DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on July 27, 2026 has been entered.
Claims 1-16, 20, 27, 31-32, and 40-41 have been canceled. Claims 17-19, 21-26, 28-30, and 33-39 are pending, Claims 19, 23, 25-26, 29, 33-34, and 36-39 have been withdrawn, and Claims 17-18, 21-22, 24, 28, 30, and 35 have been considered on the merits, insofar as they read on the elected species of hydrolase, a muramidase of SEQ ID NO: 1, a bolus addition, a fed batch fermentation process wherein the at least one enzyme is added to the fermentation broth, adding up to 500 mg/kg of the at least one enzyme per kg of fermentation broth, and a Bacillus species. All arguments have been fully considered.
Claim Rejections - 35 USC § 103
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 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102 of this title, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
Claims 17-18, 21, 24, 28, and 35 are rejected under 35 U.S.C. 103 as being unpatentable over Glanville et al (WO 2018/185048 A1; 10/11/2018.) in view of Ho (J Appl Biotechnol Bioeng. 2016;1(3):96-104.), Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) and Schnorr et al (WO 2013/076253 A1; 5/30/2013.).
The instant claims recite a submerged fermentation process of producing a polypeptide of interest, the process comprising: a) fermenting a microorganism that produces the polypeptide of interest in a stirred or agitated fermentation broth, and b) adding at least one enzyme to the fermentation broth, or co-expressing the at least one enzyme in the microorganism and secreting the at least one enzyme into the fermentation broth, in an amount sufficient to reduce the viscosity of the fermentation broth compared to when the at least one enzyme is not added or co-expressed; wherein the polypeptide of interest is a secreted enzyme; and wherein the at least one enzyme is a muramidase of the glycosyl hydrolase GH24 or GH25 family.
Glanville teaches a method for recovering a desired fermentation product from a fermentation broth (Title), comprising fermenting a microorganism that produces one or more desired fermentation products in a fermentation broth (a liquid medium – a submerged fermentation) (p.3 line 32-34, p.4 line 1-2), and adding lysozyme (muramidase) and/or hydrolyzing enzymes to the fermentation broth (p.11 line 23-24), wherein the one or more desired fermentation products is preferably selected among proteins, such as enzymes, e.g., hydrolases, isomerases, ligases, lyases, oxidoreductases, and transferases (a polypeptide of interest) (p.4 line 21-22 & 25-27), or obtained from a microorganism (p.8 line 25) such as any Bacillus cell (p.5 line 35, p.9 line 1). The method further comprises recovering said products (p.16 line 16). The method is a fed-batch fermentation (p.4 line 15). The microorganism is a Bacillus species (p.9 line 1-6).
Glanville does not teach the method wherein the fermentation broth is stirred or agitated (claim 17).
However, Glanville does teach batch submerged fermentation. Ho teaches growth conditions including agitation speed affect the outcome of enzyme production in batch submerged fermentation (p.98 col right – para 2).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to agitate a fermentation broth as a routine practice in the art, since Glanville discloses batch submerged fermentation, and Ho discloses that agitation speed affects the outcome of enzyme production in batch submerged fermentation. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference and routine practice to agitate a fermentation broth with a reasonable expectation for successfully recovering a desired fermentation product from a fermentation broth.
References cited above do not teach the method wherein the at least one enzyme is added in an amount sufficient to reduce the viscosity of the fermentation broth compared to when the at least one enzyme is not added or co-expressed (claim 17).
However, Glanville does teach the method comprises fermenting Bacillus species in a fermentation broth, and adding lysozyme to the fermentation broth. Before the effective filing date of the claimed invention, it was well-known in the art that lysozyme reduces fermentation broth viscosity. In support, Kimmenade teaches a method comprises fermenting Bacillus species in a fermentation broth (para 00127), wherein 2 µl lysozyme solution reduces fermentation broth viscosity compared to when the lysozyme solution is not added (para 00129).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to add a sufficient amount of lysozyme to a fermentation broth to reduce the viscosity of the fermentation broth, since Glanville discloses a method comprises adding lysozyme to a fermentation broth, and Kimmenade discloses that lysozyme reduces fermentation broth viscosity. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference and routine practice to add a sufficient amount of lysozyme to a fermentation broth with a reasonable expectation for successfully recovering a desired fermentation product from a fermentation broth.
References cited above do not teach the method wherein the at least one enzyme is a muramidase of the glycosyl hydrolase GH24 or GH25 family (claim 17), and the at least one enzyme comprises a GH24 or GH25 muramidase comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence shown in any of SEQ ID NO: 1 (claim 28).
However, Glanville does teach the method comprises adding one or more cell wall degrading enzymes such as lysozyme(s) (muramidase). Schnorr teaches addition of one or more lysozymes affords the breakdown of cell walls from bacteria, which is not possible using current commercial solutions, wherein said one or more lysozymes include a GH25 lysozyme having SEQ ID NO: 4 (the claimed SEQ ID NO: 1, see Result 3 in Search results filed on 07/18/2025) (p.47 line 10-12 & 22-25).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to incorporate the claimed muramidase, since Glanville discloses a method comprises adding one or more cell wall degrading enzymes such as lysozyme(s) (muramidase), and Schnorr discloses that the claimed muramidase affords the breakdown of cell walls from bacteria, which is not possible using current commercial solutions. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference to incorporate the claimed muramidase with a reasonable expectation for successfully recovering a desired fermentation product from a fermentation broth.
Claim 22 is rejected under 35 U.S.C. 103 as being unpatentable over Glanville et al (WO 2018/185048 A1; 10/11/2018.) in view of Ho (J Appl Biotechnol Bioeng. 2016;1(3):96-104.), Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) and Schnorr et al (WO 2013/076253 A1; 5/30/2013.) as applied to claims 17-18, 21, 24, 28, and 35 above, further in view of Lindskog et al (Biopharmaceutical Processing Development, Design, and Implementation of Manufacturing Processes. 2018, Chapter 31, p.625-635.).
References cited above do not teach the method wherein the at least one enzyme is added as a bolus addition (claim 22).
However, Glanville does teach fed-batch fermentation. Lindskog teaches fed-batch fermentation wherein a feed solution can be added intermittently (bolus feed), and bolus feed could be a better option when in very large-scale simplicity and robustness might need to be prioritized before sophistication (p.628 para 4).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to add an enzyme as a bolus addition, since Glanville discloses fed-batch fermentation, and Lindskog discloses fed-batch fermentation where a feed solution can be added as a bolus feed, and bolus feed could be a better option in certain situations. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference and routine practice to add an enzyme as a bolus addition with a reasonable expectation for successfully recovering a desired fermentation product from a fermentation broth.
Claim 30 is rejected under 35 U.S.C. 103 as being unpatentable over Glanville et al (WO 2018/185048 A1; 10/11/2018.) in view of Ho (J Appl Biotechnol Bioeng. 2016;1(3):96-104.), Kimmenade et al (WO 2009/102755 A1; 8/20/2009.) and Schnorr et al (WO 2013/076253 A1; 5/30/2013.) as applied to claims 17-18, 21, 24, 28, and 35 above, further in view of Robinson (Essays Biochem. 2015;59:1-41.).
References cited above do not teach the method comprises adding up to 500 mg/kg of the at least one enzyme per kg of fermentation broth (claim 30).
Robinson teaches the amount of enzyme controls reaction rates, the relationship between the reaction rate and the amount of enzyme is a simple linear relationship, and substrate concentration effects the linear relationship (p.10 para 3-5).
Thus, before the effective filing date of the claimed invention, it would have been obvious to one of ordinary skill in the art to incorporate an optimized concentration of an enzyme, since it was well-known in the art that the amount of enzyme controls reaction rates, and that substrate concentration effects the amount of enzyme used in the reaction. In other words, the optimized amount of enzyme used in fermentation broth depends on other parameters such as substrate concentration. Moreover, before the effective filing date of the claimed invention, one of ordinary skill in the art would have been motivated by the cited reference to incorporate an optimized concentration of an enzyme with a reasonable expectation for successfully recovering a desired fermentation product from a fermentation broth.
Response to Arguments
Applicant argues that the primary combination fails to render claim 17 obvious, and that secondary references do not cure the deficiencies. However, these arguments are moot in light of the new rejections above in view of applicant’s amendments.
Conclusion
No claims are allowed.
Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to LYNN Y FAN whose telephone number is (571)270-3541. The examiner can normally be reached on M-F 7am-4pm.
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/Lynn Y Fan/
Primary Examiner, Art Unit 1759
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