... determined by titration with pre-titrated bovine trypsin and the substrate l -BAPA, as previously described [ 19 ]. directed evolution protease inhibitor protein engineering protein–protein interactions (PPIs) serine proteases Extracellular proteases that are aberrantly expressed...

... 2016 10 11 2016 © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society 2017 ancestral sequence reconstruction directed evolution maximum likelihood protein engineering protein evolution thermostabilty Proteins, along with the genes...

...Verónica Sáez-Jiménez; Sandra Acebes; Eva Garcia-Ruiz; Antonio Romero; Victor Guallar; Miguel Alcalde; Francisco J. Medrano; Angel T. Martínez; Francisco J. Ruiz-Dueñas A variant of high biotechnological interest (called 2-1B) was obtained by directed evolution of the Pleurotus eryngii VP...

... of proteinaceous inhibitors, mesotrypsin presents a formidable challenge to the development of biological inhibitors. We used a powerful yeast display platform for directed evolution, employing a novel multi-modal library screening strategy, to engineer the human amyloid precursor protein Kunitz protease inhibitor...

... variants, in particular development of elegant high-throughput screening capabilities to enable optimization of desirable activities via directed evolution. These advances in enzymology have been paralleled by advances in medicinal chemistry, leading to the development of second- and third-generation...

... in Nature to perform the reactions required for modern chemical industries, many P450s show relaxed substrate specificity and exhibit some degree of activity towards non-natural substrates of relevance to applications such as drug development. Directed evolution and other protein engineering methods can...

...Eduardo Andrés; Hugo Aragunde; Antoni Planas Glycosynthases have become efficient tools for the enzymatic synthesis of oligosaccharides, glycoconjugates and polysaccharides. Enzyme-directed evolution approaches are applied to improve the performance of current glycosynthases and engineer...

... to six rounds of directed evolution, achieving a level of secretion in Saccharomyces cerevisiae (21 mg/l) as yet unseen for any ligninolytic peroxidase. The evolved variant for expression harboured four mutations and increased its total VP activity 129-fold. The signal leader processing by the STE13...

... by directed evolution using a DNA-shuffling technique. Through five repeats of DNA shuffling/screening, variants that allowed Escherichia coli C600 ( leuB − ) to grow on a minimal medium in 2 days were obtained. One of the variants LR5–1, with eight amino acid replacements, was found to possess a 65-fold...

... , 15 – 21 ]. directed evolution metal ion binding metal ion co-ordination phosphotriesterase (PTE) Metals are an essential constituent of about one-third of all proteins [ 1 ]. They can be bound to proteins in a variety of ways, from being part of a cofactor, such as a haem group...

... chaperonin CO 2 fixation directed evolution protein engineering RbcX ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) Rubisco [ribulose-P 2 (D-ribulose-1,5-bisphosphate) carboxylase/oxygenase; EC 4.1.1.39] is the enzyme employed by almost all photoautotrophic organisms to initiate...

... for CO 2 and no change in its K M for ribulose-1,5-bisphosphate or its CO 2 /O 2 selectivity. The results of the present study show that the directed evolution of the Synechococcus Rubisco in E. coli can elicit improvements in folding and catalytic efficiency. CO 2 fixation directed evolution...

... -amino acid amidohydrolase directed evolution negative charge protein folding protein solubility Although various bacterial expression systems have been established for fast and efficient expression of heterologous proteins [ 1 ], one common problem of most heterologous expression systems...

...Manuel MORILLAS; Colin E. McVEY; James A. BRANNIGAN; Andreas G. LADURNER; Larry J. FORNEY; Richard VIRDEN Two mutant forms of penicillin acylase from Escherichia coli strains, selected using directed evolution for the ability to use glutaryl- l -leucine for growth [Forney, Wong and Ferber (1989...

... the performance of a wide array of biologically active proteins. A higher operational stability is one of the most desired enzyme improvements and has been found hard to achieve by rational protein design. By directed evolution, the half-life of thermal inactivation of several mesophilic proteins has been...