Improved thermostability of AEH by combining B-FIT analysis and structure-guided consensus method

Synthetic biology for the directed evolution of protein biocatalysts: navigating sequence space intelligently

“…1068,1094 Thus, Greaves and Warwicker 1095 conclude that “charge number relates to solubility, whereas protein stability is determined by charge location”. The choice of which residues to focus on can be assisted (if a structure is available) by looking at the local flexibility via methods such as mutability 1096,1097 or via B -factors, 1062,1098,1099 or via certain kinds of mass spectrometry. 1100–1108 Constraint Network Analysis 1109 provides a useful strategy for choosing which residues might be most important for thermostability.…”

Thermostabilization of the uronate dehydrogenase from Agrobacterium tumefaciens by semi-rational design

“…Setting the B factor as a criterion was already suggested by Parthasarathy and Murthy (2000) and further successfully applied for thermal stabilization of Bacillus subtilis lipase (Reetz and Carballeira 2006) and an α-Amino ester hydrolase (Blum et al 2012). Hence, we used the B factor as our leading criterion.…”

Scope and limitations of optical pure hydantoins as chiral auxiliaries in asymmetric Mannich reactions

“…It is well known that β-amino esters are appropriate substrates for the preparation of β-lactams, which are the central constituents of a large family of antibiotics [13][14][15][16][17][18] . In our work, β-amino esters 4a and 4g-4k were treated with lithium hexamethyldisilazide(LiHMDS) at -20 °C in anhydrous tetrahydrofuran(THF) to afford the target β-lactams 5a and 5g-5k in excellent yields(Scheme 3).…”