Engineered enzyme having acetoacetyl-coa hydrolase activity, microorganisms comprising same, and methods of using same
AssigneeBraskem S.A.
InventorsAvram Michael Slovic, Iuri Estrada Gouvea, Daniel Johannes Koch, Felipe Galzerani
Life of this patent
Estimating the term…
Abstract
The disclosure provides engineered enzymes that are capable of mediating the conversion of acetoacetyl-CoA to acetoacetate that do not react with the same order of magnitude with acetyl-CoA as they do with acetoacetyl-CoA (e.g., the engineered enzymes have a specific acetoacetyl-CoA hydrolase activity at least 10x higher than its acetyl-CoA hydrolase activity). Additionally, the disclosure provides modified microorganisms that comprise the engineered enzymes disclosed herein and methods of using same.
Claims16
An engineered enzyme having acetoacetyl-CoA substrate specificity and acetoacetyl-CoA specific hydrolase activity.
An engineered enzyme having the amino acid sequence as set forth in SEQ ID NO:
- 2, SEQ ID NO:
- 4, or SEQ ID NO:
- 6.
A method of engineering an enzyme having acetoacetyl-CoA substrate specificity and acetoacetyl-CoA specific hydrolase activity, the method comprising:
- a) selecting an enzyme having acetoacetyl-CoA transferase activity, and b) substituting a glutamic acid residue to an aspartic acid residue at a position corresponding to amino acid position 51 of SEQ ID NO:
- 1 in the enzyme having acetoacetyl-CoA transferase activity to produce an engineered enzyme.
Dependencies are inferred from the claim text.