{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001839","ANALYSIS_ID":"AN002982","VERSION":"1","CREATED_ON":"June 14, 2021, 9:00 am"},

"PROJECT":{"PROJECT_TYPE":"Mass spectrometric analyses of natural products","PROJECT_SUMMARY":"With a continuous threat of antimicrobial resistance on human health worldwide, efforts for new alternatives are ongoing for the management of bacterial infectious diseases. Natural products of land and sea, being conceived to be having fewer side effects, pose themselves as a welcome relief. In this respect, we have taken a scaffolded approach to unearthing the almost unexplored chemical constituents of Malaysian red seaweed, Gracilaria edulis. Essentially, a preliminary evaluation of the ethyl acetate and acetone solvent extracts, among a series of six such, revealed potential antibacterial activity against six MDR species namely, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica, methicillin-resistant Staphylococcus aureus (MRSA), Streptococcus pyogenes, and Bacillus subtilis. Detailed analyses of the inlying chemical constituents, through LC-MS and GC-MS chromatographic separation, revealed a library of metabolic compounds. These were led for further virtual screening against selected key role playing proteins in the virulence of the aforesaid bacteria. To this end, detailed predictive pharmacological analyses added up to reinforce Eplerenone as a natural alternative from the plethora of plausible bioactives. Our work adds to the ongoing effort to re-discover and repurpose biochemical compounds to combat the antimicrobial resistance offered by the Gram-positive and the -negative bacterial species.","INSTITUTE":"Sunway University","DEPARTMENT":"Biological Sciences, Sunway University, Selangor, Malaysia","LABORATORY":"Disease Complexity","FIRST_NAME":"Chandrajit","ADDRESS":"Sunway University, No. 5, Jalan Universiti, Bandar Sunway, Petaling Jaya 47500, Selangor, Malaysia","EMAIL":"chandrajitl@sunway.edu.my","PHONE":"+60 3 7491 8622","FUNDING_SOURCE":"None","PROJECT_COMMENTS":"Ongoing","PUBLICATIONS":"Metabolite profiling of Malaysian Gracilaria edulis reveals Eplerenone as novel antibacterial compound for drug repurposing against MDR Bacteria","PROJECT_TITLE":"Exploring the Antibacterial Potentials of South-East Asian Natural Products Against Multidrug Resistant Bacteria","LAST_NAME":"Lahiri","CONTRIBUTORS":"Ali Asghar, Syafiq Asnawi"},

"STUDY":{"STUDY_TITLE":"Metabolite profiling of Malaysian Gracilaria edulis reveals Eplerenone as novel antibacterial compound for drug repurposing against MDR Bacteria","STUDY_SUMMARY":"The current study re-defines a method to reveal bioactive compounds from the crude extracts of Malaysian red seaweed Gracilaria edulis, having promising antibacterial activities against selected bacterial species. Three species of Gram-positive and - negative characters were remarkably inhibited by the sequential and direct extracts of ethyl acetate and acetone. These were further separated through chromatographic methods to reveal a plethora of chemical constituents to be considered for a downstream virtual screening against selected crucial proteins of the six bacteria.","INSTITUTE":"Sunway University","DEPARTMENT":"Biological Sciences, Sunway University, Selangor, Malaysia","LABORATORY":"Disease Complexity","LAST_NAME":"Lahiri","FIRST_NAME":"Chandrajit","ADDRESS":"Sunway University, No. 5, Jalan Universiti, Bandar Sunway, Petaling Jaya 47500, Selangor, Malaysia","EMAIL":"chandrajitl@sunway.edu.my","STUDY_TYPE":"In vitro antibacterial studies","PHONE":"+60 3 7491 8622"},

"SUBJECT":{"SUBJECT_TYPE":"Other organism","SUBJECT_SPECIES":"Gracilaria edulis","TAXONOMY_ID":"172966","GENDER":"Not applicable","SPECIES_GROUP":"Rhodophyta"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"GE-LC-EA(s)",
"Factors":{"Solvent":"Ethyl acetate","Treatment":"Sequential"}
},
{
"Subject ID":"-",
"Sample ID":"GE-LC-AC(s)",
"Factors":{"Solvent":"Acetone","Treatment":"Sequential"}
},
{
"Subject ID":"-",
"Sample ID":"GE-GC-EA(s)",
"Factors":{"Solvent":"Ethyl acetate","Treatment":"Sequential"}
},
{
"Subject ID":"-",
"Sample ID":"GE-GC-AC(s)",
"Factors":{"Solvent":"Acetone","Treatment":"Sequential"}
},
{
"Subject ID":"-",
"Sample ID":"GE-GC-EA(d)",
"Factors":{"Solvent":"Ethyl acetate","Treatment":"Direct"}
},
{
"Subject ID":"-",
"Sample ID":"GE-GC-AC(d)",
"Factors":{"Solvent":"Acetone","Treatment":"Direct"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Healthy specimens of Gracilaria edulis were collected from Pantai Morib, Selangor, Malaysia. Further, for the correct identification of G. edulis, observation of the characters was carried out according to Guiry & Guiry (2021).","SAMPLE_TYPE":"Seaweed"},

"TREATMENT":{"TREATMENT_SUMMARY":"Extracts of Gracilaria edulis were prepared through two different approaches, namely, sequential and direct, following the procedure of Subermaniam et al. (2020). For the sequential process, the solvents were used in the order of increasing polarity viz. ethyl acetate < acetone. For the direct extracts, ethyl acetate and acetone were used separately."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Essentially, for both approaches, the seaweeds were rinsed sequentially with seawater followed by normal tap and then double distilled water to eradicate dirt and impurities. Clean samples were then dried using a freeze-dryer and later crushed into fine granule powder using an electric grinder. Different fractions of extracts were prepared using 10 grams of each powder to dissolve them in 100 mL of the mentioned solvents. All the prepared mixtures were made homogeneous using a rotating shaker (Yihder LM-530D, Shaker, Taiwan) for 24 hours and finally centrifuged (Eppendorf 5810 R Centrifuge, Germany) at 4000 rpm for 10 min at 4◦C to separate the supernatant. Each of the clear supernatants of the extracts was concentrated via a Rotary evaporator (Thermo Fisher Scientific EYELA N-1200A Rotary Evaporator, Tokyo). A further concentration using a vacuum concentrator (LaboGene, Brigachtal, Germany) was done to obtain a viscous liquid for storage at 4◦C and future experiments.","PROCESSING_STORAGE_CONDITIONS":"Room temperature","EXTRACT_STORAGE":"4℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"GC-MS","CHROMATOGRAPHY_TYPE":"GC","INSTRUMENT_NAME":"Agilent 7890B","COLUMN_NAME":"Agilent HP5-MS (30m x 0.25mm, 0.25 um)","FLOW_RATE":"1.5 mL per minute","METHODS_FILENAME":"GCMS_protocol.txt","ANALYTICAL_TIME":"64 minutes","OVEN_TEMPERATURE":"80°C"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent 6520 QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"Data processing Comments: NIST Mass Spectral Search Program-2009 version 2. Software/procedures used for feature assignments: Agilent Mass-Hunter Qualitative Analysis B.05.00."},

"MS_METABOLITE_DATA":{
"Units":"Area %",

"Data":[{"Metabolite":"[2,2’-Bifuran]-3-carboxylic acid, 5’-methyl-, methyl ester","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"7.7157","GE-GC-AC(d)":""},{"Metabolite":"1,2-Benzenediol bis(trimethylsilyl) ether","GE-GC-EA(s)":"2.052","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"1-Octadecyne","GE-GC-EA(s)":"2.2","GE-GC-AC(s)":"","GE-GC-EA(d)":"0.63897","GE-GC-AC(d)":""},{"Metabolite":"26-Nor-5-cholesten-3á-ol-25-one","GE-GC-EA(s)":"","GE-GC-AC(s)":"1.725","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"2-Heptyl-1,3-dioxolane","GE-GC-EA(s)":"","GE-GC-AC(s)":"2.882","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"2-Tridecanone","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"0.66248","GE-GC-AC(d)":""},{"Metabolite":"3,8,13,18-Tetraethyl-2,7,12,17-tetramethyl-7, 8-(diacetyl) methylene-7,8-dihydro-21H,23H-porphine copper (II)","GE-GC-EA(s)":"4.035","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"3-Methyl-1,2-diazirine","GE-GC-EA(s)":"7.529","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"4-Penten-2-one, 4-methyl-","GE-GC-EA(s)":"1.628","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"á Carotene","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"0.9908","GE-GC-AC(d)":""},{"Metabolite":"Arachidonic acid","GE-GC-EA(s)":"","GE-GC-AC(s)":"1.039","GE-GC-EA(d)":"1.4028","GE-GC-AC(d)":""},{"Metabolite":"Ar-tumerone","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":"1.7063"},{"Metabolite":"Cholest-5-en-3-ol","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":"3.4662"},{"Metabolite":"Cholesterol","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"32.831","GE-GC-AC(d)":""},{"Metabolite":"Decanoic acid, ethyl ester","GE-GC-EA(s)":"1.787","GE-GC-AC(s)":"","GE-GC-EA(d)":"5.3174","GE-GC-AC(d)":"2.2412"},{"Metabolite":"Diisooctyl phthalate","GE-GC-EA(s)":"0.914","GE-GC-AC(s)":"3.878","GE-GC-EA(d)":"0.56258","GE-GC-AC(d)":"0.85802"},{"Metabolite":"Hexadecanoic acid, ethyl ester","GE-GC-EA(s)":"","GE-GC-AC(s)":"2.074","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"Molybdenum, bis[(1,2,3,4,5-ü)-1,3-bis(1,1-dimethylethyl)-2,4-cyclopentadien-1-yl] di-æ-carbonyldicarbonyldi-, (mo-mo)","GE-GC-EA(s)":"0.646","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"n-Hexadecanoic acid","GE-GC-EA(s)":"5.008","GE-GC-AC(s)":"9.078","GE-GC-EA(d)":"9.1086","GE-GC-AC(d)":"1.2408"},{"Metabolite":"Nickel tetracarbonyl","GE-GC-EA(s)":"1.797","GE-GC-AC(s)":"1.329","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"Nonadecane","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"4.034","GE-GC-AC(d)":""},{"Metabolite":"Oxalic acid, cyclobutyl octadecyl ester","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"0.15812","GE-GC-AC(d)":""},{"Metabolite":"Phenol, 2,4-bis(1,1-dimethylethyl)","GE-GC-EA(s)":"3.925","GE-GC-AC(s)":"0.771","GE-GC-EA(d)":"7.7157","GE-GC-AC(d)":"4.6668"},{"Metabolite":"Phenol, 2-methoxy-3-(2-propenyl)-","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"6.3257","GE-GC-AC(d)":"8.9936"},{"Metabolite":"Phytol","GE-GC-EA(s)":"2.3","GE-GC-AC(s)":"","GE-GC-EA(d)":"","GE-GC-AC(d)":""},{"Metabolite":"Ribo-ribo disaccharide","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"0.59025","GE-GC-AC(d)":""},{"Metabolite":"Silane, tetramethyl","GE-GC-EA(s)":"","GE-GC-AC(s)":"","GE-GC-EA(d)":"0.18473","GE-GC-AC(d)":""}],

"Metabolites":[{"Metabolite":"[2,2’-Bifuran]-3-carboxylic acid, 5’-methyl-, methyl ester","Pubchem ID":"609852"},{"Metabolite":"1,2-Benzenediol bis(trimethylsilyl) ether","Pubchem ID":"23688911"},{"Metabolite":"1-Octadecyne","Pubchem ID":"69425"},{"Metabolite":"26-Nor-5-cholesten-3á-ol-25-one","Pubchem ID":"165617"},{"Metabolite":"2-Heptyl-1,3-dioxolane","Pubchem ID":"5363203"},{"Metabolite":"2-Tridecanone","Pubchem ID":"78067"},{"Metabolite":"3,8,13,18-Tetraethyl-2,7,12,17-tetramethyl-7, 8-(diacetyl) methylene-7,8-dihydro-21H,23H-porphine copper (II)","Pubchem ID":"11622"},{"Metabolite":"3-Methyl-1,2-diazirine","Pubchem ID":"6420208"},{"Metabolite":"4-Penten-2-one, 4-methyl-","Pubchem ID":"19543"},{"Metabolite":"á Carotene","Pubchem ID":"42608244"},{"Metabolite":"Arachidonic acid","Pubchem ID":"5280489"},{"Metabolite":"Ar-tumerone","Pubchem ID":"558221"},{"Metabolite":"Cholest-5-en-3-ol","Pubchem ID":"101316738"},{"Metabolite":"Cholesterol","Pubchem ID":"5997"},{"Metabolite":"Decanoic acid, ethyl ester","Pubchem ID":"12000424"},{"Metabolite":"Diisooctyl phthalate","Pubchem ID":"97750"},{"Metabolite":"Hexadecanoic acid, ethyl ester","Pubchem ID":"3081462"},{"Metabolite":"Molybdenum, bis[(1,2,3,4,5-ü)-1,3-bis(1,1-dimethylethyl)-2,4-cyclopentadien-1-yl] di-æ-carbonyldicarbonyldi-, (mo-mo)","Pubchem ID":"11986227"},{"Metabolite":"n-Hexadecanoic acid","Pubchem ID":"628589"},{"Metabolite":"Nickel tetracarbonyl","Pubchem ID":"518772"},{"Metabolite":"Nonadecane","Pubchem ID":"12401"},{"Metabolite":"Oxalic acid, cyclobutyl octadecyl ester","Pubchem ID":"6441116"},{"Metabolite":"Phenol, 2,4-bis(1,1-dimethylethyl)","Pubchem ID":"528937"},{"Metabolite":"Phenol, 2-methoxy-3-(2-propenyl)-","Pubchem ID":"596373"},{"Metabolite":"Phytol","Pubchem ID":"5280435"},{"Metabolite":"Ribo-ribo disaccharide","Pubchem ID":"161456"},{"Metabolite":"Silane, tetramethyl","Pubchem ID":"6396"}]
}

}