Chloroform, cell disruption

Add 0.25 ml of each culture to the corresponding reaction tube. Mix vigorously in a Vortex mixer for 5 sec. The chloroform and SDS will disrupt the integrity of (solubilize) the cell membrane, allowing the ONPG substrate to come into contact with the /3-galactosidase present in the cell. 

Cells grown under autotrophic growth condition (in BM) were disrupted by sonication for 1 min five times, and the hydrophobic fraction was extracted with pure redistilled chloroform. Acetone insoluble fraction of chloroform extracts was analyzed by TOF-MS (KOMPACT MALDl 11, Shimadzu, Kyoto, Japan) and its methanolysate was analyzed by GC/MS(E1) (JMS-DX303, JEOL, Tokyo, Japan). Acetone soluble fraction was directly analyzed by GC/MS(E1). HD-1 cell extract was prepared as follows autotrophically grown cells were centrifuged, washed, freeze-dried and weighed. 

When the hydrophobic fraction was analyzed by silica 60 TLC, quite apolar substances were found at the front of the developing solvent (chloroform/methanol/watep=65 25 4). Therefore, we examined whether or not the strain HD-1 produced hydrocarbons from C02. HD-1 cells were collected from 10 liter BM culture. Non-polar substances contained in the chloroform extracts of the disrupted cells were directly analyzed by GC/MS(E1) (Fig. 3a). The characteristic fragmentation patterns and each mass number of the molecular ion peak of these... 

Carbon tetrachloride is metabolized by cytochrome P-450 to the reactive metabolites trichloromethyl free radical and trichloromethylperoxy free radical. The trichloromethyl free radical may bind directly to cellular macromolecules such as lipids and proteins, and also to DNA, disrupting cell processes and breaking down membranes. The free radical can take part in anaerobic reactions, subsequently forming such toxic compounds as chloroform, hexachloroethane, and carbon monoxide. Aerobic biotransformation of the... 

Once the cells are disrupted, the recovery of the released components can be carried out. Organic solvents like cyclohexane, hexane, chloroform, or ethanol have proven to be... 

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