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Degradation of grease waste remains a challenging task. Current work deals with the biotransformation of grease waste into fatty acids under submerged fermentation using Penicillium chrysogenum SNP5 through media formulation and artificial neural network (ANN). Fermentation media was formulated to ameliorate the uptake of hydrocarbon by enhancing alkane hydroxylase (AlkB) activity, extracellular release of fatty acids and inhibiting beta-oxidation of fatty acid by regulating transketolase. Further, the process parameters of fermentation were optimized through Artificial Neural Network (ANN) using three critical variables viz; inoculum size (spores/ml), pH, and incubation time (days) while media engineering was done with the optimal supplementation of various medium components such as glucose, YPD, MnSO4, tetrahydrobiopterin (THB) and phloretin. The maximum conversion of 66.5% of grease waste into fatty acid was achieved at optimum conditions: inoculums size 3.36 × 107 spores/ml, incubation time 11.5 days, pH 7.2 along with formulated media composed of 1% grease in czapek-dox medium supplemented with 55.5 mM glucose, 0.5% YPD, 16.6 mM hexadecane, 1 mM MnSO4, 1 mM THB, and 1 mM phloretin. The presence of long-chain fatty acids in purified extracts such as oleic acid and octadecanoic acid as end products has valued the evolved process as another source of alternative fuel.

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Octadecanoic acid-3,4-tetrahydrofuran diester is a compound with acaricidal activity isolated and extracted from neem oil. In this study, a series of derivatives were obtained by structural modification of octadecanoic acid-3,4-tetrahydrofuran diester. The acaricidal activity of these derivatives indicated that introduction of benzyloxy substitution at the 2-position of the furan ring and the formation of a benzoate at the 3,4-position of the furan ring (benzoic acid-2-benzyloxy-3,4-tetrahydrofuran diester) could enhance the acaricidal activity. At concentration of 20, 10, and 5 mg/ml, the median lethal time (LT50) values of benzoic acid-2-benzyloxy-3,4-tetrahydrofuran diester were 16.138, 47.274, and 108.122 min, respectively. The LC50 value of benzoic acid-2-benzyloxy-3,4-tetrahydrofuran diester at 60 min was 5.342 mg/ml. Transmission electron microscopy showed that after treatment with benzoic acid-2-benzyloxy-3,4-tetrahydrofuran diester, the body structure of mites was destroyed; dermal organelles were dissolved; nuclear chromatin was ablated. Further, transcriptome sequencing analysis was used to get insight into the acaricidal mechanism of benzoic acid-2-benzyloxy-3,4-tetrahydrofuran diester. The results showed that its acaricidal mechanism is related to interfering "energy metabolism" in S. scabiei, including processes such as citric acid cycle, oxidative phosphorylation pathway and fatty acid metabolism. Additionally, through the activity detection of the mitochondrial complexes of S. scabiei, it was further verified that the acaricidal mechanism of benzoic acid-2-benzyloxy-3,4-tetrahydrofuran diester was related to the energy metabolism system of S. scabiei.

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cis-9,10-Epoxy-octadecanoic acid (oleic acid epoxide, OAE) and 5α,6α-epoxy-cholesterol (ChE) are endogenous epoxides. Unlike other epoxides, the oxirane groups of OAE and ChE are relatively stable against nucleophiles. OAE lacks toxicity and mutagenicity, while ChE is considered harmful, mutagenic and cancerogenic to animals. In humans, ChE is associated with cancer. The metabolism of OAE and ChE includes hydrolysis by cytosolic and microsomal hydrolases to their diols and glutathione (GSH) conjugation by GSH S-transferases (GST) to form the GSH conjugates (R-SG; R, residue). The GST-catalyzed GSH conjugation of OAE and ChE is poorly investigated. This article reports on the GSH conjugation of OAE, its methyl ester (OAEMe) and of ChE by rat liver homogenate GST. The GSH conjugates of OAE, OAEMe and ChE, i.e., OAE-SG, OAEMe-SG and ChE-SG, respectively, were determined by pre-column derivatization with o-phthaldialdehyde (OPA)/2-mercaptoethanol, high-performance liquid chromatography (HPLC) and fluorescence detection. Complex biphasic kinetics were observed with substrate inhibition of GST activity by OAE, OAEMe and ChE, an optimum pH of about 8.3 for OAE, and no measurable chemical GSH conjugation, underlying the importance of GST for the biotransformation of these epoxides. The results confirm the substrate concentration-dependent kinetic mechanism of GST isoforms first reported by William B. Jakoby (J. Biol. Chem. 1974) for exogenous electrophiles including the epoxide 1,2-epoxy-3-(p-nitrophenoxy)propane and the organic nitrates. This mechanism allows for maximal GST activity that can be achieved under given concentrations of GSH, epoxides and other electrophiles.

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Brazil nut oil is mostly composed of unsaturated fatty acids, some of which are associated with decreased incidence of cardiovascular diseases. Vegetable proteins have been increasingly used as wall material for partial replacement of carbohydrates and whey proteins. In order to create an oil preservation method, Brazil nut oil was encapsulated with three different types of vegetable protein concentrates and gum arabic (GA): rice (RPC + GA); pea (PPC + GA); and soy (SPC + GA) .For this purpose, vegetable protein concentrates were characterized, and after the drying process the physicochemical characteristics of the microparticles were evaluated. The most stable emulsion, after seven days of evaluation, was composed of RPC + GA. RPC + GA. This treatment was also more stable based on the shelf life assessments. We concluded that RCP microparticles were the best option for encapsulating Brazil nut oil in comparison with the other particles evaluated. In addition, the product obtained is potentially capable of being included in various processed foods.

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The present manuscript has focused on the heavy metal; accumulation potential by common native plants i.e. Chenopodium album L., Ricinus communis, Ranunculus sceleratus, and Rumex dentatus growing on the disposed of pulp and paper mill effluent sludge. The sludge showed the abundance of benzene propanoic acid tert- butyldimethylsilyl ester, Octadecanoic acid, TMS, Hexadecanoic acid, TMS, cinnamic acid-α-phenyl-TMS ester, ß-sitosterol TMS, 4-mercaptobenzoic acid as residual complex organic compounds along with heavy metals Fe (98.30 mg/L-1), Zn (51.00 mg/L-1), Cu (3.21 mg/L-1), Cd (9.11 mg/L-1), Mn (18.27 mg/L-1), Ni (5.21 mg/L-1), (Hg 0.014 mg/L-1) which were above the prescribed limit of environmental standard. The complexation of organic compounds with heavy metal restricts the bioavailability of metals to plants. But the metal analysis in various parts of the plant showed a significant amount of metal accumulation. Further, histological observations of root tissue through TEM showed apparent deposition of metal granules near the cell wall and vacuole as adoption features of plants. But the variable concentration of metal accumulation in different parts by various plants indicated the variable potential of tested plants with various metals. This also indicated their metal bio-availability and movement to plant tissue. Further, their bioconcentration factor (BCF) and translocation factor (TF) > 1.0 indicated the hyperaccumulation tendency of plants Mn was accumulated maximum in leaves C. album (69.38 mg/kg-1) followed by Cu (25.75 mg/kg -1), As (23.20 mg/kg -1), Fe (20.90 mg/kg -1) and Pb was maximum accumulated (22.41 mg/kg -1) in R. cummunis leaves. The result revealed that arsenic has been accumulated in higher amount root, shoot and leaves of all tested plants. The metal accumulator plants showed phytoremediation potential also by reducing various pollution parameters after growth on sludge. These potential plants may be used as biotechnological tools for the eco-restoration of polluted sites.

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Stranded driftwood feedstocks may represent, after pretreatment with steam explosion and enzymatic hydrolysis, a cheap C-source for producing biochemicals and biofuels using oleaginous yeasts. The hydrolysis was optimized using a response surface methodology (RSM). The solid loading (SL) and the dosage of enzyme cocktail (ED) were variated following a central composite design (CCD) aimed at optimizing the conversion of carbohydrates into lipids (YL) by the yeast Solicoccozyma terricola DBVPG 5870. A second-order polynomial equation was computed for describing the effect of ED and SL on YL. The best combination (ED = 3.10%; SL = 22.07%) for releasing the optimal concentration of carbohydrates which gave the highest predicted YL (27.32%) was then validated by a new hydrolysis. The resulting value of YL (25.26%) was close to the theoretical maximum value. Interestingly, fatty acid profile achieved under the optimized conditions was similar to that reported for palm oil.

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Recently, microalgae have attracted attention as sources of biomass energy. However, fatty acids from the microalgae are mainly unsaturated and show low stability in oxygenated environments, due to oxidation of the double bonds. The branched-chain fatty acid, 10-methyl stearic acid, is synthesized from oleic acid in certain bacteria; the fatty acid is saturated, but melting point is low. Thus, it is stable in the presence of oxygen and is highly fluid. We previously demonstrated that BfaA and BfaB in Mycobacterium chlorophenolicum are involved in the synthesis of 10-methyl stearic acid from oleic acid. In this study, as a consequence of the introduction of bfaA and bfaB into the cyanobacterium, Synechocystis sp. PCC 6803, we succeeded in producing 10-methyl stearic acid, with yields up to 4.1% of the total fatty acid content. The synthesis of 10-methyl stearic acid in Synechocystis cells did not show a significant effect on photosynthetic activity, but the growth of the cells was retarded at 34 °C. We observed that the synthesis of 10-methylene stearic acid, a precursor of 10-methyl stearic acid, had an inhibitory effect on the growth of the transformants, which was mitigated under microoxic conditions. Eventually, the amount of 10-methyl stearic acid present in the sulfoquinovosyl diacylglycerol and phosphatidylglycerol of the transformants was remarkably higher than that in the monogalactosyldiacylglycerol and digalactosyldiacylglycerol. Overall, we successfully synthesized 10-methyl stearic acid in the phototroph, Synechocystis, demonstrating that it is possible to synthesize unique modified fatty acids via photosynthesis that are not naturally produced in photosynthetic organisms.

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Pre-treatment of stationary phase cells of Lactobacillus plantarum NCMIB 8826 with citric acid (pH 3 to 6) for a short period of time significantly improved subsequent cell survival in several highly acidic fruit juices namely cranberry (pH 2.7), pomegranate (pH 3.5), and lemon & lime juices (pH 2.8). Although the mechanism for this adaptation is still unclear, the analysis of the cellular fatty acid content of acid adapted cells and the reverse transcription polymerase chain reaction (RT-PCR) showed a significant increase (by ~1.7 fold) of the cellular cyclopropane fatty acid, cis-11,12-methylene octadecanoic acid (C19:0cyclow7c) and a significant upregulation (~12 fold) of cyclopropane synthase (cfa) were observed, respectively, during acid adaptation. It is likely that these changes led to a decrease in membrane fluidity and to lower membrane permeability, which prevents the cells from proton influx during storage in these low pH fruit juices.

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In living organisms, modified fatty acids are crucial for the functions of the cellular membranes and storage lipids where the fatty acids are esterified. Some bacteria produce a typical methyl-branched fatty acid, i.e., 10-methyl stearic acid (19:0Me10). The biosynthetic pathway of 19:0Me10 in vivo has not been demonstrated clearly yet. It had been speculated that 19:0Me10 is synthesized from oleic acid (18:1Δ9) by S-adenosyl-L-methionine-dependent methyltransfer and NADPH-dependent reduction via a methylenated intermediate, 10-methyelene octadecanoic acid. Although the recombinant methyltransferases UmaA and UfaA1 from Mycobacterium tuberculosis H37Rv synthesize 19:0Me10 from 18:1Δ9 and NADPH in vitro, these methyltransferases do not possess any domains functioning in the redox reaction. These findings may contradict the two-step biosynthetic pathway. We focused on novel S-adenosyl-L-methionine-dependent methyltransferases from Mycobacterium chlorophenolicum that are involved in 19:0Me10 synthesis and selected two candidate proteins, WP_048471942 and WP_048472121, by a comparative genomic analysis. However, the heterologous expression of these candidate genes in Escherichia coli cells did not produce 19:0Me10. We found that one of the candidate genes, WP_048472121, was collocated with another gene, WP_048472120, that encodes a protein containing a domain associated with flavin adenine dinucleotide-binding oxidoreductase activity. The co-expression of these proteins (hereafter called BfaA and BfaB, respectively) led to the biosynthesis of 19:0Me10 in E. coli cells via the methylenated intermediate.

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Marine Lyngbya has been proven as a potent anticancer agent by disrupting microfilament network. Lyngbya and its associated cyanobacterial compounds have been stressed for futuristic advancements in cancer research and have foreseen explicit advancements in the recent era. Moreover, compounds like lyngbyabellins, lyngbyastatins and other derived toxins are significantly studied. Therefore it is of interest to study the efficacies of Lyngbya fatty acid derivatives. Cytotoxicity and DNA laddering studies proves the efficiency and safety of marine Lyngbya. Caspase - 3 along with MMP2 and MMP9 affirms the anti-inflammatory properties. Molecular docking shows that Octadecanoic acid has strong binding affinity to MMP-2. The role of octadecanoic acid as a proapoptotic factor in emancipation of inflammation apart from inducing apoptosis is of interest to study in future.

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The project was intended to the phytochemical characterisation from the rudimentary methanolic extract of Chenopodium ambrosioides Linn., which escorts to the isolation of stigmasterol (1), ß-sitosterol (2), octadecanoic acid (3), scopoletin (4) and 1-piperoylpiperidine (5). Literature validates the medicinal authentication of these compounds extorted from other sources, while our previous findings regarding microbial activities of different solvent systems fractions are favouring the presence of medicinally important compounds in this species. Herein, however, we report these natural products for the first time from this species.

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Peroxisome proliferator activated receptor (PPAR) is a nuclear receptor that is one of the transcription factors regulating lipid and glucose metabolism. Fermented ginseng (FG) is a ginseng fermented by Lactobacillus paracasei A221 containing minor ginsenosides and metabolites of fermentation. DNA microarray analysis of rat liver treated with FG indicated that FG affects on lipid metabolism are mediated by PPAR-α. To identify a PPAR-α agonist in FG, PPAR-α transcription reporter assay-guided fractionation was performed. The fraction obtained from the MeOH extract of FG, which showed potent transcription activity of PPAR-α, was fractionated by silica gel column chromatography into 16 subfractions, and further separation and crystallization gave compound 1 together with four known constituents of ginseng, including 20(R)- and 20(S)-protopanaxadiol, and 20(R)- and 20(S)-ginsenoside Rh1. The structure of compound 1 was identified as 10-hydroxy-octadecanoic acid by (1)H- and (13)C-NMR spectra and by EI-MS analysis of the methyl ester of 1. Compound 1 demonstrated much higher transcription activity of PPAR-α than the other isolated compounds. In addition, compound 1 also showed 5.5-fold higher transcription activity of PPAR-γ than vehicle at the dose of 20 µg/mL. In the present study, we identified 10-hydroxy-octadecanoic acid as a dual PPAR-α/γ agonist in FG. Our study suggested that metabolites of fermentation, in addition to ginsenosides, contribute to the health benefits of FG.

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The aim of this study is the first time demonstration of cis-12 regio-selective linoleate double-bond hydratase. Hydroxylation of fatty acids, abundant feedstock in nature, is an emerging alternative route for many petroleum replaceable products thorough hydroxy fatty acids, carboxylic acids, and lactones. However, chemical route for selective hydroxylation is still quite challenging owing to low selectivity and many environmental concerns. Hydroxylation of fatty acids by hydroxy fatty acid forming enzymes is an important route for selective biocatalytic oxyfunctionalization of fatty acids. Therefore, novel fatty acid hydroxylation enzymes should be discovered. The two hydratase genes of Lactobacillus acidophilus were identified by genomic analysis, and the expressed two recombinant hydratases were identified as cis-9 and cis-12 double-bond selective linoleate hydratases by in vitro functional validation, including the identification of products and the determination of regio-selectivity, substrate specificity, and kinetic parameters. The two different linoleate hydratases were the involved enzymes in the 10,13-dihydroxyoctadecanoic acid biosynthesis. Linoleate 13-hydratase (LHT-13) selectively converted 10 mM linoleic acid to 13S-hydroxy-9(Z)-octadecenoic acid with high titer (8.1 mM) and yield (81%). Our study will expand knowledge for microbial fatty acid-hydroxylation enzymes and facilitate the designed production of the regio-selective hydroxy fatty acids for useful chemicals from polyunsaturated fatty acid feedstocks.

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AIM OF THE STUDY: Tetrastigma hemsleyanum (Sanyeqing) is traditionally used as a folk medicine for the treatment of cancer. However, the underlying mechanisms remain unclear. The purpose of this study was to investigate the possible mechanisms by which petroleum ether fraction (PEF) of Sanyeqing has anti-tumor activity on HeLa cells. METHODS: The chemical components of PEF were analyzed by gas chromatography-mass spectrometry. The cytotoxicity of PEF on HeLa cells was measured by MTT assay. Apoptosis was evaluated by phosphatidylserine translocation, mitochondrial membrane potential (Δψm) changes and the activation of caspase-3, caspase-8 and caspase-9. The levels in T-SOD, CAT, GSH-PX and MDA were measured. RESULTS: PEF of Sanyeqing inhibited the growth and induced apoptosis of HeLa cells in dose- and time-dependent manner. PEF triggered intrinsic apoptotic pathway indicated by the loss of mitochondrial membrane potential and the activation of caspase-9 and caspase-3. In addition, PEF activated extrinsic apoptotic pathway indicated by the activation of caspase-8. Furthermore, PEF decreased T-SOD, CAT, GSH-PX activities and increased MDA level. Chemical analysis revealed the presence of fatty acids and phytosterol in PEF. CONCLUSIONS: PEF of Tetrastigma hemsleyanum Diels et. Gilg (Sanyeqing) exhibits cytotoxic effects, triggers both extrinsic and intrinsic apoptotic pathways, and augments oxidative stress in cervical carcinoma HeLa cells. Sanyeqing has strong potential to be developed as an agent for the treatment of cervical cancer.

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Octadecanoic acid-3,4-tetrahydrofuran diester, isolated from neem (Azadirachta indica) oil, exhibited potent acaricidal activity against Sarcoptes scabiei var. cuniculi. In this paper, the acaricidal mechanism of octadecanoic acid-3,4-tetrahydrofuran diester against Sarcoptes scabiei var. cuniculi was evaluated based on pathologic histology and enzyme activities. The results showed that after compound treatment for 24h at a concentration of 20mg/mL, the lesions of mites were prominent under transmission electron microscopy. The lesions consisted of the lysis of dermis cell membranes and cell nuclear membranes, mitochondrial morphological abnormalities, the drop of spinal disorders, and mitochondrial vacuolization. The activity of superoxide dismutase (SOD), peroxidase (POD), glutathione-s-transferases (GSTs), and Ca(2+)-ATPase of mites significantly changed after treatment with octadecanoic acid-3,4-tetrahydrofuran diester compared with the control group. The activities of SOD, POD, and Ca(2+)-ATPase were significantly suppressed, whereas that of GSTs was activated. These results indicated that the mechanism of the acaricidal activity of octadecanoic acid-3,4-tetrahydrofuran diester was mainly achieved through interference with the energy metabolism of mites, thus resulting in insect death.

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