Constitutive expression of active microbial transglutaminase in Escherichia coli and comparative characterization to a known variant.

ABSTRACT:

Background

Microbial transglutaminase (mTG) is a robust enzyme catalyzing the formation of an isopeptide bond between glutamine and lysine residues. It has found use in food applications, pharmaceuticals, textiles, and biomedicine. Overexpression of soluble and active mTG in E. coli has been limited due to improper protein folding and requirement for proteolytic cleavage of the pro-domain. Furthermore, to integrate mTG more fully industrially and academically, thermostable and solvent-stable variants may be imperative.

Results

A novel expression system constitutively producing active mTG was designed. Wild-type (WT) mTG and a S2P variant had similar expression levels, comparable to previous studies. Kinetic constants were determined by a glutamate dehydrogenase-coupled assay

SUBMITTER: Javitt G 

PROVIDER: S-EPMC5331659 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature