Protein stabilization by domain insertion into a thermophilic protein

US8592192B2 · kind B2 · utility

Assignee

• Polytechnic Institute of NYU · US

Inventors

• Jin Ryoun Kim · Hicksville, US
• Brennal Pierre · Brooklyn, US
• Marc Alan Ostermeier · Baltimore, US
• Chung Sei Kim

Key dates

• Filing date: Mar 5, 2010
• Grant date: Nov 26, 2013
• Expiry date: Sep 16, 2031

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/62
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A strategy to improve protein stability by domain insertion. TEM 1 beta-lactamase (BLA) and exo-inulinase, as model target enzymes, are inserted into a hyperthermophilic maltose binding protein from Pyrococcus furiosus (PfMBP). Unlike conventional protein stabilization methods that employ mutations and recombinations, the inventive approach does not require any modification on a target protein except for its connection with a hyperthermophilic protein scaffold. For that reason, target protein substrate specificity was largely maintained, which is often modified through conventional protein stabilization methods. The insertion was achieved through gene fusion by recombinant DNA techniques.