Preparation of fatty-acylated derivatives of acyl carrier protein using Vibrio harveyi acyl-ACP synthetase.

摘要: Abstract A simple two-step purification of Vibrio harveyi fatty acyl—acyl carrier protein (acyl-ACP) synthetase, which is useful for the quantitative preparation and analysis fatty-acylated derivatives ACP, described. Acyl-ACP synthetase can be partially purified from extracts this bioluminescent bacterium by Cibacron blue chromatography Sephacryl S-300 gel filtration stable months at −20δC in presence glycerol. Incubation ACP Escherichia coli with ATP radiolabeled acids (6 to 16 carbons length) enzyme resulted conversion biologically active acylated derivatives. The reaction monitored a filter disk assay quantitate levels or sodium dodecyl sulfate—polyacrylamide electrophoresis fluorography detect cell extracts. With its broad acid chain length specificity optimal activity mild nondenaturing buffers, soluble V. acyl-ACP provides an attractive alternative current chemical enzymatic methods analysis.