Two ml cultivation medium was pelleted, washed with ddH2O, and lyophilized. Lyophilization was carried out for 2 days at − 80 °C and 0.04 mbar (VaCo 5, Zirbus Technology, Germany). Fatty acid analysis was done according to the modified protocol of Griffiths et al. [69 (link)]. Glyceryl trinonadecanoate (C19:0-TAG, 0.2 mg/mL in GC grade toluol) was added prior to the reaction as an internal standard. The dried biomass was directly converted into fatty acid methyl esters (FAME) by simultaneous extraction and transesterification of yeast lipids using 0.5 M Sodium methoxide solution in GC grade methanol and hydrogen chloride-methanol solution (Sigma, Germany) the fatty acid profiles were measured by a GC-2025 Plus gas chromatograph (Shimadzu, Japan) according to Woortman et al. [70 ].
Sodium methoxide solution
Manufactured by Merck Group
Sourced in United States, Germany
Sodium methoxide solution is a chemical compound used in various laboratory applications. It is a colorless, clear liquid that serves as a base catalyst in organic synthesis reactions. The solution contains sodium methoxide dissolved in methanol, providing a convenient and readily available source of this reagent for researchers and chemists.
Automatically generated - may contain errors
Lab products found in correlation
Chloroform, by Merck Group (2 mentions)
Hexane, by Merck Group (2 mentions)
Hydrochloric acid (hcl), by Merck Group (2 mentions)
Sodium hydroxide (naoh), by Merck Group (2 mentions)
Hydrogen chloride methanol solution, by Merck Group (1 mentions)
Gc 2025 plus gas chromatograph, by Shimadzu (1 mentions)
Potassium sodium tartrate tetrahydrate, by Merck Group (1 mentions)
Anhydrous sodium sulfate, by Merck Group (1 mentions)
Iron iii sulfate hydrate, by Merck Group (1 mentions)
Acetone, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Sulfuric acid, by Merck Group (1 mentions)
Petroleum ether, by Merck Group (1 mentions)
Lead 2 acetate trihydrate, by Merck Group (1 mentions)
Copper sulfate pentahydrate, by Merck Group (1 mentions)
Methanol, by Avantor (1 mentions)
Lc ms grade ammonium formate, by Merck Group (1 mentions)
Methyl tert butyl ether (mtbe), by Merck Group (1 mentions)
7 protocols using sodium methoxide solution
1
Fatty Acid Profiling of Microbial Biomass
2
Analytical Chemistry Reagents Protocol
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Sulfuric acid (Sigma-Aldrich Co., St. Louis, MO, USA, 95–97%), hydrochloric acid, sodium hydroxide, potassium chloride, sodium sulfate anhydrous, sodium bicarbonate, potassium sodium tartrate tetrahydrate, copper sulfate pentahydrate, lead (II) acetate trihydrate and lead (II) acetate trihydrate, acetone, and chloroform (Sigma-Aldrich Co., St. Louis, MO, USA, p.a.). Iron (III) sulfate hydrate (Sigma-Aldrich Co., St. Louis, MO, USA, ASC reagent). Potassium permanganate standard solution (Sigma-Aldrich Co., St. Louis, MO, USA, 0.02002 mol/L). Boric acid (AFT, Bratislava, Slovakia, p.a.); Kjeltabs (Velf Scientifica srl, Usmate, Italy). Petroleum ether (Sigma-Aldrich Co., St. Louis, MO, USA, b.p. 40–60 °C). Methanol (Sigma-Aldrich Co., St. Louis, MO, USA, HPLC grade). Sodium methoxide solution (Sigma-Aldrich Co., St. Louis, MO, USA, 25 wt% in Methanol). Hexane (Sigma-Aldrich Co., St. Louis, MO, USA, for pesticide residue analysis).
3
Fatty Acid Profiling by LC-MS
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Methanol was purchased from VWR (Radnor, PA, USA), and pentane was obtained from Fluka Analytical (Honeywell International Inc., Charlotte, NC, USA). Hexane and sodium methoxide solution 25 wt.% were purchased from Sigma Aldrich (St. Louise, MO, USA). Sodium sulfate was purchased from Titolchimica (Rovigo, Italy, IT) and the CPFA cis-9,10-methylene-octadecanoic acid (dihydrosterculic acid DHSA, as methyl ester, purity ≥98%) was obtained from Chem Cruz (TE Huissen, The Netherlands, NL). LC-MS grade formic acid and LC-MS grade ammonium formate were purchased from Sigma Aldrich (Steinheim, Germany). LC-MS grade Methanol and LC-MS grade acetonitrile were purchased from Honeywell (Selze, Germany), and LC-MS grade 2-propanol and methyl tert-butyl ether (MTBE) were purchased from Merck KGaA (Darmstadt, Germany) and not purified further. If not otherwise stated, Milli-Q water was employed.
4
Lipid Extraction and Methyl Esterification
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Chemicals used in this study were: Yeast Extract (Biokar Diagnostics, France); Peptone (Biokar Diagnostics, France); Glucose (Sigma, Germany); Chlormaphenicol (Biomatik, USA); Ethyl Methane Sulfonate (Sigma, Germany); Sodium Thiosulfate (Sigma, Germany); Sodium Phosphate (Sigma, Germany); Sodium Chloride (Sigma, Germany); 2,6-Di-tert-butyl-4-methylphenol (BHT) (Sigma, Germany); Methanol (Carlo-Herba, France); Hydrochloric acid 37% (Carlo Herba, France); n-Hexane (VWR chemical, France); Dichloromethane (VWR chemical, France); 2,2-dimethoxypropane (Sigma, Germany); Hydrochloric acid solution (Sigma, Germany); Sodium Methoxide solution (Sigma, Germany) and Nonadecanoic acid (Sigma, Germany).
5
Fatty Acid Methyl Ester Analysis
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Hydrochloric acid, potassium chloride, sodium sulfate anhydrous, methanol, hexane, chloroform and sodium methoxide solution were purchased from Sigma-Aldrich, (Sigma-Aldrich, Burlington, MA, USA) [49 (link)]. Supelco 37 comp. FAME mix was purchased from Supelco (Supelco Analytical, Bellefonte, PA, USA).
6
Synthesis of Phenylalanine-Based Compounds
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L-Phenylalanine methyl ester hydrochloride
(HCl·H2N-F-OMe, 98%) was purchased from Carbolution
Chemicals. Boc-L-phenylalanine monohydrate (Boc-F-OH·H2O, 99%) was received from TCI chemicals. Tris(2-carboxyethyl)phosphine
hydrochloride (TCEP, 98%), 1-hydroxybenzotriazole hydrate (HOBt, 97%),
trifluoroacetic acid (TFA, 99%), poly(ethylene glycol) (HO-PEG-OH,
average Mw = 1500 g/mol), potassium thioacetate
(KSOAc, 98%), triethylamine (Et3N, 99%), sodium methoxide
solution (NaOMe, 25 wt % in methanol), and 2-bromopropionyl bromide
(97%) were purchased from Sigma-Aldrich and used as received. Mesylchloride
(MsCl, 99%) and N,N-dicyclohexylcarbodiimide
(DCC, 99%) were received from Merck and Alfa Aesar, respectively.
Br-FFFF-OMe was adopted from our recent work published elsewhere.32 (link) Reaction solvents were dried and distilled according
to the standard procedures.
(HCl·H2N-F-OMe, 98%) was purchased from Carbolution
Chemicals. Boc-L-phenylalanine monohydrate (Boc-F-OH·H2O, 99%) was received from TCI chemicals. Tris(2-carboxyethyl)phosphine
hydrochloride (TCEP, 98%), 1-hydroxybenzotriazole hydrate (HOBt, 97%),
trifluoroacetic acid (TFA, 99%), poly(ethylene glycol) (HO-PEG-OH,
average Mw = 1500 g/mol), potassium thioacetate
(KSOAc, 98%), triethylamine (Et3N, 99%), sodium methoxide
solution (NaOMe, 25 wt % in methanol), and 2-bromopropionyl bromide
(97%) were purchased from Sigma-Aldrich and used as received. Mesylchloride
(MsCl, 99%) and N,N-dicyclohexylcarbodiimide
(DCC, 99%) were received from Merck and Alfa Aesar, respectively.
Br-FFFF-OMe was adopted from our recent work published elsewhere.32 (link) Reaction solvents were dried and distilled according
to the standard procedures.
7
Characterization of Malaysian Palm Oil
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Fresh commercial crude PMFO was obtained from a local palm oil mill in Malaysia. Sodium hydroxide (NaOH) , n-hexane, and n-heptane (HPLC grade) were purchased from Merck KGaA, Germany while 85％ phosphoric acid, ammonium monovanadate and ammonium heptamolybdate tetrahydrate were bought from Friendemann Schmidt Chemical, Germany. Natural bleaching earth (NBE) was purchased from Taiko Bleaching Earth Sdn. Bhd., Malaysia, Magnesium oxide, potassium dihydrogen phosphate, and 6N of nitric acid were acquired from Chemiz (M) Sdn. Bhd., Malaysia, Potassium hydrogen phthalate was bought from Classic Chemicals Sdn. Bhd., Malaysia. Isopropanol was purchased from Qrec (Asia) Sdn. Bhd., Malaysia, Sodium methoxide solution, tetrahydrofuran solution, and pyridine solution (HPLC grade) were purchased from Sigma-Aldrich Corporation, Germany. n-heptane (GC grade) and N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) were obtained from Acros Organics, Belgium. 1-glyceryl monononadecanoin, 1,3-glyceryl dinonadecanoin and glyceryl trinonadecanoin powders were bought from Nu-Chek Prep, Inc., USA. Analytical grade α-tocopherol (α-T, 95％) used was from Sigma-Aldrich Corporation, Germany and α-tocotrienol (α-T3, 99.1％) , β-tocotrienol (β-T3, 99.9％) , γ-tocotrienol (γ-T3, 98.3％) , and δ-tocotrienol (δ-T3, 95.5％) were from Chromadex, USA.
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