VARIANTES DE SUBTILISINA CON MULTIPLES SUSTITUCIONES.

摘要

Una variante de proteasa-subtilisina 309 que comprende un conjunto de sustituciones seleccionado del grupo: V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N252K; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R; V68A/S103A/V104I/N140D/G159D/A232V/Q236H/Q245R/N252K; N43S/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N252K; N43K/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R; N43D/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N252K; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/L257V; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D; V68A/S103A/V104I/G159D/A232V/Q236H/K237E/Q245R; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N252S; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/L257V/R275H; V68A/S103A/V104I/G159D/T224A/A232V/Q236H/Q245R/L257V; G61E/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; N43D/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S212P/A232V/Q236H/Q245R/N248D/N252K; V68A/N76D/S103A/V104I/G159D/Q236H; V68A/N76D/S103A/V104I/Q236H; V68A/N76D/S103A/V104I/G159D/Q236H/E271V; V68A/N76D/S103A/V104I/G159D/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/L217I/Q236H/E271V; V68A/N76D/S103A/V104I/G159D/Q236R; V68A/L75R/N76D/S103A/V104I/G159D/Q236H; V68A/N76D/S103A/V104I/A114V/V121I/G159D/Q236H/Q245R; Q12R/V68A/N76D/S103A/V104I/G159D/Q236H; V68A/N76D/S103A/V104I/G159D/Y209S/Q236H/T253K; V68A/N76D/S103A/V104I/N117K/G159D/N184S/Q236H; V68A/N76D/S103A/V104I/Q236H; V68A/N76D/S103A/V104I/G159D/Q236H/Q245L; V68A/N76D/S103A/V104I/N123S/G159D/Q236H/H249Y; V68A/N76D/S103A/V104I/G159D/Q236H/H249Q; V68A/N76D/S103A/V104I/G159D/Q236H/Q245R/N261D; V68A/N76D/S103A/V104I/S141N/G159D/Q236H/Q245R/T255S; V68A/N76D/S103A/V104I/G159D/Q236H/Q245R/R247H; V68A/N76D/S103A/V104I/G159D/A174V/N204D/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/N204D/Q236H/Q245R; V68A/N76D/S103A/V104I/A133V/G159D/N218D/Q236H/Q245R: V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A194I/V203A/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/T260A; V68A/N76D/S103A/V104I/G159D/P210R/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/L257V; N76D/S103A/V104I/A232V/Q236H/Q245R/L257V; N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/L257V; V68A/N76D/S103A/V104I/G159D/Y209W/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/G211R/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/G211V/A232V/Q236H/Q245R; Q12R/V68A/N76D/S103A/V104I/G159D/Y214L/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A215R/A232V/Q236H/Q245R; Q12R/V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; G20R/V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/S259G; V68A/N76D/S87R/S103A/V104I/G159D/A232V/Q236H/Q245R/T260V; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/N261G; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/N261W; N76D/S103A/V104I/A232V/Q236H/S242P/Q245R; V68A/N76D/S103A/V104I/G159D/P210L/A232V/Q236H/Q245R; Q12R/A48V/V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; N76D/S103A/V104I/A232V/Q236H/Q245R; N76D/S103A/V104I/G159D/Y192F/A232V/Q236H/Q245R; N76D/S103A/V104I/V147I/G159D/A232V/Q236H/Q245R/N248S/K251R; Q12R/V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/A272S; V68A/N76D/S103A/V104I/G159D/N183K/Q206L/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/S256R; V68A/N76D/S103A/V104I/G159D/Q206R/A232V/Q236H/Q245R; K27R/V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/N116T/G159D/R170S/N185S/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/Y209W/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/Q109R/G159D/A232V/Q236H/Q245R/N248D/N252K; G20R/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/Y209F/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/N185D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/P210R/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/N185D/P210L/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/P210L/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S212C/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S212G/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S212E/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/T213E/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/T213S/A232V/Q236H/Q245R/N248D/N252K; V68A/A103V/V104I/G159D/T213E/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/A215V/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/A215R/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S216T/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S216V/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/S216C/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/N173D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/Q206R/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252F; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252L; P55S/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252F; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/T255V; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/S256N; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/S256E; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/S256R; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/T260R; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/L257R; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/G258D; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/N261R; V68A/S103A/V104I/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/A232V/Q236H/Q245V/N248D/N252K; V68A/S103A/V104I/G159D/A228V/A232V/Q236H/Q245R/N248D/N252K; G61E/V68A/S103A/V104I/S130A/G159D/A232V/Q236H/Q245R/N248D/N252K; G61E/S103A/V104I/A133V/G159D/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248G/N252K; V68A/S103A/V104I/G159D/N218S/A232V/Q236H/Q245R/N248D/N252K; G20R/V68A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; V68A/N76D/E89D/S103A/V104I/G159D/P210L/T213R/A232V/Q236H/Q245R/T260A; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; G61E/V68A/S103A/V104I/G159D/S160V/A232V/Q236H/Q245R/N248D/N252K; S3L/G61E/V68A/N76D/S103A/V104I/A232V/Q236H/Q245R/N248D/N252K; G61E/V68A/S103A/V104I/G159D/Y167F/A232V/Q236H/Q245R/N248D/N252K; G97E/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; A98D/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S99E/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S101E/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; G102A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/S106E/G159D/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/Q109E/G159D/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K/N261R; S103A/V104I/Q109R/G159D/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/N184D/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/S166D/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/L217E/A232V/Q236H/Q245R/N248D/N252K; G20R/N62D/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/Q206R/L217E/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/G159D/Q206R/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/N184G/A232V/Q236H/Q245R/N248D/N252K; S103A/V104I/G159D/A232V/Q236H/V244T/Q245R/N248D/N252K; S103A/V104I/G159D/A232V/Q236H/V244A/Q245R/N248D/N252K; K27N/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; T38G/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K/S256R; Q12R/N62D/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/G159D/N185D/Q206E/T213R/A232V/Q236H/Q245R/N248D/N252K/E2 71Q; S101G/S103A/V104I/G159D/N185D/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/Q206E/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/T213Q/A232V/Q236H/Q245R/N248D/N252K; A98L/G102A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S101G/G102A/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; G102A/S103A/V104I/G159D/S212G/A232V/Q236H/Q245R/N248D/N252K; Q12R/G102A/S103A/V104I/G159D/S212G/A232V/Q236H/Q245R/N248D/N252K; A98L/G102A/S103A/V104I/G159D/S212G/A232V/Q236H/Q245R/N248D/N252K; S101G/G102A/S103A/V104I/G159D/S212G/A232V/Q236H/Q245R/N248D/N252K; G102A/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/Q109R/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/S130G/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/S130G/G159D/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/S128G/G159D/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/S128L/G159D/A232V/Q236H/Q245R/N248D/N252K; N62D/S101G/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/S128G/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; N62D/S103A/V104I/S128L/G159D/T213R/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/P131V/G159D/A232V/Q236H/Q245R/N248D/N252K; A98V/S101G/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S99G/S101G/S103A/V104I/G159D/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/S212G/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/Y209W/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/P210I/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/V205I/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/A230V/Q236H/Q245R; S101G/S103A/V104I/G159D/A194P/A232V/Q236H/Q245R/N248D/N252K; and N76D/S101G/S103A/V104I/G159D/A194P/A232V/Q236H/Q245R/N248D/N252K; en donde las posiciones de los restos aminoácidos corresponden a las posiciones pertinentes de la subtilisina de Bacillus amyloliquefaciens.