Question: The enzyme fatty acyl-CoA synthetase (referred to in lecture as Acyl-CoA Synthase, not to be conf…

The enzyme fatty acyl-CoA synthetase (referred to in lecture as
Acyl-CoA Synthase, not to be confused with fatty acid synthase)
plays a key role in ensuring that fatty acids are capable of
entering the β-oxidation pathway, acting as a ‘gatekeeper’ enzyme.
This ligase enzyme is responsible for the production of fatty
acyl-CoA conjugate molecules. 1. Using the molecule 10:1n-5 fatty
acid, draw out the multi-step reactions depicting the ‘activation’
of this fatty acid by fatty acyl-CoA synthetase. While working in
the research lab, you have been working on studying a disorder that
manifests from aberrant fatty acid breakdown. However, it is
unclear which step in this pathway is being affected. In order to
determine if fatty acid activation (by fatty acyl-CoA synthetase)
is the step being affected, you decide on developing an
experimental method to monitor this enzyme’s activity. 2. Based on
your answer in Question 1, identify and indicate the reagents
necessary to experimentally test fatty acyl-CoA synthetase activity
in vitro (i.e. in a test tube); assume in this case you are
continuing to use the above fatty acid. In addition, sketch out a
hypothetical Michaelis-Menten curve depicting this enzyme’s
activity as fatty acid concentration increases, fully labeling your
axes.