DETAILED CORRESPONDENCE
Status of the Application
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Applicant’s amendment to the specification, filed August 11, 2026 is acknowledged.
Claims 14-26 are pending in the application.
Restriction/Election
Applicant’s election without traverse of Group I, claims 14-22, in the reply filed on August 11, 2026 is acknowledged.
Claims 23-26 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim.
Priority
This application is filed under 35 U.S.C. 371 as a national stage of international application PCT/EP2022/055177, filed March 1, 2022.
Information Disclosure Statement
The information disclosure statements (IDSs) submitted on November 22, 2024 and October 21, 2025 are in compliance with the provisions of 37 CFR 1.97. Accordingly, the IDSs have been considered by the examiner.
Specification/Informalities
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code at p. 11, lines 11 and 20 and p. 20, line 21. Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
Claim Objections
Claims 14, 18, and 22 are objected to because of the following informalities:
Claim 14 is objected to in the recitation of the abbreviation “crp.” In the interest of improving claim form, it is suggested that the entire phrase for which the abbreviation is used (e.g., catabolite repressor protein) be recited at least once in the claim.
Claim 18 is objected to in the recitation of “as claimed claim 14” and in the interest of improving claim form, it is suggested that the noted phrase be amended to recite (with markings to show changes made) “as claimed in claim 14.”
Claim 22 is objected to in the recitation of “the expressed Crp protein is SEQ ID NO: 2” and in the interest of improving claim form, it is suggested that the noted phrase be amended to recite (with markings to show changes made) “the amino acid sequence of the expressed Crp protein is SEQ ID NO: 2.”
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.
Claims 14-22 are rejected under 35 U.S.C. 112(b) as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor regards as the invention.
Claim 14 (claims 15-22 dependent therefrom) recites the limitations “the crp gene” and “the crp promoter sequence.” There is insufficient antecedent basis for these limitations in the claim.
Claim 15 recites the limitation “the Crp protein.” There is insufficient antecedent basis for this limitation in the claim.
Claim 15 is indefinite in the recitation of “the amino acid sequence of the Crp protein is nonmutated” because it is unclear as to whether “the Crp protein” is intended to reference the Crp protein encoded by “the crp gene having a wild-type promoter” and/or “the crp gene” with a mutated crp promoter sequence. Also, the phrase “the amino acid sequence of the Crp protein is nonmutated” is indefinite because it is unclear as to the reference Crp protein to which one of skill in the art would compare in order to make a determination of whether or not “the Crp protein” is or is not mutated. In the interest of advancing prosecution, applicant may consider amending claim 15 to recite a reference Crp protein such that one of skill in the art could determine whether or not the Crp protein is or is not mutated.
Claim 22 recites the limitation “the expressed Crp protein.” There is insufficient antecedent basis for this limitation in the claim. Also, it is unclear as to whether “the expressed Crp protein” is intended to reference the Crp protein encoded by “the crp gene having a wild-type promoter” and/or “the crp gene” with a mutated crp promoter sequence.
Claim Rejections - 35 USC § 102/103
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.
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, 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 14-19 and 22 are rejected under 35 U.S.C. 102(a)(1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Zhou et al. (Microorganisms 8:517, 2020, 13 pages; cited on Form PTO-892 filed June 18, 2026; hereafter “Zhou”) as evidenced by UniProt Accession Number A0AAU8PUA3 (June 2026, 2 pages; cited on the attached Form PTO-892; hereafter “UniProt A0AAU8PUA3”).
Applicant’s attention is directed to MPEP 2112.III regarding a rejection under 35 U.S.C. 102 and 103 when the prior art product seems to be identical except that the prior art is silent as to an inherent characteristic.
Applicant’s attention is also directed to MPEP 2131.01.III regarding a multiple reference 35 U.S.C. 102 rejection with an extra reference to show an inherent characteristic of the thing taught by the primary reference, the critical date of extrinsic evidence showing a universal fact need not antedate the filing date.
Claims 14-19 and 22 are drawn to a microorganism strain having a deregulated cysteine biosynthesis pathway that is thereby suitable for fermentative production of at least one substance selected from L-cysteine, L-cystine and thiazolidine, characterized in that the relative expression of the crp gene is reduced in relation to the expression of the crp gene having a wild-type promoter sequence as a result of mutation of the crp promoter sequence.
Given a broadest reasonable interpretation, the phrase “mutation of the crp promoter sequence” in claim 14 encompasses any deletion(s), insertion(s), or substitution(s) of the crp promoter sequence.
Regarding claim 14, Zhou teaches a gram-negative bacterium with a replacement of the crp gene promoter with a PBAD promoter (ΔPcrp), which, in the absence of arabinose, does not express the crp gene (p. 1, Abstract; p. 10, bottom).
Zhou does not teach ΔPcrp exhibits a “deregulated cysteine biosynthesis pathway that is thereby suitable for fermentative production of at least one substance selected from L-cysteine, L-cystine and thiazolidine.” However, according to MPEP 2112.01.I, when the structure recited in the reference is substantially identical to that of the claims, claimed properties or functions are presumed to be inherent. Since the ΔPcrp of Zhou does not express the crp gene in the absence of arabinose, it is presumed that in the absence of arabinose, the ΔPcrp of Zhou has a deregulated cysteine biosynthesis pathway that is thereby suitable for fermentative production of at least one substance selected from L-cysteine, L-cystine and thiazolidine. Since the Office does not have the facilities for examining and comparing applicants’ microorganism strain with the ΔPcrp of Zhou, the burden is on the applicant to show a novel or unobvious difference between the claimed microorganism strain and the ΔPcrp of Zhou. See MPEP 2112.V.
Regarding claim 15, as stated above, claim 15 does not recite a reference Crp protein such that one of ordinary skill in the art could make a determination of whether or not the Crp is or is not mutated. Also, Zhou does not mutate the Crp protein of ΔPcrp.
Regarding claim 16, Zhou teaches the crp promoter (Pcrp) and its 5’ and 3’ flanking regions were replaced by the PBAD promoter in constructing ΔPcrp (p. 3, bottom).
Regarding claim 17, Zhou teaches the crp promoter (Pcrp) and its 5’ and 3’ flanking regions were replaced by the PBAD promoter in constructing ΔPcrp (p. 3, bottom). Given that the entire crp promoter was deleted from ΔPcrp, it follows that at least nucleotides 565-624 are deleted from the crp promoter sequence in ΔPcrp.
Regarding claim 18, Zhou teaches the crp promoter (Pcrp) and its 5’ and 3’ flanking regions were replaced by the PBAD promoter in constructing ΔPcrp (p. 3, bottom). Given that the entire crp promoter was deleted from ΔPcrp, it follows that the crp promoter sequence is completely deleted in ΔPcrp.
Regarding claim 19, Zhou teaches ΔPcrp is derived from Edwardsiella piscicida, which belongs to the Enterobacteriaceae family (p. 10, bottom).
Regarding claim 22, Zhou does not teach the amino acid sequence of the CRP expressed from ΔPcrp. However, given that ΔPcrp comprises an Edwardsiella piscicida crp gene, it follows that ΔPcrp express an E. piscicida CRP protein. Evidentiary reference UniProt A0AAU8PUA3 is cited to show that the amino acid sequence of E. piscicida CRP protein is identical to instant SEQ ID NO: 2 (see Appendix A for sequence alignment).
Therefore, claims 14-19 and 22 are anticipated by or, in the alternative, are obvious over Zhou.
Claims 14-17 and 19-22 are rejected under 35 U.S.C. 102(a)(1) as anticipated by or, in the alternative, under 35 U.S.C. 103 as obvious over Ishizuka et al. (EMBO J. 13:3077-3082, 1994; cited on the attached Form PTO-892; hereafter “Ishizuka”) as evidenced by UniProt Accession Number P0ACJ8 (June 2026, 15 pages; cited on the attached Form PTO-892; hereafter “UniProt P0ACJ8”).
Applicant’s attention is directed to MPEP 2112.III regarding a rejection under 35 U.S.C. 102 and 103 when the prior art product seems to be identical except that the prior art is silent as to an inherent characteristic.
Applicant’s attention is also directed to MPEP 2131.01.III regarding a multiple reference 35 U.S.C. 102 rejection with an extra reference to show an inherent characteristic of the thing taught by the primary reference, the critical date of extrinsic evidence showing a universal fact need not antedate the filing date.
Regarding claims 14, 16, and 19-21, Ishizuka teaches a crp– strain of Escherichia coli referred to as PP47 (p. 3077, column 2, bottom; p. 079, column 2, middle). Ishizuka teaches PP47 cells transformed with plasmids comprising the crp gene with point mutations and/or deletions of the crp promoter (paragraph bridging pp. 3079-3080; p. 3081, Figure 7). Ishizuka teaches that when the deletion extends into the CRP binding site II, the crp expression was significantly reduced (sentence bridging pp. 3079; p. 3081, Figure 8A) and with pHA10m, which carries point mutations in the CRP binding site II, the CRP level was clearly reduced (p. 3080, column 1, top; p. 3081, Figure 8B).
Ishizuka does not teach the PP47 cells transformed with plasmids comprising the crp gene with mutations and/or deletions of the crp promoter (PP47 transformants) exhibit a “deregulated cysteine biosynthesis pathway that is thereby suitable for fermentative production of at least one substance selected from L-cysteine, L-cystine and thiazolidine.” However, according to MPEP 2112.01.I, when the structure recited in the reference is substantially identical to that of the claims, claimed properties or functions are presumed to be inherent. Since the PP47 transformants of Ishizuka exhibit reduced expression of CRP, it is presumed that the PP47 transformants of Ishizuka have a deregulated cysteine biosynthesis pathway that is thereby suitable for fermentative production of at least one substance selected from L-cysteine, L-cystine and thiazolidine. Since the Office does not have the facilities for examining and comparing applicants’ microorganism strain with the PP47 transformants of Ishizuka, the burden is on the applicant to show a novel or unobvious difference between the claimed microorganism strain and the PP47 transformants of Ishizuka. See MPEP 2112.V.
Regarding claim 15, as stated above, claim 15 does not recite a reference Crp protein such that one of ordinary skill in the art could make a determination of whether or not the Crp is or is not mutated. Also, Ishizuka does not mutate the Crp protein of the plasmids comprising the crp gene with mutations and/or deletions of the crp promoter.
Regarding claim 17, the CRP binding site II shown in Figure 7B of Ishizuka (p. 3081) corresponds to nucleotides 628-649 of SEQ ID NO: 1. As such, PP47 transformed with plasmids pHA101 and pHA102, which have a partial 5’ deletion or a complete deletion of the CRP binding site II (p. 3081, Figure 7A), have at least nucleotides 565-624 deleted from the crp promoter sequence.
Regarding claim 22, Ishizuka does not teach the amino acid sequence of the CRP expressed from the plasmids comprising the crp gene with point mutations and/or deletions of the crp promoter. However, given that the plasmids comprising the crp gene with point mutations and/or deletions of the crp promoter were derived from pHA5 (p. 3081, Figure 7A), which comprises an E. coli crp gene, it follows that the plasmids comprising the crp gene with point mutations and/or deletions of the crp promoter express an E. coli CRP protein. Evidentiary reference UniProt P0ACJ8 is cited to show that the amino acid sequence of E. coli CRP protein is identical to instant SEQ ID NO: 2 (see Appendix B for sequence alignment).
Therefore, claims 14-17 and 19-22 are anticipated by or, in the alternative, are obvious over Ishizuka.
Claim Rejections - 35 USC § 103
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, 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 14-22 are rejected under 35 U.S.C. 103 as being unpatentable over Tanaka et al. (J. Gen. Appl. Microbiol. 65:234-239, 2019; cited on the attached Form PTO-892; hereafter “Tanaka”) in view of Cossart et al. (Nucleic Acids Res. 10:1363-1378, 1982; cited on the attached Form PTO-892; hereafter “Cossart”) and Ishizuka.
Regarding claims 14, 16, and 18-21, Tanaka teaches the production of cysteine (Cys) and its derivatives by fermentation is important in terms of its applications. In the processes of E. coli fermentative overproduction, H2S gas is generated from the cells, which is often considered as a serious risk to human life in the manufacturing environment due to the H2S toxicity. Therefore, it is important to understand the potential mechanisms of H2S production in E. coli (p. 235, column 1, middle). Tanaka investigates and reports findings on H2S production in E. coli (see whole document). Tanaka teaches their results suggest that deletion of the crp gene is a novel strategy to produce a sulfur-containing organic compound while minimizing H2S generation, which can be a risk for human life and product yield (p. 237, sentence bridging columns 1-2).
The difference between Tanaka and claims 14, 16, and 18-21 is that Tanaka does not explicitly teach mutation of the crp promoter sequence to reduce expression of the crp gene.
Cossart teaches cloning and sequence of the crp gene of Escherichia coli K12 (p. 1363, title). Cossart teaches the whole crp region within a 4 kilobase fragment including the crp promoter was cloned into a plasmid for sequencing (p. 1370, middle; p. 1371, Figure 2).
In view of the combined teachings of Tanaka and Cossart, it would have been obvious to one of ordinary skill in the art before the effective filing date to delete the whole crp region – including the crp promoter – of an E. coli for the fermentative production of L-cysteine and its derivatives. One would have been motivated and would have expected success to do this because Tanaka teaches that fermentative overproduction of cysteine and its derivatives by E. coli produces H2S gas, which is considered to be a serious risk to human life; Tanaka suggests deletion of the crp gene to produce a sulfur-containing organic compound while minimizing H2S generation; and Cossart teaches the whole crp region includes the crp promoter.
Regarding claim 15, as stated above, the recitation of “the amino acid sequence of the Crp protein is nonmutated” is indefinite because it is unclear as to whether “the Crp protein” is intended to reference the Crp protein encoded by “the crp gene having a wild-type promoter” and/or “the crp gene” with a mutated crp promoter sequence. In the interest of advancing prosecution, claim 15 is interpreted as meaning the amino acid sequence of the Crp protein encoded by “the crp gene having a wild-type promoter” is nonmutated while the Crp protein of the “the crp gene” with a mutated crp promoter sequence can be mutated by any deletion(s), insertion(s), or substitution(s). In view of this interpretation, an E. coli with a deletion of the whole crp region is encompassed by claim 15.
Regarding claim 17, E. coli with a deletion of the whole crp region – including the crp promoter – has a deletion of at least nucleotides 565-624 from the crp promoter sequence.
Regarding claim 22, as stated above, it is unclear as to whether “the expressed Crp protein” is intended to reference the Crp protein encoded by “the crp gene having a wild-type promoter” and/or “the crp gene” with a mutated crp promoter sequence. In the interest of advancing prosecution, claim 22 is interpreted as meaning the amino acid sequence of the Crp protein encoded by “the crp gene having a wild-type promoter” is SEQ ID NO: 2 while the Crp protein of the “the crp gene” with a mutated crp promoter sequence can be mutated by any deletion(s), insertion(s), or substitution(s). In view of this interpretation, an E. coli with a deletion of the whole crp region is encompassed by claim 22.
Therefore, the microorganism strain of claims 14 and 16-22 would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention.
Conclusion
Status of the claims:
Claims 14-26 are pending.
Claims 23-26 are withdrawn from further consideration.
Claims 14-22 are rejected.
No claim is in condition for allowance.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to DAVID J STEADMAN whose telephone number is (571)272-0942. The examiner can normally be reached Monday to Friday, 7:30 AM to 4:00 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, MANJUNATH N RAO can be reached on 571-272-0939. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/David Steadman/Primary Examiner, Art Unit 1656
APPENDIX A
ALIGNMENT:
Query Match 100.0%; Score 1073; Length 210;
Best Local Similarity 100.0%;
Matches 210; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MVLGKPQTDPTLEWFLSHCHIHKYPSKSTLIHQGEKAETLYYIVKGSVAVLIKDEEGKEM 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MVLGKPQTDPTLEWFLSHCHIHKYPSKSTLIHQGEKAETLYYIVKGSVAVLIKDEEGKEM 60
Qy 61 ILSYLNQGDFIGELGLFEEGQERSAWVRAKTACEVAEISYKKFRQLIQVNPDILMRLSAQ 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ILSYLNQGDFIGELGLFEEGQERSAWVRAKTACEVAEISYKKFRQLIQVNPDILMRLSAQ 120
Qy 121 MARRLQVTSEKVGNLAFLDVTGRIAQTLLNLAKQPDAMTHPDGMQIKITRQEIGQIVGCS 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 MARRLQVTSEKVGNLAFLDVTGRIAQTLLNLAKQPDAMTHPDGMQIKITRQEIGQIVGCS 180
Qy 181 RETVGRILKMLEDQNLISAHGKTIVVYGTR 210
||||||||||||||||||||||||||||||
Db 181 RETVGRILKMLEDQNLISAHGKTIVVYGTR 210
APPENDIX B
ALIGNMENT:
Query Match 100.0%; Score 1073; Length 210;
Best Local Similarity 100.0%;
Matches 210; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MVLGKPQTDPTLEWFLSHCHIHKYPSKSTLIHQGEKAETLYYIVKGSVAVLIKDEEGKEM 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MVLGKPQTDPTLEWFLSHCHIHKYPSKSTLIHQGEKAETLYYIVKGSVAVLIKDEEGKEM 60
Qy 61 ILSYLNQGDFIGELGLFEEGQERSAWVRAKTACEVAEISYKKFRQLIQVNPDILMRLSAQ 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 ILSYLNQGDFIGELGLFEEGQERSAWVRAKTACEVAEISYKKFRQLIQVNPDILMRLSAQ 120
Qy 121 MARRLQVTSEKVGNLAFLDVTGRIAQTLLNLAKQPDAMTHPDGMQIKITRQEIGQIVGCS 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 MARRLQVTSEKVGNLAFLDVTGRIAQTLLNLAKQPDAMTHPDGMQIKITRQEIGQIVGCS 180
Qy 181 RETVGRILKMLEDQNLISAHGKTIVVYGTR 210
||||||||||||||||||||||||||||||
Db 181 RETVGRILKMLEDQNLISAHGKTIVVYGTR 210
Prosecution Insights