Compositions and methods comprising variant microbial proteases

Abstract

The present invention provides variant subtilisins and compositions comprising at least one variant subtilisin set forth herein, as well as methods for using these variants and compositions. In some embodiments, the present invention provides variant proteases suitable for laundry cleaning applications.

What is claimed is: 
     
         1 . A subtilisin variant comprising at least one modification, wherein said modification is selected from: G100C, S101D, S101H, Q103D, M124G, L126G, S132H, S161N, Q275Z, and/or at least set of modifications selected from A001T/L096V/G097S/A098D/D099I, A001V/G097S/A098R/D099G, V004M/A098D/D099G, V004M/D099R/G100S/Z100.01L, V004M/G127C/G128S/P129R, V004M/M119C/D120S, V008I/A098G/D099R, V008I/A098N, V008I/M119C/D120G, Q010L/G097R/A098C/D099S, K012N/A098R/D099V, K012R/G097D/A098H/D099G, A013T/S101R/Q103S, A013V/G097N/A098H/D099G/G160S, A013V/G097S/Z097.01R/A098R, A013V/G100R/S101G, A013V/N118G/M119I, A013V/P129Z/S130H/S132G, P014L/N062D/S101R/Q103S, P014S/A098S/D099G, A015T/V068N/A069G, L016M/G128R/P129R/Z129.01G, H017Q/G127R/P129G, H017Y/A098R/D099I, H017Y/A098R/D099Z/S101G, H017Y/P129R, H017Y/S130Y/G131S/S132L, Q019H/G128R/Z128.01R/P129D, Q019H/Z128.01G/Z128.02G, G020D/G097R/A098G/D099C, G020D/P129Z/S130G/G131R/S132C, N025Y/S101L/Q103N, A029V/G097R/A098S, S033I/T066G/H067R/V068D/A216E, S038F/A098R/D099S, V044I/G097C/A098D/D099G, V044I/G097R/A098R, V044I/G100S/S101G, A045V/G097C/A098R/D099S, A048V/G097L/A098G/D099S, A048V/G100R/S101G, A048V/G128S/P129L/Z129.01G, M050I/A098G, S053F/S130I/S132G, S053T/D099H/S101Z, E054D/A098R/D099G, T055I/S101R/Q103R, T055S/T066I/Z066.01G/Z066.02G, Q059H/D099S/S101G, Q059H/M119S/D120R, Q059K/M119V/V121C, Q059L/P129G, N061D/G128R/P129V/Z129.01G, S063T/G097D/A098R/D099S, G065R/Z101.01R/Z101.02G, T066S/H067S, V068C/A069G, V068C/A069G/H238Q, V068G/A069G, V068G/A069G/V150I, V068G/A069S, V068I/A069G/A144V, V068I/G097D/A098G/D099Z/G100R, V068L/A069G, V068L/A069S, V068S/A069G, V068S/A069G/A116T, A069G/T071G, A069G/T071I, L075G/N076G/N077Z, L075R/N076G/N077Z, N076D/D099R/Z100.01G, 1079F/A098G, V081A/S101R/Q103V, V081I/G097R/A098G/D099C, V084D/G097S/A098G/D099N, V084I/G127C/P129G, V084I/G127L, A085V/A098G, A085V/G097R/A098S/D099N, A085V/G128S/P129R/Z129.01G, A085V/N218I, A085V/S101G, A085V/S204P, A085V/Z099.01H/S101R, P086S/G131D/S132G, P086T/S101D/Q103S/V147I, A088S/S101R/Q103V, A088T/A098R/D099R, A088T/D099G/Z100.01G, A088T/G127N/P129R, A088T/G127S/P129R/Q206R/A273S, A088T/P129S/G146D, A088T/S101G, A088T/S130R/G131H/S132N, A088V/G100Z/S101C/Q103R, A088V/P129Z, A092G/S249R, A092G/V093C, A092G/V093I, A092G/V093L, A092L/V093G/K094Z, A092S/K094V, A092S/V093C, A092S/V093G, A092S/V093S, A092V/V093L, V093I/G128D/P129R, V093I/P129Z, V093R/Z093.01G/K094R, K094E/V095D, K094S/V148I, L096I/N118R/M119C/D120G/A151V, Z096.01D/A098R, Z096.01H/A098G, Z096.01N/A098H, Z096.01R/A098C, Z096.01R/A098G, Z096.01R/A098G/V192I, Z096.01R/A098R, Z096.01R/A098R/V192A, Z096.01R/A098S, Z096.01R/A098S/G157S, Z096.01R/Z096.02G, Z096.01R/Z096.02G/V147I, Z096.01S/A098G, Z096.01S/A098N, G097C/A098D/D099S, G097C/A098G/D099G, G097C/A098G/D099N, G097C/A098G/D099S, G097C/A098G/D099S/G100Z, G097C/A098G/D099V, G097C/A098N/D099S, G097C/A098R, G097C/A098R/D099G, G097C/A098R/D099H, G097C/A098R/D099H, G097C/A098R/D099N, G097C/A098R/D099R, G097C/A098R/D099R/N109D/A114S, G097C/A098S/D099H/A151V, G097C/A098V/D099S, G097C/Z097.01G/A098H/T253I, G097C/Z097.01S/A098G, G097D/A098C, G097D/A098G/D099G, G097D/A098G/D099R, G097D/A098H/D099R, G097D/A098H/D099V, G097D/A098R/D099G, G097D/A098R/D099R, G097D/A098R/D099S, G097D/A098R/D099Z, G097D/G100C/S101R, G097D/G100R/S101G, G097H/A098C/D099G, G097H/A098D/D099N, G097H/A098G, G097H/A098G/D099G, G097H/A098G/D099R, G097H/A098G/D099Z, G097H/A098G/D099Z/A151V, G097H/A098G/D099Z/G100R, G097H/A098G/D099Z/G100V, G097H/A098G/D099Z/S101N/G166S, G097H/A098R, G097H/A098R/D099G, G097H/A098R/D099L, G097H/A098R/D099N, G097H/A098S/D099G, G097H/A098S/D099I, G097H/A098S/D099L, G097H/A098S/D099S/K237E, G097L/A098C/D099S, G097L/A098G/D099S, G097L/A098G/D099S/G100Z, G097L/A098G/D099Z, G097L/A098H, G097L/A098R/D099N, G097L/A098V, G097N/A098G/D099C, G097N/A098G/D099R, G097N/A098G/D099R/V270I, G097N/A098G/D099S, G097N/A098G/D099V, G097N/A098G/D099V/A151V, G097N/A098G/D099V/S101F, G097N/A098H/D099N, G097N/A098R/D099R, G097N/A098R/D099S, G097N/A098R/D099V, G097N/A098S/D099S, G097R/A098C, G097R/A098C/D099S, G097R/A098F/A200V, G097R/A098G, G097R/A098G/D099C, G097R/A098G/D099G, G097R/A098G/D099G/A273T, G097R/A098G/D099S, G097R/A098G/D099S/A133D, G097R/A098G/D099V, G097R/A098H/D099C/S182N, G097R/A098H/D099G, G097R/A098N/D099G, G097R/A098N/D099Z, G097R/A098R, G097R/A098R/D099C, G097R/A098R/D099G, G097R/A098R/D099S, G097R/A098S/D099C, G097R/A098S/D099G, G097R/A098S/D099L, G097R/A098S/D099R, G097R/A098S/D099S, G097R/A098S/D099V, G097R/A098V/D099G, G097R/Z097.01H/A098C, G097R/Z097.01R/A098G, G097R/Z097.01R/A098G/V147I, G097R/Z097.01R/A098S, G097R/Z097.01R, G097R/Z097.01S/A098G, G097R/Z097.01S/A098R, G097S/A098C, G097S/A098C/D099G, G097S/A098C/D099H, G097S/A098C/D099L, G097S/A098C/D099R, G097S/A098C/D099S, G097S/A098C/D099Z, G097S/A098G/D099F, G097S/A098G/D099G, G097S/A098G/D099G, G097S/A098G/D099S, G097S/A098G/D099Z, G097S/A098G/D099Z/G100R, G097S/A098H/D099C, G097S/A098H/D099G, G097S/A098N/D099G, G097S/A098R, G097S/A098R/D099C, G097S/A098R/D099G, G097S/A098R/D099L, G097S/A098R/D099V, G097S/A098R/D099Y, G097S/A098S/D099G, G097S/A098S/D099G/A133V/A153V/S236Y, G097S/A098S/D099G/A153V, G097S/A098S/D099Z, G097S/Z097.01D/A098G/A144T/Q271H, G097S/Z097.01G/A098V, G097S/Z097.01S/A098G, G097S/Z097.01S/A098G/A273T, G097S/Z097.01Y, G097V/A098R, G097V/A098R/D099G, G097V/A098S/D099S, G097Y/A098H/D099N, G097Z/A098S/D099H, Z097.01D/A098R/D099G, Z097.01D/A098S/D099L, Z097.01D/A098V/D099G, Z097.01G, Z097.01G/A098G, Z097.01G/A098G/D099S, Z097.01G/A098R, Z097.01G/A098R/D099C, Z097.01G/A098R/D099G, Z097.01G/A098R/D099S, A098C/D099G, A098C/D099G/A116T, A098C/D099H, A098C/D099L, A098C/D099L/K213R, A098C/D099L, A098C/D099N, A098C/D099R, A098C/D099S, A098C/D099S/P194L, A098D/D099C, A098D/D099G, A098D/D099G/G100L, A098D/D099H/G100S, A098D/D099H/H238Y, A098D/D099R, A098D/D099R/G100D, A098D/D099R/G160S, A098F/D099S/G100L, A098G/D099C A098G/D099G, A098G/D099G/A228T, A098G/D099G/G100H, A098G/D099G/N243D, A098G/D099G/S101P, A098G/D099G/T244S, A098G/D099G, A098G/D099H, A098G/D099L, A098G/D099R, A098G/D099R/A116T, A098G/D099R/G100D, A098G/D099R/V148I, A098G/D099S, A098G/D099S/L267M, A098G/D099V, A098G/D099Z, A098H/D099C, A098H/D099G, A098H/D099G/A216T, A098H/D099G/G100D, A098H/D099G/G100N/L267M, A098H/D099G/G100S, A098H/D099G/V143I, A098H/D099R, A098H/D099R/G100S, A098H/D099S/G100C, A098H/D099S/G100N, A098H/D099Y, A098I/D099G, A098I/D099S, A098L/D099G, A098L/D099S, A098N/D099G, A098N/D099L, A098N/D099S, A098R/A144T, A098R/A274V, A098R/D099C, A098R/D099G, A098R/D099G/G100C, A098R/D099G/G100D, A098R/D099G/G100L/S145T, A098R/D099G/G100N, A098R/D099G/G100R, A098R/D099G/G100S, A098R/D099G/V150I, A098R/D099H, A098R/D099I, A098R/D099L, A098R/D099N, A098R/D099N/G100L, A098R/D099R, A098R/D099R/G100D, A098R/D099R/S101Z, A098R/D099R/S130F, A098R/D099R, A098R/D099S, A098R/D099S/G100L/A153V, A098R/D099S, A098R/D099V, A098R/D099Z/S101G, A098R/G100C/A137T, A098R/G100S, A098S/D099C, A098S/D099G, A098S/D099G/G100C, A098S/D099G/G100D, A098S/D099G/G100R/L250I, A098S/D099G/G100S, A098S/D099G/Z099.01R, A098S/D099H, A098S/D099H/M124V, A098S/D099H/Z099.01R/G169A/V180A, A098S/D099R, A098S/D099R/G100L, A098S/D099R/G154S, A098S/D099R/G166S, A098S/D099R/Z100.01G, A098S/D099S, A098S/D099S/Z099.01R, A098S/D099V, A098T/P129Z/S130R/S132G, A098T/S101G, A098V/D099G, A098V/D099G/G100D, A098V/D099G/G100D/A153V, A098V/D099G/G100S, A098Y/D099R/G100L, A098Y/D099R/G100Z/S101Z/G102D, A098Z/S101R, Z098.01N, Z098.01R, Z098.01R/G100R, D099C/K136N, D099C/S101D, D099C/S101R, D099C/S101Z, D099C/Z099.01S, D099C/Z100.01G, D099F/S101Z, D099G/G100C, D099G/G100D/S101R, D099G/G100H, D099G/G100R/S101V, D099G/G100S/S101R, D099G/S101G, D099G/S101H, D099G/S101V, D099G/S101Z, D099G/S101Z/N269D, D099G/Z099.01N, D099G/Z099.01R/G100S, D099G/Z099.01S, D099G/Z100.01G, D099G/Z100.01G/S183N/Q275K, D099H/S101Z, D099H/S101Z/V150I, D099H/Z100.01G/A273V, D099H/Z100.01G/Z100.02G, D099I/Z099.01S, D099L/N118D, D099L/S101G, D099L/S101V, D099L/Z100.01G, D099N/A116T, D099N/G100R/S101V, D099N/S101G, D099N/S101H, D099N/S101Z, D099N/Z099.01S, D099N/Z100.01G, D099R/S101D, D099R/S101G, D099R/S101V, D099R/S101Z, D099R/S101Z/A133T, D099R/Z099.01N, D099R/Z099.01S, D099R/Z100.01G, D099R/Z100.01N, D099S/S101G, D099S/S101I, D099S/S101Z, D099S/Z099.01H, D099S/Z099.01L, D099S/Z099.01V, D099S/Z100.01G, D099S/Z100.01G/Z100.02G, D099V/S101D, D099V/S101V, D099V/S101Z, Z099.01H/S101L, Z099.01H/Z099.02R, Z099.01N/S101R, Z099.01S, Z099.01S/S101V, Z099.01S/Z099.02G/A228T, G100D/S101G, G100D/S101R, G100H/G102V, G100H/S101G, G100H/S101R, G100L/S101G, G100L/S101R, G100N/S101D, G100N/S101L, G100N/S101R, G100R/L267R/I268V, G100R/S101G, G100R/S101G/G160S/S182N, G100R/S101G/Q103Z/Y104Z/S161N, G100R/S101G/S163N, G100R/S101N, G100R/S101R, G100R/S101R/Q103Z/V270I, G100R/S101Z/G102C/Q103D, G100S/S101C, G100/S101G, G100S/S101R, G100S/S101R, G100S/S101V, G100S/S101V/A231T, Z100.01G, Z100.01L, Z100.01N, Z100.01R, Z100.01R/S101G, S101C/Q103I, S101C/Q103S, S101D/Q103C, S101D/Q103D, S101D/Q103F, S101D/Q103H, S101D/Q103H, S101D/Q103R, S101D/Q103R/H238Y, S101D/Q103S, S101D/Q103V, S101F/Q103G, S101G/A151V, S101G/A179V, S101G/G102S, S101G/Q103G, S101G/Q103L, S101G/Q103N, S101G/Q103R, S101G/Q103R/A179T, S101G/Q103R/G157D, S101G/Q103S, S101G/Q103S/L233S, S101G/Q103S/L233S, S101G/Q245H, S101G/S5249N, S101G/V139I, S101G/Z101.01R, S101G/Z101.01R/A200T, S101G/Z102.01G, S101H/G102S, S101H/Q103D, S101H/Q103G, S101H/Q103G/E251Q, S101H/Q103H, S101H/Q103I, S101H/Q103R, S101H/Q103R/A274Z/Q275Z, S101H/Q103S, S101H/Q103V, S101I/Q103R, S101L/G102S/Q103R, S101L/Q103D, S101L/Q103G, S101L/Q103R, S101L/Q103R/A138T, S101L/Q103R/V121I/K213E, S101L/Q103S, S101L/Q103V, S101L/Z101.01R, S101N/Q103R, S101N/Z102.01G, S101R/A151V, S101R/A273V, S101R/E156G, S101R/G102L/Q103G, S101R/G102S, S101R/G102S/Q103R, S101R/G102S/Q103V, S101R/G131D, S101R/Q103C, S101R/Q103D, 5101R/Q103D/S248R, S101R/Q103G, S101R/Q103G/A116I, S101R/Q103G/V147I/A153V, S101R/Q103G, S101R/Q103L, S101R/Q103N, S101R/Q103R, S101R/Q103R/S161N, S101R/Q103R/V148I, S101R/Q103S, S101R/Q103S/L126F, S101R/Q103V, S101V/Q103G, S101V/Q103H, S101V/Q103N/Z275.01K, S101V/Q103R, S101V/Q103V, S101Y/G102S/Q103R, S101Y/Q103L, S101Y/Q103S, S101Y/Q103V, S101Z/G102I/Q103H/Y104G/L135F, S101Z/G102V/Q103R/Y104L, S101Z/Q103D, S101Z/Q103F, S101Z/Q103L, S101Z/Q103R, G102C/Q103L/Y104S, G102R/Q103C/Y104C/V192I, G102R/Q103G/Y104R, G102R/Q103R/Z103.01G, G102Z/Q103S, Z102.01G/Q103R, Z102.01G/Q103R/L267V, Z102.01G/Z102.02R/Z102.03G, Q103H/Y104N/Q275Z, Q103N/E156D, Q103R/I122L/N123S/M124V, Q103R/Y104F, Q103Y/S182N, S105G/P129S/S130H/G131H, W106C/A116D/N117S/N118C, N109S/M119C/D120G/V121L, A116G/N117Z/N118R/M119C, A116H/N117G/N118R, N117C/N118G/M119S, N117C/N118G/M119S/A216V, N117G/N118G/M119C, N117G/N118R/M119C, N117H/N118C/M119C, N117H/N118D/M119S, N117H/N118G/M119C, N117H/N118G/M119L, N117H/N118G/M119S, N117H/N118G/M119V, N117H/N118H/M119V, N117H/N118R/M119C, N117H/N118R/M119S, N117H/N118S/M119C, N117H/N118S/M119S, N117H/N118S/M119V, N117I/N118G/M119V, N117L/N118G/M119V, N117R/N118G/M119C, N117R/N118G/M119I/V270I, N117R/N118G/M119S, N117R/N118H/M119V, N117R/N118R/M119C, N117R/N118R/M119I, N117R/N118R/M119S, N117R/N118R/M119V, N117R/N118S/M119I, N117R/N118S/M119L, N117S/N118G/M119C, N117S/N118G/M119S, N117S/N118H/M119V, N117S/N118R/M119V, N117V/N118G/M119C, N117V/N118H/M119C, N117Y/N118G/M119S, N117Y/N118G/M119V, Z117.01G/N118R/M119C, N118C/M119V/D120S, N118C/Z118.01G/M119C, N118D/M119C/D120L, N118D/M119S/D120R, N118G/M119C/D120G, N118G/M119C/D120L, N118G/M119I/D120R, N118G/M119N, N118G/M119S/D120G, N118G/M119S/D120R, N118G/M119V/D120G, N118G/M119V/D120S/S182N, N118H/M119C/D120R, N118H/M119I/D120G, N118H/M119N/V148I, N118H/M119S/D120G, N118H/M119V/D120G, N118R/M119C, N118R/M119C/D120G, N118R/M119C/D120R, N118R/M119S, N118R/M119S/D120G, N118R/M119S/D120R, N118R/M119S/D120S, N118R/M119V/D120C, N118R/M119V/D120G, N118R/M119V/D120G/A232T, N118S/M119C/D120R, M119C/D120G, M119C/D120G/V121I, M119C/D120G/V121L, M119C/D120H, M119C/D120H/A187V, M119C/D120R, M119H/V121I, M119I/D120C, M119I/D120G, M119I/D120R, M119L/D120G, M119S/D120R, M119S/D120S/V121I, M119S/V121I, M119V/D120C, M119V/D120G, M119V/D120R, M119V/D120S/V198L, D120G/I122C, D120G/I122C/L217S, D120G/I122G, D120G/I122V, D120H/I122L, D120H/V121C/I122V, D120N/V121L/I122V, D120R/A151V, D120R/I122V, D120R/V121I, D120R/V121I/I122C, D120R/V121I/I122C/A228T, D120S/I122C/A133S, V121C/I122C/N123C, V121C/I122G/N123V, V121C/I122S/N123V, V121C/I122V/N123S, V121I/I12C/N123C, V121I/I122C/V150I, V121I/I122G/N123C, V121I/I122S/N123C, V121I/I122S/N123I, V121I/I122V/N123S, V121L/I122S/N123V, V121L/N123C, I122C/N123C, I122C/N123S/M124L, I122D/N123V/M124L, I122G/N123C/M124L, I122G/N123G/Z123.01C, I122H/N123V/M124LI122S/N123S, I122V/N123G/M124C/A138V, I122V/N123S/M124I, I122V/S183G, N123C/M124S, N123C/M124V, N123S/M124L, N123S/M124S, M124C/L126S, M124C/L126S/T244I, M124C/L126V, M124I/L126H, L126F/P129Z, L126F/P129Z/S182N, L126I/P129Z, L126R/G127R, Z126.01S, G127C/G128S/P129R, G127C/P129G, G127C/P129G/A200T, G127C/P129R, G127C/P129R/A153T, G127D/P129G, G127D/P129S, G127H/P129G, G127H/P129R, G127L/P129R, G127N/P129C, G127N/P129G, G127N/P129G/A133T, G127N/P129G/A151V/K213N, G127N/P129R, G127N/P129S/A273V, G127N/P129V, G127R/A230V, G127R/P129D, G127R/P129D/L217S, G127R/P129G, G127R/P129R, G127R/P129S, G127S/P129D, G127S/P129F, G127S/P129G, G127S/P129G/G166D, G127S/P129G/I175T, G127S/P129R, G127S/P129S, G127S/P129V, G127V/P129G, G127V/P129R, Z127.01G/Z127.02H/P129D, Z127.01G/Z127.02H/P129G, Z127.01G/Z127.02H/P129R, Z127.01H/G128C/P129C/A230V, Z127.01H/P129G, Z127.01H/P129G/V148I, Z127.01H/P129R, Z127.01N/G128C/P129V, Z127.01N/G128S/P129S, Z127.01N/G128S/P129V, Z127.01N/P129R, Z127.01R/G128S/P129S, Z127.01R/P129G, Z127.01S/P129D, Z127.01S/P129G, Z127.01S/P129R, G128D/P129G, G128D/P129R/Z129.01R, G128D/P129S/G154S, G128H/P129C, G128H/P129H/Z129.01G, G128H/P129H/Z129.01R, G128H/P129R, G128H/P129R/Z129.01G, G128H/P129R/Z129.01R, G128H/P129S/Z129.01G, G128H/P129S/Z129.01R, G128H/P129S/Z129.01R/S248N, G128H/P129Y, G128H/Z128.01R/P129D, G128H/Z128.01R/P129S, G128H/Z128.01R/P129S/A144T, G128H/Z129.01R, 128N/P129R/Z129.01G, G128N/P129R/Z129.01R, G128N/P129S/Z129.01D, G128N/Z128.01R/P129S, G128R/P129G, G128R/P129H/Z129.01G, G128R/P129L/Z130.01S, G128R/P129R, G128R/P129R/Z129.01G, G128R/P129R/Z129.01G/A216V, G128R/P129S, G128R/P129S/Z129.01G, 128R/P129Z/S130R/S132G, G128R/Z128.01R/P129D, G128R/Z128.01R/P129F, G128S/P129C, G128S/P129C/Z129.01R, G128S/P129D, G128S/P129D/S248R, G128S/P129D/T244N, G128S/P129G, G128S/P129G/A187V, G128S/P129H/Z129.01R, G128S/P129R/L209F, G128S/P129R/Z129.01G, G128S/P129S/Z129.01D, G128S/P129S/Z129.01R, G128S/P129V, G128Y/P129R/Z129.01G, Z128.01G, Z128.01G/P129D, Z128.01G/P129R, Z128.01G/P129S/A134T, Z128.01G/Z128.02G, Z128.01R/P129D, Z128.01R/Z128.02G/P129S, Z128.01R/Z128.02H/P129G/G160S, Z128.01R/Z128.02R/P129D, Z128.01Y/Z128.02H/P129R, P129C/S130C/S132Z, P129D/Z129.01G/Z129.02D/G160D, P129D/Z129.01G/Z129.02G, P129G/G131Z, P129G/S130G/Z131.01G, P129G/S130H/S132Z, P129G/S130R/S132Z, P129G/V270I, P129G/Z129.01G, P129G/Z129.01R/Z129.02G, P129H/G131Z, P129H/G131Z/S132C, P129H/G131Z/S132H, P129H/S132Z, P129H/Z130.01D/Z130.02S, P129R/G131Z/S132H, P129R/S130D/S132Z, P129R/S130N/G131Z, P129R/S132T, P129R/S132Z, P129R/Z129.01G, P129R/Z129.01G/Z129.02G, P129R/Z129.01G/Z129.02G/A144T, P129R/Z129.01G/Z129.02G/G146D, P129R/Z129.01G/Z129.02R, P129R/Z129.01G, P129R/Z129.01R/Z129.02G, P129S/G131ZP129S/S130H/S132Z, P129S/Z129.01C, P129S/Z129.01R/Z129.02S, P129S/Z130.01N/Z130.02S, P129V/S132Z, P129Z, P129Z/G131R/S132C, P129Z/S130G, P129Z/S130G/G131H/S132H, P129Z/S130G/G131V, P129Z/S130H, P129Z/S130H/S132G, P129Z/S130H/S132N, P129Z/S130L, P129Z/S130R, P129Z/S130R/G131C, P129Z/S130R/G131C, P129Z/S130R/G131D/S132C, P129Z/S130R/G131H/A144E, P129Z/S130R/G131H/S132D, P129Z/S130R/G131R/S132D, P129Z/S130R/L267M, P129Z/S130R/S132C, P129Z/S130R/S132G, P129Z/S130V/S132G, P129Z/S130Z/G131N/S132G, S130D/G131C/S132D, S130D/G131R, S130D/G131R/S132D, S130D/G131Z, S130D/G131Z/S132H, S130F/G131N/S132V, S130F/G131S/S132V, S130F/S132V, S130G/G131D/S132C, S130G/G131H/S132D, S130G/G131S/S132I, S130G/S132G, S130H/G131C/S132G, S130H/G131H/S132R, S130H/G131R/S132C, S130H/G131S, S130H/G131S/S132C, S130H/S132Z, S130I/G131S/S132H, S130I/S132G, S130N/G131S/S132C, S130R/G131R/S132C, S130R/G131S/S132C/S161N, S130R/S132G, S130R/S132G/S183N, S130R/S132Z, S130V/G131D/S132I, S130V/G131H/S132C, S130V/S132N, S130V/S132Z, S130Y/G131H/S132C, S130Y/S132G, S130Z/G131H/S132H, G131H/S132L/W241R, G131N/S132C, G131S/S132G, G131Y/S132G, and A142V/G211V, wherein the positions correspond to the positions of BPN′ subtilisin of SEQ ID NO:2. 
     
     
         2 . A subtilisin variant comprising at least one modification, wherein said modification is selected from: V072G/G097A/G128A/Y217Q, G097A/G128S/Y217Q, G097A/G128S/P129D/Y217Q, G097A/G128A/A134T/Y217Q, M124V/L126A/Y217Q, G097A/G128A/P129D/Y217Q/A232P, G097A/G128A/P129D/Y217Q, G097A/G128S/P129V/A134T/Y217Q, G097A/G128S/P129V/Y217Q, V068I/G097A/G128S/P129D/Y217Q, G097A/S101D/G128S/S204F/Y217Q/S236F/T254P, G097A/S101D/G128S/Y217Q, G097A/G128A/P129V/Y217Q, G097A/S101D/G128A/Y217Q, G097A/S101D/G128S/P129D/Y217Q, G097A/S101D/G128A/A134T/Y217Q, G097A/Z099.01S/A114S/G128S/Y217Q, G097A/S101D/G128A/P194L/Y217Q, G097A/Z099.01S/G128S/Y217Q, G097A/S101D/G128A/P129D/Y217Q, Q019R/G097A/Z099.01S/G128S/P129D/Y217Q, G097A/G100S/G128S/Y217Q, G097A/S101D/G128S/P129V/Y217Q, G097A/Z099.01S/G128A/A134T/Y217Q, G097A/G100S/I107V/G128S/P129D/Y217Q, G097A/G100S/G128S/Y217Q/A230V, G097A/G100S/G128A/Y217Q/K237N, G097A/Z099.01S/G128S/P129D/Y217Q, G097A/G100S/G128S/P129D/Y217Q, Z096.01D/G097A/A098R/G128A/Y217Q, G097A/Z099.01S/I122S/N123G/G128A/Y217Q, G097A/Z099.01S/I122S/N123G/G128A/Y217Q/T220A/A232T, P005S/G097A/Z099.01S/S101D/G128A/Y217Q, G097A/S101D/G128A/P129V/Y217Q, V093Z/K094S/V095C/L096S/G097A/G128A/Y217Q, G097A/G100S/S101D/G128A/Y217Q/A232P, Z096.01D/G097A/A098R/S101D/G128A/Y217Q, and G097A/Z099.01S/G100S/G128A/Y217Q, wherein the positions correspond to the positions of BPN′ subtilisin of SEQ ID NO:2. 
     
     
         3 . A subtilisin variant comprising at least two modifications, wherein said modifications are selected from: G097A/G128S/P129T/Y217Q, G097A/G128A/P129S/Y217Q, G097A/G128S/P129C/Y217Q, G097A/G128S/P129Q/Y217Q, G097A/G128S/P129E/Y217Q, G097A/P129S/Y217Q, G097A/G128S/P129A/Y217Q, G097A/G128S/P129K/Y217Q, G097A/P129N/Y217Q, G097A/P129E/Y217Q, G097A/G128S/P129K/G131S/Y217Q, G097A/G128S/P129R/Y217Q, G097A/G128S/P129V/Y217Q, G097A/G128S/P129Y/Y217Q, G097A/G128A/P129R/Y217Q, G097A/G128H/P129S/Y217Q, G097A/G128N/P129S/Y217Q/V270A, G097A/G128S/P129L/Y217Q, G097A/P129R/Y217Q, G097A/P129W/Y217Q, G097A/P129Z/S130A/Y217Q, G097A/G128H/P129R/Y217Q, G097A/G128H/P129T/Y217Q, G097A/P129G/Y217Q, G097A/G128N/P129A/Y217Q, G097A/G128H/Y217Q, G097A/G128S/P129W/Y217Q, G097A/P129Z/Y217Q, G097A/G128N/Z128.01A/P129S/Y217Q, G097A/G128N/Y217Q, G097A/G128A/P129L/Y217Q, G097A/G128T/P129N/Y217Q, G097A/G128H/P129A/Y217Q, G097A/G128M/P129R/Y217Q, G097A/G128Q/P129K/Y217Q, G097A/G128T/P129Q/Y217Q, G097A/G128T/P129H/Y217Q, G097A/G128T/P129R/Y217Q, G097A/G128D/P129E/Y217Q, G097A/G128T/P129T/Y217Q, G097A/G128T/P129A/Y217Q, G097A/G128Z/P129Z/Y217Q, G097A/G128S/Z128.01A/P129S/Y217Q, G097A/G128N/P129M/Y217Q, G097A/G128N/P129V/Y217Q, G097A/G128E/P129K/Y217Q, G097A/G128N/Z128.01A/P129F/Y217Q, G097A/G128C/P129R/Y217Q, G097A/G128T/P129M/Y217Q, G097A/G128R/Z128.01A/P129S/Y217Q, G097A/G128S/Z128.01A/P129A/Y217Q, G097A/G128D/Y217Q, G097A/G128T/P129Y/Y217Q, G097A/G128R/P129S/Y217Q, G097A/G128R/Z128.01A/P129T/Y217Q, G097A/G128R/Z128.01A/P129A/Y217Q, G097A/G128T/P129W/Y217Q, G097A/G128Y/Z128.01A/P129R/Y217Q, G097A/G128C/P129D/Y217Q, V084L/G097A/P129Z/Y217Q, G097A/G128R/P129D/Y217Q, G097A/G128A/Z128.01A/P129H/E195A/Y217Q, G097A/G128S/Z128.01A/P129Y/Y217Q, G097A/G128N/P129L/Y217Q, G097A/G128C/P129H/Y217Q, G097A/G128T/Z128.01A/P129T/Y217Q, G097A/G128C/Z128.01A/P129A/Y217Q, G097A/G128S/Z128.01A/P129I/Y217Q, G097A/G128S/Z128.01A/P129E/Y217Q/Q271H, G097A/G128S/Z128.01A/P129L/Y217Q, G097A/G128Y/Z128.01A/P129S/Y217Q, G097A/G128S/Z128.01A/P129M/R186H/Y217Q, G097A/G128E/Z128.01A/P129D/Y217Q, G097A/G128T/P129L/Y217Q, G097A/G128F/P129D/Y217Q, G097A/G128V/Z128.01A/P129R/Y217Q, G097A/G128L/Z128.01A/P129S/Y217Q, G097A/G128Y/P129T/Y217Q, G097A/G128C/P129Y/Y217Q, G097A/G128C/P129T/Y217Q, wherein the positions correspond to the positions of BPN′ subtilisin of SEQ ID NO:2. 
     
     
         4 . A subtilisin variant comprising at least two modifications, wherein said modifications are selected from: G097A/G128A/Y217Q/M222Q, M124V/L126A/P129V/Y217Q, G128A/P129V/Y217Q G097A/G128A/P129V/Y217Q, N123G/P129V/Y217Q, V30I/Z096.01D/G097A/A098R/G128A/Y217Q, Z096.01D/A098R/G128A/Y217Q, Z096.01D/A098R/M124V/L126A/Y217Q, G097A/Z098.01S/G128A/Y217Q, Z096.01D/A098R/N123G/Y217Q, Z096.01D/G097A/A098R/G128A/Y217Q/S236F, Z098.01S/M124V/L126A/Y217Q, Z098.01S/G128A/Y217Q, Z096.01D/G097A/A098R/G128A/Y217Q, V093I/Z098.01S/M124V/L126A/Y217Q, A073V/Z098.01S/N123G/Y217Q, G065D/Z098.01S/G128A/Y217Q, I011L/Z098.01S/G128A/Y217Q, L082F/Z096.01D/A098R/N123G/Y217Q, Z098.01S/N123G/A138V/Y217Q, Z098.01S/N123G/Y217Q, wherein the positions correspond to the positions of BPN′ subtilisin of SEQ ID NO:2. 
     
     
         5 . A subtilisin variant comprising the substitution Y217L and further comprising at least one modification set forth in any of  claims 1 - 4 , and wherein the positions correspond to the positions of BPN′ subtilisin of SEQ ID NO:2. 
     
     
         7 . A subtilisin variant comprising the substitutions G97A/G128A/Y217Q and further comprising at least one modification set forth in any of  claims 1 - 4 , and wherein the positions correspond to the positions of BPN′ subtilisin of SEQ ID NO:2. 
     
     
         8 . A cleaning composition comprising at least one subtilisin variant of any of  claims 1  to  7 . 
     
     
         9 . The cleaning composition of  claim 8 , wherein said cleaning composition is a laundry detergent. 
     
     
         10 . The laundry detergent of  claim 9 , wherein said laundry detergent is a heavy duty liquid laundry detergent. 
     
     
         11 . The cleaning composition of  claim 8 , wherein said cleaning composition is a dish detergent. 
     
     
         12 . The cleaning composition of any of  claims 8 - 11 , further comprising one or more additional enzymes or enzyme derivatives selected from the group consisting of hemicellulases, peroxidases, proteases, cellulases, xylanases, lipases, phospholipases, esterases, cutinases, pectinases, keratinases, reductases, oxidases, phenol oxidases, lipoxygenases, ligninases, pullulanases, tannases, pentosanases, malanases, β-glucanases, arabinosidases, hyaluronidase, chondroitinase, laccase, and amylases, or mixtures thereof. 
     
     
         13 . The cleaning composition of any of  claims 8 - 12 , further comprising at least one stabilizing agent. 
     
     
         14 . A cleaning composition comprising at least 0.0001 weight percent of at least one subtilisin variant of any of  claims 1 - 7 , and optionally, at least one suitable adjunct ingredient. 
     
     
         15 . A method of cleaning, said method comprising the steps of:
 a) contacting a surface and/or an article comprising a fabric with the cleaning composition of any of  claims 8 - 14 ; and   b) optionally washing and/or rinsing said surface or article.   
     
     
         16 . An animal feed comprising at least one subtilisin variant of any of  claims 1 - 7 . 
     
     
         17 . A food processing composition comprising at least one subtilisin variant of any of  claims 1 - 7 .