VARIANTES DE SUBTILISINA CON MÚLTIPLES SUSTITUCIONES.

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

Tipo: Patente Europea. Resumen de patente/invención. Número de Solicitud: E05006461.

Solicitante: Danisco US Inc.
THE PROCTER & GAMBLE COMPANY
.

Nacionalidad solicitante: Estados Unidos de América.

Dirección: 925 PAGE MILL ROAD PALO ALTO, CA 94304 ESTADOS UNIDOS DE AMERICA.

Inventor/es: POULOSE, AYROOKARAN, J., SCHELLENBERGER, VOLKER, PAECH, CHRISTIAN, NADHERNY, JOANNE, NAKI, DONALD, P., CALDWELL, ROBERT, M., COLLIER, KATHERINE, D., BAECK, ANDRE, C., Kellis,James T.

Fecha de Publicación: .

Fecha Solicitud PCT: 23 de Octubre de 1998.

Clasificación Internacional de Patentes:

  • A23G4/12M
  • A61K8/66 NECESIDADES CORRIENTES DE LA VIDA.A61 CIENCIAS MEDICAS O VETERINARIAS; HIGIENE.A61K PREPARACIONES DE USO MEDICO, DENTAL O PARA EL ASEO (dispositivos o métodos especialmente concebidos para conferir a los productos farmacéuticos una forma física o de administración particular A61J 3/00; aspectos químicos o utilización de substancias químicas para, la desodorización del aire, la desinfección o la esterilización, vendas, apósitos, almohadillas absorbentes o de los artículos para su realización A61L; composiciones a base de jabón C11D). › A61K 8/00 Cosméticos o preparaciones similares para el aseo. › Enzimas.
  • A61Q11/00 A61 […] › A61Q USO ESPECIFICO DE COSMETICOS O DE PREPARACIONES SIMILARES PARA EL ASEO.Preparaciones para el cuidado de los dientes, la cavidad bucal o la dentadura, p.ej. dentífricos o pasta dental; Colutorios.
  • C11D3/386 QUIMICA; METALURGIA.C11 ACEITES, GRASAS, MATERIAS GRASAS O CERAS ANIMALES O VEGETALES; SUS ACIDOS GRASOS; DETERGENTES; VELAS.C11D COMPOSICIONES DETERGENTES; UTILIZACION DE UNA SOLA SUSTANCIA COMO DETERGENTE; JABON O SU FABRICACION; JABONES DE RESINA; RECUPERACION DE LA GLICERINA.C11D 3/00 Otros compuestos que entran en las composiciones detergentes cubiertas por C11D 1/00. › Preparaciones que contienen enzimas.
  • C12N9/54 C […] › C12 BIOQUIMICA; CERVEZA; BEBIDAS ALCOHOLICAS; VINO; VINAGRE; MICROBIOLOGIA; ENZIMOLOGIA; TECNICAS DE MUTACION O DE GENETICA.C12N MICROORGANISMOS O ENZIMAS; COMPOSICIONES QUE LOS CONTIENEN; PROPAGACION, CULTIVO O CONSERVACION DE MICROORGANISMOS; TECNICAS DE MUTACION O DE INGENIERIA GENETICA; MEDIOS DE CULTIVO (medios para ensayos microbiológicos C12Q 1/00). › C12N 9/00 Enzimas, p. ej. ligasas (6.); Proenzimas; Composiciones que las contienen (preparaciones para la limpieza de los dientes que contienen enzimas A61K 8/66, A61Q 11/00; preparaciones de uso médico que contienen enzimas A61K 38/43; composiciones detergentes que contienen enzimas C11D ); Procesos para preparar, activar, inhibir, separar o purificar enzimas. › siendo las bacterias del género Bacillus.

Clasificación PCT:

  • A23K1/165

Clasificación antigua:

  • A23G3/30
  • A61K7/28
  • A61K7/48
  • C11D3/386 C11D 3/00 […] › Preparaciones que contienen enzimas.
  • C12N9/54 C12N 9/00 […] › siendo las bacterias del género Bacillus.

Países PCT: Austria, Bélgica, Suiza, Alemania, Dinamarca, España, Francia, Reino Unido, Grecia, Italia, Liechtensein, Luxemburgo, Países Bajos, Suecia, Mónaco, Portugal, Irlanda, Finlandia, Rumania, Chipre.

PDF original: ES-2368718_T3.pdf

 


Fragmento de la descripción:

Las serinaproteasas son un subgrupo de carbonilhidrolasas. Las mismas comprenden una clase diversa de enzimas que tienen una extensa gama de especificidades y funciones biológicas. Stroud, R. Sci. Amer., 131: 7488. A pesar de su diversidad funcional, la maquinaria catalítica de las serinaproteasas ha sido abordada por al menos dos familias de enzimas genéticamente distintas: 1) las subtilisinas y 2) las serinaproteasas de mamífero afines a la quimotripsina y serinaproteasas bacterianas homólogas (v.g., tripsina y S. gresius tripsina). Estas dos familias de serinaproteasas exhiben mecanismos de catálisis notablemente similares. Kraut, J. (1977), Annu. Rev. Biochem., 46: 331358. Adicionalmente, aunque la estructura primaria no está emparentada, la estructura terciaria de estas dos familias de enzimas reúne una tríada catalítica conservada de aminoácidos constituida por serina, histidina y aspartato. Las subtilisinas son serinaproteasas (PM aprox. 27.500) que son secretadas en grandes cantidades por una gran diversidad de especies de Bacillus y otros microorganismos. La secuencia proteínica de la subtilisina ha sido determinada a partir de al menos nueve especies diferentes de Bacillus. Markland, F.S., et al. (1983), Hoppe Seyler's Z., Physiol. Chem., 364: 15371540. La estructura cristalográfica tridimensional de las subtilisinas de Bacillus amyloliquefaciens, Bacillus licheniformis y varias variantes naturales de B. lentus ha sido consignada. Estos estudios indican que aunque la subtilisina no está emparentada genéticamente con las serinaproteasas de mamífero, tiene una estructura de sitio activo similar. Las estructuras cristalinas de rayos X de inhibidores peptídicos unidos covalentemente que contienen subtilisina (Robertus, J.D., et al. (1972), Biochemistry, 11: 24392449) o complejos de productos (Robertus. J.D., et al. (1976), J. Biol. Chem., 251: 10971103) han proporcionado también información concerniente al sitio activo y a la hendidura supuesta de unión al sustrato de la subtilisina. Adicionalmente, se ha llevado a cabo un gran número de mutagénesis cinéticas y químicas (Biol. Chem., 244: 5333 5338) y orientadas extensas (Wells y Estell (1988) TIBS 13: 291297). La publicación internacional WO 91/00345 (Novo Nordisk A/S) describe subtilisina-proteasas mutadas, que tienen una carga electrostática neta, para usar en composiciones detergentes. La publicación internacional WO 95/10615 (Genencor International, Inc.) describe variantes de subtilisina que tienen una pluralidad de sustituciones de aminoácidos, incluyendo en la posición 76 y en una o más de las posiciones 99, 101, 103, 104, 107, 123, 27, 105, 109, 126, 128, 135, 156, 166, 195, 204, 206, 210, 216, 217, 218, 222, 260 y/o 274 Sumario de la Invención La invención proporciona una variante de proteasa subtilisina 309 que comprende un conjunto de sustituciones que se seleccionan 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/VN68A/S103AN104I/G159D/A232V/Q236H/Q245R/N252K; N43K/V68AS103AN104I/G159D/A232V/Q236H/Q245R; ES 2 368 718 T3 N43D/V68A/S103AN104I/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/G1159D/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; 2 V68A/S103A/V104I/G159D/S212P/A232V/Q236H/Q245R/N248D/N252K; V68A/N76D/S103A/V104I/G1159D/Q236H; V68A/N76D/S103A/V104I/Q236H; V68A/N76D/S103A/V104I/G159D/Q236H/E271V; V68A/V76D/S103A/V104I/G159D/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/L217I/Q236H/E271V; V68A/N76D/S103AN104I/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/G 159D/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/S03A/V104I/G159D/Q236H/Q245R/R247H; V68A/N76D/S103AN104I/G159D/A174V/N204D/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/N204D/Q236H/Q245R; V68A/N76D/S103A/V104I/A133V/G 159D/N218D/Q236H/Q245R; V68A/N76D/S103A/V104I/G 159D/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A194I/V203A/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/T213R/A232V/Q236H/Q245R/T260A; V68A/N76D/S103AN104I/G159D/P210R/A232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/L257V; N76D/S103A/V104I/A232V/Q236H/Q245R/L257V; ES 2 368 718 T3 N76D/S103AN104I/G159D/A232V/Q236H/Q245R/L257V; V68A/N76D/S1103A/V104I/G59D/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/S103AN104I/G159D/Y214UA232V/Q236H/Q245R; V68A/N76D/S103A/V104I/G159D/A215R/A232V/Q236H/Q245R; Q12R/V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; G20R/V68A/N76D/S103AN104I/G159D/A232V/Q236H/Q245R/S259G; V68A/N76D/S87R/S103A/V104I/G159D/A232V/Q236H/Q245R/T260V; 3 V68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R/N261G; V68A/N76D/S103AN104I/G159D/A232V/Q236H/Q245R/N261W; N76D/S103A/V104I/A232V/Q236H/S242P/Q245R; V68A/N76D/S103A/V104I/G159D/P210L/A232V/Q236H/Q245R; Q12R/A48VN68A/N76D/S103A/V104I/G159D/A232V/Q236H/Q245R; N76D/S103A/V104I/A232V/Q236H/Q245R; ES 2 368 718 T3 N76D/S103AN104I/G159D/Y192F/A232V/Q236H/Q245R; N76D/S103A/V104I/V1471/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/S1103A/V104I/G1159D/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/N248D1N252K; 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/S103AN104I/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/V1104I/G159D/T213E/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V1104I/G 159D/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/V1104I/G 159D/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; 4 ES 2 368 718 T3 V68A/S103A/V104I/G159D/Q206R/A232V/Q236H/Q245R/N248D/N252K; V68A/S103A/V1104I/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/V1104I/G159D/A232V/Q236H/Q245R/N248D/N252K/T255V; V68A/S1103A/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/N261 R; 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;... [Seguir leyendo]

 


Reivindicaciones:

1. 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; ES 2 368 718 T3 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; 51 52 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; ES 2 368 718 T3 53 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; ES 2 368 718 T3 ES 2 368 718 T3 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; 54 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. 1. 2. Un DNA que codifica una variante de proteasa subtilisina 309 de la reivindicación 1. 3. Un vector de expresión que comprende el DNA de la reivindicación 2. 4. Una célula hospedadora transformada con el vector de expresión de la reivindicación 3. 5. Una composición de limpieza que comprende la variante de proteasa subtilisina 309 de la reivindicación 6. Una comida para animales que comprende la variante de proteasa subtilisina 309 de la reivindicación 1. 7. Una composición para tratar un producto textil que comprende la variante de proteasa subtilisina 309 de la reivindicación 1. 8. Un método para producir una variante de proteasa subtilisina de Bacillus que tiene unas características de lavado mejores que las de un precursor de proteasa subtilisina de Bacillus en al menos una formulación detergente y/o en al menos un conjunto de condiciones de lavado, que comprende introducir en dicho precursor de proteasa subtilisina de Bacillus 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; ES 2 368 718 T3 56 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; ES 2 368 718 T3 57 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; ES 2 368 718 T3 58 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/Q248R/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/Q248R/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; ES 2 368 718 T3 ES 2 368 718 T3 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; 59 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/Gl59D/V205I/A232V/Q236H/Q245R/N248D/N252K; S101G/S103A/V104I/G159D/A230V/Q236H/Q245R; ES 2 368 718 T3 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 de aminoácidos corresponden a las posiciones pertinentes de la subtilisina de Bacillus amyloliquefaciens. 9. El método de la reivindicación 8, en el que un DNA que codifica el precursor de la proteasa subtilisina de Bacillus se modifica para producir una variante de DNA que codifica la variante de la proteasa subtilisina de Bacillus. ES 2 368 718 T3 61 ES 2 368 718 T3 62 ES 2 368 718 T3 63 ES 2 368 718 T3 64 ES 2 368 718 T3 RESTOS QUE SE CONSERVAN EN SUBTILISINAS PROVENIENTES DE COMPARACIÓN DE SECUENCIAS DE SUBTILISINA PROVENIENTE DE: ES 2 368 718 T3 66 ES 2 368 718 T3 67

 

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