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
Claims 2-3, 7-8, 11, 13, 15, 17 and 19-24 are canceled. Claims 1, 4-6, and 9-10, 12, 14, 16, and 18 are pending and under consideration in this action.
Priority
The instant claims are entitled to the effective filing date of 04/13/2021.
Continued Examination Under 37 CFR 1.114
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 05/05/2026 has been entered.
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 1, 4, 6, 9, 14 and 16 are rejected under 35 U.S.C. 103 as being unpatentable over Shrestha (Microb Cell Fact, 2019, 18, 7) in view of Reddy, (Metabolic engineering, 2007, 9(1), 95-111), and University of Georgia (NCBI, accession number ASM479914v2, direct submission 2019, April 17).
Regarding claims 1 and 6, Shrestha teaches transforming E. coli with plasmids containing anthocyanidin synthase from Petunia hybrida (PhANS) and flavonoid 3-O-glycosyltransferase from Arabidopsis thaliana (At3GT) genes for cyanidin 3-O-glucoside (C3G) production. The PhANS and At3GT genes are expressed under either the Pt7, Ptrc, or PlacUV5 promoters. See the “construction of plasmids for cyanidin 3-O-glucoside production” section on page 13. The highest C3G production is observed in strain S12 when 2mM (+)-catechin is fed the strain. See the paragraph spanning the left and right columns on page 11, see figure 7 C3G production levels and see figure 1 for the transformation of catechin to C3G. The S12 strain carries At3GT and PhANS under the Ptrc promoter. See table 1.
Shrestha does not teach the overexpression of anthocyanin synthase.
Reddy discloses that anthocyanin synthase (ANS) mediates multiple dioxygenase activities in the flavonoid pathway, its overexpression in transgenic plants may lead to the accumulation of a mixture of flavonoids with enhanced antioxidant activity. See the last four lines in the last full paragraph on page 96. Reddy discloses that the overexpression of ANS in rice results in a mixture of flavonoids and enhanced antioxidant potential. See the abstract.
It would have been obvious to a person of ordinary skill in the art prior to the effective filing date of the instantly claimed invention to overexpress Shrestha’s anthocyanidin synthase from Petunia hybrida (PhANS) based on Reddy’s suggestion. One of ordinary skill in the art would have been motivated to do so because Reddy suggests that overexpressing anthocyanin synthase may lead to an accumulation of flavonoids. There would have been a reasonable expectation of success because Shrestha demonstrates expressing the PhANS under a promoter, which controls gene expression.
Shrestha and Reddy do not teach an anthocyanin synthase that has an amino acid sequence that is at least 95% identical to SEQ ID NO: 69.
University of Georgia teaches the whole-genome assembly of Neltuma alba isolate B1F8T4 on 04/17/2019, which includes gene ID 114743801 encoding protein id XP 028787346.1. See page 1. The protein id XP 028787346.1 is a leucoanthocyanidin dioxygenase-like protein (i.e. anthocyanin synthase), which is 100% identical to instant SEQ ID NO: 69. See page 2 of the University of Georgia reference and see the sequence alignment in the office action appendix.
It would have been obvious to a person of ordinary skill in the art prior to the effective filing date of the instantly claimed invention to substitute University of Georgia’s anthocyanin synthase (protein id XP_028787846.1) for the PhANS of Shrestha. One of ordinary skill in the art would have been motivated to do so because Shrestha suggests that anthocyanins are of particular interest because of their applications in food and their health beneficial properties (see page 9, the second sentence of the discussion section). There would have been a reasonable expectation of success because the leucoanthocyanidin dioxygenase-like protein taught by the University of Georgia reasonably serves the same function as the PhANS of Shrestha, as leucoanthocyanidin dioxygenase is synonymous with anthocyanin synthase.
Regarding claims 4 and 9, Shrestha teaches transforming E. coli with plasmids containing PhANS and flavonoid 3-O-glycosyltransferase from Arabidopsis thaliana (At3GT) genes for cyanidin 3-O-glucoside production. See the “construction of plasmids for cyanidin 3-O-glucoside production” section on page 13. The PhANS and At3GT genes are expressed under either the Pt7, Ptrc, or PlacUV5 promoters. See the “construction of plasmids for cyanidin 3-O-glucoside production” section on page 13.
It would have been obvious to a person of ordinary skill in the art prior to the effective filing date of the instantly claimed invention to overexpress the anthocyanidin synthase, as discussed above, and in the process overexpress the flavonoid-3-glycosyl transferase (i.e. the A3GT), because Shrestha teaches expressing the flavonoid-3-glycosyl transferase under the same promoter as the anthocyanin synthase.
Regarding claims 14 and 16, University of Georgia teaches protein id XP 028787346.1, which is 100% identical to instant SEQ ID NO: 69. See page 2 of the University of Georgia reference and see the sequence alignment in the office action appendix.
Claims 5, 10, 12 and 18 are rejected under 35 U.S.C. 103 as being unpatentable over Shrestha (Microb Cell Fact, 2019, 18, 7) in view of Reddy, (Metabolic engineering, 2007, 9(1), 95-111), and University of Georgia (NCBI, accession number ASM479914v2, direct submission 2019, April 17), as applied to claims 1, 4, 6, 9, 14 and 16 above, and further in view of Hall (Phytochemistry, 2012, 74, 90-99).
Regarding claim 5, Shrestha teaches transforming E. coli with plasmids containing anthocyanidin synthase from Petunia hybrida (PhANS) and flavonoid 3-O-glycosyltransferase from Arabidopsis thaliana (At3GT) genes for cyanidin 3-O-glucoside (C3G) production. See the “construction of plasmids for cyanidin 3-O-glucoside production” section on page 13.
Shrestha, Reddy and the University of Georgia do not teach a flavonoid-3-glucosyl transferase that is selected from a group consisting of: (i) the flavonoid-3-glucosyl transferase in Vitis labrusca that encodes SEQ. ID NO: 14; (ii) the flavonoid-3-glucosyl transferase has an amino acid sequence that is at least 80% identical to SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, or SEQ ID NO: 73; (iii) the flavonoid-3-glucosyl transferase has an amino acid sequence at least 80% identical to SEQ ID NO: 14, and (iv) any combinations thereof.
Hall teaches the flavonoid 3-O-glucosyltransferase (3Gt) of Vitis vinifera (VV3GT) and Vitis labrusca (VL3GT). Hall discloses that VL3GT glucosylates favonols and anthocyanidins in vitro. See the last paragraph of the introduction section. In a substrate specificity study, recombinant VL3GT glucosylated several anthocyanidins including cyanidin. See the first three lines of section 2.2 on page 92. The VL3GT of Hall is a 100% identity match to instant SEQ ID NO: 14. See the alignment in the office action appendix attached of the non-final rejection mailed 11/18/2024.
It would have been obvious to a person of ordinary skill in the art prior to the effective filing date of the instantly claimed invention to substitute Hall’s flavonoid-3-glucosyl transferase of Vitis labrusca (VL3GT) for Shrestha’s flavonoid 3-O-glycosyltransferase from Arabidopsis thaliana (At3GT). One of ordinary skill in the art would have been motivated to do so because Hall suggests that VL3GT preferentially glucosylates anthocyanidin, such as cyanidin; and Shrestha suggests that cyanidin glucosylation affects cyanidin 3-O-glucoside production (see fig. 1 of Shrestha). There would have been be a reasonable expectation of success because the VL3GT of Hall would reasonably serve the same 3GT function in the method of Shrestha.
Regarding claim 10, 12 and 18, Hall teaches VL3GT, which is a 100% identity match to instant SEQ ID NO: 14. See the alignment in the office action appendix attached of the non-final rejection mailed 11/18/2024.
Response to Arguments
Applicant's arguments filed 05/05/2026 have been fully considered but they do not apply to the new grounds for rejection discussed above.
Conclusion
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/LOUISE W HUMPHREY/Supervisory Patent Examiner, Art Unit 1657
/K.C.B./Examiner, Art Unit 1657
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