Engineering Thermostability in Artificial Metalloenzymes to Increase Catalytic Activity

Citations Over TimeTop 10% of 2021 papers

Abstract

Protein engineering has shown widespread use in improving the industrial application of enzymes and broadening the conditions they are able to operate under by increasing their thermostability and solvent tolerance. Here, we show that protein engineering can be used to increase the thermostability of an artificial metalloenzyme. Thermostable variants of the human steroid carrier protein 2L, modified to bind a metal catalyst, were created by rational design using structural data and a 3DM database. These variants were tested to identify mutations that enhanced the stability of the protein scaffold, and a significant increase in melting temperature was observed with a number of modified metalloenzymes. The ability to withstand higher reaction temperatures resulted in an increased activity in the hydroformylation of 1-octene, with more than fivefold improvement in turnover number, whereas the selectivity for linear aldehyde remained high up to 80%.

Related Papers

• → Engineering proteins for thermostability: the use of sequence alignments versus rational design and directed evolution(2001)290 cited
• → Recent advances in simultaneous thermostability-activity improvement of industrial enzymes through structure modification(2023)124 cited
• → Protein engineering of Bacillus acidopullulyticus pullulanase for enhanced thermostability using in silico data driven rational design methods(2015)55 cited
• → Engineering a high-performance, metagenomic-derived novel xylanase with improved soluble protein yield and thermostability(2014)44 cited
• → Introduction to Directed Evolution and Rational Design as Protein Engineering Techniques(2023)3 cited