Enzymatic Reagent

Enzymatic Reagent

Modality

Tag Removal Proteases

To improve the solubility and simplify the purification process of recombinant proteins, researchers usually add fusion tags, of which His-tag, maltose-binding protein (MBP), and glutathione-S-transferase (GST) are commonly used.

While tagged as an extra sequence of protein, it should be removed to maintain biological activity in the pharmaceutical industry. The removal of fusion tags requires site-specific proteases, such as enterokinase (EK), thrombin, tobacco etch virus protease (TEVp), human rhinovirus protease 3C (HRV3C), small ubiquitin modifying protein (SUMO) protease, tobacco vein mottling virus (TVMV) protease, and carboxypeptidase A/B (CPA/CPB).

Type	Enzyme	Recognition Site
Tag removal endoproteases	Enterokinase (EK), Enteropeptidase	DDDDK↓
	Thrombin	LVPR↓GS
	TEV protease	ENLYFQ↓G
	HRV3C protease	LEVLFQ↓GP
	SUMO protease	SUMO tertiary structure
	TVMV protease	ETVRFQG↓S
Tag removal exoproteases	Carboxypeptidase A (CPA)	C-terminal amino acids, except Pro, Lys and Arg
Tag removal exoproteases	Carboxypeptidase B (CPB)	C-terminal Lys and Arg

Other Proteases

Application	Enzyme	Function
Other proteases	Protease K	A serine protease that digests proteins by hydrolyzing peptide bonds.
	IdeS, IgG protease	IgG degrading enzymes (Ides) that cleave at a specific site of immunoglobulin G (IgG), generating Fab and Fc fragments
	IgA1 protease	A proteolytic enzyme that cleaves specific site in the human immunoglobulin A1 (IgA1) hinge region sequence.

Nuclease

Application	Enzyme	Function
Nuclease	Nuclease	Cleaves nucleic acids (DNA or RNA) by hydrolyzing the phosphodiester bonds.
Nuclease	Restrict enzyme	An endonuclease that cleaves DNA at or near specific sites.

Amidase

Application	Enzyme	Function
Amidase	PNGase F	Cleaves between the innermost GlcNAc and asparagine residues of high mannose, hybrid, and complex oligosaccharides.

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Enzymatic Reagent

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