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Enhanced Recombinant Protein Production of Soluble, Highly Active and Immobilizable PNGase F.

ABSTRACT: High resolution analysis of N-glycans can be performed after their endoglycosidase mediated removal from proteins. N-glycosidase F peptide (PNGase F) is one the most frequently used enzyme for this purpose. Because of the significant demand for PNGase F both in basic and applied research, rapid and inexpensive methods are of great demand for its large-scale production, preferably in immobilizable form to solid supports or surfaces. In this paper, we report on the high-yield production of N-terminal 6His-PNGase F enzyme in a bacterial Escherichia coli SHuffle expression system. The activity profile of the generated enzyme was compared to commercially available PNGase F enzymes, featuring higher activity for the former. The method described here is thus suitable for the cost-effective production of PNGase F in an active, immobilizable form.

SUBMITTER: Kovacs N

PROVIDER: S-EPMC9259526 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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Enhanced Recombinant Protein Production of Soluble, Highly Active and Immobilizable PNGase F.

Kovács Noémi N Farsang Róbert R Szigeti Márton M Vonderviszt Ferenc F Jankovics Hajnalka H

Molecular biotechnology 20220304 8

High resolution analysis of N-glycans can be performed after their endoglycosidase mediated removal from proteins. N-glycosidase F peptide (PNGase F) is one the most frequently used enzyme for this purpose. Because of the significant demand for PNGase F both in basic and applied research, rapid and inexpensive methods are of great demand for its large-scale production, preferably in immobilizable form to solid supports or surfaces. In this paper, we report on the high-yield production of N-termi ...[more]

PMID: 35244857

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