efficient production of terpenoids by engineered microorganisms.In this article,the role of oxygen in regulating terpenoid bioproduction is overviewed from the viewpoints of cellular carbon metabolism,energy metabolism and terpenoid anabolism.Strategies on adjusting oxygen availability to microorganisms,including genetic modification of cellular metabolism related with oxygen utilization,are summarized and discussed,to provide helpful information for further improvement of terpenoid biosynthesis by microbes.

The constitutions of terpenoids and hormones of leaves in red-flesh citrus mutants and their wild types

To study the profiles and relationships of terpenoids and phytohormones in leaves of red-flesh citrus,we examined four red-flesh mutants that accumulate lycopene in the fruit and their corresponding wild types.We compared their contents of monoterpenoid and sesquiterpenoid volatiles,triterpenoid limonoid aglycones,and tetraterpenoid carotenoids.The types and concentrations of these terpenoids in the leaves varied among genotypes,and the leaf terpenoid profiles differed between red-fleshmutants and theirwild types.However,lycopenewas not detected in citrus leaves of all eight varieties,including the fourmutants.According to phytohormone accumulation in the leaves,the citrus varieties could be classified into a low-phytohormone group(‘Red Anliu’sweet orange,‘Anliu’sweet orange,and‘Cara Cara’navel orange),a high-phytohormone group(‘Seike’navel orange),and a high-jasmonic acid group(‘Red-flesh Guanxi’pummelo,‘Guanxi’pummelo,‘Chuhong’pummelo,and‘Feicui’pummelo).The contents of terpenoid volatiles were very low in leaves of‘Red-flesh Guanxi’and‘Guanxi’pummelo;therefore,they are ideal materials for verifying the functions of genes related to terpenoid volatiles in overexpression analyses.Sesquiterpenoid volatiles were positively correlated with phytohormones of abscisic acid,jasmonic acid,and salicylic acid.Taken together,lycopene was undetected in the leaves of redflesh citrus mutants with abundant lycopene in their juice sacs,which suggested a tissue-specific accumulation pattern of lycopene in citrus red-flesh mutants.Furthermore,leaf volatile profiles could be suitable to develop fingerprint chromatograms of citrus resources,since leaf volatile profiles were dominated by several compounds that varied among genotypes in their combinations and concentrations.

Recent Advances in Microbial Production of Terpenoids from Biomass-derived Feedstocks

Terpenoids are a diverse class of natural products widely used as pharmaceuticals,perfumes,flavors,and biofuels.Traditionally,terpenoids are obtained from natural sources,such as plants,but their production is limited by the insufficiency of resources and low yields of extraction.Microbial production of terpenoids has emerged as a promising alternative due to that it is sustainable and easy to scale up.This review aims to summarize recent advances in microbial production of terpenoids from inexpensive biomass-derived feedstocks.Metabolic pathways and key enzymes involved in terpenoid biosynthesis are introduced.Microorganisms that can utilize low-cost lignocellulosic feedstocks for terpenoid production are highlighted.The challenges and prospects faced by microbial terpenoid production are proposed.We believe that continuous progress in the fields of biomass transformation and synthetic biology will ultimately achieve industrial production of microbial terpenoids.

Clavukoelloids A—M,Thirteen Rare N-Containing Nardosinane Type Sesquiterpenoids from the Soft Coral Clavularia koellikeri

Thirteen new nitrogen-containing nardosinane-type sesquiterpenoids,clavukoelloids A-M(1-13)with unique acrylamide fragments vs.the reported general nardosinane-type sesquiterpenoids containing lactone,were isolated from the soft coral Clavularia koellikeri collected from the South China Sea.The structures and absolute configurations of new compounds were unambiguously determined by analysis of spectroscopic data,CP3,DP4+,ECD spectra,and X-ray diffraction.Compound 6 showed anti-inflammatory activities and hepatoprotective activities in transgenic fluorescent zebrafish.

Meroterpenoids Possessing Diverse Rearranged Skeletons with Anti-inflammatory Activity from the Mangrove-Derived Fungus Penicillium sp. HLLG-122

Fifteen new meroterpenoids, littoreanoids A—O (1—15), including three rearranged skeleton meroterpenoids (1—3), were isolated from the mangrove-derived fungus Penicillium sp. HLLG-122. Compound 1 was a novel berkeleyacetal-derived meroterpenoid featuring an unusual spirocyclic 2-oxaspiro[5.5]undeca-4,7-dien-3-one moiety. Compound 2 possessed an unusual 6/6/6/6/6 pentacyclic system with a novel 1-hydroxy-7,7-dimethyl-2-oxabicyclo[2.2.2]octan-5-yl acetate moiety. Compound 3 was an unusual 6/7/6/5/6/5/4 polycyclic systems containing a β-lactone ring. The structures and absolute configurations of new compounds were elucidated by HRESIMS, NMR spectroscopy, single crystal X-ray diffraction analysis, and electronic circular dichroism calculations. The plausible biosynthetic pathways of 1—3 were also proposed. Compounds 6 and 11 exhibited anti-inflammatory effects with IC_(50) values of 30.41 and 19.02 μmol/L, respectively. The bioactive compound 11 was selected for the investigation of preliminary mechanism using molecular docking and Western blotting experiments. Compound 11 could suppress the levels of TNF-α and IL-6, and down-regulate the protein expression of iNOS and COX-2 in RAW 264.7 cells.

Five new terpenoids from Viburnum odoratissimum var.sessiliflorum

Five new terpenoids,including two vibsane-type diterpenoids(1,2)and three iridoid allosides(3-5),together with eight known ones,were isolated from the leaves and twigs of Viburnum odoratissimum var.sessiliflorum.Their planar structures and relative configurations were determined by spectroscopic methods,especially 2D NMR techniques.The sugar moieties of the iridoids were confirmed asβ-D-allose by GC analysis after acid hydrolysis and acetylation.The absolute configurations of neovibsanin Q(1)and dehydrovibsanol B(2)were determined by quantum chemical calculation of their theoretical electronic circular dichroism(ECD)spectra and Rh2(OCOCF_(3))_(4)-induced ECD analysis.The anti-inflammatory activities of compounds 1,3,4,and 5 were evaluated using an LPS-induced RA W264.7 cell model.Compounds 3 suppressed the release of NO in a dose-dependent manner,with an IC_(50) value of 55.64μmol·L^(-1).The cytotoxicities of compounds 1-5 on HCT-116 cells were assessed and the results showed that compounds 2 and 3 exhibited moderate inhibitory activities with IC_(50) values of 13.8 and 12.3μmol·L^(-1),respectively.

Modified Atmospheric Packaging and Its Effect on Postharvest Cannabis Quality

Cannabis sativa L. is used as fiber, food, and medicine in several countries. Though it is illegal for recreational use in most of the world, there are some countries that have legalized production and sale. There is a lot of research on production of cannabis, but less so on storage technologies. Cannabis contains several high value compounds, such as cannabinoids and terpenoids, that are susceptible to degradation via light, temperature, and oxygen. Several studies have explored temperature and light, and industry has adjusted accordingly. However, less is known about oxygen-induced degradation. Biochemical studies have demonstrated oxidative degradation of high value compounds, and many producers use some form of modified atmospheric packaging (MAP) for storage. However, the efficacy of MAP is unclear. The objective of this paper is to review our current understanding of MAP in postharvest cannabis storage and identify avenues where additional research is needed.

Plant Terpenoids： Biosynthesis and Ecological Functions

Among plant secondary metabolites terpenolds are a structurally most diverse group; they function as phytoalexins In plant direct defense, or as signals In Indirect defense responses which involves herbivores and their natural enemies. In recent years, more and more attention has been paid to the Investigation of the ecological role of plant terpenolds. The biosynthesis pathways of monoterpenes, sesquiterpenes, and diterpenes Include the synthesis of C5 precursor isopentenyl diphosphate （IPP） and Its allylic isomer dlmethylallyl dlphosphate （DMAPP）, the synthesis of the immediate diphosphate precursors, and the formation of the diverse terpenoids. Terpene synthases （TPSs） play a key role In volatile terpene synthesis. By expression of the TPS genes, significant achievements have been made on metabolic engineering to Increase terpenoid production. This review mainly summarizes the recent research progress In elucidating the ecological role of terpenoids and characterization of the enzymes Involved in the terpenold biosynthesis. Spatial and temporal regulations of terpenoids metabolism are also discussed.

Effects of Gibberellic Acid on Primary Terpenoids and △^9 -Tetrahydrocannabinol in Cannabis sativa at Flowering Stage

Plants synthesize an astonishing diversity of isoprenoids, some of which play essential roles in photosynthesis, respiration, and the regulation of growth and development. Two independent pathways for the biosynthesis of isoprenoid precursors coexist within the plant cell： the cytosolic mevalonic acid （MVA） pathway and the plastidial methylerythritol phosphate （MEP） pathway. However, little is known about the effects of plant hormones on the regulation of these pathways. In the present study we investigated the effect of gibberellic acid （GA3） on changes in the amounts of many produced terpenoids and the activity of the key enzymes, 1-deoxy-D-xylulose 5-phosphate synthase （DXS） and 3-hydroxy-3-methylglutaryl coenzyme A reductase （HMGR）, in these pathways. Our results showed GA3 caused a decrease in DXS activity in both sexes that it was accompanied by a decrease in chlorophylls, carotenoids and Ag-tetrahydrocannabinol （THC） contents and an increase in α-tocopherol content. The treated plants with GA3 showed an increase in HMGR activity. This increase in HMGR activity was followed by accumulation of stigmasterol and β-sitosterol in male and female plants and campestrol in male plants. The pattern of the changes in the amounts of sterols was exactly similar to the changes in the HMGR activity. These data suggest that GA3 can probably influence the MEP and MVA pathways oppositely, with stimulatory and inhibitory effects on the produced primary terpenoids in MVA and DXS pathways, respectively.

Anti-proliferation activity of terpenoids isolated from Euphorbia kansui in human cancer cells and their structure-activity relationship

Euphorbia kansui is a commonly used traditional Chinese medicine for the treatment of edema,pleural effusion,and asthma,etc.According to the previous researches,terpenoids in E.kansui possess various biological activities,e.g.,anti-virus,anti-allergy,antitumor effects.In this work,twenty five terpenoids were isolated from E.kansui,including thirteen ingenane-and eight jatrophane-type diterpenoids(with two new compounds,kansuinin P and Q)and four triterpenoids.Eighteen of them were analyzed by MTS assay for in vitro anticancer activity in five human cancer cell lines.Structure-activity relationship for 12 ingenane-type diterpenoids in colorectal cancer Colo205 cells were preliminary studied.Significant anti-proliferation activities were observed in human melanoma cells breast cancer MDA-MB-435 cells and Colo205 cells.More than half of the isolated ingenane-type diterpenoids showed inhibitory activities in MDA-MB-435 cells.Eight ingenane-and one jatrophane-type diterpenoids possessed much lower IC_(50) values in MDA-MB-435 cells than positive control staurosporine.Preliminary structure-activity relationship analysis showed that substituent on position 20 was important for the activity of ingenane-type diterpenoids in Colo205 cells and substituent on position 3 contributed more significant biological activity of the compounds than that on position 5 in both MDA-MB-435 and Colo205 cells.

RNA-seq reveals mechanisms of SlMX1 for enhanced carotenoids and terpenoids accumulation along with stress resistance in tomato

Improving nutritional fruit quality and impacts important agro-traits such as biotic or abiotic stresses are extremely important for human civilization.Our previous study reported that manipulation of Sl MX1 gene enhanced carotenoids accumulation and drought resistance in tomato.Here,RNA-Seq analysis proved to be a very useful tool to provide insights into the regulatory mechanisms of Sl MX1 involved in stress resistance and enhanced secondary metabolites.Physiological analysis showed that overexpression of Sl MX1 results in substantially increased broad-spectrum tolerance to a wide-range of abiotic and biotic(fungus,bacteria,virus and insects) stresses in tomato.This research appears to be of remarkable interest because enhanced terpenoids content has been achieved by increasing trichome density.In addition,we reported two types of trichome which seems to be aberrant types in tomato.This study unravels the mechanism of regulation of Sl MX1,which simultaneously modulates resistance and metabolic processes through regulating key structural and regulatory genes of the corresponding pathways.

Virtual Evaluation on the Activities of Phthalides and Terpenoids from Angelica sinensis

Objective To elucidate potential activities of phthalides and terpenoids from Angelica sinensis by theoretical docking calculation. Methods Eleven components of phthalides and terpenoids were selected as ligand. The crystalline structures of targets related to common diseases were used as the receptors for calculation. The calculations were conducted with Schr?dinger software package. The grading standard of selectivity was developed according to G-score between ligands and receptors. Results Selective targets of phthalides and terpenoids were related to nevous system diseases, cancer, pain, diabetes, cardiovascular disease, liver cirrhosis, nephrotic syndrome, inflammatory diseases, rheumatoid arthritis, dermatosis, leukemia, microbial inflections, immune diseases, and hypercholesterolemia. In addition to the medical treatments reported in the literature, our research also indicated that these two classes of compounds may be used for tumor, diabetes, rheumatoid arthritis, dermatosis, leukemia, liver cirrhosis, and nephrotic syndrome. According to our research, the effects of phthalides and terpenoids may be not so strong. Conclusion The effects of phthalides and terpenoids on diseases founded through virtual evaluation accord greatly with those reported in experiment and clinic. The combination of computer-aided drug evaluation technique and experiment is definitely an important and fast way to investigate the effects and mechanisms of traditional Chinese medicine.

Discovery of β-Dihydroagarofuran-Type Sesquiterpenoids from the Leaves of Tripterygium wilfordii with Neuroprotective Activities

Nine undescribed dihydro-β-agarofuran sesquiterpenoid derivatives(1—9),along with a known analogue(10),were obtained from the leaves of Tripterygium wilfordii.Their gross structures were determined via extensive spectroscopic data,and the absolute configurations were elucidated by means of single-crystal X-ray diffraction analysis and electron circular dichroism(ECD)techniques,which include ECD exciton chirality,octant rule of saturated cyclohexanone and comparison between the experimental and calculated ECD spectra.All the isolated compounds were tested for their neuroprotective activities against H2O2-induced cell injury in human neuroblastoma SH-SY5Y cells.Compounds 5 and 6 improved cell viability by 16.15%and 15.12%compared with the H2O2 treated group at 25μmol·L^(-1),respectively.

Terpenoids of Sinularia soft corals:chemistry and bioactivity

Soft corals of the genus Sinularia are one of the most widespread soft corals.They are a rich source of bioactive su bstances with intriguing and unique structural features.The present paper reviews the latest progress in the chemistry and pharmacological activities of terpenoids from Sinularia soft corals and provides a perspective on future areas of research interest.

Anisotanols A—D,Four Norsesquiterpenoids with an Unprece-dented Sesquiterpenoid Skeleton from Anisodus tanguticus

Anisotanols A—D(1-4),four new compounds possessing an unprecedented sesquiterpenoid skeleton with a congested tricyclic 6/3/5 ring system,were obtained from Anisodus tanguticus.Their structures were elucidated by comprehensive spectroscopic techniques,and the absolute configurations were confirmed via ECD calculations and single-crystal X-ray diffractions.A putative biosynthetic pathway for these compounds was proposed.Biological evaluation disclosed that compound 3 showed anti-angiogenic activity by inhibiting the viability,migration,and tube formation in HUVECs.