Search

Santalenes and santalols from Santalum album are the main components of the valuable spice sandalwood essential oil, which also has excellent pharmacological activities such as antibacterial, anti-inflammatory, and antitumor. Firstly, we constructed biosynthesis pathways of santalenes by synthetic biology strategy. The assembled biosynthetic cassettes were integrated into the multiple copy loci of δ gene in S. cerevisiae BY4742 with assistance of pDi-CRISPR, and 94.6 mg/L santalenes was obtained by shake flask fermentation of engineered yeast. Secondly, a selected optimized P450-CPR redox system was integrated into the chromosome of the santalenes-producing strain with a single copy, and 24.6 mg/L santalols were obtained. Finally, the yields of santalenes and santalols were increased to 164.7 and 68.8 mg/L, respectively, by downregulating ERG9 gene. This is the first report on the de novo synthesis of santalols by P450-CPR chimera in S. cerevisiae. Meanwhile, the optimized chimeric CYP736A167opt-46tATR1opt exhibits higher activity to oxidize santalenes into santalols. It would provide a feasible solution for the optimal biosynthesis of santalols. KEY POINTS: • First-time de novo synthesis of santalols by P450-CPR chimera in S. cerevisiae. • Truncated 46tATR1 has higher activity than that of CPR2. • Yields of santalenes and santalols were increased by downregulating ERG9 gene.

The profitability of the olive flounder (Paralichthys olivaceus) aquaculture industry in Korea depends on high production and maintenance of flesh quality, as consumers prefer to eat raw flounders from aquaria and relish the raw muscles as ‘sashimi’. For sustaining high production, easy-to-deliver and efficient vaccination strategies against serious pathogens, such as viral hemorrhagic septicemia virus (VHSV), is very important as it cause considerable losses to the industry. Whereas, a safe and non-invasive vaccine formulation that is free from unacceptable side-effects and does not devalue the fish is needed to maintain flesh quality. We previously developed a squalene–aluminium hydroxide (Sq + Al) adjuvanted VHSV vaccine that conferred moderate to high protection in flounder, without causing any side effects when administered through the intraperitoneal (IP) injection route. However, farmers often demand intramuscular (IM) injection vaccines as they are relatively easy to administer in small fishes. Therefore, we administered the developed vaccine via IP and IM routes and investigated the safety and persistency of the vaccine at the injection site. In addition, we conducted a comparative analysis of vaccine efficacy and serum antibody response. The clinical and histological observation of the IM and IP groups showed that our vaccine remained persistence at the injection sites for 10–17 weeks post vaccination (wpv), without causing any adverse effects to the fish. The relative percentage of survival were 100% and 71.4% for the IP group and 88.9% and 92.3% for the IM group at 3 and 17 wpv, respectively. Thus, considering the persistency period (24 wpv) and both short and long-term efficacy of our vaccine, the present study offers an option to flounder farmers in selecting either IM or IP delivery strategy according to their cultured fish size and harvesting schedule — IM vaccination for small-sized fish and IP vaccination for table-sized fish.

Transcriptional downregulation is widely used for metabolic flux control. Here, marO, a cis-element of Escherichia coli mar operator, was explored to engineer promoters of Saccharomyces cerevisiae for downregulation. First, the ADH1 promoter (PADH1) and its enhanced variant PUADH1 were engineered by insertion of marO into different sites, which resulted in decrease in both gfp5 transcription and GFP fluorescence intensity to various degrees. Then, marO was applied to engineer the native ERG1 and ERG11 promoters due to their importance for accumulation of value-added intermediates squalene and lanosterol. Elevated squalene content (4.9-fold) or lanosterol content (4.8-fold) and 91 or 28% decrease in ergosterol content resulted from the marO-engineered promoter PERG1(M5) or PERG11(M3), respectively, indicating the validity of the marO-engineered promoters in metabolic flux control. Furthermore, squalene production of 3.53 g/L from cane molasses, a cheap and bulk substrate, suggested the cost-effective and promising potential for squalene production.

Heavy metal toxicity and drought stresses are two growing concerns of the global population as these have a disastrous effect on the agriculture sector, creating oxidative stress in plants and leads to deleterious effects, which end up causing a major decline in yield. Hence, carving out the best candidate for combating both these stresses have become the prime objective of researchers. Plectranthus amboinicus (Lour.) Spreng., a medicinal plant, is regarded as one such candidate that can tolerate both drought and zinc stressors by the elicitation of metabolic changes. Variation in the composition of primary and secondary metabolites of P. amboinicus was evaluated in the leaf tissues of the plants subjected to drought and ZnSO4 (4 mM) treatments. Drought stress resulted in the accumulation of amino acids and sugars in the leaves of plants exposed to this stress. Similarly, zinc stress exhibited a remarkable impact on the synthesis of secondary metabolites like alkaloids, phenols, and flavonoids. Further, on carrying out GC–MS profiling, the compositional variation of secondary metabolites produced in P. amboinicus implicated its inherent potential to survive environments of these two stresses. Compounds like 2-methoxy-4-ethyl-6-methylphenol, gamma-sitosterol, hexadecanoic acid, alpha-amyrin, and ethyl linalool were some of the major secondary metabolites developed in leaves of plants under drought, whereas during ZnSO4 treatment, the major compounds developed were trans-alpha-bergamotene, squalene, 1,5-dimethyl-1-vinyl-4-hexenyl isovalerate, and spathulenol. This in turn makes the plant more compatible with the stressful environmental conditions, aiding it with better survival and protection. At the same time, the enhancement in the content of these metabolites in this medicinal plant under the influence of these stressors may have applications in the pharmaceutical industry, necessary for the development of novel drugs.

PURPOSE: The enzyme Cyclooxygenases (COX-1 and COX-2) catalyze the formation of prostaglandin, a mediator of the inflammatory pathway. Inflammation related pathological conditions may be alleviated by targeting the Cox enzymes.COX-2 inhibitors that are currently available in the market causes undesirable side effects. Our present study focuses on the in-silico inhibition of COX -2 enzyme by the phytocompounds from Albizia amara and Phyla nodiflora. METHODS: The phytochemicals present in Albizia amara and Phyla nodiflora were analyzed for their COX-2 inhibition potential. Eight compounds from Albizia amara and eleven compounds from Phyla nodiflora obtained from GC–MS analysis was used for the current study. Molecular docking was performed using AutoDock vina. The crystal structure of COX-2 (PDB ID: 5IKR) was obtained from Protein data bank. PyMol was used to remove any solvent, organic and inorganic molecules. Energy minimization of the protein was carried out using SPDBV software. Geometrical optimizations of the ligands were performed using Avogadro software. Celecoxib was used as the positive control. ADMET properties of the compounds were analyzed using SwissADME and ProtoxII online servers. Molecular mechanics/generalized born surface area (MM/GBSA) calculations were performed to evaluate the binding efficiency. Molecular dynamics of the protein and protein–ligand complex was studied for about 100 ns using Desmond package of Schrodinger suite. RESULTS: Among the eighteen compounds, Squalene present in both the plants showed a better binding energy of -7.7 kcal/mol, when compare to other phytocompounds present in the extract. The control celecoxib showed a binding energy of about – 9.4 kcal/mol. The toxicity and ADMET properties of squalene indicated that it is non-toxic and followed Lipinski’s rule. Molecular Dynamics (MD) analysis showed that the binding of squalene to the enzyme was stable. CONCLUSION: Squalene could potentially inhibit COX2 and o wing to its properties, squalene can be formulated in gels/creams and could be possibly used for external edema and inflammation GRAPHICAL ABSTRACT: [Image: see text] SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s40199-021-00408-6.

Clerodendrum inerme is a perennial shrub belonging to the family Lamiaceae and is used in many of the herbal preparations of Siddha and Ayurveda in India. Ethnomedicinally, the leaves of this plant are used to cure various ailments. In spite of enormous ethnomedicinal implications, proper chemical profiling and characterization of isolated compounds of C. inerme still remains unaddressed. Present study involves the characterization and chemical profiling of phytoconstituents from leaf extract which promise to be of immense therapeutic significance. Two compounds namely squalene and linolenic acid methyl ester were isolated from the leaves of C. inerme employing column chromatography technique. The structures were elucidated by spectroscopic methods including FTIR, 1H NMR and 13C NMR. Subsequently, antioxidant and antimicrobial activities of the isolated compounds were estimated in-vitro. Pertaining results were validated with in silico molecular docking-based investigations which revealed that both the compounds displayed high binding affinities with the target proteins of the antioxidant pathway commonly targeted for drug development.

The oil palm (Elaeis guineensis) fruit is a source of vegetable oil and various phytonutrients. Phytochemical compounds present in palm oil include tocotrienols, carotenoids, phytosterols, squalene, coenzyme Q10, and phospholipids. Being a fruit, the oil palm is also a rich source of water-soluble phytonutrients, including phenolic compounds. Extraction of phytonutrients from the oil palm vegetation liquor of palm oil milling results in a phenolic acid-rich fraction termed Water-Soluble Palm Fruit Extract (WSPFE). Pre-clinical in vitro, ex vivo, and in vivo studies carried out using various biological models have shown that WSPFE has beneficial bioactive properties, while clinical studies in healthy volunteers showed that it is safe for human consumption and confers antioxidant and anti-inflammatory effects. The composition, biological properties, and relevant molecular mechanisms of WSPFE discovered thus far are discussed in the present review, with a view to offer future research perspectives on WSPFE for health and non-health applications.

Panax quinquefolius L., a medicinal plant of Araliaceae family, is used in traditional Chinese medicine. Arbuscular mycorrhizal fungi (AMF) has been shown to promote plant growth and the accumulation of secondary metabolites in many plants. Our group has found a significant positive correlation between AMF infection rate and the accumulation of ginsenoside, while the effects and underlying mechanism of AMF on the growth and accumulation of ginsenoside in P. quinquefolius have been poorly studied. In this study, the effects on the biomass, photosynthesis and expressions of genes involved in biosynthesis of ginsenoside: hydroxymethylglutaryl-CoA reductase (HMGR), geranylgeranyl pyrophosphate synthase (GPS), farnesyl diphosphate synthase (FPS), squalene synthase (SS), squalene epoxidase (SE), dammarendiol synthase (DS) and cytochrome P450 enzyme (CYP716A47, CYP716A53v2) in P. quinquefolius during AMF inoculation were investigated. Plant biomass and photosynthesis of P. quinquefolius were stimulated by AMF treatment. Chlorophyll concentration in P. quinquefolius leaves treated by R. intraradices was higher than those in non-inoculated leaves. The concentration of Rb1 and Re was markedly increased by 26.15% and 22.38% (P

Azadirachta indica is used to insects repellent, control diabetes, and combat with cancer. In this study, the effect of different concentrations of chitosan and sampling times on cell suspension culture and SQS1 and MOF1 genes expression were studied and response surface methodology was used to construct the prediction models for azadirachtin, mevalonic acid and squalene accumulation and production. The highest fresh and dry cell weight and azadirachtin accumulation were 726.21 g/L, 20.76 g/L, and 11.74 mg/g DW obtained 6 days after using 50 mg/L chitosan, respectively. The highest azadirachtin production was 165.50 mg/L achieved by using 50 mg/L chitosan for 8 days. Maximum mevalonic acid accumulation (1.87 mg/g DW) and production (22.50 mg/L) were observed 2 and 4 days after the addition of 50 mg/L chitosan and control condition, respectively. The highest amount of squalene accumulation (0.319 mg/g DW) and production (3.56 mg/L) were obtained 10 days after using 50 mg/L chitosan. Also, the prediction results showed the highest azadirachtin accumulation (10.76 mg/g DW) and production (136.83 mg/L), mevalonic acid accumulation (1.869 mg/g DW) and production (20.576 mg/L) and squalene accumulation (0.217 mg/g DW) and production (2.23 mg/L) by using 71.50 mg/L chitosan for 5.24 days, 66.25 mg/L chitosan for 9.1 days, 2.5 mg/L chitosan for 2 days, without chitosan for 2 days, 98.50 mg/L chitosan for 2 days and 80.25 mg/L chitosan for 6.08 days, respectively. The qRT-PCR analysis indicated the maximum relative expression of SQS1 and MOF1 genes by using 25 and 50 mg/L chitosan for 6 days. For the first time, chitosan concentrations and sampling times optimized by response surface methodology for azadirachtin, mevalonic acid and squalene accumulation and production and investigated SQS1 and MOF1 genes expression.

Background Wrightia tinctoria R.Br. (Apocyanaceae) is known as a biologically effective plant for the treatment of jaundice in the Indian traditional system of medicine. It is a wild medicinal tree possessing anti-inflammatory, antidiabetic, antinociceptive, hepatoprotective, antibacterial, antifungal, antiviral, antipsoriatic, anticancerous, anthelmintic, aphrodisiac, analgesic, and antipyretic activities. Its constituents are of utmost interest to pharmaceutical industries owing to their many actions and biological activities. Method Methanolic extract of W. tinctoria (MEWT) was investigated by gas chromatography-mass spectroscopy and provided affirmative results assisting in the identification and characterization of therapeutic claims regarding this species in the traditional system. The antioxidant activity of MEWT was determined by the most suitable DPPH method. Results The basic compounds found in MEWT were β-caryophyllene (0.22%), mome inositol (12.02%), neophytadiene (1.61%), eicosanoic acid methyl ester (0.32%), 8,11,14-eicosatrienoic acid methyl ester (0.60%), phytol (0.94%), phytol palmitate (1.37%), squalene (1.57%), flavone 4-OH, 5-OH, 7-di-O-glucoside (29.34%), γ-tocopherol (0.49%), stigmast-5-en-3-ol (3.14%), methyl commate B (1.76%), methyl commate A (5.20%), and 24-norursa-3,12-diene (20.36%). The obtained results in the analysis of antioxidant activity of MEWT exhibited considerable free radical scavenging capacity against DPPH-generated free radicals. Conclusions This study expands the knowledge of MEWT chemical composition and provides evidence to substantiate ethno-medicinal use of the plant by exploring antioxidant activity. The substantial antioxidant activity of MEWT could be due to presence of terpenes, flavonoids, vitamin E, and other reported compounds. Highlights This study includes identification of phytochemicals and antioxidant potential of methanolic extract of Wrightia tinctoria, assisting in therapeutic claims regarding this species in the traditional system.

Sabkhas in Kuwait are unique hypersaline marine environments under-explored for bacterial community composition and bioprospecting. The 16S rRNA sequence analysis of 46 isolates with distinct morphology from two Kuwait sabkhas recovered 11 genera. Phylum Firmicutes dominated these isolates, and Bacillus (32.6%) was recovered as the dominant genera, followed by Halococcus (17.4%). These isolates were moderately halophilic, and some of them showed tolerance and growth at extreme levels of salt (20%), pH (5 and/or 11), and temperature (55 °C). A higher percentage of isolates harbored protease (63.0), followed by DNase (41.3), amylase (41.3), and lipase (32.6). Selected isolates showed antimicrobial activity against E. faecalis and isolated Halomonas shengliensis, and Idiomarina piscisalsi harbored gene coding for dNDP-glucose 4,6-dehydratase (Glu 1), indicating their potential to produce biomolecules with deoxysugar moieties. Palmitic acid or oleic acid was the dominant fatty acid, and seven isolates had some polyunsaturated fatty acids (linolenic or γ-linolenic acid). Interestingly, six isolates belonging to Planococcus and Oceanobacillus genus produced squalene, a bioactive isoprenoid molecule. Their content increased 30-50% in the presence of Terbinafine. The potential bioactivities and extreme growth conditions make this untapped bacterial diversity a promising candidate for future bioprospecting studies.

Anaerobically treated molasses-based distilleries spent wash is a major source of terrestrial and aquatic pollution due to the mixture of the various unknown organometallic compounds in discharged waste. The nature of pollutants present in effluent has to be characterized prior to the evaluation of their fate in the environment at various tropic levels. The absorption maxima (λmax) were obtained at 295 nm of pollutants present in spent wash which indicated that melanoidins are a major colorant along with other organic compounds. Abundantly identified compounds with GC–MS in effluent were found: 1,2,4,5-tetrahydro-2-methyl-3H-2-benzazepine-3-one, 6-(trifluoromethyl)-1H-imidazole [4,5-c] pyridine, 4,6-di-t-butyl-1H, 3H-thieno[3,4-c] thiophene, butanoic acid, trimethylsilyl ester, silane, (dodecyloxy)trimethylsilyl ester, 4-styryl [2,2] paracyclophane, 4-(p-cumylphenoxy) phthalonitrile, hexadecanoic acid, trimethylsilyl ester, butyl ester, and squalene. Some of these compounds are known with mutagenic and androgenic properties. The toxicity test of post-methanated distillery effluent (PMDE) showed direct toxic effects on catfish (Heteropneustes fossilis) even at 1% (v/v) concentration and caused degeneration of primary and secondary lamellae of the gill and the epithelial layer. Further, higher concentrations between 5 and 10% of PMDE showed loss of primary and secondary lamellae of gills, and liver catalase activity was also increased dramatically in H. fossilis, indicating the generation of oxidative free radicals. Similarly, inhibition of α-amylase activities in germinating Zea mays L. (Maize) seeds was also noted at 10% PMDE. This study confirmed the environmental toxicity of effluent with freshwater fish and Zea mays seeds.

Neem is the medicinal plant used as antimalarial, antibacterial, antiviral, insecticide, and antimicrobial. This study aimed to investigate and prediction the effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression by response surface methodology. The highest fresh and dry cell weights were 580.25 g/L and 21.01 g/L obtained 6 days after using 100 mg/L yeast extract. The highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained 2 and 4 days after using 25 mg/L yeast extract. Maximum mevalonic acid accumulation and production were 1.75 mg/g DW and 23.77 mg/L observed 2 days after application of 50 mg/L yeast extract. The highest amount of squalene accumulation (0.22 mg/g DW) and production (4.53 mg/L) were achieved 4 days after using 50 mg/L yeast extract. Prediction results showed that the highest azadirachtin accumulation (13.61 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.50 mg/g DW) and production (5.57 mg/L), and squalene accumulation (0.30 mg/g DW) obtained by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, without yeast extract for 6.54 days and 4 days, respectively. Also, predicted that the highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. The qRT-PCR analysis showed the maximum relative gene expression of SQS1 and MOF1 obtained by using 150 mg/L yeast extract for 4 days and 25 mg/L yeast extract for 2 days, respectively.

Volatile terpenoids are a large group of important secondary metabolites and possess many biological activities. The acyclic sesquiterpene trans-nerolidol is one of the typical representatives and widely used in cosmetics and agriculture. Here, the accumulation of volatile terpenes in different tissues of Celastrus angulatus was investigated, and two trans-nerolidol synthases, CaNES1 and CaNES2, were identified and characterized by in vitro enzymatic assays. Both genes are differentially transcribed in different tissues of C. angulatus. Next, we constructed a Saccharomyces cerevisiae cell factory to enable high-level production of trans-nerolidol. Glucose was the sole carbon source to sequentially control gene expression between the competitive squalene and trans-nerolidol pathways. Finally, the trans-nerolidol production of recombinant strain LWG003-CaNES2 was 7.01 g/L by fed-batch fermentation in a 5 L bioreactor. The results clarify volatile terpenoid biosynthesis in C. angulatus and provide a promising potential for industrial production of trans-nerolidol in S. cerevisiae.

Objective: The aim of this study was to investigate the chemical compound of the antioxidant activity of the extract from andaliman fruit (Zanthoxylum acanthopodium DC.) and the quality parameters and the total flavonoid determination of the highest antioxidant activity. Methods: The andaliman fruits were extracted using a solvent such as n-hexane, ethyl acetate and 70% ethanol obtained extracts, follow the antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. The quality parameters and total flavonoid determination were performed from highest antioxidant activity. Fractionated was done using vacuum liquid chromatography (VLC) with the mobile phase chloroform-methanol (9:1 ~1:9). Purification with Thin Layer Chromatography (TLC) preparative, identification using UV-spectrophotometry, FTIR, Gas chromatography Mass Spectrometry (GCMS) analysis. Results: The 70% of ethanol has the highest antioxidant activity with IC50 value 84.11±0.47 ppm; the quality parameters in this study meet the quality requirements, both specific and nonspecific parameters. Total flavonoid content of the 70% ethanol extract was 2.37±0.02%. The results of the GCMS studies showed the presence of 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester; squalene, methyl palmitate, methyl isopalmitate, n-tetracosanol-1 and 4-isopropyl chalcone. Conclusion: The results indicate that the 70% ethanol extracts of andaliman fruits contains 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester; squalene, methyl palmitate, methyl isopalmitate, n-tetracosanol-1 and 4-isopropyl chalcone have antioxidant activity with IC50 value 51.35 ppm.

Legume seeds (Fabaceae) of seven species Cyamopsis tetragonoloba (guar), Glycine max (soybean), Lablab purpureus (lablab-bean), Macrotyloma uniflorum (kulthi bean), Phaseolus vulgaris (common bean), Trigonella foenum-graecum (fenugreek) and Vigna unguiculata (cowpea) were studied. The oil yield ranged from 1.2 to 20.2% dw, in the lablab-bean and soybean, respectively. The polyunsaturated fatty acids (PUFA) constituted the largest part (46-78%) of total fatty acids in studied legumes. γ-Tocopherol was the predominant tocopherol (T) homologue (61-95%) in most of the tested legumes with the exception of fenugreek (α-T, 97%) and cowpea (γ-T and δ-T, nearly equal). The β-sitosterol was the main sterol (51-56%) in most legumes. While in cowpea, lablab-bean and kulthi bean the main sterols were β-sitosterol and Δ5-stigmasterol (28-37% and 14-42%, respectively). Squalene was detected only in kulthi bean and lablab-bean (58 and 284 mg/100 g oil). The total concentration of carotenoids, tocochromanols, and sterols in the studied legumes was 0.2-9.2, 12.4-276.0, and 350-8,542 mg/100 g oil, respectively. Based on the levels of minor lipophilic compounds of this study, C. tetragonoloba, T. foenum-graecum and G. max seem to have a better nutritional value compared to P. vulgaris, V. unguiculata, L. purpureus, and M. uniflorum.

The traditional drying method of Camellia oleifera seeds is sun drying and shade drying. In order to further improve the oil content and production efficiency, light and no light treatment were carried out and the oil quality was compared. The results show that the oil contents of these seeds (light treatment, 37.17 ± 0.68%; no light, 36.15 ± 0.57%) were higher than those obtained from sun-dried (33.15 ± 1.23%) or shade-dried (34.29 ± 0.86%) seeds. Increased oil content after light and no-light treatment was attributed to saturated fatty acids and linoleic acid. Squalene content was 2.82–3.68 times higher than that in Camellia oleifera seed oil treated by sun drying and shade drying. Principal component analysis ranking of the comprehensive score was: sun-dried > no-light-treated > light-treated > shade-dried. Heatmap analysis showed two categories of [sun drying + shade drying] and [light treatment + no-light treatment]. Random forest analysis showed that the strongest factors on seed oil quality were: Δ-7-stigmasterol, α-tocopherol, and β-tocopherol.

Among many relevant issues dealing with fish farming, microbial infections are a major problem. There are different viral infections, which are continuously creating problems in fish farming and among these viral infections Betanoda viral infection is a foremost problem. The Betanodavirus is an important, emerging group of viruses known to infect around 40 species worldwide. The major target of this virus is the central nervous system and retina of fishes especially in Barramundi species. Viral Nervous Necrosis (VNN) is now a serious problem for different fish species which is yet to be resolved through strong antiviral compounds. The In-silico screening of potential phytochemicals as a drug molecule with low or no side effects against viral nervous necrosis in barramundi is the major objective of the study. The present study discusses the molecular interaction studies carried out between virtually screened phytochemicals and MX protein of barramundi fish. Findings based on virtual screening, calculation of molecular properties and bioactivity score showed that among 101 compounds, the hypogallic acid, cineole, eugenol, linalool, camphene, oligonol, azulene, caravacrol, pistol and squalene are the active phytochemicals against the selected MX protein. Further intense screening showed that Camphene is the best screened phytochemical with the lowest binding energy in complex with MX protein of Barramundi. Further molecular dynamic simulation study at 100ns (Nano seconds) proved the importance, stability and establishment of camphene as better natural prophylactic and therapeutic approaches to overcome or reduce the problem of viral nervous necrosis in barramundi.

The chemical investigation of the nonpolar constituents of a methanol extract of the bulbs of Boophone haemanthoides yielded ten known compounds as minor constituents. The compounds were identified as stigmast-4-ene-3,6-dione (1); cholest-4-en-3-one (2); (22E)-stigmast-4,22-dien-3-one (3); stigmast-4-en-3-one (4); 6β-hydroxystigmast-4-en-3-one (5); 6β-hydroxycholest-4-en-3-one (6); cycloartenol (7); acetovanillone (8); tyrosol (10) and 3-hydroxy-1-(4-hydroxyphenyl)-1-propanone (11). The isolation of compound 7 with other cholestane (2, 6) and stigmastane (1, 3–5) derivatives aligned with the biosynthetic pathway of plant steroids through 7 from squalene 2,3-epoxide. This is the first report on the isolation and identification of these compounds from B. haemanthoides and 1–6 and 11 for the family Amaryllidaceae.

The coconut mite, Aceria guerreronis Keifer (Acari: Eriophyidae), is a major pest of coconut plantations (Cocos nucifera L.) worldwide. Here, we conducted a bioguided phytochemical approach using toxicity and repellency bioassays of nonpolar extract and its fractions of Vitex gardneriana Schauer (Lamiaceae) leaves to this pest. Nonpolar crude extract was fractionated by column chromatography using solvents with increased polarity and binary mixtures, resulting in five semipurified groups. The biomonitoring bioassay provided active fractions and led to the isolation and characterization of the bioactive compound squalene, a biosynthetic precursor of 20-hydroxyecdysone, which plays an important role in plant defense against arthropods. The LC50 of the crude extract of V. gardneriana for A. guerreronis was estimated to be 0.185 mg·mL-1 and LC80 = 4.123 mg·mL-1. Also, the extract was highly repellent to this pest for up to 24 h. The fractions of V. gardneriana, and also squalene, caused mortality to A. guerreronis. The potential of V. gardneriana fractions/squalene as biopesticides for controlling A. guerreronis in coconut plantations is discussed herein.

The seedlings ofOroxylum indicumwere inoculated with plant growth promoting microbes (PGPMs) mainly,Glomus mosseae,Trichoderma harzianumandPseudomonas putidaboth alone and consortium. The GCMS analysis of the methanolic root extract of inoculated seedlings ofO. indicumshowed that seedlings treated with mixed consortium of mycorrhizal fungi, bacteria and fungus showed the presence of maximum number of phytocompounds. The GC-MS analysis of control seedlings showed presence of 55 compounds where three new compounds were found i.e. 2-Cyclobutene-1-Carboxamide; Tetradecanoic Acid, 10, 13-dimethyl-, methyl ester; 1-methylene-2b-hydroxymethyl-3, 3-dimethyl-4b-(3-methylbut-2-enyl)-cy. 53 compounds were found in seedlings treated with mycorrhizae i.e.,Glomus mosseae, and three new compounds were found i.e., 1-Ethyl-2-Hydroxymethylimidazole; Octadecanoic Acid, 11-Methyl-, methyl ester; 4-Methyl-1, 4-Heptadiene. The seedlings treated with bacteria i.e.Pseudomonas putidashowed the presence of 52 compounds and three new compounds were found i.e. Meso-4, 5-octanediol; 1-ethyl-2-hydroxymethylimidazole; 2, 5-cyclohexadiene-1, 4-dione, 2, 5-dihydroxy-3-methyl-6-(1-methylethyl) - . A total of 56 compounds were present in seedlings treated with fungus i.e.Trichoderma harzianumand five new compounds were found i.e. 2-CyclohexeN-1-one, 2-Butyl-3-Methoxy; Methyl 12, 13-Tetradecadienoate; Methyl 6, 9, 12-hexadecatrienoate; 1, 9-Decadiyne; 1, 4-Naphthalenedione. The seedlings treated with dual consortium of mycorrhizae and bacteria showed the presence of 88 compounds and five new compounds were found i.e., N-(1-Methoxycarbonyl-1-methylethyl)-4-methyl-2-aza-1,3-dioxane;1-ethyl-2 hydroxy methylimidazole; Methyl 8-methyl-nonanoate; Naphthalene, 1,2,3,4,4a,5,6,8a-octahydro-4a,8-dimethyl; Methyl 12,13-tetradecadienoate. 152 compounds were present in seedlings treated with dual consortium of mycorrhizal fungi and fungus and ten new compounds were found to be present i.e. 1,9-Decadiyne; 3,7,11-Trimethyl-3-hydroxy-6,10-dodecadien-1-yl acetate; 3-Heptyne, 7-chloro; 3-Methyl-4-(methoxycarbonyl) hexa-2,4-dienoic acid; Benzo[c]cinnolin-2-amine ; Tetradecanoic acid, 10,13-dimethyl-,Methyl ester; Cis,cis-4,6-octadienol; 2-Cyclohexen-1-one, 2-butyl-3-methoxy; Methyl 12,13-tetradecadienoate; 2-Aminopyridazino(6,1-b) quinazolin-10-one. A total of 36 compounds were present in seedlings treated with dual consortium of bacteria and fungi and two new compounds were found i.e. [1,4] Dioxino [2,3-b]-1,4-dioxin, hexahydro-2,3,6,7 ; 1-Ethyl-2-hydroxymethylimidazole. The seedlings inoculated with mixed consortium of mycorrhizae, bacteria and fungus showed the presence of 213 compounds and fourteen new compounds were found i.e. 3,7,11-Tridecatrienenitrile, 4,8,12-Trimethyl; 1,9-Decadiyne; 2,6,10,14,18,22-Tetracosahexaene, 2,6,10,15,19,23-Hexamethyl-, (ALL-E) ; 1-Methylene-2b-hydroxymethyl-3,3-dimethyl-4b-(3-methylbut-2-enyl)-cy; 1,9-Decadiyne, Cyclobutane, 1,2-bis(1-methylethenyl)-, trans-, 3,7,11-Trimethyl-3-hydroxy-6,10-dodecadien-1-yl acetate, 5-Hydroxy-4-hydroxymethyl-1-(1-hydroxy-1-isopropyl)cyclohex-3-ene, 5,8,11,14-Eicosatetraenoic acid, methyl ester, (all-z)-, 1-Cyclohexyl-2-buten-1-ol (c,t) , 1-Oxetan-2-one, 4,4-diethyl-3-methylene-, Tetradecanoic acid, 10,13-dimethyl-, methyl ester, 2-Cyclohexen-1-one, 2-butyl-3-methoxy-, Methyl 12,13-tetradecadienoate, Heptacosanoic acid, 25-methyl-, methyl ester Hexadecanoic Acid, Methyl Ester; 2-Chloroethyl Linoleate; 9,12-Octadecadienoic Acid, Methyl Ester, (E,E); Butanoic acid, methyl ester; 4A,5,6,7,8,8A(4H) HexahydroBenzopyran-3-Carboxamide, 8A-Methoxy-4A-M,; Octadecanoic acid; Farnesene; Squalene; Myrcene; Naphthalene; Tetradecanoic Acid, Methyl Ester; Octadecanoic Acid, Methyl Ester; 1H-Cycloprop[E] Azulene, Decahydro-1,1,4,7-Tetramethyl-, [1AR-(1A].Alph ; Cyclohexane, 1-methyl-4-(1-methylethenyl)-, trans (Elemene); Cyclohexene, 1-methyl-4-(1-methylethenyl)-, (s)- (Limonene); were found to be present in this treatment.

Biodiesel and natural products derived from microalgae require a smaller land area and have higher production rates compared to plants and animals and has recently attracted considerable interest. However, biodiesel production from microalgal triacylglycerol is still far from commercial realization due to its high production cost. One way to overcome this obstacle is to improve the triacylglycerol accumulation and couple its production with other high-value compounds. Of particular interest is the sterol biosynthetic pathway with squalene as an intermediate due to its close relationship with triacylglycerol and carotenoid biosynthetic pathways. Besides, both squalene and carotenoids are isoprenoid lipids that have health benefits. Perturbation of one pathway has been suggested to affect other pathways. Three terbinafine-sensitive mutants of the green microalga Chlamydomonas reinhardtii were isolated using terbinafine, a drug that inhibits squalene epoxidase, leading to squalene accumulation. One of the mutants, tfs2, accumulated twice the amount of wild-type triacylglycerol. As well as squalene accumulation, the presence of terbinafine further increased the triacylglycerol content. The level of prenyl lipid carotenoid and chlorophyll was also more significant than that of the wild type. Growth and photosynthesis were not compromised in this mutant. This is the first study that has demonstrated a mutant screening method to improve the co-production of TAG and isoprenoid lipids in a green microalgae.

Abroma augusta L plant traditionally was used to treat swellings, cuts, sores, and bruises. In the province of Jambi, A. augusta is used in folk medicine to treat wounds. This study aims to isolate the steroid compound from the root of A. augusta L and determine its anti-inflammatory activities. Extraction and fractionation have been done with graded maceration using solvents with different polarities, which are n-hexane, ethyl acetate, and methanol. The separation was performed by column chromatography, followed by preparative thin-layer chromatography. The characterization of the isolate was carried out using UV-Vis spectrophotometry and infrared spectrophotometry, GC-MS. The anti-inflammatory activities of methanol extract and isolate of A. augusta was performed in this study was designed to evaluate the dose-response relationship of the anti-inflammatory activity in rat models of chronic inflammation chromatography to obtain isolate 2.1.1 that characterize and showed maximum absorbance at 265. The result of IR showed the presence of functional groups, -C=C-H, -C=H, -CH, CH3, CH2, and –CO belongs to the steroid compound. The results of the GC-MS show that isolates contain squalene compounds with a value of m/z 410, Isolate and crude extract showed an anti-inflammatory activity that almost approached the positive control of sodium 4-chlorophenolate. It could be concluded that isolate and extract provide good anti-inflammatory activity, that promise for new drug candidate squalene-based A. augusta.

The review gives an insight into amaranth, a very promising crop for the whole world. This crop has a long history dating back to the days of the Aztecs and Incas, for whom it was one of the most important crops, along with corn. However, unlike the latter, amaranth lost its nutritional value after being introduced into Europe. Only in the 20th century, largely thanks to Nikolai Vavilov, amaranth attracted great interest as a food and feed crop. Amaranth is a plant unique in its nutritional properties. It is characterized by a high content of protein saturated with essential amino acids, especially lysine, and a large number of bioactive compounds, such as vitamin C, amaranthine, rutin, carotenoids, etc. Of particular value is grain oil saturated with various lipid compounds: squalene, vitamin E, phytosterols, and fatty acids. These lipid compounds have a number of important properties from the point of view of functional nutrition: as natural antioxidants they bind free radicals, normalize lipid metabolism, and help to decrease blood cholesterol levels. The review focuses on genes that determine the content of the main, valuable biochemical components: squalene, vitamin C, and lysine. The genetic pathways that control the biosynthesis of these components have been studied in detail in various model plant objects. The presence of the complete genomic sequence of Amaranthus hypochondriacus L. makes it possible to identify orthologs of key biosynthetic genes. At the moment, only few genes in amaranth have been identified, including the squalene synthase (SQS) gene, the VTC2 ascorbic acid synthesis gene, and the key genes for lysine synthesis – AK and DHDPS. The article discusses the prospects and trends of marker-assisted selection of this crop as well as the difficulties of its systematization and genotyping, which have to be overcome to successfully solve plant breeding problems.

A mixture experimental design resulted in water-ethanol mixture to replace toxic extractants used to date and enhance greenness of extraction procedure. Oleuropein (OLP) and tyrosol (TYR) contents, in leaves from five olive Egyptian cultivars (North Sinai – Egypt) at different times representing summer, winter, and spring, respectively, were determined by high-performance liquid chromatography (HPLC) attached with photo diode array detector (DAD). The results showed that the concentration of OLP and TYR in leaf samples collected in spring and winter was higher than in summer seasons in different varieties. Tyrosol (TYR) and squalene (SQL) contents in oil, from different six olive oil commercial products that are popularly used in El-Arish, were bought from El-Arish local market and were determined as well using another HPLC-DAD method. This work can help nutraceutical companies attempting to manufacture medical products from olive leaves selecting best cultivar and optimum collection season of olive leaves.

Physalis angulata is an annual herb which has tremendous medicinal uses. The antioxidant activity of the whole plant extract of Physalis angulata was investigated using DPPH radical scavenging acti...

Jacaranda ulei Bureau & K.Schum., Bignoniaceae, is an evergreen shrub used to treat skin diseases, rheumatism, urinary tract disorders, and dysentery. This study aimed to perform a morphoanatomical study of J. ulei and analyse the composition and chemical variability of the volatile oils from its leaves over 11 months. The identification of the volatile oil components from the leaves was performed by gas chromatography coupled with mass spectrometry (GC-MS). The major chemical components of the volatile oils from the leaves were γ-amorphene (1.3–27.1%), n-heneicosane (0.4–59.5%), ethyl hexadecanoate (0.4–8.1%), eicosane-3-phenyl (0.9–18.5%), squalene (March—66.4%, May—8.3%, August—17.6%, September—0.64%, and November—6.47%), and 9-octadecenamide (50.9% in the June sample). Two clusters were identified: one was characterized by the vegetative period and the other by the flowering period of the species, showing that the chemical compounds of the volatile oils varied according to phenological factors. The anatomical description contributed to the taxonomic identification of the species. This is the first report on the volatile oil obtained from J. ulei

The efficient bioproduction of squalene-type triterpenoids (STs) has attracted considerable attention due to their significant biological activities. In a previous study, we constructed a recombinant Saccharomyces cerevisiae capable of producing three STs; 4,8-dihydroxy-22,23-oxidosqualene (ST-1), 8-hydroxy-2,3;22,23-squalene dioxide (ST-2), and 2,3;22,23-squalene dioxide (ST-3). Here, we first evaluated the effects of these STs on the growth of human non-small cell lung cancer (NSCLC) cells, and found that ST-3 exhibited the greatest potency compared to the other two STs. To further enhance the bioproduction of ST-3, we adopted a tunable system to balance the expression of the Ganoderma lucidum cytochrome P450 gene CYP505D13 in S. cerevisiae, which significantly improved the ST-3 production titer. The most effective strain produced 78.61 mg/L of ST-3 after 62 h fermentation, which was 6.43 times higher than that of our previous study. The present study demonstrated that ST-3 effectively inhibits the proliferation of NSCLC cells, and provides insight into its efficient bioproduction.

Carica papaya Linn. is well known for its nutritional and therapeutic properties. Growing, successfully in Egypt environment, we analyzed content of essential minerals, aqueous ethanol extract of leaves for lipoid constituents and main alkaloidal component, examination of in-vivo potential as antioxidant and hepatoprotective, anticancer and antimicrobial activity. Leaves and fruits are good source of micro and macro essential minerals. Squalene, 2-hexadecen-1-ol, β- sitosterol, campesterol and palmitic acid were the major identified lipoid constituents. Carpaine, alkaloid was isolated and identified by different spectroscopic means. 80% Ethanol extract of leaves retained reduced glutathione which were highly compatible to those receiving vitamin E, it retained liver serum levels to which were compatible to the results of those receiving silymarin and demonstrated improvement in deleterious effect induced by CCl4. The extract had moderate effect, while carpaine showed high activity against ovary carcinoma. It had moderate antibacterial activity against Escherichia coli. This plant has highly economic importance.

Economically feasible photosynthetic cultivation of microalgal and cyanobacterial strains is crucial for the biological conversion of CO2 and potential CO2 mitigation to challenge global warming. To overcome the economic barriers, the production of value-added chemicals was desired by compensating for the overall processing cost. Here, we engineered cyanobacteria for photosynthetic squalene production and cultivated them in a scalable photobioreactor using industrial flue gas. First, an inducer-free gene expression system was developed for the cyanobacteria to lower production const. Then, the recombinant cyanobacteria were cultivated in a closed photobioreactor (100 L) using flue gas (5% CO2) as the sole carbon source under natural sunlight as the only energy source. Seasonal light intensities and temperatures were analyzed along with cyanobacterial cell growth and squalene production in August and October 2019. As a result, the effective irradiation hours were the most critical factor for the large-scale cultivation of cyanobacteria. Thus, an automated photobioprocess system will be developed based on the regional light sources.

This study was aimed at evaluating thein vitroeffect of the edible mushroom (EM)Pleurotus eryngiiagainst the eggs and larvae (L3) ofHaemonchus contortus. The evaluation included acetone (AE) and hydroalcoholic (HA) extracts of the following strains: ECS-1138, ECS-1156, ECS-1255, ECS-1258, ECS-1261, ECS-1282, and ECS-1292. The HA extract of the ECS-1255 strain showed the highest effect on mortality rates of L3 (18.83%) at 20 μg/mL. After subjecting this HA extract to a normal phase chromatography column, five fractions were obtained; fraction F5 (100% MeOH) was the most effective against eggs, with hatching inhibition percentages of 88.77 and 91.87% at 20 and 40 mg/mL, respectively. Gas chromatography-mass spectrometry (GC-MS) subjected this fraction to an acetylation reaction to determine the content of the secondary metabolites. The GC-MS analysis showed that the F5 fraction was composed of trehalose CAS: 6138-23-4, polyols (L-iditol CAS: 488-45-9, galactitol CAS: 608-66-2, D-mannitol CAS: 69-65-8, D-glucitol CAS: 50-70-4, and myoinositol CAS: 87-89-8), adipic acid CAS: 124-04-9, stearic acid CAS: 57-11-4, squalene CAS: 111-02-4, andβ-sitosterol CAS: 83-46-5.

Background Metabolomics of Camellia japonica leaf has been studied to identify the terpenoids present in it and their interrelations regarding biosynthesis as most of their pathways are closely situated. Camellia japonica is famous for its anti-inflammatory activity in the field of medicines and ethno-botany. In this research, we intended to study the metabolomics of Camellia japonica leaf by using gas chromatography-mass spectroscopy technique. Results A total of twenty-nine anti-inflammatory compounds, occupying 83.96% of total area, came out in the result. Most of the metabolites are terpenoids leading with triterpenoids like squalene, lupeol, and vitamin E. In this study, the candidate molecules responsible for anti-inflammatory activity were spotted out in the leaf extract and biosynthetic relation or interactions between those components were also established. Conclusion Finding novel anticancer and anti-inflammatory medicinal compounds like lupeol in a large amount in Camellia japonica leaf is the most remarkable outcome of this gas chromatography-mass spectroscopy analysis. Developing probable pathway for biosynthesis of methyl commate B is also noteworthy.

Transcriptome mining is a powerful tool for gene discovery in plant sciences. Biosynthetic enzyme transcripts of specialized metabolites have been isolated using transcriptome datasets of non-model plants; however, this methodology has not been applied to fern metabolites. Herein, we attempted to isolate transcripts of squalene cyclases from the Lygodium japonicum transcriptome database. A BLAST search revealed the presence of four isotigs (12872, 12873, 33018, and 33763) annotated as squalene cyclase, two of which contained full-length open-reading frame sequences. However, both isotigs failed to be amplified; our investigation revealed them to be chimeric sequences due to incorrect assembly. We also revealed that part of isotigs 12873 and 33018 constituted a single gene, designated LJH. Conversely, homology-based PCR resulted in the isolation of a squalene cyclase homolog, LjSCx, distinct from the four isotigs. Biochemical characterization of LJH and LjSCx revealed that LJH is a hopene synthase gene and LjSCx is a pseudogene. These findings highlight the usefulness and limitations of transcriptome datasets, especially with respect to genes comprising a gene family.

Tea oil obtained fromCamellia oleifera is a high-quality vegetable oil rich in squalene, a triterpenoid bioactive substance with several demonstrated health benefits. WRKY transcription factors in ...

BackgroundAlong with swift economic evolution and continuous amelioration of lifestyle, people at present are paying more attention to health issues. Synthetic drugs will be compensated with other natural ones that belong to natural origin. Plants have always been considered as sources of several compounds that are used in many fields, especially human and animal health, starting from boosting immunity to the treatment of infectious diseases caused by some pathogenic microbes such as bacteria, fungi as well as viruses. This study aimed to incorporate some types of plants within the antimicrobial portfolio through the examination of different six plants which wereCichorium intybus,Cinnamomum camphora,Commiphora myrrha,Foeniculum vulgare,Nerium oleander, andSpartium junceum. As well, attempting to identify the active constituents of their extracts using GC–MS.Materials and methodsAll selected plants were analyzed to determine their phytochemical composition such as phenolics, alkaloids, flavonoids, terpenoids, and so on. The extraction step was done by sophisticated equipment called supercritical fluid extractor SFE through adjustment of specific conditions include temperature, time, flow rate and pressure to change the behavior of CO2. Testing the antimicrobial activity of each plant extract via agar well diffusion method through the formation of clear zones against a wide range of test microorganisms including both Gram-positive and Gram-negative bacteria as well as yeasts. Finally, attempting to primarily identify the constituents of each plant extract using GC–MS.Results and discussionThe crude extract ofF. vulgareshowed the highest potency againstC. albicans,E. faecalisandS. typhimurium, it contains some unique compounds such as squalene, eugenol and isoeugenol while, Extract ofC. intybusshowed a moderate activity especially againstC. lipolyticaand MRSA and it includes Vitamin A like compound which indicates antioxidant property.ConclusionConclusively, fennel gave a promising result as a good wide spectrum antimicrobial agent because it contains some compounds act as antimicrobial agents such as eugenol which was used as food preservatives in addition to squalene which acts as an antioxidant and antimycotic agent so, it will be useful especially while it was used in highly purified form excluding all undesirable subcomponents.

The olive tree is a member of the Oleaceae family (Olea europaea). Olea europaea is commonly known as Zaitoon. The ancient Egyptians, Greeks, Romans and other Mediterranean nations cultivated olives for its edible fruits and to obtain oil from them. Leaves of the tree became important when olive leaf extract was reported to be potent in reducing incidence of degenerative diseases, particularly coronary heart disease (CHD) and cancers of the breast, skin, and colon. Olive leaves have the highest antioxidant power among the different parts of the olive tree. While in olive fruit, phenols (e.g. TYR) and terpenoid hydrocarbon (squalene) are the major components found in the oil. Much research has been carried out on the medical applications of olive. The choice of the plant was based on the good previous biological study of Olea europaea. Interest to choose this plant, may have been due to the widespread use of olive species medicinally, food industry and cosmetics.

Poverty and lack of access to health facilities had necessitated the less privileged to depend on herbal remedies to treat a number of diseases including malaria. The successes recorded from the discovery of novel antimalarials from plant sources had paved the way for the search of antimalarial compounds from traditional pharmacopeia. Phytochemical analysis as well as GC-MS analysis of crude leaves extract of Senna siamea was conducted, with a view to identifying compounds with biological activities. Further antimalarial assessment of the crude extracts was also performed in an in vitro assay. The phytochemicals; phenols, tannins, anthraquinones, alkaloid and flavonoids were detected from hexane and methanol extracts. GC-MS analysis revealed the presence of Saponins specifically, triterpenoids (Lupeol, α-amyrin), Sesquiterpene (Octadecane), Diterpenes (Eicosane), Esters of pthalic acids (Diethyl phthalate) Squalene, α-Tocopherol (a fat soluble Vitamin E) and hexadecanoic acid butyl ester. A dose dependent suppression of parasite growth was observed for all extracts, with methanolic extract showing less antimalarial potency (IC50= 3.74 μg/mL) when compared to the hexane extract (IC50=4.349 μg/mL). Extracts and compounds detected from the leaves of this plant could be used as novel lead compounds to develop new drugs. Keywords: Phytochemical analysis; GC-MS; antimalarial; Senna siamea; medicinal plants.

Plant extracts prompting free radical formation causing DNA damage among especially symbiotic microorganisms of Aedes aegypti has not been extensively examined in previous studies. Here, investigated whether: (1) it was possible to reduce the vectorial capacity by eliminating the microbial consortia of Aedes aegypti larvae, (2) the excess formation of free radical could induce damages of genomic DNA and alter the morphological appearance of Aedes aegypti larvae, and (3) which are the probable mechanisms driving the death of mosquitoes that has been treated with phytochemicals. The toxicity of Catunaregam spinosa seed extracts on the microbial community of Aedes aegypti larvae was determined as previously described. The formation of free radicals was confirmed using the ammonium molybdate method. While the GCMS method was used to assess the phytochemical analysis and the DNA lysing potential. The petroleum ether, ethanol, acetone and water extracts of Catunaregam spinosa has exhibited potent toxicity towards Aedes aegypti larvea’s gut bacterial microbiota, including Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumonia, and Proteus mirabilis with strong MIC values ranging from 0.07 mg/ml to 5.9 mg/ml. Plant extract induced free radical formations and oxidative damage to DNA. The presence of the bioactive element squalene justifies the insecticidal proprieties of Catunaregam spinosa extract. This study reflects the probable mechanisms underlying the lethal effect of Catunaregam spinosa extract on Aedes aegypti, and it potential as a novel biological vector control approach.

Artocarpus heterophyllus (jackfruit) leaves (JL) are a waste product that is commonly used as livestock feed. Jackfruit leaves have been revealed to possess many medicinal values such as antioxidant and anti-inflammatory properties. In this study, different drying treatments (shade (SD), sun (SN), and oven (OV)) and ethanol/water ratios (E/W) were investigated to evaluate the impact on drying kinetics, color, and antioxidant properties of jackfruit leaves. Results showed that the Newton model was the best fitted mathematical model for the JL drying kinetics. The moisture effective diffusivities ranged from 2.920 × 10-10 to 6.814 × 10-10 m2/s over the temperature range studied. Shade drying was able to preserve the green pigment better than OV and SN drying treatments. Treatment with ethanol/water ratio at 80% and oven-dried (OV80) revealed the highest phenolic content (195.05 ± 1.21 mg gallic acid equivalent (GAE)/g extract weight (EW)), flavonoid content (11.02 ± 0.17 mg artocarpin equivalent (AE)/g EW), and antioxidant activities (90% scavenging activity and reducing power of 1043.84 ± 5.28 µM trolox equivalent (TE)/g EW) compared to SD and SN treatments. The OV80 also possessed the highest artocarpin, squalene, and β-sitosterol contents determined. The OV80 was selected for improving antioxidant and colour stability, and has the potential to be developed into functional biopolymer production.

The present study investigated the phytochemical constituents of Cnidoscolus aconitifolius leaves extracts and its anti-hypercholesterolemia potentials using standard analytical methods. Forty five male albino rats weighing (115-121 g), divided into nine groups of five rats were used. Group I served as the control while the other groups were administered 200, 400, 600 and 800 mg/kg body weight of aqueous and ethanol leaf extracts. GC-MS analysis showed 3,7,1,5-tetramethyl-2-hexadecen-1-ol, farnesyl bromide, β–sitosterol, squalene, β-amyrin, 1-heptatriacotanol, hexadecanoic acid, methyl ester, 2-pentadecanone, 6,10,14-trimethyl- ,n-hexadecanoic acid, 9,12-octadecadienoyl chloride, (Z,Z, δ- tocopherol, Ergosta-5,22-dien-3-ol acetate, (3β,22E)-, 9,10-secocholesta-5,7,10(19)-triene-3,24,25-triol, (3β,5Z,7E)-acetamide, N-methyl-N-[4-(3-hydroxypyrrolidinyl)-2-butynyl]-, 1-gala-I-ido-octose, 10-methyl-E-11-tridecen-1-ol propionate, dodecanoic acid, 2-(acetyloxy)-1-[(acetyloxy)methyl]ethyl ester, 11,14-octadecadienoic acid, methyl ester, cyclopentaneundecanoic acid and methyl ester. Lipid profile showed significant reduction in TC, LDL and TG with increase in HDL in dose dependent ratio. This shows that extracts of this plant could be useful in treatment of coronary heart diseases. Key words: Phytochemicals,hypercholesterolemia, Cnidoscolus aconitifolius, potentials.

Squalene synthase (SQS) catalyzes the conversion of two farnesyl pyrophosphates to squalene, an important intermediate in between isoprene and valuable triterpenoids. In this study, we have constructed a novel biosynthesis pathway for squalene in Bacillus subtilis and performed metabolic engineering aiming at facilitating further exploitation and production of squalene-derived triterpenoids. Therefore, systematic studies and analysis were performed including selection of multiple SQS candidates from various organisms, comparison of expression vectors, optimization of cultivation temperatures, and examination of rate-limiting factors within the synthetic pathway. We were, for the first time, able to obtain squalene synthesis in B. subtilis. Furthermore, we achieved a 29-fold increase of squalene yield (0.26-7.5 mg/L) by expressing SQS from Bacillus megaterium and eliminating bottlenecks within the upstream methylerythritol-phosphate pathway. Moreover, our findings showed that also ispA could positively affect the production of squalene.

Therapeutic properties of the medicinal plant are due to the presence of phytochemical constituents. The phytochemical constituents of Plumeria alba flower were investigated by phytochemical screening assays and gas chromatography-mass spectrometry (GC-MS). The phytochemical screening of hexane, dichloromethane, ethyl acetate, butanol, and aqueous extracts of P. alba flower showed it contains a various concentration of saponins, flavonoids, tannins, steroids, volatile oil and phenolic compounds. Several major chemical constituents that were identified is squalene, bis(2-ethylhexyl) phthalate, methyl (methyl 4-O-methyl-α-d-mannopyranoside) uronate and tricyclo [7.2.0.0(2,6)] undecan-5-ol, 2,6,10,10-tetramethyl- (isomer 2) by using GC-MS technique.

Squalene has wide applications in the food and pharmaceutical industries. Engineering microbes to produce squalene is a promising alternative for traditional production approaches. In this study, squalene production was enhanced to 978.24 mg/L through stepwise overexpression of the enzymes that catalyze acetyl-CoA to squalene. Subsequently, to increase the activity of HMG-CoA reductase and alleviate the high dependence on NADPH, the HMG-CoA reductase (NADH-HMGR) from Silicibacter pomeroyi, highly specific for NADH, was introduced, which increased squalene production to 1086.31 mg/L. Native ethanol dehydrogenase ADH2 and acetaldehyde dehydrogenase ADA from Dickeya zeae were further overexpressed, which enhanced the capability to utilize ethanol for squalene synthesis and endowed the engineered strain with greater adaptability to high ethanol concentrations. Finally, a remarkable squalene production of 9472 mg/L was obtained from ethanol via carbon source-controlled fed-batch fermentation. This study will greatly accelerate the process of developing microbial cell factories for squalene production.

In this paper, a mixture of methanol, ethanol, benzene and ethanol were used to extract the leaves of Albizia kalkora (Roxb.) Prain. Then, using Fourier transform infrared (FT-IR) spectroscopy, gas chromatography-mass spectrometry (GC-MS), pyrolysis as chromatography - mass spectrometry, and thermogravimetric analysis (TGA) and other advanced instruments Albizia kalkoraleaves were analyzed to determine their composition. According to the test data, Albizia kalkoraleaves contain epinephrine, trans squalene, stachydrine and other chemical components with high medicinal value, indicating that Albizia kalkoraleaves have potential applications in the pharmaceutical industry. This study has expanded the utilization value of Albizia kalkoratrees and provided technical support for the resource utilization of Albizia kalkoratrees.

Made available in DSpace on 2020-12-12T01:54:11Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-28 Scheelea phalerata Mart. ex Spreng (Arecaceae) is a palm tree found in the Brazilian cerrado. There are no topics related to volatile oils from S. phalerata leaves in the literature. This work determines its chemical composition and evaluates the biological activity under two different seasonal conditions (dry and rainy seasons). The dry essential oil yield was 0.034 ± 0.001% and the rainy essential oil yield was 0.011 ± 0.003%. Both essential oils presented different qualitative and quantitative compositions (99.4 and 98.5%). The main constituents of the dry essential oil were phytol (36.7%), nonadecane (9.7%), linolenic acid (9.1%), (Z)-hex-3-en-1-ol (4.2%), and squalene (4.0%). The main constituents of the rainy essential oil were phytol (26.1%), palmitic acid (18.7%), hexan-1-ol (15.6%), (Z)-hex-3-en-1-ol (9.7%), and oleic acid (4.0%). The antileishmanial activity against promastigotes of Leishmania amazonensis was observed only for the rainy season essential oil (IC50 value of 165.05 ± 33.26 μg mL-1). A molecular docking study showed that alcohols exert a paramount efficacy and that the action of some essential oil compounds may be similar to that of amphotericin B. Still, only the essential oil from the dry season showed moderate antibacterial activity against S. sanguinis (MICs 200-400 μg mL-1). The cytotoxicity against Vero cells was identical (>512) for both essential oils. The novel data here for both chemical characterization and biological activity, in particular, evidence that the action of these compounds is similar to that of amphotericin B, provide valuable information to the drug-discovery field. Nucleus of Research in Natural Products (NuPPeN) Institute of Chemistry Federal University of Uberlândia Campus Santa Mônica, Av. João Naves de ávila, 2121 Institute of Biosciences of Botucatu Department of Microbiology and Immunology Unesp-São Paulo State University, 250 Distrito de Rubião Junior Institute of Biosciences of Botucatu Department of Physics and Biophysics Unesp-São Paulo State University, 250 Distrito de Rubião Junior Nucleus of Research in Sciences and Technology Laboratory of Research in Applied Microbiology (LaPeMA) University of Franca, 201 Parque Universitário Institute of Biomedical Sciences Laboratory of Trypanosomatids Federal University of Uberlândia Campus Umuarama, Av. Pará 1720 Bloco 2B Nucleus of Bioprospecting in Natural Products (NuBiProN) Chemistry Department Federal Institute of the Triângulo Mineiro, 4000 Distrito Industrial I Institute of Biosciences of Botucatu Department of Microbiology and Immunology Unesp-São Paulo State University, 250 Distrito de Rubião Junior Institute of Biosciences of Botucatu Department of Physics and Biophysics Unesp-São Paulo State University, 250 Distrito de Rubião Junior

Taxodisones A and B, C30-terpenes with an unprecedented tetracyclo[12.4.0.0.2,709,14]octodecane core, were isolated from the seeds of Taxodium distichum. Their structures, including their configurations, were unambiguously determined. Their biomimetic synthesis suggests that they stem from diterpenes and monoterpenes, and not from squalene or oxidosqualene. In addition, their bioactivities were also evaluated.

Basella rubra (Malabar spinach) is a commonly consumed green leafy vegetable in southern parts of India. The chemical composition, nutraceuticals characterization, squalene Nuclear Magnetic Resonance (NMR), in vitro antioxidant activities and cytotoxicity of B. rubra seed oil (33.08%) was investigated. Gas chromatography-mass spectrometry (GC/MS) analysis revealed the presence of palmitic (27.21 μmol%), oleic (33.83 μmol%) and linoleic acid (26.02 μmol%) with a total of 64.38 μmol% unsaturated fatty acids respectively. HPLC nutraceutical characterization showed a major constituent of gallic acid (11.23 mg%), γ-tocopherols (17.74 mg%), cycloartenylferulate (1.7 mg%), and squalene (1 g%). Squalene was further recovered (98%), purified (99.9%), and confirmed through 1H and 13C NMR. The in vitro antioxidant activities recorded by using 2,2-diphenyl-1-picrylhydrazyl (EC50 = 6 mg mL−1), ferric reducing antioxidant power (361.85 mM of Trolox Eq./100 g) and 2,2′-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (EC50 = 56.19 mg mL−1) scavenging activity. In vitro cytotoxicity assessed on 3T3-L1 showed good cell survival without any toxicity (upto 400 μg mL−1). B. rubra seed oil has proven nutraceuticals and antioxidant potentials with least toxicity which can be recommended for functional foods applications.

Background: Periodontitis is an infection in tooth-supporting tissues caused by a specific microorganism, Porphyromonas Gingivalis (PG), which can trigger collagen destruction. Generally, periodontal therapy employs a combination of mechanical (scaling root planning/SRP) and chemical (antibiotics) remedies, the latter of which can cause bacterial resistance. On the other hand, shark liver oil contains active natural ingredients such as alkylglycerols, squalene, squalamine, and omega-3, which have antibacterial and antioxidant effects. Purpose: This study aims to determine the impact of shark liver oil on fibroblasts and collagen density in the periodontal ligament of Wistar rats induced with PG. Methods: This study represents a laboratory experiment with post-test only control group design. The research subjects consisted of 35 Wistar rats divided into five groups, namely; a negative control group (K-); a positive control group with PG induction (K+); and three treatment groups induced with PG and shark liver oil once a day for seven days at varying doses of 0.2 g/gBB (P1), 0.3 g/gBB (P2), and 0.4 g/gBB (P3). Following treatment, the subjects were euthanized. The number of fibroblasts was then histologically examined with Hematoxylin Eosin (HE). Meanwhile, the collagen density was histologically analyzed with Masson’s Tricrome. Fibroblast cells were observed through a microscope at 400x magnification. Data was statistically analyzed with a one-way ANOVA and post hoc LSD. Collagen Density scoring was then performed. The results were analyzed with a non-parametric Kruskal-Wallis test (p=0.05), and subsequently with a Mann-Whitney U test (p

We cloned two squalene epoxidases and five oxidosqualene cyclases, and identified their function using CRISPR/Cas9 tool and yeast heterologous expression. Triterpenes are the main active ingredients of Tripterygium wilfordii Hook.f., a traditional Chinese medicinal plant with many encouraging preclinical applications. However, the biosynthetic pathways of triterpenes in this plant are poorly understood. Here, we report on the isolation and identification of two squalene epoxidases (SQE6 and SQE7) and five oxidosqualene cyclases (OSC4-8) from T. wilfordii. Yeast complementation assays showed that TwSQE6 and TwSQE7 can functionally complement an erg1 yeast mutant that was constructed using the CRISPR/Cas9 system. The putative OSC genes were functionally characterized by heterologous expression in yeast. GC/MS analysis of the fermentation products of the transgenic yeast showed that both TwOSC4 and TwOSC6 are cycloartenol synthases, while TwOSC8 is a β-amyrin synthase. The discovery of these genes expands our knowledge of key enzymes in triterpenoid biosynthesis, and provides additional target genes for increasing the production of triterpenes in T. wilfordii tissue cultures by disrupting competing pathways, or in chassis cells by reconstituting the triterpenoid biosynthetic pathway.