Structural enzymological studies of multifunctional enzyme, type-1 (MFE1) with the 2E-decenoyl-CoA and 2E,4E-decadienoyl-CoA substrates: The regeneration of the dehydrogenase catalytic site is the rate limiting step of its combined reactions.

Key Points

• This research aims to investigate the structural and enzymatic behavior of multifunctional enzyme type-1 (MFE1) with specific CoA substrates.
• Conducted crystallographic binding studies of RnMFE1 with substrates 2E-decenoyl-CoA and 2E,4E-decadienoyl-CoA.
• Analyzed the enzyme's kinetic behavior and regeneration of the HAD active site post-dehydrogenation.
• Examined enzyme surface properties affecting substrate channeling through electrostatic interactions.
• Regeneration of the dehydrogenase catalytic site was identified as the rate-limiting step of the combined reactions.
• Crystallographic studies showed distinct binding modes for the enzyme with its substrates, impacting kinetic efficiency.
• Positively charged surface patches were found to enhance substrate transfer between the ECH and HAD active sites, implying a structured channeling mechanism.