Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
DETAILED ACTION
1. Preliminary amendment and claims 1-22 filed 3/12/25 are present and under consideration.
2. Priority
Acknowledgment is made of applicant's claim for foreign priority based on an application filed in Bangladesh on 12/15/2016. The priority document was received in USSN: 17328003.
3. Applicants’ Claims Priority from Provisional Application 62435340 , filed 12/16/2016, is also acknowledged.
4. Drawings
The drawings filed on 11/7/24 are acknowledged.
5. Specification
The specification has not been checked to the extent necessary to determine the presence of all possible minor errors. Applicant's cooperation is requested in correcting any errors of which applicant may become aware in the specification.
6. IDS filed 11/7/24 is acknowledged. A signed copy of the IDS is provided with this Office Action.
7. Abstract
*This application does not contain an abstract of the disclosure as required by 37 CFR 1.72(b). An abstract on a separate sheet is required.
*The abstract should be in narrative form and generally limited to a single paragraph within the range of 50 to 150 words [in length since the space provided for the abstract on the computer tape by the printer is limited]. The form and legal phraseology often used in patent claims, such as "means" and "said", should be avoided in the abstract. The abstract should sufficiently describe the disclosure to assist readers in deciding whether there is a need for consulting the full patent text for details. MPEP 608.01(b).
Lines 3, 5, 6 & 7 of the abstract recite legal phraseology “said” which must be deleted. Also in line 2, correction is required for the spelling of ‘have’.
8. Sequence Rules
The instant specification, drawings (Figures 2A-2H, 3A-3B & 4A-4B) present alignment of amino acids sequences that are encompassed by the definitions for nucleotide and/or amino acid sequences set forth in 37 CFR 1.821(a)(1) and (a)(2), but fails to comply with the requirements. According to 37 CFR 1.821-825, every disclosed amino acid sequence of four or more residues or 10 or more nucleotides must be identified by a SEQ ID NO? The amino acid sequences presented require having a sequence identifier. In order to comply with the sequence rules Applicants must identify the sequence by providing SEQ ID Nos, and where required provide a new version of the sequence listing and disk.
Applicant must submit a CRF copy and paper copy of the Sequence Listing, a statement that the content of the paper and computer readable copies are the same and where applicable include no new matter as required by 37 C.F.R. j 1.821(e) or 1.821(9 or 1.821(g) or 1 .825(d), as well as an amendment directing its entry into the specification.
Note: If the amino acid sequences are already part of the sequence listing and the CRF, Applicants may amend the specification (legend to the Figures) by providing the appropriate SEQ ID Nos:, following the sequence.
Applicant's cooperation is requested in correcting other unidentified sequences of which applicant may become aware of in the specification.
9. 35 U.S.C. § 112, first paragraph (Written Description)
Claims 1-22 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
Claims 1-22 are directed to the following genus claims:
1. A cellulase variant, or an active fragment thereof, comprising an amino acid sequence comprising two or more mutations at two or more positions corresponding to SEQ ID NO:1 positions selected from: (i) 4,23,29, 32, 34, 36, 44, 51, 65, 77, 80, 87, 90, 97, 98, 99, 102, 112, 116, 119, 135, 136, 142, 153, 154, 156, 157, 161, 163, 178, 192, 194, 202, 204, 205, 206, 208, 210, 212, 217, 221, 222, 225, 227, 232, 233, 236, 237, 238, 241, 247, 249, and 258; (ii) 4, 29, 32, 36, 51, 77, 80, 87, 90, 102, 112, 116, 135, 136, 142, 153, 154, 161, 163, 192, 204, 210, 217, 221, 225, and 227; (iii) 4V, 23L, 29W, 32D/G/Y, 34D, 36T, 44V, 51T, 65G, 77K/M, 80S, 87A, 90A, 97S, 98G/Y, 99E/H/Y, 102K, 112S/T/V, 116V, 119L, 135T, 136E/K/S, 142E/P, 153D, 154E, 156G, 157D, 161E/P, 163V, 178H, 192V, 194S, 202E, 204S, 205D, 206S, 208H/K, 210V, 212S, 217G/M, 221L/M, 222A, 225K, 227R, 232T, 233A/S, 236A, 237D, 238D, 241A/L/R, 247T, 249G/P, and 258K; (iv) 4V, 29W, 32D, 36T, 51T, 77K/M, 80S, 87A, 90A, 102K, 112S/T, 116V, 135T, 136E, 142P, 153D, 154E, 161P, 163V, 192V, 204S, 210V, 217G, 221L, 225K, and 227R; (v) K4, 523, F29, 532, N34, Q36, K44, S51, D65, 577, N80, G87, E90, P97, V98, A99, T102, G112, T116, F119, S135, P136, A142, S153, Q154, D156, S157, A161, K163, N178, L192, A194, D202, G204, N205, F206, V208, T210, P212, S217, S221, S222, S225, K227, 5232, T233, T236, 5237, T238, T241, A247, S249, and G258; (vi) K4, F29, S32, Q36, S51, S77, N80, G87, E90, T102, G112, T116, S135, P136, A142, S153, Q154, A161, K163, L192, G204, T210, S217, S221, S225, and K227; (vii) K4V, S23L, F29W, S32D/G/Y, N34D, Q36T, K44V, S51T, D65G, S77K/M, N80S, G87A, E90A, P97S, V98G/Y, A99E/H/Y, T102K, G112S/T/V, T116V, F119L, S135T, P36E/K/S, A142E/P, S153D, Q154E, D156G, S157D, A161E/P, K163V, N178H, L192V, A194S, D202E, G204S, N205D, F206S, V208H/K, T210V, P212S, S217G/M, S221L/M, S222A, S225K, K227R, S232T, T233A/S, T236A, S237D, T238D, T241A/L/R, A247T, S249G/P, and G258K; or (viii) K4V, F29W, S32D, Q36T, S51T, S77K/M, N80S, G87A, E90A, T102K, G112S/T, T116V, S135T, P136E, A142P, S153D, Q154E, A161P, K163V, L192V, G204S, T210V, S217G, S221L, S225K, and K227R; wherein said variant has endoglucanase activity, and wherein the amino acid positions of the variant, or active fragment thereof, are numbered by correspondence with the amino acid sequence of SEQ ID NO:1.
2. The cellulase variant, or active fragment thereof, of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO:1 positions are selected from: (i) 142+161, 4+142, 36+142, 4+77, 77+142, 87+142, 87+161, 142+204, 4+161, 161+204, 102+161, 142+192, 142+227, 153+161, 4+102, 4+210, 29+153, 32+142, 36+161, 36+221, 36+225, 87+204, 90+142, 116+161, 135+142, 135+154, 135+161, 136+161, 204+221, 4V+80S, 4+153, 4+221, 32+36, 32+142, 77+142, 77+153, 77+161, 77+221, 80+142, 87+153, 87+192, 87+221, 102+136, 102+142, 102+154, 112+135, 112+142, 142+153, 142+217, 142+221, 153+210, 154+161, 161+192, and 161+221; (ii) 142P+161P, 4V+142P, 36T+142P, 4V+77K, 77K+142P, 87A+142P, 87A+161P, 142P+204S, 4V+161P, 161P+204S, 102K+161P, 142P+192V, 142P+227R, 153D+161P, 4V+102K, 4V+210V, 29W+153D, 32D+142P, 36T+161P, 36T+221L, 36T+225K, 87A+204S, 90A+142P, 116V+161P, 135T+142P, 135T+154E, 135T+161P, 136E+161P, 204S+221L, 4V+80S, 4V+153D, 4V+221L, 32D+36T, 32Y+142P, 77M+142P, 77K+153D, 77K+161P, 77K+221L, 80S+142P, 87A+153D, 87A+192V, 87A+221L, 102K+136E, 102K+142P, 102K+154E, 112T+135T, 112S+142P, 142P+153D, 142P+217G, 142P+221L, 153D+210V, 154E+161P, 161P+192V, and 161P+221L; or (iii) A142P+A161P, K4V+A142P, Q36T+A142P, K4V+S77K, S77K+A142P, G87A+A142P, G87A+A161P, A142P+G204S, K4V+A161P, A161P+G204S, T102K+A161P, A142P+L192V, A142P+K227R, S153D+A161P, K4V+T102K, K4V+T210V, F29W+S153D, S32D+A142P, Q36T+A161P, Q36T+S221L, Q36T+S225K, G87A+G204S, E90A+A142P, T116V+A161P, S135T+A142P, S135T+Q154E, S135T+A161P, P136E+A161P, G204S+S221L, K4V+N80S, K4V+S153D, K4V+S221L, S32D+Q36T, S32Y+A142P, S77M+A142P, S77K+S153D, S77K+A161P, S77K+S221L, N80S+A142P, G87A+S153D, G87A+L192V, G87A+S221L, T102K+P136E, T102K+A142P, T102K+Q154E, G112T+S135T, G112S+A142P, A142P+S153D, A142P+S217G, A142P+S221L, S153D+T210V, Q154E+A161P, A161P+L192V, and A161P+S221L.
3. The cellulase variant, or active fragment thereof, of claim 1, wherein said variant further comprises one or more mutations selected from: (i) a deletion of one or more amino acid residues at one or more positions corresponding to any SEQ ID NO:1 position between positions 213 and 257; (ii) a deletion of one or more amino acid residues at one or more positions corresponding to SEQ ID NO:1 positions selected from D177, S232, T233, S234, T235, T236, S237, T238, K239, A240, T241, S242, and T243; (iii) a deletion of one or more amino acid residues at one or more positions corresponding to SEQ ID NO:1 positions selected from D177, S232, T233, S234, T235, T236, S237, T238, K239, A240, T241, S242, and T243, wherein said deletions are selected from D177Z, T233Z, T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, and S232Z-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z; (iv) an insertion of one, two, three, four, five, six, or more amino acid residues after a position corresponding to position 233 of SEQ ID NO:1; (v) an insertion of one, two, three, four, five, six, or more amino acid residues after a position corresponding to position 233 of SEQ ID NO:1, wherein said insertion is selected from Z233.01T-Z233.02T-Z233.03S-Z233.04T-Z233.05S-Z233.06T, Z233.01T-Z233.02A-Z233.03S-Z233.04T-Z233.05K-Z233.06A-Z233.07T-Z233.08S-Z233.09T-Z233.10T, Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05D-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, Z233.01S-Z233.02T-Z233.03T-Z233.04D-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, and Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T; and (vi) a combination of (i) to (v).
4. The cellulase variant, or active fragment thereof, of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO:1 positions are selected from: K4V-Q36T-S77M-T102K-Q154E-N178H-S225K-S232Z-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, K4V-K44V-S77K-E90A-S135T-A142P-Q154E-L192V-V208H-S221L-S232Z-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z-A247T, K4V-N80S-G87A-Q154E-A161P-G204S-S221L-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, S32Y-T102K-A142P-G204S-V208H-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z-A247T, K4V-K44V-S77K-N80S-T102K-G112T-S135T-A142P-L192V-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, S77M-A142P-K163V-V208H-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z-A247T, S77M-G87A-A99H-G112T-A142P-S232T-T233Z-T236A, S23L-A142P-D202E-S232T-T233Z, Q36T-S77K-G87A-A142P-S153D-A161P-G204S-S221L-S237D, K4V-F29W-S77K-T102K-P136E-S153D-A161P-T210V, A99Y-G112T-S135T-S153D-K227R, S23L-S32Y-G112T-A142P-A161P-G204S-S217G-K227R-S237D, F29W-Q36T-K44V-T116V-S135T-Q154E-A161P-S217G-S221L-S225K-T238D, K4V-Q36T-S77K-N80S-A142P-L192V-G204S-T210V-S221L, F29W-G87A-E90A-A99Y-A142P-S153D-K227R-S237D, K4V-S77M-Q154E-A161P-L192V-V208H-S217G-T233A-S237D, E90A-G112T-S135T-A161P-G204S, S51T-S77M-P136E-K163V-G204S-T210V-K227R, K4V-F29W-S51T-T116V-S135T-S221L-S225K, S77M-T102K-T116V-S135T-S153D-A161P-T238D, K4V-S23L-S32Y-N80S-T102K-A142P-A161P-S237D, K44V-G87A-Q154E-A161P, S32Y-A142P-Q154E-K163V-S217G-K227R-T233A, N34D-S51T-G87A-T102K-T116V-P136E-Q154E-A161P-L192V-T233A-T238D, K4V-S77K-E90A-A142P-S157D-G214Z-S217G-T233A-T238D, K4V-Q36T-S51T-N80S-S135T-S157D-T233A, K4V-S51T-V98G-S135T-Q154E-T233A, T102K-A142P-Q154E-G204S-K227R, S51T-G87A-A99Y-S135T-N205D-T238D, G87A-A142P-A161P-L192V-G204S-T241A, K4V-S77K-E90A-A142P-S157D-S217G-T233A-T238D, K4V-P136E-S153D-A161P-T210V-K227R, S51T-N80S-A142P-L192V-V208H, K4V-S23L-Q36T-S77M-G112T-S157D-A161P-L192V-S221L, Q36T-V98Y-A142P-A161E-S225K, Q36T-P136E-A142P-S225K-S249P, A99Y-P136E-A142P-A161E-S221L, S32D-Q36T-S221L-S225K, S32D-A99Y-A142P-A161E-S249P, K44V-A142P-A161P-K163V, S32D-Q36T-G112S-A142P, S32D-Q36T-S77M-S135T-A142P-A161P, F29W-S77M-A99E-A142P, G112S-P136K-A142E, S135T-A142P-K163V, K4V-GI 12S-S135T-A142P-A161P, S32D-Q36T-A99E-P136K-A142P, G112S-P136K-A142P-S217M-S221M-T241L-G258K, K4V-S77K-GI 12S-A142P-A194S-S249G-G258K, P97S-G112V-P136S-A142P-A161E-K163V-P212S-S222A, P97S-G112V-P136S-S153D-A161E-S225K-T241R, P97S-G112V-P136S-S153D-S225K-T241R, S32D-A142E-V208K-S225K-T241R, S32D-A142E, S51T-A99Y-F119L-P136E-Q154E-D156G-S217G, K44V-A142P-A161P-K163V-K227R-T236A, G112T-S135T-Q154E-T233S-Z233.01T-Z233.02T-Z233.03S-Z233.04T-Z233.05S-Z233.06T-S237D, Q36T-S51T-V98G-T116V-A142P-A161P-T233S-Z233.01T-Z233.02A-Z233.03S-Z233.04T-Z233.05K-Z233.06A-Z233.07T-Z233.08S-Z233.09T-Z233.10T, Q36T-S77K-G87A-A142P-S153D-A161P-D177Z-G204S-S221L-S237D, K4V-F29W-S51T-D65G-S77M-T116V-S153D-K163V-G204S-F206S-T210V-S221L-Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, S77M-S135T-Q154E-K163V-K227R-Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05D-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T-S237D, K4V-S32G-Q36T-S77M-G87A-T116V-A142P-A161P-Z233.01S-Z233.02T-Z233.03T-Z233.04D-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, and S77M-G87A-P136E-S157D-S221L-Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T-T236A-T238D-K286E.
5. The cellulase variant or active fragment of claim 1, with the proviso that one or more of said mutations is non-naturally occurring.
6. The cellulase variant, or active fragment thereof, of claim 1, wherein said variant comprises (i) an amino acid sequence corresponding to position 1-295 of SEQ ID NO:1 and having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or less than 100% amino acid sequence identity to the amino acid sequence of SEQ ID NO:1, 70, 71, 72, 73, 74, or 75; (ii) an amino acid sequence corresponding to position 1-211 of SEQ ID NO:1 and having at least 70%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or less than 100% amino acid sequence identity to an amino acid sequence as shown in positions 1-211 of SEQ ID NO:1, 70, 71, 72, 73, 74, or 75 set forth in FIG. 2; (iii) an amino acid sequence corresponding to position 212-258 of SEQ ID NO:1 and having at least 70%, 75%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to an amino acid sequence as shown in positions 212-258 of SEQ ID NO:1, 70, 71, 72, 73, 74, or 75 set forth in FIG. 3; (iv) an amino acid sequence corresponding to position 259-295 of SEQ ID NO:1 and having at least 60%, 67%, 68%, 70%, 73%, 75%, 76%, 80%, 81%, 83%, 84%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% amino acid sequence identity to an amino acid sequence as shown in positions 259-295 of SEQ ID NO:1, 70, 71, 72, 73, 74, or 75 set forth in FIG. 4; or (v) a combination of (i) to (iv).
7. The cellulase variant, or active fragment thereof, of claim 1, wherein said variant has at least one improved property selected from improved thermostability, stability in the presence of one or more protease, and stability in the presence of one or more protease and one or more other detergent component when compared to a parent or reference polypeptide.
8. The cellulase variant, or active fragment thereof, of claim 1, wherein said variant is derived from a parent or reference polypeptide selected from SEQ ID NOs:1, 70, 71, 72, 73, 74, and 75.
9. The cellulase variant, or active fragment thereof, of claim 1, wherein the improved property is (i) improved thermostability and wherein said variant, or active fragment thereof, has a thermal PI that is greater than 1 or ≥1.1; (ii) improved stability in the presence of one or more protease and wherein said variant, or active fragment thereof, has a PI that is greater than 1 or ≥1.1, 1.2, 1.3, 1.4, 1.5, or 2.0 when the stability of said variant, or active fragment thereof is tested in the presence of said protease; and/or (iii) improved stability in the presence of one or more protease and one or more other detergent component, and wherein said variant, or active fragment thereof, has a PI that is greater than 1 or ≥1.1, 1.5, 2.0, or 2.5 when the stability of said variant, or active fragment thereof is tested in the presence of said protease and said other detergent component.
10. The cellulase variant, or active fragment thereof, of claim 9, wherein the PI is measured in accordance with the Cellulase Activity Assay of Example 1.
11. The cellulase variant, or active fragment thereof, of claim 9, wherein the other detergent component is a surfactant.
12. The cellulase variant, or active fragment thereof, of claim 1, wherein said variant, or active fragment thereof, is a family GH45 cellulase.
13. A composition comprising the cellulase variant, or active fragment thereof, of claim 1.
14. The composition of claim 13, wherein said composition is selected from an enzyme composition, detergent composition, and fabric care composition.
15. The composition of claim 13, further comprising (i) one or more other enzymes selected from the group consisting of acyl transferases, amylases, alpha-amylases, beta-amylases, alpha-galactosidases, arabinases, arabinosidases, aryl esterases, beta-galactosidases, beta-glucanases, carrageenases, catalases, chondroitinases, cutinases, endo-beta-mannanases, exo-beta-mannanases, esterases, exo-mannanases, galactanases, glucoamylases, hemicellulases, hyaluronidases, keratinases, laccases, lactases, ligninases, lipases, lipolytic enzymes, lipoxygenases, mannanases, metalloproteases, oxidases, oxidoreductases, pectate lyases, pectin acetyl esterases, pectinases, pentosanases, perhydrolases, peroxidases, phenoloxidases, phosphatases, phospholipases, phytases, polyesterases, polygalacturonases, proteases, pullulanases, reductases, rhamnogalacturonases, second cellulase, beta-glucanases, tannases, transglutaminases, xylan acetyl-esterases, xylanases, and xylosidases; (ii) one or more surfactants; (iii) one or more ions selected from calcium and zinc; (iv) one or more adjunct ingredients; (v) one or more stabilizers; (vi) from about 0.001% to about 5.0 weight % of said cellulase variant, or active fragment thereof of any one of claims 1-15; (vii) one or more bleaching agents; and/or (viii) combinations thereof.
16. The composition of claim 13, wherein said composition is a laundry detergent.
17. The composition of claim 13, wherein the composition is in a form selected from a liquid, a powder, a granulated solid, a tablet, a sheet, and a unit dose.
18. The composition of claim 13, wherein said composition contains phosphate or is phosphate-free and/or contains boron or is boron-free.
19. A polynucleotide comprising a nucleic acid sequence encoding the cellulase variant, or active fragment thereof of claim 1.
20. An expression vector comprising the polynucleotide of claim 19.
21. A host cell comprising the expression vector of claim 20.
22. A method for producing the cellulase variant, or active fragment thereof comprising: (a) stably transforming a host cell of with the expression vector of claim 20; (b) cultivating said transformed host cell under conditions suitable for said host cell to produce said cellulase variant or active fragment thereof; and (c) recovering said cellulase variant or active fragment thereof.
In University of California v. Eli Lilly & Co., 43 USPQ2d 1938, the Court of Appeals for the Federal Circuit has held that “A written description of an invention involving a chemical genus, like a description of a chemical species, ‘requires a precise definition, such as by structure, formula, [or] chemical name,’ of the claimed subject matter sufficient to distinguish it from other materials”. As indicated in MPEP § 2163, the written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, reduction to drawings, or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show that Applicant was in possession of the claimed genus. In addition, MPEP § 2163 states that a representative number of species means that the species which are adequately described are representative of the entire genus. Thus, when there is substantial variation within the genus, one must describe a sufficient variety of species to reflect the variation within the genus.
The specification, however, only provides description of a 53 mutant species of SEQ ID NO: 1 including the elected one consisting of the variant at position 161 in the instant specification. The specification thus provide description of about 17.96% modification of the sequence of SEQ ID NO: 1 (295 amino acids in length) or a sequence that is about 82% identical to the sequence of SEQ ID NO: 1 with a substitution at position 161.
The specification does not contain any disclosure or description of the structure and function of all cellulase variant sequences from any source and active fragments thereof, or having a sequence homology of at least 60%, 70%, 75%, 80%, identical to the amino acid sequence of SEQ ID NO: 1 and has endoglucanase activity.
The 53 variant species disclosed from Staphylotrichum coccosporum are not representative of the genus claimed. According to MPEP 2163, to satisfy the written description requirement, a patent specification must describe the claimed invention in sufficient detail that one skilled in the art can reasonably conclude that the inventor had possession of the claimed invention. See, e.g., Moba, B.V. v.Diamond Automation, Inc., 325 F.3d 1306, 1319, 66 USPQ2d 1429, 1438 (Fed.Cir. 2003); Vas-Cath, Inc. v. Mahurkar, 935 F.2d at 1563, 19 USPQ2d at 1116.
The scope of each genus includes many members of cellulase enzymes with widely differing structural, chemical, and physical characteristics. Furthermore, each genus is highly variable because a significant number of structural differences between genus members exit. The specification does not describe and define any structural features and amino acid sequences commonly possessed by each genus. There is no art-recognized correlation between any structure of a cellulase (and/or fragment thereof) and sequences having varying sequence homology, i.e., less than 82% of SEQ ID NO: 1. One of skill in the art would not be able to identify without further testing what specific cellulase sequences would have endoglucanase activity. There is no teaching regarding which 18% (or more) of the sequence that can be varied and still result in a protein having endoglucanase activity. An important consideration is that structure is not necessarily a reliable indicator of function. The instant specification provides no disclosure relating similarity or identity of structure to conservation of function. General knowledge in the art provides guidance to modification of some amino acids that are tolerated without losing a protein’s tertiary structure.
An important consideration is that structure is not necessarily a reliable indicator of function. In this example, there is no disclosure relating similarity of structure to conservation of function. General knowledge in the art included the knowledge that some amino acid variations are tolerated without losing a protein’s tertiary structure. The results of amino acid substitutions have been studied so extensively that amino acids are grouped in so-called “exchange groups” of similar properties because substituting within the exchange group is expected to conserve the overall structure. For example, the expectation from replacing leucine with isoleucine would be that the protein would likely retain its tertiary structure. On the other hand, when non-exchange group members are substituted, e.g., proline for tryptophan, the expectation would be that the substitution would not likely conserve the protein’s tertiary structure. Given what is known in the art about the likely outcome of substitutions on structure, those in the art would have likely expected the applicant to have been in possession of a genus of proteins having a tertiary structure similar to SEQ ID NO: 1 although the claim is not so limited. However, conservation of structure is not necessarily a surrogate for conservation of function. In this case, there is no disclosed correlation between structure and function. There is no disclosure of the active site amino acid residues responsible for the catalytic activity. While general knowledge in the art may have allowed one of skill in the art to identify other proteins expected to have the same or similar tertiary structure, in this case there is no general knowledge in the art about similar proteins to SEQ ID NO: 1 to suggest that general similarity of structure confers the activity. Accordingly, one of skill in the art would not accept the disclosure of SEQ ID NO: 1 and 53 mutants as representative of other proteins or fragments thereof having endoglucanase activity. The specification, taken with the pre-existing knowledge in the art of amino acid substitution and the genetic code, fails to satisfy the written description requirement of 35 U.S.C. 112, first paragraph.
10. Claim(s) 1-3 & 5-22 is/are rejected under 35 U.S.C. 102(a)(1)/(a)(2) as being anticipated by Lai et al. [US 20190106690 A1 (November 16, 2015)]. Lai et al. teach cellulase variants and polynucleotides encoding the variants. And provided are nucleic acid constructs, vectors, and host cells comprising the polynucleotides. Furthermore provided are methods of using the variants (See abstract). The cellulase variants, comprising an alteration at one or more (e.g., several) positions corresponding to positions 292, 274, 266, 265, 255, 246, 237, 224 and 221, wherein each alteration is independently a substitution, insertion or deletion and the variant has cellulase activity, are disclosed among others.
The following sequence present various additional positional differences which read upon the various mutations or variants claimed and are derived from the alignment of Applicants’ SEQ ID NO: 1 and the SEQ ID NO: 2 of US20190106690A1 (Lai et al.). SEQ ID NO: 2 being 90.1% identical to Applicants’ SEQ ID NO: 1.
These cellulase variants include:
K4Q, S23A, F29Y, S32D, K44Q, S77A, N80S, E90A, T116S, P136S, A161P, L192I and A142P. Also Q36 can also be mutated with Q36Q unless mutated with a different amino acid, and such a recitation is lacking.
Result 86
US-15-775-120-2
; Sequence 2, Application US/15775120
; Publication No. US20190106690A1
; GENERAL INFORMATION
; APPLICANT: Novozymes A/S
; TITLE OF INVENTION: CELLULASE VARIANTS AND POLYNUCLEOTIDES ENCODING SAME
; FILE REFERENCE: 13134-US-PCT[2]
; CURRENT APPLICATION NUMBER: US/15/775,120
; CURRENT FILING DATE: 2018-05-10
; PRIOR APPLICATION NUMBER: PCT/CN2016/105927
; PRIOR FILING DATE: 2016-11-15
; PRIOR APPLICATION NUMBER: PCT/CN2015/094695
; PRIOR FILING DATE: 2015-11-16
; NUMBER OF SEQ ID NOS: 79
; SOFTWARE: PatentIn version 3.5
; SEQ ID NO 2
; LENGTH: 318
; TYPE: PRT
; ORGANISM: Artificial Sequence
; FEATURE:
; OTHER INFORMATION: Synthetic Construct
US-15-775-120-2
Query Match 90.1%; Score 1478; DB 18; Length 318;
Best Local Similarity 88.1%;
Matches 258; Conservative 24; Mismatches 11; Indels 0; Gaps 0;
Qy 3 GKSTRYWDCCKPSCSWPGKASVNQPVFACSANFQRISDPNVKSGCDGGSAYACADQTPWA 62 Q36Q
|:||||||||||||:|||||:|:|||:|| |||||:|| ||:|||:|||||:||||||||
Db 26 GQSTRYWDCCKPSCAWPGKAAVSQPVYACDANFQRLSDFNVQSGCNGGSAYSCADQTPWA 85
Qy 63 VNDNFSYGFAATSISGGNEASWCCGCYELTFTSGPVAGKTMVVQSTSTGGDLGTNHFDLA 122
|||| :||||||||:||:|:|||| || |||||||||||||||||||||||||:||||:|
Db 86 VNDNLAYGFAATSIAGGSESSWCCACYALTFTSGPVAGKTMVVQSTSTGGDLGSNHFDIA 145
Qy 123 MPGGGVGIFDGCSPQFGGLAGDRYGGVSSRSQCDSFPAALKPGCYWRFDWFKNADNPTFT 182 A61P
|||||||||:||| ||||| | :|||:||| ||||||| ||||| ||||||:||||||||
Db 146 MPGGGVGIFNGCSSQFGGLPGAQYGGISSRDQCDSFPAPLKPGCQWRFDWFQNADNPTFT 205
Qy 183 FRQVQCPSELVARTGCRRNDDGNFPVFTPPSGGQSSSSSSSSSAKPTSTSTSTTSTKATS 242
|:|||||:|:|||:||:|||||||||||||||||||||||||||||||||||||||||||
Db 206 FQQVQCPAEIVARSGCKRNDDGNFPVFTPPSGGQSSSSSSSSSAKPTSTSTSTTSTKATS 265
Qy 243 TTSTASSQTSSSTGGGCAAQRWAQCGGIGFSGCTTCVSGTTCNKQNDWYSQCL 295
|||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 266 TTSTASSQTSSSTGGGCAAQRWAQCGGIGFSGCTTCVSGTTCNKQNDWYSQCL 318
Having the claimed structure, the improved property such as improved thermostability and wherein said variant, or active fragment thereof, has a thermal PI that is greater than 1 or 1.1, will be considered the inherent property of the cellulase variant of Lai et al.
The reference further teaches addition of protease from various sources such as Aspergillus oryzae alkaline protease, Aspergillus oryzae triose phosphate isomerase, Fusarium oxysporum trypsin-like protease (WO 96/00787), Fusarium venenatum amyloglucosidase (WO 00/56900), Fusarium venenatum Daria (WO 00/56900), Fusarium venenatum Quinn (WO 00/56900). [0429] Scouring is used to remove impurities from the fibers, to swell the fibers and to remove seed coat. It is one of the most critical steps. The main purposes of scouring is to a) uniformly clean the fabric, b) soften the motes and other trashes, c) improve fabric absorbency, d) saponify and solubilize fats, oils, and waxes, and e) minimize immature cotton. Sodium hydroxide scouring at about boiling temperature is the accepted treatment for 100% cotton, while calcium hydroxide and sodium carbonate are less frequently used. Synthetic fibers are scoured at much milder conditions. Surfactant and chelating agents are essential for alkaline scouring. Enzymatic scouring has been introduced, wherein cellulase, hemicellulase, pectinase, lipase, and protease are all reported to have scouring effects. Also Provided are cellulase variants and polynucleotides encoding the variants. Also provided are nucleic acid constructs, vectors, and host cells comprising the polynucleotides. Furthermore provided are methods of using the variants. The reference anticipates the claims.
11. Double Patenting Rejection
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the claims at issue are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); and In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on a nonstatutory double patenting ground provided the reference application or patent either is shown to be commonly owned with this application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP §§ 706.02(l)(1) - 706.02(l)(3) for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
Effective January 1, 1994, a registered attorney or agent of record may sign a terminal disclaimer. A terminal disclaimer signed by the assignee must fully comply with 37 CFR 3.73(b).
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/forms/. The filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to http://www.uspto.gov/patents/process/file/efs/guidance/eTD-info-I.jsp.
Claims 1-22 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-22 of U.S. Patent No. 11,053,488. Although the claims at issue are not identical, they are not patentably distinct from each other because of the following reason(s).
Instant claims are drawn to as shown above in paragraph 9.
Claims 1-22 of U.S. Patent No. 11,053,488 are drawn to as follows.
1. A cellulase variant comprising an amino acid sequence comprising two or more mutations at two or more positions corresponding to SEQ ID NO:1 wherein said two or more mutations comprise a mutation at position 161 and at least one additional mutation at a position selected from 4, 23, 29, 32, 34, 36, 44, 51, 65, 77, 80, 87, 90, 97, 98, 99, 102, 112, 116, 119, 135, 136, 142, 153, 154, 156, 157, 163, 178, 192, 194, 202, 204, 205, 206, 208, 210, 212, 217, 221, 222, 225, 227, 232, 233, 236, 237, 238, 241, 247, 249, and 258, wherein said variant has endoglucanase activity, wherein said variant has an amino acid sequence having at least 91% amino acid sequence identity to the amino acid sequence of SEQ ID NO:1, and wherein the amino acid positions of the variant, are numbered by correspondence with the amino acid sequence of SEQ ID NO:1.
2. The cellulase variant, of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO:1 positions are selected from: (i) 142+161, 87+161, 4+161, 161+204, 102+161, 153+161, 36+161, 116+161, 135+161, 136+161, 77+161, 154+161, 161+192, and 161+221; (ii) 142P+161P, 87A+161P, 4V+161P, 161P+204S, 102K+161P, 153D+161P, 36T+161P 116V+161P, 135T+161P, 136E+161P, 77K+161P, 154E+161P, 161P+192V, and 161P+221L; or (iii) A142P+A161P, G87A+A161P, K4V+A161P, A161P+G204S, T102K+A161P, S153D+A161P, T116V+A161P, S135T+A161P, P136E+A161P, S77K+A161P, Q154E+A161P, A161P+L192V, and A161P+S221L.
3. The cellulase variant, of claim 1, wherein said variant further comprises one or more mutations selected from: (i) a deletion of one or more amino acid residues at one or more positions corresponding to any SEQ ID NO:1 position between positions 213 and 257; (ii) a deletion of one or more amino acid residues at one or more positions corresponding to SEQ ID NO:1 positions selected from D177, S232, T233, S234, T235, T236, S237, T238, K239, A240, T241, S242, and T243; (iii) a deletion of one or more amino acid residues at one or more positions corresponding to SEQ ID NO:1 positions selected from D177, S232, T233, S234, T235, T236, S237, T238, K239, A240, T241, S242, and T243, wherein said deletions are selected from D177Z, T233Z, T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, and S232Z-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z; (iv) an insertion of one, two, three, four, five, six , or more amino acid residues after a position corresponding to position 233 of SEQ ID NO:1; (v) an insertion of one, two, three, four, five, six , or more amino acid residues after a position corresponding to position 233 of SEQ ID NO:1, wherein said insertion is selected from Z233.01T-Z233.02T-Z233.03S-Z233.04T-Z233.05S-Z233.06T, Z233.01T-Z233.02A-Z233.03S-Z233.04T-Z233.05K-Z233.06A-Z233.07T-Z233.08S-Z233.09T-Z233.10T, Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05D-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, Z233.01S-Z233.02T-Z233.03T-Z233.04D-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, and Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T; and (vi) a combination of (i) to (v).
4. The cellulase variant of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO:1 positions are selected from: K4V-N80S-G87A-Q154E-A161P-G204S-S221L-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, Q36T-S77K-G87A-A142P-S153D-A161P-G204S-S221L-S237D, K4V-F29W-S77K-T102K-P136E-S153D-A161P-T210V, S23L-S32Y-G112T-A142P-A161P-G204S-S217G-K227R-S237D, F29W-Q36T-K44V-T116V-S135T-Q154E-A161P-S217G-S221L-S225K-T238D, K4V-S77M-Q154E-A161P-L192V-V208H-S217G-T233A-S237D, E90A-G112T-S135T-A161P-G204S, S77M-T102K-T116V-S135T-S153D-A161P-T238D, K4V-S23L-S32Y-N80S-T102K-A142P-A161P-S237D, K44V-G87A-Q154E-A161P, N34D-S51T-G87A-T102K-T116V-P136E-Q154E-A161P-L192V-T233A-T238D, G87A-A142P-A161P-L192V-G204S-T241A, K4V-P136E-S153D-A161P-T210V-K227R, K4V-S23L-Q36T-S77M-G112T-S157D-A161P-L192V-S221L, Q36T-V98Y-A142P-A161E-S225K, Q36T-P136E-A142P-S225K-S249P, A99Y-P136E-A142P-A161E-S221L, S32D-A99Y-A142P-A161E-S249P, K44V-A142P-A161P-K163V, S32D-Q36T-G112S-A142P, S32D-Q36T-S77M-S135T-A142P-A161P, K4V-G112S-S135T-A142P-A161P, P97S-G112V-P136S-A142P-A161E-K163V-P212S-S222A, P97S-G112V-P136S-S153D-A161E-S225K-T241R, K44V-A142P-A161P-K163V-K227R-T236A, Q36T-S51T-V98G-T116V-A142P-A161P-T233S-Z233.01T-Z233.02A-Z233.03S-Z233.04T-Z233.05K-Z233.06A-Z233.07T-Z233.08S-Z233.09T-Z233.10T, Q36T-S77K-G87A-A142P-S153D-A161P-D177Z-G204S-S221L-S237D, and K4V-S32G-Q36T-S77M-G87A-T116V-A142P-A161P-Z233.01S-Z233.02T-Z233.03T-Z233.04D-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T.
5. The cellulase variant of claim 1, with the proviso that one or more of said mutations is non-naturally occurring.
6. The cellulase variant of claim 1, wherein said variant has at least one improved property selected from improved thermostability, stability in the presence of one or more protease, and stability in the presence of one or more protease and one or more other detergent component when compared to a parent or reference polypeptide.
7. The cellulase variant of claim 1, wherein said variant is derived from a parent or reference polypeptide selected from SEQ ID NOs:1, 70, 71, 72, 73, 74, and 75.
8. The cellulase variant of claim 1, wherein the improved property is (i) improved thermostability and wherein said variant, has a thermal PI that is greater than 1 or ≥1.1; (ii) improved stability in the presence of one or more protease and wherein said variant, has a PI that is greater than 1 or ≥1.1, 1.2, 1.3, 1.4, 1.5, or 2.0 when the stability of said variant, is tested in the presence of said protease; and/or (iii) improved stability in the presence of one or more protease and one or more other detergent component, and wherein said variant, has a PI that is greater than 1 or ≥1.1, 1.5, 2.0, or 2.5 when the stability of said variant, is tested in the presence of said protease and said other detergent component.
9. The cellulase variant of claim 8, wherein the PI is measured in accordance with the Cellulase Activity Assay of Example 1.
10. The cellulase variant of claim 8, wherein the other detergent component is a surfactant.
11. The cellulase variant of claim 1, wherein said variant, is a family GH45 cellulase.
12. A composition comprising the cellulase variant of claim 1.
13. The composition of claim 12, wherein said composition is selected from an enzyme composition, detergent composition, and fabric care composition.
14. The composition of claim 12, further comprising (i) one or more other enzymes selected from the group consisting of acyl transferases, amylases, alpha-amylases, beta-amylases, alpha-galactosidases, arabinases, arabinosidases, aryl esterases, beta-galactosidases, beta-glucanases, carrageenases, catalases, chondroitinases, cutinases, endo-beta-mannanases, exo-beta-mannanases, esterases, exo-mannanases, galactanases, glucoamylases, hemicellulases, hyaluronidases, keratinases, laccases, lactases, ligninases, lipases, lipolytic enzymes, lipoxygenases, mannanases, metalloproteases, oxidases, oxidoreductases, pectate lyases, pectin acetyl esterases, pectinases, pentosanases, perhydrolases, peroxidases, phenoloxidases, phosphatases, phospholipases, phytases, polyesterases, polygalacturonases, proteases, pullulanases, reductases, rhamnogalacturonases, second cellulase, beta-glucanases, tannases, transglutaminases, xylan acetyl-esterases, xylanases, and xylosidases; (ii) one or more surfactants; (iii) one or more ions selected from calcium and zinc; (iv) one or more adjunct ingredients; (v) one or more stabilizers; (vi) from about 0.001% to about 5.0 weight % of said cellulase variant; (vii) one or more bleaching agents; and/or (viii) combinations thereof.
15. The composition of claim 12, wherein said composition is a laundry detergent.
16. The composition of claim 12, wherein the composition is in a form selected from a liquid, a powder, a granulated solid, a tablet, a sheet, and a unit dose.
17. The composition of claim 12, wherein said composition contains phosphate or is phosphate-free and/or contains boron or is boron-free.
18. A polynucleotide comprising a nucleic acid sequence encoding the cellulase variant, of claim 1.
19. An expression vector comprising the polynucleotide of claim 18.
20. A host cell comprising the expression vector of claim 19.
21. A method for producing the cellulase variant, comprising: (a) cultivating a transformed host cell of claim 20 under conditions suitable for said host cell to produce said cellulase variant; and (b) recovering said cellulase variant.
22. The cellulase variant, of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO:1 positions comprise a mutation A161E/P, and at least one additional mutation selected from the group consisting of K4V, S23L, F29W, S32D/G/Y, N34D, Q36T, K44V, S51T, D65G, S77K/M, N80S, G87A, E90A, P97S, V98G/Y, A99E/H/Y, T102K, G112S/T/V, T116V, F119L, S135T, P36E/K/S, A142E/P, S153D, Q154E, D156G, S157D, K163V, N178H, L192V, A194S, D202E, G204S, N205D, F206S, V208H/K, T210V, P212S, S217G/M, S221L/M, S222A, S225K, K227R, S232T, T233A/S, T236A, S237D, T238D, T241A/L/R, A247T, S249G/P, and G258K.
Given the fact pattern of the instant case as well as the patent the species claims of the patent anticipates the instant genus (or broad) claims.
12. Claims 1-22 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-21 of U.S. Patent No. 12,180,522. Although the claims at issue are not identical, they are not patentably distinct from each other because of the following reason(s).
Instant claims are drawn to as shown above in paragraph 9.
Claims of the US Patent 12,180,522 are drawn to as follows.
1. A cellulase variant, comprising an amino acid sequence comprising two or more mutations at two or more positions corresponding to SEQ ID NO:1 wherein said two or more mutations comprise a threonine at position 36 and at least one additional mutation at a position selected from 4, 23, 29, 32, 34, 44, 51, 65, 77, 80, 87, 90, 97, 98, 99, 102,112, 116, 119, 135, 136, 142, 153, 154, 156, 157, 161, 163, 178, 192, 194, 202, 204, 205, 206, 208, 210, 212, 217, 221, 222, 225, 227, 232, 233, 236, 237, 238, 241, 247, 249, and 258, wherein said variant has endoglucanase activity, wherein said variant has an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence of SEQ ID NO:1, and wherein the amino acid positions of the variant, are numbered by correspondence with the amino acid sequence of SEQ ID NO:1.
2. The cellulase variant of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO: 1 positions are selected from: Q36T+A142P, Q36T+A161P, Q36T+S221L, Q36T+S225K, S32D+Q36T.
3. The cellulase variant, of claim 1, wherein said variant further comprises one or more mutations selected from: (i) a deletion of one or more amino acid residues at one or more positions corresponding to any SEQ ID NO:1 position between positions 213 and 257; (ii) a deletion of one or more amino acid residues at one or more positions corresponding to SEQ ID NO: 1 positions selected from D177, S232, T233, S234, T235, T236, S237, T238, K239, A240, T241, S242, and T243; (iii) a deletion of one or more amino acid residues at one or more positions corresponding to SEQ ID NO: 1 positions selected from D177, S232, T233, S234, T235, T236, S237, T238, K239, A240, T241, S242, and T243, wherein said deletions are selected from D177Z, T233Z, T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, and S232Z-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z; (iv) an insertion of one, two, three, four, five, six, or more amino acid residues after a position corresponding to position 233 of SEQ ID NO:1; (v) an insertion of one, two, three, four, five, six, or more amino acid residues after a position corresponding to position 233 of SEQ ID NO:1, wherein said insertion is selected from Z233.01T-Z233.02T-Z233.03S-Z233.04T-Z233.05S-Z233.06T, Z233.01T-Z233.02A-Z233.03S-Z233.04T-Z233.05K-Z233.06A-Z233.07T-Z233.08S-Z233.09T-Z233.10T, Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05D-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, Z233.01S-Z233.02T-Z233.03T-Z233.04D-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T, and Z233.01S-Z233.02T-Z233.03T-Z233.04S-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T; and (vi) a combination of (i) to (v).
4. The cellulase variant of claim 1, wherein said two or more mutations at two or more positions corresponding to two or more SEQ ID NO: 1 positions are selected from: K4V-Q36T-S77M-T102K-Q154E-N178H-S225K-S232Z-T233Z-S234Z-T235Z-T236Z-S237Z-T238Z-K239Z-A240Z-T241Z-S242Z-T243Z, Q36T-S77K-G87A-A142P-S153D-A161P-G204S-S221L-S237D, F29W-Q36T-K44V-T116V-S135T-Q154E-A161P-S217G-S221L-S225K-T238D, K4V-Q36T-S77K-N80S-A142P-L192V-G204S-T210V-S221L, K4V-Q36T-S51T-N80S-S135T-S157D-T233A, K4V-S23L-Q36T-S77M-G112T-S157D-A161P-L192V-S221L, Q36T-V98Y-A142P-A161E-S225K, Q36T-P136E-A142P-S225K-S249P, S32D-Q36T-S221L-S225K, S32D-Q36T-G112S-A142P, S32D-Q36T-S77M-S135T-A142P-A161P, Q36T-S51T-V98G-T116V-A142P-A161P-T233S-Z233.01T-Z233.02A-Z233.03S-Z233.04T-Z233.05K-Z233.06A-Z233.07T-Z233.08S-Z233.09T-Z233.10T, Q36T-S77K-G87A-A142P-S153D-A161P-D177Z-G204S-S221L-S237D, and K4V-S32G-Q36T-S77M-G87A-T116V-A142P-A161P-Z233.01S-Z233.02T-Z233.03T-Z233.04D-Z233.05T-Z233.06K-Z233.07A-Z233.08T-Z233.09S-Z233.10T-Z233.11T.
5. The cellulase variant of claim 1, with the proviso that one or more of said mutations is non-naturally occurring.
6. The cellulase variant of claim 1, wherein said variant has at least one improved property selected from improved thermostability, stability in the presence of one or more protease, and stability in the presence of one or more protease and one or more other detergent component when compared to a parent or reference polypeptide.
7. The cellulase variant of claim 1, wherein said variant is derived from a parent or reference polypeptide selected from SEQ ID NOs: 1, 70, 71, 72, 73, 74, and 75.
8. The cellulase variant of claim 1, wherein the improved property is (i) improved thermostability and wherein said variant has a thermal PI that is greater than 1 or ≥1.1; (ii) improved stability in the presence of one or more protease and wherein said variant has a PI that is greater than 1 or ≥1.1, 1.2, 1.3, 1.4, 1.5, or 2.0 when the stability of said variant is tested in the presence of said protease; and/or (iii) improved stability in the presence of one or more protease and one or more other detergent component, and wherein said variant has a PI that is greater than 1 or ≥1.1, 1.5, 2.0, or 2.5 when the stability of said variant, is tested in the presence of said protease and said other detergent component.
9. The cellulase variant of claim 8, wherein the PI is measured in accordance with the Cellulase Activity Assay.
10. The cellulase variant of claim 8, wherein the other detergent component is a surfactant.
11. The cellulase variant of claim 1, wherein said variant is a family GH45 cellulase.
12. A composition comprising the cellulase variant of claim 1.
13. The composition of claim 12, wherein said composition is selected from an enzyme composition, detergent composition, and fabric care composition.
14. The composition of claim 12, further comprising (i) one or more other enzymes selected from the group consisting of acyl transferases, amylases, alpha-amylases, beta-amylases, alpha-galactosidases, arabinases, arabinosidases, aryl esterases, beta-galactosidases, beta-glucanases, carrageenases, catalases, chondroitinases, cutinases, endo-beta-mannanases, exo-beta-mannanases, esterases, exo-mannanases, galactanases, glucoamylases, hemicellulases, hyaluronidases, keratinases, laccases, lactases, ligninases, lipases, lipolytic enzymes, lipoxygenases, mannanases, metalloproteases, oxidases, oxidoreductases, pectate lyases, pectin acetyl esterases, pectinases, pentosanases, perhydrolases, peroxidases, phenoloxidases, phosphatases, phospholipases, phytases, polyesterases, polygalacturonases, proteases, pullulanases, reductases, rhamnogalacturonases, second cellulase, beta-glucanases, tannases, transglutaminases, xylan acetyl-esterases, xylanases, and xylosidases; (ii) one or more surfactants; (iii) one or more ions selected from calcium and zinc; (iv) one or more adjunct ingredients; (v) one or more stabilizers; (vi) from about 0.001% to about 5.0 weight % of said cellulase variant; (vii) one or more bleaching agents; and/or (viii) combinations thereof.
15. The composition of claim 12, wherein said composition is a laundry detergent.
16. The composition of claim 12, wherein the composition is in a form selected from a liquid, a powder, a granulated solid, a tablet, a sheet, and a unit dose.
17. The composition of claim 12, wherein said composition contains phosphate or is phosphate-free and/or contains boron or is boron-free.
18. A polynucleotide comprising a nucleic acid sequence encoding the cellulase variant of claim 1.
19. An expression vector comprising the polynucleotide of claim 18.
20. A host cell comprising the expression vector of claim 19.
21. A method for producing the cellulase variant, comprising: (a) stably transforming a host cell with the expression vector of claim 19; (b) cultivating said transformed host cell under conditions suitable for said host cell to produce said cellulase variant; and (c) recovering said cellulase variant.
Given the fact pattern of the instant case as well as the patent the species claims of the patent anticipates the instant genus (or broad) claims.
13. No claim is allowed.
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/TEKCHAND SAIDHA/
Primary Examiner, Art Unit 1652
Recombinant Enzymes, Hoteling
Telephone: (571) 272-0940
Fax: (571) 273-0940
Prosecution Insights