Menaquinones is produced by Flavobacterium meningosepticum via submerged fermentation with 60% working volume in 30-L fermentation. To obtain the crude menaquinones, the fermentation medium is centrifuged at 12,000 rpm for 15 minutes. The vacuum-freeze dried particles are crushed and collected. After that the samples are treated with methanol for 20 minutes. To remove polar lipid compounds from bacterial cells, HZ816 macroporous adsorption resin column is used with methanol-dichloromethane (l:1) as mobile phase and flow rate of 0.75 mL/min. Rotary evaporation at 500 C is carried out to concentrate the eluent. After that, the concentrated sample is purified with Bio-BeadsTM S-X3 to remove components without UV absorption. To obtain the homolog purified menaquinones, RP-C18 column is used. Recrystallization is carried out to obtain pure crystalized menaquinones. Answer the following: Indicate how to obtain the sample in the methanol solvent. Three different chromatography are used in the purification process. Indicate the type of each column used.
Menaquinones is produced by Flavobacterium meningosepticum via submerged fermentation with 60% working volume in 30-L fermentation. To obtain the crude menaquinones, the fermentation medium is centrifuged at 12,000 rpm for 15 minutes. The vacuum-freeze dried particles are crushed and collected. After that the samples are treated with methanol for 20 minutes. To remove polar lipid compounds from bacterial cells, HZ816 macroporous adsorption resin column is used with methanol-dichloromethane (l:1) as mobile phase and flow rate of 0.75 mL/min. Rotary evaporation at 500 C is carried out to concentrate the eluent. After that, the concentrated sample is purified with Bio-BeadsTM S-X3 to remove components without UV absorption. To obtain the homolog purified menaquinones, RP-C18 column is used. Recrystallization is carried out to obtain pure crystalized menaquinones.
Answer the following:
- Indicate how to obtain the sample in the methanol solvent.
- Three different chromatography are used in the purification process. Indicate the type of each column used.
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