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
This application is a 371 of PCT/EP2021/073869.
The amendment filed on June 8, 2026 has been entered.
Election/Restrictions
Applicant elected with traverse of Group I with a species election of (1) SEQ ID NO:1 as the parent protease and (2) X215K/R/Q/N/T and at least three alterations selected from X3T, X4l, X9E, I35ID, X43R, X76D, X99D, X99F, X101E, X101L, X103A, X108T, X104I, X120D, X160S, X195E, X205I, X206L, X209W, X235L, X259D, X261W, and X262E in the reply filed on September 29, 2025.
Claims 16-19 and 24 are withdrawn from further consideration pursuant to 37 CFR 1.142(b), as being drawn to a nonelected invention, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on September 29, 2025.
Applicant’s request for rejoinder of Groups III-IV have been noted.
A search and examination of protease variants having A215K and at least three amino acid substitutions selected from the group consisting of X3T, S3T, S9E, N43R, N76D, S99D, S101E, H120D, G195E, Q206L, Y209W, K235L, S259D, and L262E are disclosed in the prior art. See the rejection under 102 below. Therefore, examination has not been extended to a subsequent species.
Status of Claims
Claims 1-22 and 24 are pending.
Claims 16-19 and 24 are withdrawn.
Claims 1-15 and 20-22 are under examination.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on December 23, 2025 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner.
Response to Amendments/Arguments
Claim Rejections - 35 USC § 112(b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Withdrawn Rejections
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claims 1, 7, and 13-14 and claims 2-6, 8-12, 15 and 20-22 depending therefrom have been fully considered and are persuasive. Claims 1, 7, and 14 have been amended to delete the phrase “e.g.”. Therefore, the rejection of claims 1, 7, and 13-14 and claims 2-6, 8-12, 15 and 20-22 depending therefrom under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claims 7 and 9-11 and claims 7-8 depending therefrom have been fully considered and are persuasive. Claims 6 and 9-11 have been amended to delete the phrases “preferably” and “most preferably”. Therefore, the rejection of claims 7 and 9-11 and claims 7-8 depending therefrom under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claim 6 and claims 7-12 depending therefrom have been fully considered and are persuasive. Claim 6 has been amended to delete the use of a narrow numerical range that falls within a broader range. Therefore, the rejection of claim 6 and claims 7-12 depending therefrom under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claims 9 and 11 have been fully considered and are persuasive. Claims 9 and 11 have been amended to delete the use of a narrow numerical range that falls within a broader range. Therefore, the rejection of claims 9 and 11 under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claims 10-11 have been fully considered and are persuasive. Claims 10-11 have been amended to delete the use of a narrow numerical range that falls within a broader range. Therefore, the rejection of claims 10-11 under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claim 12 have been fully considered and are persuasive. Claim 12 has been amended to delete the recitation of “Example 1”. Therefore, the rejection of claim 12 under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claims 13 and 15 have been fully considered and are persuasive. Claims 13 and 15 has been amended to delete the indefinite limitation “on par”. Therefore, the rejection of claims 13 and 15 under 35 U.S.C. 112(b) has been withdrawn.
Applicant’s arguments, see page 2 of the Remarks, filed June 8, 2026, with respect to claim 13 have been fully considered and are persuasive. Claim 13 has been amended to delete the use of a narrow numerical range that falls within a broader range. Therefore, the rejection of claim 13 under 35 U.S.C. 112(b) has been withdrawn.
Maintained Rejection
Claims 2-5 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 2-5 recite the phrase “preferably”. The phrase renders the claim indefinite because it is unclear whether the limitation(s) following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Applicant's arguments filed June 8, 2026 have been fully considered but they are not persuasive.
Applicant argues that all claims have been amended to delete use of “preferably”.
This is not found persuasive.
Claims 2-5 have not been amended to delete the phrase “preferably”. Therefore, the rejection has been maintained.
Claim Rejections - 35 USC § 112(a)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Withdrawn Rejection
Applicant’s arguments, see Section III at pages 2-4 of the Remarks, filed June 8, 2026, with respect to claims 6-12 have been fully considered and are persuasive. Applicant argues that “claims 1 and 6-12 recite an explicit description of the invention as protease variants in which substitution at position 215 confers improved solubility relative to an otherwise identical parent enzyme…All variants of the invention possess a substitution at position 215 and the functionality of improved solubility.”. Therefore, the rejection of claims 6-12 under 35 U.S.C. 112(a) as lacking written description has been withdrawn.
Modified Rejection due to amendment of claim 1
Claims 1-15 and 20-22 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a protease variant of SEQ ID NO:1, wherein the variant consists of A215K/Q/N/T/S and wherein the protease variant has having improved solubility at 20°C and pH 4-5, does not reasonably provide enablement any protease variant having at least 80% but less than 100% sequence identity and having the amino acid substitutions identified above, wherein the protease variant has improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, has an improved solubility at 10-30°C, an improved solubility at a pH of 3-9, and has an improved solubility at 15-25°C and pH 4-6. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims.
Factors to be considered in determining whether undue experimentation is required are summarized in In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988). They include (1) the quantity of experimentation necessary, (2) the amount of direction or guidance presented, (3) the presence or absence of working examples, (4) the nature of the invention, (5) the state of the prior art, (6) the relative skill of those in the art, (7) the predictability or unpredictability of the art, and (8) the breadth of the claims.
The breadth of the claims.
MPEP 2111.01 states that ''[d]uring examination, the claims must be interpreted as broadly as their terms reasonably allow.'' In this case, the claims have been broadly interpreted to encompass any protease variants having at least 80% but less than 100% sequence identity to SEQ ID NO:1 and a X215K/R/Q/N/S/T amino acid substitution and at least three amino acid substitutions recited in claim 1 (amino acid position corresponding to SEQ ID NO:2), wherein the protease variant has improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, has an improved solubility at 10-30°C, an improved solubility at a pH of 3-9, and has an improved solubility at 15-25°C and pH 4-6. Therefore, the claims are drawn to any protease variant having at least 80% but less than 100% sequence identity and having the amino acid substitutions identified above, wherein the protease variant has an improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, has an improved solubility at 10-30°C, an improved solubility at a pH of 3-9, and has an improved solubility at 15-25°C and pH 4-6.
The claims are not commensurate with the enablement provided by the disclosure with regard to the extremely large number of polypeptides having improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, having an improved solubility at 10-30°C, an improved solubility at a pH of 3-9, and having an improved solubility at 15-25°C and pH 4-6. In the instant case, the specification is limited to a protease variant of SEQ ID NO:1, wherein the variant consists of A215K/Q/N/T/S and wherein the protease variant has having improved solubility at 20°C and pH 4-5.
The quantity of experimentation required to practice the claimed invention based on the teachings of the specification.
While enzyme isolation techniques, recombinant and mutagenesis techniques were known in the art at the time of the invention, e.g. mutagenesis, and it is routine in the art to screen for variants comprising multiple substitutions or multiple modifications as encompassed by the instant claims, the specific amino acid positions within the protein's sequence where amino acid modifications can be made with a reasonable expectation of success in obtaining the desired activity/utility are limited in any protein and the result of such modifications is unpredictable. In addition, one skilled in the art would expect any tolerance to modification for a given protein to diminish with each further and additional modification, e.g. multiple substitutions.
In the absence of: (a) rational and predictable scheme for making proteases having improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6 and (b) a correlation between structure and the function of having improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6, the specification provides insufficient guidance as to which of the essentially infinite possible choices is likely to be successful. One of skill in the art would have to test these infinite possible polypeptides to determine which proteases have improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6. While enablement is not precluded by the necessity for routine screening, if a large amount of screening is required, as is the case herein, the specification must provide a reasonable amount of guidance which respect to the direction in which the experimentation should proceed so that a reasonable number of species can be selected for testing. In view of the fact that such guidance has not been provided in the instant specification, it would require undue experimentation to enable the full scope of the claims.
The state of prior art, the relative skill of those in the art, and predictability or unpredictability of the art.
Since the amino acid sequence of the mutant determines its structural and functional properties, predictability of which changes can be tolerated in a protein's amino acid sequence and obtain the desired activity requires a knowledge of and guidance with regard to which amino acids in the protein's sequence, if any, are tolerant of modification and which are conserved (i.e. expectedly intolerant to modification), and detailed knowledge of the ways in which the proteins' structure relates to its function. In the instant case, neither the specification or the art provide a correlation between structure and activity such that one of skill in the art can envision the structure of any polypeptides having deoxyribose-phosphate aldolase activity or predict said function of a polypeptide from its primary structure. In addition, the art does not provide any teaching or guidance as to (1) which amino acids within the polypeptide of SEQ ID NO:1 other than A215 that can be modified and which ones are conserved such that one of skill in the art can make the recited protease variants having improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6, (2) which segments of the polypeptide of SEQ ID NO:1 that are essential for having improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6, and (3) the general tolerance of the polypeptide of SEQ ID NO:1 to structural modifications and the extent of such tolerance. The art clearly teaches that changes in a protein's amino acid sequence to obtain the desired activity without any guidance/knowledge as to which amino acids in a protein are required for that activity is highly unpredictable. At the time of the invention there was a high level of unpredictability associated with altering a polypeptide sequence with an expectation that the polypeptide will maintain the desired activity. For example, Studer (Residue mutations and their impact on protein structure and function: detecting beneficial and pathogenic changes. Biochem. J. (2013) 449, 581–594. – form PTO-892) teach that (1) protein engineers are frequently surprised by the range of effects caused by single mutations that they hoped would change only one specific and simple property in enzymes, (2) the often surprising results obtained by experiments where single mutations are made reveal how little is known about the rules of protein stability, and (3) the difficulties in designing de novo stable proteins with specific functions.
Bacillus lentus protease was known in the art. Babe CN 109715791 – form PTO-892 and English Translation of CN 109715791. Retrieved on December 3, 2025 – form PTO-892) discloses a Bacillus lentus having 100% sequence identity to SEQ ID NO:1 of the instant application (Figure 1 and see the sequence alignment below). Although Babe discloses protease variants having improved protease activity and improved thermostability (2nd paragraph at page 89 of the English Translation), neither Babe, the prior art, nor the instant specification teach Bacillus lentus protease variants having at least 80% but less than 100% sequence identity to SEQ ID NO:1, a X215K/R/Q/N/S/T amino acid substitution, and at least three amino acid substitutions recited in claim 1, wherein the protease variants have improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, has an improved solubility at 10-30°C, an improved solubility at a pH of 3-9, and has an improved solubility at 15-25°C and pH 4-6.
Fransceus (J Ind Microbiol Biotechnol. 2017 May;44(4-5):687-695. – form PTO-892) reviews protein engineering techniques, such as random mutagenesis and recombination, directed evolution and iterative or combinatory saturation “hotspots”. Fransceus states that “a recurring problem, however, is choosing which amino acid positions should be targeted. Answering this question is not an easy feat and requires substantial insight in the relationship between an enzyme’s sequence or structure and its properties.” Sanavia (Computational and Structural Biotechnology Journal, Volume 18, 2020, Pages 1968-1979. – form PTO-892) discloses challenges in the prediction of protein stability in the occurrence of multiple mutations. “Multiple-point mutations are common variations of the protein sequence that may be needed in protein engineering when a single-point mutation is not enough to yield the desired stability change. Dealing with multiple-site variations adds another level of complexity beyond the prediction of the effect of a single variant on protein stability, since it requires the learning of many types of combinatorial effects”.
The amount of direction or guidance presented and the existence of working examples.
The specification is limited to a protease variant of SEQ ID NO:1, wherein the variant consists of A215K/Q/N/T/S and wherein the protease variant has having improved solubility at 20°C and pH 4-5. However, the speciation fails to provide any information as to (1) specific substrates associated with any protease variants having at least 80% sequence identity to SEQ ID NO:1 and having improved solubility or improved solubility of at least 4% compared to SEQ ID NO:1 not having the X215K/R/Q/N/S/T amino acid substitution, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6 or (2) structural elements required in a polypeptide having improved solubility of at least 4% to at least 500% or more compared to SEQ ID NO:1, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6. No correlation between structure and function of having improved solubility of at least 4% to at least 500% or more compared to SEQ ID NO:1, improved solubility at 10-30°C, improved solubility at a pH of 3-9, and improved solubility at 15-25°C and pH 4-6 has been presented.
Thus, in view of the overly broad scope of the claims, the lack of guidance and working examples provided in the specification, the high level of unpredictability of the prior art in regard to structural changes and their effect on function and the lack of knowledge about a correlation between structure and function, an undue experimentation would be necessary one having ordinary skill in the art to make and use the claimed invention in a manner reasonably correlated with the scope of the claims. The scope of the claims must bear a reasonable correlation with the scope of enablement (In re Fisher, 166 USPQ 19 24 (CCPA 1970)). Without sufficient guidance, determination of polypeptides having the desired biological characteristics recited in the claims are unpredictable and the experimentation left to those skilled in the art is unnecessarily, and improperly, extensive and undue. See In re Wands 858 F.2d 731, 8 USPQ2nd 1400 (Fed. Cir, 1988).
Applicant has not submitted any rebuttal. Therefore, the rejection has been maintained.
Claim Rejections - 35 USC § 102
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Withdrawn Rejections
Applicant’s arguments, see Section IV at pages 4-5 of the Remarks, filed June 8, 2026, with respect to claims 1-4 and 13 have been fully considered and are persuasive. Claim 1 has been amended to require X262E as one of the at least three further alterations, which is not taught by Masui (Rational design for stabilization and optimum pH shift of serine protease AprN. Journal of Fermentation and Bioengineering, Volume 85, Issue 1,1998, Pages 30-36 – form PTO-892). Therefore, the rejection of claims 1-4 and 13 under 35 U.S.C. 102(a)(1) as being anticipated by Masui has been withdrawn.
Applicant’s arguments, see Section V at pages 5-6 of the Remarks, filed June 8, 2026, with respect to claims 1-4, 6-8, 13-15, and 20-22 have been fully considered and are persuasive. Claim 1 has been amended to require X262E as one of the at least three further alterations, which is not taught by Babe (CN 109715791 – form PTO-892 and English Translation of CN 109715791. Retrieved on December 3, 2025 – form PTO-892). Therefore, the rejection of claims 1-4, 6-8, 13-15, and 20-22 under 35 U.S.C. 102(a)(1) as being anticipated by Babe has been withdrawn.
Maintained Rejections
Claim(s) 1-4, 6-8, 13-15, and 20-22 is/are rejected under 35 U.S.C. 102(a)(2) as being anticipated by Knotzel (US 2022/0145220 – form PTO-892).
Knotzel discloses a Bacillus lentus protease of SEQ ID NO:1 which as 100% sequence identity to SEQ ID NO:1 of the instant application (Figure 1 and see the sequence alignment below). Knotzel discloses protease variants of SEQ ID NO:1, wherein the position numbers are based on the numbering of the Bacillus amyloliquefaciens protease of SEQ ID NO:2, which has 100% sequence identity to the protease of SEQ DI NO:2 of the instant application (Figure 2 and see the sequence alignment below).
Regarding claims 1-4, Knotzel disclose a protease variant of SEQ ID NO:1, wherein the variant has A215K amino acid substitution and one or more S9E, N43R, N76D, S99D, S101E, H120D, G195E, K235L, S259D, Y209W, and L262E amino acid substitutions ([0088]-[0089] and [0104]), A215K amino acid amino acid substitution and S9E+N43R+N76D+V205I+Q206L+Y209W+S259D+N261W+L262E ([0089] and [0096], and N62D+G97D+S101E+V1771+Y209W+A215K+L262E ([0110] and claims 7-8 and 17). Since the protease variant of Knotzel has up to 12 amino acid substitutions, the protease variant of Knotzel has at least 80% sequence identity to SEQ ID NO:1 of the instant application.
Regarding claims 1 and 6-8, the protease variant of Knotzel has improved solubility, as evidenced by Lenhard (US 2023/0323330 – form PTO-892). Lenhard discloses that the X215K amino acid substitution in the protease of SEQ ID NO:1 (which has 100% sequence identity to the protease of SEQ ID NO:1 of the instant application, see the sequence alignment below) confers improved solubility and improves solubility by at least 100% to at least 500% compared to the parent protease of SEQ ID NO:1 (page 13, lines 14-16 and page 57 of the reference specification). MPEP 2112. II. states that “[t]here is no requirement that a person of ordinary skill in the art would have recognized the inherent disclosure at the relevant time, but only that the subject matter is in fact inherent in the prior art reference.”.
Regarding claim 5, Knotzel disclose a protease variant of SEQ ID NO:1, wherein the variant has A215K amino acid amino acid substitution and S9E+N43R+N76D+V205I+Q206L+Y209W+S259D+N261W+L262E ([0089] and [0096]).
Regarding claim 13, Knotzel discloses that the protease variants has improved washing performance and therefore, the protease variants have a protease activity that is equal with the protease activity of the parent protease of SEQ ID NO:1 ([0009] and [0016]).
Regarding claims 14-15, the parent protease of Knotzel is identical to the parent protease of SEQ ID NO:1 of the instant application and does not have the amino acid substitutions recited in claim 14 (Figures 1-2 and see the sequence alignment below).
Regarding claims 20-22, Knotzel discloses a detergent composition in the form of a gel or liquid comprising the protease variants and one or more detergent components ([0384]-[0385] and [0389] and claims 7-8 and 17).
Therefore, the reference of Knotzel anticipates claims 1-8, 13-15, and 20-22.
The applied reference has a common assignee with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2). This rejection under 35 U.S.C. 102(a)(2) might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C. 102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B) if the same invention is not being claimed; or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed in the reference and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement.
Applicant's arguments filed June 8, 2026 (Section VI at pages 6-7) have been fully considered but they are not persuasive.
Applicant argues that the claims are not anticipated by Knotzel because Knotzel does not disclose protease variants with improved solubility and the evidentiary reference of Lenhard does not teach that or offer any indication of how a different position (different from A215K) might affect solubility.
This is not found persuasive. Lenhard discloses that introducing X215K amino acid substitution in the protease of SEQ ID NO:1 (which has 100% sequence identity to the protease of SEQ ID NO:1 of the instant application) confers improved solubility and improves solubility to the protease variants by at least 100% to at least 500% compared to the parent protease of SEQ ID NO:1 (page 13, lines 14-16 and page 57 of the reference specification). Lenhard discloses a protease variant comprising X215K and S9E+N43R+N76D+V205I+Q206L+Y209W+S259D+N261W+L262E (page 57 of reference specification), which is one of the protease variants taught by Knotzel. The specification of the instant application discloses that a protease variant having A215K and at least three further amino acid substitutions, such as S9E, N43R, N76D, V205I, Q206L, Y209W, S259D, N261W, and L262E recited in claim 5, has improved solubility compared to the parent not having the 215K amino acid substitution (page 11, lines 9). Therefore, the protease variants of Knotzel inherently has improved solubility compared to the parent protease not having the A215K amino acid substitution. Further, Applicant at page 3 of the Remarks filed on June 8, 2026 states that “claims 1 and 6-12 recite an explicit description of the invention as protease variants in which substitution at position 215 confers improved solubility relative to an otherwise identical parent enzyme…All variants of the invention possess a substitution at position 215 and the functionality of improved solubility.”. Therefore, according to Applicant’s statement, a protease variant having 215K amino acid substitution confers improved solubility to said protease variant having at least three amino acid substitutions recited in the claims, wherein one of the at least three substitutions is X262E. Hence the rejection has been maintained.
Claim Rejections - 35 USC § 103
Applicant’s arguments, see Section VII at pages 7-8 of the Remarks, filed June 8, 2026, with respect to claims 20-22 have been fully considered and are persuasive. Claim 1 has been amended to require X262E as one of the at least three further alterations and a protease variant having improved solubility, which is not taught by Babe (CN 109715791 – form PTO-892 and English Translation of CN 109715791. Retrieved on December 3, 2025 – form PTO-892) and Babe (CN 109715791 – form PTO-892 and English Translation of CN 109715791. Retrieved on December 3, 2025 – form PTO-892). Therefore, the rejection of claims 20-22 under 35 U.S.C. 103 as being unpatentable over Masui and Babe has been withdrawn.
Claim Rejections - 35 USC § 101
Applicant’s arguments, see Section VIII at pages 8-9 of the Remarks, filed June 8, 2026, with respect to claims 1-4 and 20-22 have been fully considered and are persuasive. Claim 1 has been amended to require X262E as one of the at least three further alterations and a protease variant having improved solubility, which no longer reads on a product of nature. Therefore, the rejection of claims 1-4 and 20-22 under 35 U.S.C. 101 has been withdrawn.
Double Patenting
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 conflicting claims 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); 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 nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined 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 § 2146 et seq. 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).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
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Claims 1-4, 6-8, 13-15, and 20-22 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim claims 1, 4, 10-13, 15, and 18-25 of copending Application No. 17/435,555 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because the claims of the instant application and the calims of the reference application are both directed to protease variants of SEQ ID NO:1. The protease of SEQ ID NO:1 of the instant application is identical to the protease of SEQ ID NO:1 of the reference application. The protease of SEQ ID NO:2 of the instant application is identical to the protease of SEQ ID NO:2 of the reference application.
Regarding claims 1-4 of the instant application, claim 1 of the reference application recites a detergent composition comprising a protease variant of SEQ ID NO:1, wherein the variant has N62D+G97D+S101E +V177I+ Y209W+A215K+L262E, N76D+G97D+N140D+S156D+ Y209W+A215K+L262E, N76D+G97D+S101E+T180A+Y209W+A215K+L262E, S9E+G97D+S101E+Y209W+A215K+L262E, S9E+N43R+N76D+S188E+Q191N+A194P+Q206L+Y209W+A215K+S259D+L262E, and S9R+K27M+N43R+N76D+V205I+Q206L+Y209W+A215K+Q245R+S259D+N261W+L262E. Since the protease variant of the reference application has 11 amino acid substitutions, the protease variant of the reference application has at least 80% sequence identity to SEQ ID NO:1 of the instant application. Therefore, claims 1-4 of the instant application are anticipated by claim 1 of the reference application.
Regarding claims 1 and 6-8 of the instant application, the protease variant of the reference application has improved solubility, as evidenced by Lenhard (US 2023/0323330 – form PTO-892). Lenhard discloses that the X215K amino acid substitution in the protease of SEQ ID NO:1 (which has 100% sequence identity to the protease of SEQ ID NO:1 of the instant application, see the sequence alignment below) confers improved solubility and improves solubility by at least 100% to at least 500% compared to the parent protease of SEQ ID NO:1 (page 13, lines 14-16 and page 57 of the reference specification). MPEP 2112. II. states that “[t]here is no requirement that a person of ordinary skill in the art would have recognized the inherent disclosure at the relevant time, but only that the subject matter is in fact inherent in the prior art reference.”.
Regarding claim 13 of the instant application, claim 13 of the reference application recites that the protease variant has improved washing performance and therefore, the protease variant has a protease activity that is on par with the protease activity of the parent protease of SEQ ID NO:1 ([0009] and [0016]). Therefore, claim 13 of the instant application is anticipated by claim 1 of the reference application.
Regarding claims 14-15, of the instant application the parent protease of the reference application is identical to the parent protease of SEQ ID NO:1 of the instant application and does not have the amino acid substitutions recited in claim 14. Therefore, claims 14-15 of the instant application are anticipated by claim 1 of the reference application.
Regarding claims 20-22 of the instant application, claim 23 of the reference application recite a detergent composition having an improved wash performance comprising the protease variant of claim 1 and claim 25 recites that the detergent composition comprises one or more components. Therefore, claims 20-22 of the instant application is anticipated by claims 23 and 15 of the reference application.
Therefore, the conflicting claims are not patentably distinct from each other.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Applicant has requested that the rejection be held in abeyance until indication of allowable subject matter.
Claims 1-8 and 13-15 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim claims 1-19 and 21-22 of copending Application No. 18/043,515 (reference application). Although the claims at issue are not identical, they are not patentably distinct from each other because the claims of the instant application and the claims of the reference application are both directed to protease variants of SEQ ID NO:1. The protease of SEQ ID NO:1 of the instant application is identical to the protease of SEQ ID NO:1 of the reference application. The protease of SEQ ID NO:2 of the instant application is identical to the protease of SEQ ID NO:2 of the reference application.
Regarding claims 1-5 of the instant application, claim 1 and claim 5 of the reference application recites a protease variant of SEQ ID NO:1, wherein the variant has S9E, N43R, N76D, S99F, S101L, S103T, V104I, X127I, X194P, V205I, Q206L, Y209W, A215K, S259D, N261W, and L262E. Since the protease variant of the reference application has 16 amino acid substitutions, the protease variant of the reference application has at least 80% sequence identity to SEQ ID NO:1 of the instant application. Therefore, claims 1-5 of the instant application are anticipated by claims 1 and 5 of the reference application.
Regarding claims 1 and 6-8 of the instant application, the protease variant of the reference application has improved solubility by at least 100% to at least 500% compared to the parent protease of SEQ ID NO:1 (page 57 of the reference specification). The reference specification also discloses that the X215K amino acid substitution confers improved solubility (page 13, lines 14-16). Therefore, claims 6-8 of the instant application are anticipated by claim 1 of the reference application.
Regarding claim 13 of the instant application, claim 9 of the reference application recites that the protease variant has improved thermostability, which reads on improved protease activity at high temperatures. Therefore, claim 13 of the instant application is anticipated by claims 1, 5, and 9 of the reference application.
Regarding claims 14-15 of the instant application, the parent protease of the reference application is identical to the parent protease of SEQ ID NO:1 of the instant application and does not have the amino acid substitutions recited in claim 14. Therefore, claims 14-15 of the instant application are anticipated by claims 1, 5, and 9 of the reference application.
Therefore, the conflicting claims are not patentably distinct from each other.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Applicant has requested that the rejection be held in abeyance until indication of allowable subject matter.
Claims 1-8, 13-15, and 20-22 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim claims 1-19 and 21-22 of copending Application No. 18/043,515 (reference application) in view of Babe (CN 109715791 – form PTO-892 and English Translation of CN 109715791. Retrieved on December 3, 2025 – form PTO-892. The English Translation of CN 109715791 is used for specific passages). Although the claims at issue are not identical, they are not patentably distinct from each other because the claims of the instant application and the claims of the reference application are both directed to protease variants of SEQ ID NO:1. The protease of SEQ ID NO:1 of the instant application is identical to the protease of SEQ ID NO:1 of the reference application. The protease of SEQ ID NO:2 of the instant application is identical to the protease of SEQ ID NO:2 of the reference application.
Regarding claims 1-5 of the instant application, claim 1 and claim 5 of the reference application recites a protease variant of SEQ ID NO:1, wherein the variant has S9E, N43R, N76D, S99F, S101L, S103T, V104I, X127I, X194P, V205I, Q206L, Y209W, A215K, S259D, N261W, and L262E. Since the protease variant of the reference application has 16 amino acid substitutions, the protease variant of the reference application has at least 80% sequence identity to SEQ ID NO:1 of the instant application. Therefore, claims 1-5 of the instant application are anticipated by claim 1 of the reference application.
Regarding claims 6-8 of the instant application, the protease variant of the reference application has improved solubility by at least 100% to at least 500% compared to the parent protease of SEQ ID NO:1 (page 57 of the reference specification). The reference specification also discloses that the X215K amino acid substitution confers improved solubility (page 13, lines 14-16). Therefore, claims 6-8 of the instant application are anticipated by claim 1 of the reference application.
Regarding claim 13 of the instant application, claim 9 of the reference application recites that the protease variant has improved thermostability, which reads on improved protease activity at high temperatures. Therefore, claim 13 of the instant application is anticipated by claims 1, 5, and 9 of the reference application.
Regarding claims 14-15 of the instant application, the parent protease of the reference application is identical to the parent protease of SEQ ID NO:1 of the instant application and does not have the amino acid substitutions recited in claim 14. Therefore, claims 14-15 of the instant application are anticipated by claims 1, 5, and 9 of the reference application.
The claims of the reference application do not recite a detergent composition comprising the protease variant.
Regarding claims 20-22, Babe discloses a detergent composition in the form of a powder or liquid comprising Bacillus protease variants of and one or more detergent components (2nd and 3rd paragraphs at page 94).
Therefore, it would have been obvious to one having ordinary skill to modify the claims of the reference application by making a liquid or powder detergent composition comprising the protease variant and at least one detergent component. One of ordinary skill in the art would have been motivated to do so because use of a protease in a detergent composition was known in the art and the protease variant of the reference claim has improved protease activity and improved solubility compared to the parent protease. One of ordinary skill in the art would have had a reasonable expectation of success since the claims of the reference application recites a protease and Babe discloses a liquid or powder detergent comprising a protease and at least one detergent component. The rationale to support that the claim would have been obvious is that all the claimed elements were known in the prior art and one skilled in the art could have combined the elements as claimed by known methods with no change in their respective functions, and the combination yielded nothing more than predictable results to one of ordinary skill in the art.
Therefore, the conflicting claims are not patentably distinct from each other.
This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Applicant has requested that the rejection be held in abeyance until indication of allowable subject matter.
Conclusion
Claims 1-22 and 24 are pending.
Claims 16-19 and 24 are withdrawn.
Claims 1-15 and 20-22 are rejected.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to YONG D PAK whose telephone number is (571)272-0935. The examiner can normally be reached M-Th: 5:30 am - 3:30 pm.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Robert Mondesi can be reached on 408-918-7584. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/YONG D PAK/Primary Examiner, Art Unit 1652
Sequence alignment between the protease of SEQ ID NO:1 of the instant application (“Qy”) and protease of Masui (“Db”)
O66153_BACSP
ID O66153_BACSP Unreviewed; 379 AA.
AC O66153;
DT 01-AUG-1998, integrated into UniProtKB/TrEMBL.
DT 01-AUG-1998, sequence version 1.
DT 05-FEB-2025, entry version 98.
DE SubName: Full=AprN {ECO:0000313|EMBL:BAA25184.1};
OS Bacillus sp.
OC Bacteria; Bacillati; Bacillota; Bacilli; Bacillales; Bacillaceae; Bacillus.
OX NCBI_TaxID=1409 {ECO:0000313|EMBL:BAA25184.1};
RN [1] {ECO:0000313|EMBL:BAA25184.1}
RP NUCLEOTIDE SEQUENCE.
RC STRAIN=B21-2 {ECO:0000313|EMBL:BAA25184.1};
RA Masui A., Fujiwara N., Yamamoto K., Takagi M., Imanaka T.;
RT "Rational design for stabilization and optimum pH shift of serine protease
RT AprN.";
RL J. Ferment. Bioeng. 85:30-36(1998).
CC -!- COFACTOR:
CC Name=Ca(2+); Xref=ChEBI:CHEBI:29108;
CC Evidence={ECO:0000256|ARBA:ARBA00001913};
CC -!- SUBCELLULAR LOCATION: Secreted {ECO:0000256|ARBA:ARBA00004613}.
CC -!- SIMILARITY: Belongs to the peptidase S8 family.
CC {ECO:0000256|ARBA:ARBA00011073, ECO:0000256|PROSITE-ProRule:PRU01240,
CC ECO:0000256|RuleBase:RU003355}.
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DR EMBL; AB005792; BAA25184.1; -; Genomic_DNA.
DR AlphaFoldDB; O66153; -.
DR SMR; O66153; -.
DR MEROPS; S08.157; -.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004252; F:serine-type endopeptidase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0006508; P:proteolysis; IEA:UniProtKB-KW.
DR CDD; cd07477; Peptidases_S8_Subtilisin_subset; 1.
DR Gene3D; 3.30.70.80; Peptidase S8 propeptide/proteinase inhibitor I9; 1.
DR Gene3D; 3.40.50.200; Peptidase S8/S53 domain; 1.
DR InterPro; IPR000209; Peptidase_S8/S53_dom.
DR InterPro; IPR036852; Peptidase_S8/S53_dom_sf.
DR InterPro; IPR023827; Peptidase_S8_Asp-AS.
DR InterPro; IPR022398; Peptidase_S8_His-AS.
DR InterPro; IPR023828; Peptidase_S8_Ser-AS.
DR InterPro; IPR050131; Peptidase_S8_subtilisin-like.
DR InterPro; IPR015500; Peptidase_S8_subtilisin-rel.
DR InterPro; IPR010259; S8pro/Inhibitor_I9.
DR InterPro; IPR037045; S8pro/Inhibitor_I9_sf.
DR InterPro; IPR034202; Subtilisin_Carlsberg-like.
DR PANTHER; PTHR43806:SF11; CEREVISIN-RELATED; 1.
DR PANTHER; PTHR43806; PEPTIDASE S8; 1.
DR Pfam; PF05922; Inhibitor_I9; 1.
DR Pfam; PF00082; Peptidase_S8; 1.
DR PRINTS; PR00723; SUBTILISIN.
DR SUPFAM; SSF54897; Protease propeptides/inhibitors; 1.
DR SUPFAM; SSF52743; Subtilisin-like; 1.
DR PROSITE; PS51892; SUBTILASE; 1.
DR PROSITE; PS00136; SUBTILASE_ASP; 1.
DR PROSITE; PS00137; SUBTILASE_HIS; 1.
DR PROSITE; PS00138; SUBTILASE_SER; 1.
PE 3: Inferred from homology;
KW Calcium {ECO:0000256|ARBA:ARBA00022837};
KW Hydrolase {ECO:0000256|ARBA:ARBA00022801, ECO:0000256|PROSITE-
KW ProRule:PRU01240};
KW Protease {ECO:0000256|ARBA:ARBA00022670, ECO:0000256|PROSITE-
KW ProRule:PRU01240}; Secreted {ECO:0000256|ARBA:ARBA00022525};
KW Serine protease {ECO:0000256|ARBA:ARBA00022825, ECO:0000256|PROSITE-
KW ProRule:PRU01240}; Signal {ECO:0000256|SAM:SignalP}.
FT SIGNAL 1..27
FT /evidence="ECO:0000256|SAM:SignalP"
FT CHAIN 28..379
FT /evidence="ECO:0000256|SAM:SignalP"
FT /id="PRO_5004159498"
FT DOMAIN 34..110
FT /note="Inhibitor I9"
FT /evidence="ECO:0000259|Pfam:PF05922"
FT DOMAIN 133..370
FT /note="Peptidase S8/S53"
FT /evidence="ECO:0000259|Pfam:PF00082"
FT ACT_SITE 142
FT /note="Charge relay system"
FT /evidence="ECO:0000256|PROSITE-ProRule:PRU01240"
FT ACT_SITE 172
FT /note="Charge relay system"
FT /evidence="ECO:0000256|PROSITE-ProRule:PRU01240"
FT ACT_SITE 325
FT /note="Charge relay system"
FT /evidence="ECO:0000256|PROSITE-ProRule:PRU01240"
SQ SEQUENCE 379 AA; 38961 MW; EB6918B44172829D CRC64;
Query Match 85.3%; Score 1161; Length 379;
Best Local Similarity 82.8%;
Matches 222; Conservative 23; Mismatches 23; Indels 0; Gaps 0;
Qy 2 QSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGNG 61
|:|||||:||||| | :|| ||:||:||||||||| | || ||||||||||||: ||||
Db 112 QTVPWGINRVQAPIAQSRGFTGTGVRVAVLDTGISNHADLRIRGGASFVPGEPNISDGNG 171
Qy 62 HGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVAN 121
|||||||||||||||||||||||: :|| |||||||||||:| |||||:|| |||||:||
Db 172 HGTHVAGTIAALNNSIGVLGVAPNVDLYGVKVLGASGSGSISGIAQGLQWAANNGMHIAN 231
Qy 122 LSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNRA 181
:|||| : |||:||||| ||: ||||||||||||||:: :||||||||||||||||||||
Db 232 MSLGSSAGSATMEQAVNQATASGVLVVAASGNSGAGNVGFPARYANAMAVGATDQNNNRA 291
Qy 182 SFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQIR 241
|||||||||||||||| |||| ||: |:| |||||||||||| |||||||||||||||||
Db 292 SFSQYGAGLDIVAPGVGVQSTVPGNGYSSFNGTSMATPHVAGVAALVKQKNPSWSNVQIR 351
Qy 242 NHLKNTATSLGSTNLYGSGLVNAEAATR 269
||||||||:||:|| :||||||||||||
Db 352 NHLKNTATNLGNTNQFGSGLVNAEAATR 379
Sequence alignment between the protease of SEQ ID NO:2 of the instant application (“Qy”) and protease of Masui (“Db”)
Title: US-18-043-532-2
Perfect score: 1391
Sequence: 1 AQSVPYGVSQIKAPALHSQG..........KLGDSFYYGKGLINVQAAAQ 275
Scoring table: BLOSUM62
Gapop 10.0 , Gapext 0.5
Searched: 1 seqs, 379 residues
Total number of hits satisfying chosen parameters: 1
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 1 summaries
Database : AASEQ2_12032025_085931.pep:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 824 59.2 379 1 AASEQ2_12032025_085931
ALIGNMENTS
RESULT 1
AASEQ2_12032025_085931
Query Match 59.2%; Score 824; DB 1; Length 379;
Best Local Similarity 56.2%;
Matches 154; Conservative 54; Mismatches 60; Indels 6; Gaps 3;
Qy 2 QSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQDN 61
|:||:|:::::|| |:|:||: |:|||:|:|| |:| ||:: |||| || | | |
Db 112 QTVPWGINRVQAPIAQSRGFTGTGVRVAVLDTGI-SNHADLRIRGGASFVPGEPN-ISDG 169
Qy 62 NSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIA NNMDV 121
| ||||||||:||||||||||||||: || |||||| ||| | | |::|| | | :
Db 170 NGHGTHVAGTIAALNNSIGVLGVAPNVDLYGVKVLGASGSGSISGIAQGLQWAANNGMHI 229
Qy 122 INMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAVD 181
||||| :||| :: ||::| ||||:||||:|| | : ||:| :| : :|||| |
Db 230 ANMSLGSSAGSATMEQAVNQATASGVLVVAASGNSG----AGNVGFPARYANAMAVGATD 285
Qy 182 SSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPNW 241
:| ||||| | ||::|||| :|||:||| | ::||||||:||||| |||: |:|:|
Db 286 QNNNRASFSQYGAGLDIVAPGVGVQSTVPGNGYSSFNGTSMATPHVAGVAALVKQKNPSW 345
Qy 242 TNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ 275
:| |:|: |:|| | ||:: :| ||:| :|| :
Db 346 SNVQIRNHLKNTATNLGNTNQFGSGLVNAEAATR 379
Sequence alignment between the protease of SEQ ID NO:1 of the instant application (“Qy”) and protease of SEQ ID NO:2 of the instant application (“Db”)
Title: US-18-043-532-1
Perfect score: 1361
Sequence: 1 AQSVPWGISRVQAPAAHNRG..........SLGSTNLYGSGLVNAEAATR 269
Scoring table: BLOSUM62
Gapop 10.0 , Gapext 0.5
Searched: 1 seqs, 275 residues
Total number of hits satisfying chosen parameters: 1
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 1 summaries
Database : US-18-043-532-2.pep:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 851 62.5 275 1 US-18-043-532-2 Protease variants
ALIGNMENTS
RESULT 1
US-18-043-532-2
Query Match 62.5%; Score 851; DB 1; Length 275;
Best Local Similarity 60.0%;
Matches 165; Conservative 45; Mismatches 59; Indels 6; Gaps 3;
Qy 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGI-STHPDLNIRGGASFVPGEPST-QD 58
|||||:|:|:::||| |::| ||| |||||:|:|| |:|||| : |||| || | : ||
Db 1 AQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQD 60
Qy 59 GNGHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMH 118
| ||||||||:|||||||||||||||| ||||||||| ||| | | |:||| | |
Db 61 NNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIA NNMD 120
Qy 119 VANLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSG----AGSISYPARYANAMAVGAT 174
| |:||| || || |: ||: | : ||:||||:|| | : :: || :| : :||||
Db 121 VINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAV 180
Qy 175 DQNNNRASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPS 234
| :| ||||| | ||::||||::||| ||: | : ||||||:|||||||||: |:|:
Db 181 DSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPN 240
Qy 235 WSNVQIRNHLKNTATSLGSTNLYGSGLVNAEAATR 269
|:| |:|: |:|| | || : || ||:| :|| :
Db 241 WTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ 275
Sequence alignment between the protease of SEQ ID NO:1 of the instant application (“Qy”) and the protease of SEQ ID NO:1 of Babe (“Db”)
Title: US-18-043-532-1
Perfect score: 1361
Sequence: 1 AQSVPWGISRVQAPAAHNRG..........SLGSTNLYGSGLVNAEAATR 269
Scoring table: BLOSUM62
Gapop 10.0 , Gapext 0.5
Searched: 1 seqs, 269 residues
Total number of hits satisfying chosen parameters: 1
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 1 summaries
Database : AASEQ2_12032025_115153.pep:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 1361 100.0 269 1 AASEQ2_12032025_115153
ALIGNMENTS
RESULT 1
AASEQ2_12032025_115153
Query Match 100.0%; Score 1361; DB 1; Length 269;
Best Local Similarity 100.0%;
Matches 269; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGN 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGN 60
Qy 61 GHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 GHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVA 120
Qy 121 NLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNR 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 NLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNR 180
Qy 181 ASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQI 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 ASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQI 240
Qy 241 RNHLKNTATSLGSTNLYGSGLVNAEAATR 269
|||||||||||||||||||||||||||||
Db 241 RNHLKNTATSLGSTNLYGSGLVNAEAATR 269
Sequence alignment between the protease of SEQ ID NO:2 of the instant application (“Qy”) and the protease of SEQ ID NO:2 of Babe (“Db”)
Title: US-18-043-532-2
Perfect score: 1391
Sequence: 1 AQSVPYGVSQIKAPALHSQG..........KLGDSFYYGKGLINVQAAAQ 275
Scoring table: BLOSUM62
Gapop 10.0 , Gapext 0.5
Searched: 1 seqs, 275 residues
Total number of hits satisfying chosen parameters: 1
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 1 summaries
Database : AASEQ2_12032025_115533.pep:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 1384 99.5 275 1 AASEQ2_12032025_115533
ALIGNMENTS
RESULT 1
AASEQ2_12032025_115533
Query Match 99.5%; Score 1384; DB 1; Length 275;
Best Local Similarity 99.6%;
Matches 274; Conservative 0; Mismatches 1; Indels 0; Gaps 0;
Qy 1 AQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQD 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQD 60
Qy 61 NNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIA NNMD 120
|||||||||||||||||||||||||||||||||||||| |||||||||||||||||||||
Db 61 NNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGAOGSGQYSWIINGIEWAIA NNMD 120
Qy 121 VINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAV 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 VINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAV 180
Qy 181 DSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPN 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 DSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPN 240
Qy 241 WTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ 275
|||||||||||||||||||||||||||||||||||
Db 241 WTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ 275
Sequence alignment between the protease of SEQ ID NO:1 of the instant application (“Qy”) and the protease of SEQ ID NO:1 of Knotzel (“Db”)
US-17-435-555-1
Filing date in PALM: 2021-09-01
Sequence 1, US/17435555
Publication No. US20220145220A1
GENERAL INFORMATION
APPLICANT: Novozymes A/S
TITLE OF INVENTION: DETERGENT COMPOSITIONS COMPRISING TWO PROTEASES
FILE REFERENCE: 14991-WO-PCT
CURRENT APPLICATION NUMBER: US/17/435,555
CURRENT FILING DATE: 2021-09-01
NUMBER OF SEQ ID NOS: 2
SEQ ID NO 1
LENGTH: 269
TYPE: PRT
ORGANISM: Bacillus lentus
Query Match 100.0%; Score 1361; Length 269;
Best Local Similarity 100.0%;
Matches 269; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGN 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGN 60
Qy 61 GHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 GHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVA 120
Qy 121 NLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNR 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 NLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNR 180
Qy 181 ASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQI 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 ASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQI 240
Qy 241 RNHLKNTATSLGSTNLYGSGLVNAEAATR 269
|||||||||||||||||||||||||||||
Db 241 RNHLKNTATSLGSTNLYGSGLVNAEAATR 269
Sequence alignment between the protease of SEQ ID NO:2 of the instant application (“Qy”) and the protease of SEQ ID NO:2 of Knotzel (“Db”)
US-17-435-555-2
Filing date in PALM: 2021-09-01
Sequence 2, US/17435555
Publication No. US20220145220A1
GENERAL INFORMATION
APPLICANT: Novozymes A/S
TITLE OF INVENTION: DETERGENT COMPOSITIONS COMPRISING TWO PROTEASES
FILE REFERENCE: 14991-WO-PCT
CURRENT APPLICATION NUMBER: US/17/435,555
CURRENT FILING DATE: 2021-09-01
NUMBER OF SEQ ID NOS: 2
SEQ ID NO 2
LENGTH: 275
TYPE: PRT
ORGANISM: Bacillus amyloliquefaciens
Query Match 100.0%; Score 1391; Length 275;
Best Local Similarity 100.0%;
Matches 275; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQD 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AQSVPYGVSQIKAPALHSQGYTGSNVKVAVIDSGIDSSHPDLKVAGGASMVPSETNPFQD 60
Qy 61 NNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIA NNMD 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 NNSHGTHVAGTVAALNNSIGVLGVAPSASLYAVKVLGADGSGQYSWIINGIEWAIA NNMD 120
Qy 121 VINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAV 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 VINMSLGGPSGSAALKAAVDKAVASGVVVVAAAGNEGTSGSSSTVGYPGKYPSVIAVGAV 180
Qy 181 DSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPN 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 DSSNQRASFSSVGPELDVMAPGVSIQSTLPGNKYGAYNGTSMASPHVAGAAALILSKHPN 240
Qy 241 WTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ 275
|||||||||||||||||||||||||||||||||||
Db 241 WTNTQVRSSLENTTTKLGDSFYYGKGLINVQAAAQ 275
Sequence alignment between the protease of SEQ ID NO:1 of the instant application (“Qy”) and the protease of SEQ ID NO:1 of Lenhard (“Db”)
US-18-043-515-1
Filing date in PALM: 2023-02-28
Sequence 1, US/18043515
Publication No. US20230323330A1
GENERAL INFORMATION
APPLICANT: Novozymes A/S
TITLE OF INVENTION: Polyester degrading protease variants
FILE REFERENCE: 15178-WO-PCT
CURRENT APPLICATION NUMBER: US/18/043,515
CURRENT FILING DATE: 2023-02-28
NUMBER OF SEQ ID NOS: 91
SEQ ID NO 1
LENGTH: 269
TYPE: PRT
ORGANISM: Bacillus clausii
Query Match 100.0%; Score 1361; Length 269;
Best Local Similarity 100.0%;
Matches 269; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGN 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AQSVPWGISRVQAPAAHNRGLTGSGVKVAVLDTGISTHPDLNIRGGASFVPGEPSTQDGN 60
Qy 61 GHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVA 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 GHGTHVAGTIAALNNSIGVLGVAPSAELYAVKVLGASGSGSVSSIAQGLEWAGNNGMHVA 120
Qy 121 NLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNR 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 NLSLGSPSPSATLEQAVNSATSRGVLVVAASGNSGAGSISYPARYANAMAVGATDQNNNR 180
Qy 181 ASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQI 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 ASFSQYGAGLDIVAPGVNVQSTYPGSTYASLNGTSMATPHVAGAAALVKQKNPSWSNVQI 240
Qy 241 RNHLKNTATSLGSTNLYGSGLVNAEAATR 269
|||||||||||||||||||||||||||||
Db 241 RNHLKNTATSLGSTNLYGSGLVNAEAATR 269
Prosecution Insights