Enhancing Catecholase Activity of a Recombinant Human Tyrosinase Through Multiple Strategies.

ABSTRACT:

Background

Tyrosinases are copper-containing enzymes that initiate the melanin synthesis. They catalyze the direct oxidation of L-tyrosine or L-DOPA into L-DOPAquinone.

Objectives

In present study, we aimed to obtain a recombinant tyrosinase with enhanced catecholase activity through site-directed mutagenesis.

Materials and methods

The coding sequence of human tyrosinase along with native signal sequence was cloned into pET-28a (+). BL-21 was used as expression host and recombinant protein was purified by Ni-NTA resins. Site-directed mutagenesis was performed on M374 residue to achieve four mutants: M374D, M374T, M374K and M374R. Chloride ions (Cl^-) were removed from all solutions, and an extra amount of Cu²⁺ ions was added to recombinant tyrosin

SUBMITTER: Shahrisa A 

PROVIDER: S-EPMC7856396 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature