Chemical profiling of bioactive compounds in the methanolic extract of wild leaf and callus of Vitex negundo using gas chromatographymass spectrometry

BACKGROUND The investigation of plant-based therapeutic agents in medicinal plants has revealed their presence in the extracts and provides the vision to formulate novel techniques for drug therapy.Vitex negundo(V.negundo),a perennial herb belonging to the Varbanaceae family,is extensively used in conventional medication.AIM To determine the existence of therapeutic components in leaf and callus extracts from wild V.negundo plants using gas chromatography-mass spectrometry(GCMS).METHODS In this study,we conducted GC-MS on wild plant leaf extracts and correlated the presence of constituents with those in callus extracts.Various growth regulators such as 6-benzylaminopurine(BAP),2,4-dichlorophenoxyacetic acid(2,4-D),α-naphthylacetic acid(NAA),and di-phenylurea(DPU)were added to plant leaves and in-vitro callus and grown on MS medium.RESULTS The results clearly indicated that the addition of BAP(2.0 mg/L),2,4-D(0.2 mg/mL),DPU(2.0 mg/L)and 2,4-D(0.2 mg/mL)in MS medium resulted in rapid callus development.The plant profile of Vitex extracts by GC-MS analysis showed that 24,10,and 14 bioactive constituents were detected in the methanolic extract of leaf,green callus and the methanolic extract of white loose callus,respectively.CONCLUSION Octadecadienoic acid,hexadecanoic acid and methyl ester were the major constituents in the leaf and callus methanolic extract.Octadecadienoic acid was the most common constituent in all samples.The maximum concentration of octadecadienoic acid in leaves,green callus and white loose callus was 21.93%,47.79%and 40.38%,respectively.These findings demonstrate that the concentration of octadecadienoic acid doubles in-vitro compared to in-vivo.In addition to octadecadienoic acid;butyric acid,benzene,1-methoxy-4-(1-propenyl),dospan,tridecanedialdehyde,methylcyclohexenylbutanol,chlorpyrifos,n-secondary terpene diester,anflunine and other important active compounds were also detected.All these components were only available in callus formed in-vitro.This study showed that the callus contained additional botanical characteristics compared with wild plants.Due to the presence of numerous bioactive compounds,the medical use of Vitex for various diseases has been accepted and the plant is considered an important source of therapeutics for research and development.

Production and Purification of Cutinase from <i>Fusarium oxysporum</i>Using Modified Growth Media and a Specific Cutinase Substrate

Cutinases are hydrolytic enzymes used by phytopathogenic fungi to gain entry into plants by breaking down the cuticular barrier of higher plants. Cutinase displayed hydrolytic activity not only towards cutin, the main component of the plant cuticle, but also towards a variety of soluble synthetic esters, insoluble triglycerides and polyesters. Therefore, cutinase was evaluated for use in the chemical, food and cotton bio-scouring industry and for synthetic fibers modification. This research examined the production and purification of extracellular cutinase from the phytopathogenic fungus Fusarium oxysporum. The addition of apple cutin or its hydrolysate to the fungus growth medium resulted in an enhanced secretion of cutinase into the extracellular fluid. Testing 1-hexadecanol as an alternative to natural cutin to induce cutinase production resulted in a high process yield under modified growth conditions. Cutinase enzyme production was followed by an optimized purification method for enzyme preparation using high-performance liquid chromatography and high-specificity 4-nitrophenyl (16-methyl sulfide ester) hexadecanoate (pNMSEH) cutinase substrate. Electrophoresis sodiumdodecyl sulfate-polyacrylamide and isoelectric focusing gels enabled the final separation and identification of the protein. The purified cutinase had an approximate molecular weight of 20 kDa and an isoelectric point of 4.7. The method presented here could be modified and used for cutinase production and purification in other microorganisms that exhibit cutinolytic activity.

Hexadecanoic acid-enriched extract of Halymenia durvillei induces apoptotic and autophagic death of human triple-negative breast cancer cells by upregulating ER stress

Objective:To investigate the effect of the hexane solvent fraction of Halymenia durvillei(HDHE)on triple-negative breast cancer.Methods:The phytochemical profile of HDHE was investigated by GC-MS.The cytotoxicity of HDHE against MDA-MB-231 cells was determined.The apoptotic and autophagic effects of HDHE were analyzed.The expression of molecular markers controlling apoptosis,autophagy,DNA damage,and endoplasmic reticulum(ER)stress was determined.Results:HDHE contains a mixture of fatty acids,mainly hexadecanoic acid.HDHE at a cytotoxic concentration induced apoptotic death of MDA-MB-231 cells through mitochondrial membrane dysfunction,and induction of apoptosis markers,and increased the expression of proteins related to DNA damage response.HDHE also induced the expression of LC-3,a marker of autophagic cell death at a cytotoxic concentration.Moreover,HDHE modulated the expression of ER stress genes.Conclusions:The hexadecanoic acid-enriched extract of Halymenia durvillei promotes apoptosis and autophagy of human triple-negative breast cancer cells.This extract may be further explored as an anticancer agent for triple-negative breast cancer.

Chemical profiling of Elaeocarpus serratus L.by GC-MS

Objective:To determine the possible bioactive components of the ethanolic extract of leaves of Elaeocarpus serratus(E.serratus).Methods:The present research was carried out by using GC-MS analysis,while mass spectra of the compounds found in the extract was matched with the National Institute of Standards and Technology and Wiley library.Results:Thirty components from leaves of the above said plant were identified.The active principles with their retention time,molecular formula,molecular weight and concentration(%)in the ethanol extracts of leaf of E.serratus are obtained.Conclusions:This is the first report of documentation of active constituents from leaves of E.serratus.The research reveals the potential of E.serratus leaves as a good source of bioactive compounds such as fatty acid esters,alcohols,hydrocarbons,aldehydes,alkenes,fatty acids and amides that justify the use of this plant for its various ailments by traditional practitioners.