Part:BBa_K256028

placI:CHE

Generator of CHE (Cholesteryl esterase)

Usage and Biology

Promoted by pLacI, the part is inducible by IPTG and/or Lactose. It works better in Lactose.

CHE catalyzes the hydration reactions of Cholesteryl Esters into free cholesterol and fatty acids with high specificity.

'Cholesteryl Ester + H2O → cholesterol + RCOOH

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 928
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Applications of BBa_K256028

Cholesteryl esterase (CHE) is produced under the promotion of an inducible promoter, pLacI.

CHE can degrade cholesteryl esters into free cholesterols and fatty acids. The acitivity of the enzyme is tested by cholesterol assay, which measures the cholesterol levels when cholesteryl esters are used as the substrate. The amount of cholesterol in the sample is proportional to the fluorescence generated.

Chracterization Data

Cells expressing CHE degrades more choleteryl esters into free choleterol than cells not expressing CHE. Negative control denotes the fluorescence emitted when cells are tested under no substrate conditions.

As protein purification was not carried out, data shown was obtained using soluble portion of sonicated cells. The assay is non-specific, so negative control also gives significant signals. Cholesteryl Linoleate was used as the enzyme substrate.