DETAILED ACTION
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 .
Applicants’ amendment and response of 10/6/2025 is acknowledged.
Claim Amendments
3. Applicants’ amendment of 10/6/2025 is acknowledged. Claims 1, 2, 3, 11 have been amended. Claims 5-8 and 27 have been canceled. Claims 9 and 22-26 have been previously canceled.
Status of the Claims
4. Claims 1, 2, 3, 4, and 10-21 are pending. Claims 1, 2, 3 and 11 have been amended. Claims 5-8 and 27 have been canceled. Claims 9 and 22-26 have been previously canceled. Claims 1, 2, 3, 4, 10,11,12, 16, 17,18 and 19 are under consideration. Claims 13-15 and 20-21 are withdrawn from further consideration as being drawn non-elected invention.
Claim Rejections - 35 USC § 103 Moot
5. Rejection of claims 5-8 and 27 under 35 U.S.C. 103 as being unpatentable over Mijts et al. (WO 2018079887 A1 filed 4/5/2018) in view of Fan (CN 116065411 English translation CN 116065411) and Fukui et al. (WO 20060574450 A1–and UNIPROT A0A1D81TKD3 (2/18/2017), is moot in view of cancellation of said claims.
Claim Rejections - 35 USC § 103 Maintained
6. Rejection of claims 1-4, 10-12 and 16-19 under 35 U.S.C. 103 as being unpatentable over Mijts et al. (WO 2018079887 A1 filed 4/5/2018) in view of Fan (CN 116065411 English translation CN 116065411) and Fukui et al. (WO 20060574450 A1–and UNIPROT A0A1D81TKD3 (2/18/2017), is moot in view of cancellation of said claims.
The rejection is as stated below:
Claims 1-4, 10-12, 16-19 is/are rejected under 35 U.S.C. 103 as being unpatentable over Mijts et al. (WO 2018079887 A1 filed 4/5/2018) in view of Fan (CN 116065411 English translation CN 116065411) and Fukui et al. (WO 20060574450 A1–and UNIPROT A0A1D81TKD3 (2/18/2017).
The amended claims are drawn to:
A microorganism comprising genes coding for enzymes having a function of an argininosuccinate lyase, an ornithine carbamoyltransferase, an argininosuccinate synthetase and a carbamoylphosphate synthase and comprising at least one heterologous gene coding for a protein having a function an activity of a L-arginine:glycine amidinotransferase, wherein said genes coding for enzymes having
the function of said argininosuccinate lyase, said ornithine carbamoyltransferase, said
argininosuccinate synthetase and said carbamoylphosphate synthase, and the at least one heterologous gene coding for the protein having the function of said L-arginine:glycine amidinotransferase, have increased activity compared to respective activity in a wildtype microorganism.
WO 2018079687 A1 discloses a microorganism (corynebacterium) comprising genes encoding an enzyme ( see abstract, claims 2, 11 and paragraphs 0004, 0006,0015, 0017, 0022, 0057, 0062, 0064 ). WO 2018079687 A1 para [0017] recites: “ It is a further aspect of the present invention to provide the method as described above, wherein the activity of the L-cysteine biosynthesis enzyme is increased by in-creasing the expression of a gene encoding the L-cysteine biosynthesis enzyme.”
WO 2018079687 A1 discloses L-arginine :glycine amidino transferase ( see para 0091). WO 2018079687 A1 discloses Corynebacterium, E.coli and wildtype microorganisms( see claims 17, 18, 19, 20 , 50 and para 0072-0073). WO 2018079687 A1 discloses the possibility to produce creatine from arginine and glycine using L- arginine biosynthesis enzymes, glycine biosynthesis enzymes and enzymes that catalyze the conversion of L-arginine and glycine into creatine. L-arginine and glycine can be combined to generate guanidinoacetate and ornithine by the action of arginine: glycine amidino transferase (AGAT, EC 2.1.4.1); and guanidinoacetate can be methylated to generate creatine by the action of guanidinoacetate N-methyltransferase (GAMT, EC 2.1.1.2), using SAM as the methyl donor, see par. [0091]. Enzymes for arginine biosynthesis are disclosed in WO 2018079687 in the preceding par. [0090] including carbamoyl phosphate synthetase (carAB), ornithine carbamoyl transferase (argF, argl), arginine succinate synthetase (argG), arginine succinate lyase (argH) etc. Although par.[0090] of WO 2018079687 is directed to production of polyamines, the content is recognized by the skilled person as generally disclosing L-arginine biosynthesis enzymes, one of them being carbamoyl phosphate synthetase (carAB). Since the following par. [0091] which is directed to creatine and guanidino acetate production and ornithine by the action of arginine :glycine amidino transferase (AGAT) and also refers to L-arginine biosynthesis enzymes, it is clear for the skilled person that all genes of both closely linked par. can be combined which also applies to combining AGAT with any of the L-arginine biosynthesis enzymes including carbamoyl phosphate synthetase (carAB).WO 2018079687 further discloses Corynebacteria and Escherichia, see pages 8-11.
Mijts et al. WO 2018079687 A1 disclose that, the Enzymes for arginine biosynthesis are disclosed in the preceding para. [0090] including carbamoyl phosphate synthetase (carAB), ornithine carbamoyl transferase (argF, argl), arginine succinate synthetase (argG), arginine succinate lyase (argH) etc ( see page 22. Para.[0090] of WO 2018079687 is recognized by the skilled person as generally disclosing L-arginine biosynthesis enzymes, one of them being carbamoyl phosphate synthetase (carAB). Since the following para. [0091] which is directed to the action of arginine :glycine amidino transferase (AGAT) and also refers to L-arginine biosynthesis enzymes, it is clear for the skilled person that all genes of both closely linked par. can be combined which also applies to combining AGAT with any of the L-arginine biosynthesis enzymes including carbamoyl phosphate synthetase (carAB).WO 2018079687 further discloses, limitations of claims 16-19 (Corynebacteria glutamicum, Pseudomonas putida and Escherichia) see pages 8-11 and para 0205, 0236, 0238. WO 2018079687 teaches gene overexpression see para 0139, 0143, 0147, 0252. WO 2018079687 teaches limitations of claim 1 argininosuccinate ( see para 0090).
WO 2018079687 teaches the host cell expressing the claimed genes and also teach the recombinant host strain, source of the enzymes, and expression and mutations of the different enzymes ( see pages 28, 40, 42, 45,46).
Additionally, regarding claim 1, Fan discloses a microorganism comprising a gene encoding a glycine amidinotransferase (EC 2.1.4.1), (English Translation - page 2, last two paragraphs through page 3, 1st full paragraph and claims 1-2). Glycine amidinotransferase (EC 2.1.4.1) is also known as a L-arginine:glycine amidinotransferase, catalyzing L-arginine + glycine → guaninoacetic acid.
Regarding claims 11, Fan discloses that the microorganism overexpresses glycine amidinotransferase (English Translation - page 3, 1st full paragraph).
Regarding claims 16-17, Fan discloses that the microorganism is a Corynebacterium glutamicum (English Translation – page 3, third full paragraph and claims 1-2).
As to limitations claim 12, SEQ ID 2, UNITPROT database teach sequences 100% identity to SEQ ID NO:2. See sequence alignment below:
As to claim 10, mutations, deletions and ArgR , an increased activity by mutation and over expression, these are all taught by Fukui et al. ( see pages 7, 8, 14, 16 and 19).
Therefore, in combining the teachings of the above references, it would have been obvious to one having ordinary skill in the art at the time the claimed invention was effectively filed to modify the microorganism/C. glutamicum of Fan by overexpressing a gene encoding a glyoxylate aminotransferasem because WO 2018079687 a microorganism (corynebacterium) comprising genes encoding an enzyme ( see abstract, claims 2, 11 and paragraphs 0004, 0006,0015, 0017, 0022, 0057, 0062, 0064 ). WO 2018079687 A1 para [0017] recites: “ It is a further aspect of the present invention to provide the method as described above, wherein the activity of the L-cysteine biosynthesis enzyme is increased by in-creasing the expression of a gene encoding the L-cysteine biosynthesis enzyme.”
And Fan discloses a microorganism comprising a gene encoding a glycine amidinotransferase. One of ordinary skill in the art would have had a reasonable expectation of success since Fan discloses a C. glutamicum overexpressing a gene encoding a glycine amidinotransferase (L-arginine:glycine amidinotransferease) in C. glutamicum,
Since this enzyme was known from UNIPROT A0A1D81TKD3 which discloses a glycine amidinotransferase, which is 100% identical to SEQ ID NO:2 of the present application, the obvious combination of Mijts et al. WO 2018079687 A1 with UNIPROT renders the subject matter of claim 12 obvious for a skilled person trying to solve the technical problem in comparison to the closest prior art Mijts et al. WO 2018079687 A1 of providing an alternative AGAT enzyme.
The ArgR repressor deletion was well known in the art in the context of arginine operon use, see e.g. Fukui et al., page 10 rendering the subject matter of claim 12 obvious.
The microorganisms of claims 16-19 were well known in the art in the context of microbial production methods which renders the subject matter of claims 16-19 not
Therefore, the above references render claims prima facie obvious.
RESULT 1
A0A1D8TKD3_9CYAN
ID A0A1D8TKD3_9CYAN Unreviewed; 381 AA.
AC A0A1D8TKD3;
DT 15-FEB-2017, integrated into UniProtKB/TrEMBL.
DT 15-FEB-2017, sequence version 1.
DT 05-FEB-2025, entry version 21.
DE SubName: Full=Glycine amidinotransferase {ECO:0000313|EMBL:AOW98079.1};
GN ORFNames=BJP34_00300 {ECO:0000313|EMBL:AOW98079.1};
OS Moorena producens PAL-8-15-08-1.
OC Bacteria; Bacillati; Cyanobacteriota; Cyanophyceae; Oscillatoriophycideae;
OC Oscillatoriales; Oscillatoriaceae; Moorena.
OX NCBI_TaxID=1458985 {ECO:0000313|EMBL:AOW98079.1, ECO:0000313|Proteomes:UP000177870};
RN [1] {ECO:0000313|Proteomes:UP000177870}
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=PAL-8-15-08-1 {ECO:0000313|Proteomes:UP000177870};
RA Leao T., Castelao G., Korobeynikov A., Monroe E.A., Podell S., Glukhov E.,
RA Allen E., Gerwick W.H., Gerwick L.;
RT "Comparative genomics uncovers the prolific and rare metabolic potential of
RT the cyanobacterial genus Moorea.";
RL Submitted (OCT-2016) to the EMBL/GenBank/DDBJ databases.
CC -!- SIMILARITY: Belongs to the amidinotransferase family.
CC {ECO:0000256|ARBA:ARBA00006943}.
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DR EMBL; CP017599; AOW98079.1; -; Genomic_DNA.
DR RefSeq; WP_070390602.1; NZ_CP017599.1.
DR AlphaFoldDB; A0A1D8TKD3; -.
DR STRING; 1458985.BJP34_00300; -.
DR KEGG; mpro:BJP34_00300; -.
DR OrthoDB; 258252at2; -.
DR Proteomes; UP000177870; Chromosome.
DR GO; GO:0015068; F:glycine amidinotransferase activity; IEA:TreeGrafter.
DR GO; GO:0006601; P:creatine biosynthetic process; IEA:TreeGrafter.
DR Gene3D; 3.75.10.10; L-arginine/glycine Amidinotransferase, Chain A; 1.
DR InterPro; IPR033195; AmidinoTrfase.
DR PANTHER; PTHR10488; GLYCINE AMIDINOTRANSFERASE, MITOCHONDRIAL; 1.
DR PANTHER; PTHR10488:SF1; GLYCINE AMIDINOTRANSFERASE, MITOCHONDRIAL; 1.
DR SUPFAM; SSF55909; Pentein; 1.
PE 3: Inferred from homology;
KW Reference proteome {ECO:0000313|Proteomes:UP000177870};
KW Transferase {ECO:0000256|ARBA:ARBA00022679, ECO:0000313|EMBL:AOW98079.1}.
FT ACT_SITE 197
FT /evidence="ECO:0000256|PIRSR:PIRSR633195-1"
FT ACT_SITE 248
FT /evidence="ECO:0000256|PIRSR:PIRSR633195-1"
FT ACT_SITE 356
FT /note="Amidino-cysteine intermediate"
FT /evidence="ECO:0000256|PIRSR:PIRSR633195-1"
SQ SEQUENCE 381 AA; 44861 MW; 47E28C1B8DCF8A70 CRC64;
Query Match 100.0%; Score 2094; Length 381;
Best Local Similarity 100.0%;
Matches 381; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MSEKIVNSWNEWDELEEMVVGIADYASFEPKEPGNHPKLRNQNLAEIIPFPSGPKDPKVL 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MSEKIVNSWNEWDELEEMVVGIADYASFEPKEPGNHPKLRNQNLAEIIPFPSGPKDPKVL 60
Qy 61 EKANEELNGLAYLLKDHDVIVRRPEKIDFTKSLKTPYFEVANQYCGVCPRDVMITFGNEI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 EKANEELNGLAYLLKDHDVIVRRPEKIDFTKSLKTPYFEVANQYCGVCPRDVMITFGNEI 120
Qy 121 MEATMSKRARFFEYLPYRKLVYEYWNKDEHMIWNAAPKPTMQDSMYLENFWELSLEERFK 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 MEATMSKRARFFEYLPYRKLVYEYWNKDEHMIWNAAPKPTMQDSMYLENFWELSLEERFK 180
Qy 181 RMHDFEFCITQDEVIFDAADCSRLGKDILVQESMTTNRTGIRWLKKHLEPRGFRVHPVHF 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 RMHDFEFCITQDEVIFDAADCSRLGKDILVQESMTTNRTGIRWLKKHLEPRGFRVHPVHF 240
Qy 241 PLDFFPSHIDCTFVPLRPGLILTNPERPIREEEEKIFKENGWELITVPQPTCSNDEMPMF 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 PLDFFPSHIDCTFVPLRPGLILTNPERPIREEEEKIFKENGWELITVPQPTCSNDEMPMF 300
Qy 301 CQSSKWLSMNVLSISPTKVICEEREKPLQELLDKHGFEVFPLPFRHVFEFGGSFHCATWD 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 CQSSKWLSMNVLSISPTKVICEEREKPLQELLDKHGFEVFPLPFRHVFEFGGSFHCATWD 360
Qy 361 IRRKGECEDYLPNLNYQPICG 381
|||||||||||||||||||||
Db 361 IRRKGECEDYLPNLNYQPICG 381
Applicants’ Arguments
7. Applicant's arguments filed 10/6/2025 have been fully considered but they are not persuasive. Applicants argue:
Mijts in paragraph 90 is focused on producing polyamines which involves a large
number of enzymes including those that convert L-arginine or L-ornithine into a polyamine: L-arginine can be converted to agmatine by the action of arginine decarboxylase, then to putrescine by action of agmatine ureohydrolase; L-ornithine can be converted to putrescine by action of ornithine decarboxylase; putrescine can be converted to spermidine by action of spermine synthase; agmatine can be converted to aminopropylagmatine by action of agmatine/triamine aminopropyl transferase, and then to spermidine by action of aminopropylagmatine ureohydrolase. The combination of references relies on Mijts, which cannot be separated from its reliance on producing polyamines no matter the other references being relied upon. Otherwise, this amounts to impermissible hindsight by picking and choosing from isolated disclosures in the references. This section on page 90 of Mijts does not discuss an organism designed to have increased activity of certain genes, let alone all of argininosuccinate lyase, an ornithine carbamoyltransferase, an argininosuccinate synthetase and a carbamoylphosphate synthase, and at least one heterologous gene coding for the protein having the function of said L-arginine:glycine amidinotransferase, as recited in amended claim 1. Mijts is focused on producing polyamines as discussed above and would employ some or all of the other enzymes listed above in making polyamines, which is not what claim 1 is directed to. Therefore, the features of claim 1 are not disclosed or suggested by Mijts.
Paragraph 91 of Mijts does not remedy the deficiencies of paragraph 90. It does not disclose increased activity of any enzyme. It also does not disclose carbamoylphosphate synthase, or that it has increased activity, as recited in present claim 1.
Mijts also fails to disclose the features of amended claim 11. This claim recites a
rmcroorgan1sm comprising genes coding for enzymes of a biosynthetic pathway of L-ornithine and L-arginine, wherein the genes coding for enzymes of the biosynthetic pathway of L-orniithine and L-arginine comprise gdh, argJ, argB, argC and argD coding for a glutamate dehydrogenase, for an ornithine acetyltransferase, for an acetylglutamate kinase, for an acetylglutamylphosphate reductase, and for an acetylornithine aminotransferase, respectively, and a gene coding for an enzyme having the function of a carbamoylphosphate synthase, and at least one heterologous gene coding for a protein having a function of a L-arginine:glycine amidinotransferase.
This claim 11 also recites wherein the genes coding for the enzymes of the biosynthetic pathway of L-ornithine and L-arginine, including the gene coding for the enzyme having the function of the carbamoylphosphate synthase, and the at least one heterologous gene coding for the protein having the function of the L-arginine:glycine
amidinotransferase, have increased activity compared to respective activity in a wildtype
microorganism.
As discussed above, paragraphs 90 and 91 of Mijts fail to disclose increased activity for any enzyme, let alone those claimed by Applicants. In addition, paragraph 90 of Mijts requires producing polyamines, which is not what the organism of present claim 11 is directed to.
Therefore, the rejection of Claims 1-8, 10-12, 16-19 and 27 under 35 U.S.C. §103 as being unpatentable over Mijts et al. in view of Wenchao Fan and Fukui et al. and UNIPROT is believed to be unsustainable as the present invention is neither anticipated nor obvious and withdrawal of this rejection is respectfully requested.
Office Response
8. Applicant's arguments filed 10/6/2025 have been fully considered but they are not persuasive.
In response to applicants’ arguments’ it is the examiner’s position that the applicants mainly argue Mijts WO 2018079687 reference para 90 and 91.
It is the examiner’s position that the combination of references teach the claimed invention. See below:
Mijts WO 2018079687 A1 discloses a microorganism (corynebacterium) comprising genes encoding an enzyme ( see abstract, claims 2, 11 and paragraphs 0004, 0006,0015, 0017, 0022, 0057, 0062, 0064 ). WO 2018079687 A1 para [0017] recites: “ It is a further aspect of the present invention to provide the method as described above, wherein the activity of the L-cysteine biosynthesis enzyme is increased by in-creasing the expression of a gene encoding the L-cysteine biosynthesis enzyme.”
WO 2018079687 A1 discloses L-arginine :glycine amidino transferase ( see para 0091). WO 2018079687 A1 discloses Corynebacterium, E.coli and wildtype microorganisms( see claims 17, 18, 19, 20 , 50 and para 0072-0073). WO 2018079687 A1 discloses the possibility to produce creatine from arginine and glycine using L- arginine biosynthesis enzymes, glycine biosynthesis enzymes and enzymes that catalyze the conversion of L-arginine and glycine into creatine. L-arginine and glycine can be combined to generate guanidinoacetate and ornithine by the action of arginine: glycine amidino transferase (AGAT, EC 2.1.4.1); and guanidinoacetate can be methylated to generate creatine by the action of guanidinoacetate N-methyltransferase (GAMT, EC 2.1.1.2), using SAM as the methyl donor, see par. [0091]. Enzymes for arginine biosynthesis are disclosed in WO 2018079687 in the preceding par. [0090] including carbamoyl phosphate synthetase (carAB), ornithine carbamoyl transferase (argF, argl), arginine succinate synthetase (argG), arginine succinate lyase (argH) etc. Although par.[0090] of WO 2018079687 is directed to production of polyamines, the content is recognized by the skilled person as generally disclosing L-arginine biosynthesis enzymes, one of them being carbamoyl phosphate synthetase (carAB). Since the following par. [0091] which is directed to creatine and guanidino acetate production and ornithine by the action of arginine :glycine amidino transferase (AGAT) and also refers to L-arginine biosynthesis enzymes, it is clear for the skilled person that all genes of both closely linked par. can be combined which also applies to combining AGAT with any of the L-arginine biosynthesis enzymes including carbamoyl phosphate synthetase (carAB).WO 2018079687 further discloses Corynebacteria and Escherichia, see pages 8-11.
Mijts et al. WO 2018079687 A1 disclose that, the Enzymes for arginine biosynthesis are disclosed in the preceding para. [0090] including carbamoyl phosphate synthetase (carAB), ornithine carbamoyl transferase (argF, argl), arginine succinate synthetase (argG), arginine succinate lyase (argH) etc ( see page 22)..
Para.[0090] of WO 2018079687 is recognized by the skilled person as generally disclosing L-arginine biosynthesis enzymes, one of them being carbamoyl phosphate synthetase (carAB). Since the following para. [0091] which is directed to the action of arginine :glycine amidino transferase (AGAT) and also refers to L-arginine biosynthesis enzymes, it is clear for the skilled person that all genes of both closely linked par. can be combined which also applies to combining AGAT with any of the L-arginine biosynthesis enzymes including carbamoyl phosphate synthetase (carAB).WO 2018079687 further discloses, limitations of claims 16-19 (Corynebacteria glutamicum, Pseudomonas putida and Escherichia) see pages 8-11 and para 0205, 0236, 0238. WO 2018079687 teaches gene overexpression see para 0139, 0143, 0147, 0252. WO 2018079687 teaches limitations of claim 1 argininosuccinate ( see para 0090).
WO 2018079687 teaches the host cell expressing the claimed genes and also teach the recombinant host strain, source of the enzymes, and expression and mutations of of the different enzymes ( see pages 28, 40, 42, 45,46).
And Fan discloses a microorganism comprising a gene encoding a glycine amidinotransferase.
The combination of references makes the claimed invention obvious. Therefore, the rejection is maintained.
Conclusion
9. No claims are allowed.
10. 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 KHATOL S SHAHNAN SHAH whose telephone number is (571)272-0863. The examiner can normally be reached on Mon-Tue, Thu-Fri 12pm-8pm.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Daniel Kolker can be reached on 571-272-3181. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/Khatol S Shahnan-Shah/
Examiner, Art Unit 1645
February 19, 2026
/JANA A HINES/Primary Examiner, Art Unit 1645