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 .
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and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.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 11/10/2025 has been entered.
Priority
The instant application filed on 10/07/2022 is a 371 of PCT/JP2021/014958 filed on
04/08/2021 and which claims priority to JP2020-071054 filed on 04/10/2020. The certified
copy of foreign priority application for JP2020-071054 is not in English; therefore, the
effective filing date of the instant claims is 04/08/2021.
Should applicant desire to obtain the benefit of foreign priority under 35 U.S.C. 119(a)-(d) prior to declaration of an interference, a certified English translation of the foreign application must be submitted in reply to this action. 37 CFR 41.154(b) and 41.202(e).
Failure to provide a certified translation may result in no benefit being accorded for the non-English application.
Withdrawal of Rejections
The response and amendments filed on 11/10/2025 are acknowledged. Any previously applied minor objections and/or minor rejections (i.e., formal matters), not explicitly restated here for brevity, have been withdrawn necessitated by Applicant’s formality corrections and/or amendments. For the purposes of clarity of the record, the reasons for the Examiner’s withdrawal, or maintaining, if applicable, of the substantive or essential claim rejections are detailed directly below and/or in the Examiner’s response to arguments section.
Briefly, the previous claim rejections under 35 U.S.C. 112(b) for indefiniteness have been withdrawn necessitated by Applicant’s amendments. The previous claim rejections under 35 U.S.C. 103 for obviousness have been withdrawn necessitated by Applicant’s amendments; however, new grounds of rejection are set forth below.
The following rejections and/or objections are either reiterated or newly applied. They constitute the complete set presently being applied to the instant application.
Claim Rejections - 35 USC § 103, Obviousness
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 1-3, 8, 10, 19-21, 23-24, and 26-27 are rejected under 35 U.S.C. 103 as being unpatentable over Fukui (US 2013/0071892; Date of Publication: March 21, 2013 – previously cited) in view of Riedel (Recovery of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from Ralstonia eutropha cultures with non-halogenated solvents; 2013 – previously cited), Maruyama (US 2008/0038801; Date of Publication: February 14, 2008 – previously cited), and Clemente (U.S. Patent No. 5,849,894; Date of Publication: December 15, 1998 – newly cited).
Fukui’s general disclosure relates to production of “poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] with a high 3-hydroxyhexanoic acid fraction using a vegetable oil as a basic raw material” (see, e.g., Fukui, abstract). Moreover, Fukui discloses production of a microorganism that produces [P(3HB-co-3HHx)] with a high 3-hydroxyhexanoic acid fraction using a vegetable oil as a basic raw material by “introducing a gene encoding an R-hydratase that converts a fatty acid β-oxidation system intermediate into a monomer, (R)-3-hydroxyacyl-CoA [R-3HA-CoA], into a recombinant Cupriavidus necator strain that was conferred an ability of producing P(3HB-co-3HHx)” (see, e.g., Fukui, abstract).
Regarding claim 1 pertaining to producing polyhydroxyalkanoate copolymer mixture, Fukui teaches a method of producing poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) [P(3HB-co-3HHx)] (see, e.g., Fukui, abstract). Moreover, Fukui teaches culturing C. necator to produce P(3HB-co-3HHx) (see, e.g., Fukui, [0023], [0117]-[0118], [0120]).
Regarding claim 2 pertaining to the weight percentage, Fukui teaches a weight percentage of 50-90% (see, e.g., Fukui, [0004], [0024], [0056]).
Regarding claim 19 pertaining to the (R)-specific enoyl-CoA hydratase, Fukui teaches a gene encoding an (R)-form-specific enoyl-CoA hydratase (see, e.g., Fukui, [0024]).
Regarding claim 20 pertaining to the carbon source, Fukui teaches vegetable oil as a carbon source (see, e.g., Fukui, [0118]).
Regarding claim 21 pertaining to the carbon source, Fukui teaches using octanoic acid as a carbon source (see, e.g., Fukui, [0006]), which is a middle-chain fatty acid that has 8 carbon atoms.
Regarding claims 23-24 pertaining to the microorganism, Fukui teaches that the microorganism used is Cupriavidus necator (see, e.g., Fukui, abstract).
However, Fukui does not teach: wherein the microorganism comprises gene (A) and gene (B), wherein gene (A) encodes an amino acid sequence having a sequence identity of 90% to 100% with the amino acid sequence of SEQ ID NO: 2 or 3, and wherein gene (B) encodes an amino acid sequence having a sequence identity of 90 to 100% with the amino acid sequence of SEQ ID NO: 6 (claims 1, 8, 10, and 12); polyhydroxyalkanoate fraction (I) comprising a polyhydroxyalkanoate copolymer having a 3-hydroxybutyrate structural unit and a 3-hydroxyhexanoate structural unit, wherein the average 3-hydroxyhexanoate unit ratio is 20 mol% or more (claim 1); or a polyhydroxyalkanoate fraction (II) comprising a polyhydroxyalkanoate having a 3-hydroxybutyrate structural unit, wherein the average 3-hydroxyhexanoate unit ratio is 0 to 15 mol% (claim 1); or wherein the polyhydroxyalkanoate copolymer mixture has an average 3-hydroxyhexanoate unit ratio of 22 mol% or less (claim 1); or wherein the average 3-hydroxyhexanoate unit ratio of the polyhydroxyalkanote copolymer mixture is from 10 to 22 mol% (claim 3).
Riedel’s general disclosure pertains to a process design for “effective recovery of the copolymer poly(hydroxybutyrate-co-hydroxyhexanoate) (P(HB-co-HHx)) containing high levels of HHx (>15 mol%) from Ralstonia eutropha biomass using non-halogenated solvents” (see, e.g., Riedel, abstract). Moreover, Riedel discloses that the higher HHx content makes the polymer more soluble (see, e.g., Riedel, “Discussion”, pg. 468), meanwhile the lower HHx content forms a gel-like phase (see, e.g., Riedel, abstract).
Regarding claim 1 pertaining to polyhydroxyalkanoate fraction (I), Riedel teaches a PHA fraction having 3-hydroxybutyrate and 3-hydroxyhexanoate, with higher HHx content (17-30 mol%) (see, e.g., Riedel, abstract & Introduction, pg. 462).
Regarding claims 1 and 26 pertaining to polyhydroxyalkanoate fraction (II), Riedel teaches a PHA fraction having 3-hydroxybutyrate and 3-hydroxyhexanoate, with lower HHx content (11-16 mol%) (see, e.g., Riedel, abstract & Introduction, pg. 462). Moreover, the Examiner has interpreted the limitations recited after “optionally” to not be required as part of the claimed invention. Moreover, the Examiner has interpreted the 3-hydoxyhexanoate structural unit in fraction (II) to be optional since the average 3-hydroxyhexanoate unit ratio can be 0%.
Regarding claims 1, 3, and 27 pertaining to the average 3-hydroxyhexanoate unit ratio, Riedel teaches an average HHx content of 22 mol% (see, e.g., Riedel, “Larger Scale PHA Recovery”, pg. 468).
Maruyama’s general disclosure relates to “a recombinant microbial strain capable of stably producing a polyhydroxyalkanoic acid (PHA) at a high production rate in an industrial fermentation process” (see, e.g., Maruyama, abstract). Moreover, Maruyama’s SEQ ID NO: 9, encodes “the exogenous polyhydroxyalkanoic acid synthase gene codes for a mutant enzyme derived from Aeromonas caviae represented by the amino acid sequence shown under SEQ ID NO:9, which is prepared by substituting serine for asparagine of the 149th amino acid and substituting glycine for aspartic acid of the 171st amino acid” (see, e.g., Maruyama, [0041]). Furthermore, Maruyama discloses that the polyhydroxyalkanoic acid synthase encoded by Maruyama’s SEQ ID NO: 9 gives a microorganism that expresses this synthase the ability to accumulate polyhydroxyalkanoic acid stably at a high level (see, e.g., Maruyama, [0021]).
Regarding claims 1 and 8 pertaining to gene (A) encoding the amino acid sequence of SEQ ID NO: 2, Maruyama teaches SEQ ID NO: 9, which encodes for a exogenous polyhydroxyalkanoic acid synthase gene derived from Aeromonas caviae, and which has 100% sequence similarity to instant SEQ ID NO: 2 (see, e.g., Maruyama, [0041] & Office Action Appendix from final office action mailed 08/08/2025). Moreover, one of ordinary skill in the art would readily understand that a microorganism can contain a gene that encodes for this amino acid sequence since the amino acid sequence is taught by Maruyama.
Clemente’s general disclosure relates to “Isolated DNA fragments encoding a Rhodospirillum rubrum (ATCC 25903) polyhydroxyalkanoate (PHA) synthase, or biologically functional equivalents thereof” (see, e.g., Clemente, abstract). Moreover, Clemente discloses “The present invention relates to poly-β-hydroxyalkanoate (PHA) synthases, such as that from Rhodospirillum rubrum, which exhibit flexible substrate specificity. These synthases can be expressed in transformed microorganisms and plants to produce poly-β-hydroxyalkanoates (PHAS) possessing varied physical properties depending upon the monomers incorporated therein (see, e.g., Clemente, col. 1, lines 9-16).
Regarding claims 1 and 10 pertaining to instant SEQ ID NO: 6, Clemente teaches SEQ ID NO: 5, which encodes a PHA synthase from Alcaligenes eutrophus, and which has 78.3% sequence identity to instant SEQ ID NO: 6 (see, e.g., Office Action Appendix). Moreover, the transitional phrase “having” is synonymous to “comprising”; therefore, this is open-ended and does not exclude additional, unrecited elements (see, e.g., MPEP 2111.03(I)).
It would have been first obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce Fukui’s polyhydroxyalkanoate copolymer mixture comprising a polyhydroxyalkanoate fraction (I) with higher a HHx content (17-30 mol%), as taught by Riedel. One would have been motivated to do so because Riedel teaches that the higher HHx content makes the polymer more soluble (see, e.g., Riedel, “Discussion”, pg. 468), meanwhile the lower HHx content forms a gel-like phase (see, e.g., Riedel, abstract). Furthermore, Fukui teaches a C. necator strain that produces P(3HB-co-3HHx) and contains a high 3HHx fraction (see, e.g., Fukui, [0023]). Additionally, Fukui teaches “the 3HHx fraction may preferably be 7-15 mol % in order to make polymers having a feasible flexibility” (see, e.g., Fukui, [0004]). Therefore, based on the teachings of Fukui and Riedel, it would have been obvious to produce a polyhydroxyalkanoate copolymer mixture with a higher HHx content in order to increase polymer solubility, as well as produce a polymer with increased flexibility. Furthermore, if one of ordinary skill in the art wanted to produce a gel-like polymer substance, based on the teachings of Fukui and Riedel, it would have been obvious to produce a polymer with a lower HHx content.
It would have been secondly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce Fukui’s polyhydroxyalkanoate copolymer mixture comprising an average HHx content of 22 mol%, as taught by Riedel. One would have been motivated to do so because Riedel teaches fractionation in multiple phases (i.e., aqueous phase, gel phase, solution phase, and residual cell mass phase) with an average HHx content of 22 mol% (see, e.g., Riedel, “Larger Scale PHA Recovery”, pg. 468). Moreover, Riedel teaches that the higher HHx content makes the polymer more soluble (see, e.g., Riedel, “Discussion”, pg. 468), meanwhile the lower HHx content forms a gel-like phase (see, e.g., Riedel, abstract). Furthermore, Fukui teaches that the amount of HHx affects polymer flexibility (see, e.g., [0003]-[0004]). Therefore, based on the teachings of Fukui and Riedel, it would have been obvious to produce a polyhydroxyalkanoate copolymer mixture with an average HHx unit ratio of 22 mol % or more because this would result in altered flexibility of the polymer composition. One would have expected success because Fukui and Riedel both teach polyhydroxyalkanoate copolymer mixtures with high HHx fractions.
It would have been thirdly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce Fukui’s polyhydroxyalkanoate copolymer mixture by a microorganism, wherein the microorganism expresses instant SEQ ID NO: 2, as taught by Maruyama. One would have been motivated to do so because Maruyama teaches that SEQ ID NO: 9, which encodes instant SEQ ID NO: 2, encodes “the exogenous polyhydroxyalkanoic acid synthase gene codes for a mutant enzyme derived from Aeromonas caviae represented by the amino acid sequence shown under SEQ ID NO: 9, which is prepared by substituting serine for asparagine of the 149th amino acid and substituting glycine for aspartic acid of the 171st amino acid” (see, e.g., Maruyama, [0041]). Moreover, Maruyama teaches that the polyhydroxyalkanoic acid synthase encoded by Maruyama’s SEQ ID NO: 9 gives a microorganism that expresses this synthase the ability to accumulate polyhydroxyalkanoic acid stably at a high level (see, e.g., Maruyama, [0021]). Furthermore, Fukui teaches “a recombinant strain of a hydrogen-oxidizing bacterium Cupriavidus necator (C. necator) that accumulates P(3HB-co-3HHx) with a high content (70-80% by weight) from vegetable oils by introducing phaCAC, a PHA synthase (polymerase) derived from A. caviae, into the PHB-4 strain, a PHA accumulation-deficient strain of C. necator that efficiently produces P(3HB)” (see, e.g., Fukui, [0004]). Therefore, based on the teachings of Fukui and Maruyama, it would have been obvious to express Maruyama’s SEQ ID NO: 9 in C. necator PHB-4, as taught by Fukui, because this would result in the strain being able to express PHA synthase stably and accumulate PHA at high levels. One would have expected success because Fukui and Maruyama both teach the expression of PHA synthase, as well as PHA production in microbial cells.
It would have been fourthly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce Fukui’s polyhydroxyalkanoate copolymer mixture by a microorganism, wherein the microorganism expresses instant SEQ ID NO: 6, as taught by Clemente. One would have been motivated to do so because Clemente teaches that SEQ ID NO: 5, which has 78.3% sequence similarity to instant SEQ ID NO: 6, is derived from A. eutrophus and “The most extensively studied PHA biosynthetic pathway is that of Alcaligenes eutrophus (Peoples et al. (1989) J. Biol. Chem. 264:15298 and Valentin et. al. (1995) Eur. J. Biochem. 227:43). This organism is capable of forming either a homopolymer of C4 (polyhydroxybutyrate, PHB) or a co-polymer of C4-C5 (PHB-PHV, polyhydroxybutyrate-polyhydroxyvalerate) (Koyama and Doi (1995) Biotechnol. Lett. 17:281). Hence, A. eutrophus is classified as a scl PHA organism (see, e.g., Clemente, col. 1, lines 37-46). Furthermore, Fukui teaches “a recombinant strain of a hydrogen-oxidizing bacterium Cupriavidus necator (C. necator) that accumulates P(3HB-co-3HHx) with a high content (70-80% by weight) from vegetable oils by introducing phaCAC, a PHA synthase (polymerase) derived from A. caviae, into the PHB-4 strain, a PHA accumulation-deficient strain of C. necator that efficiently produces P(3HB)” (see, e.g., Fukui, [0004]). Therefore, based on the teachings of Fukui and Clemente, it would have been obvious to alter the PHA synthase derived from A. eutrophus to utilize the PHA synthase derived from A. caviae. One would have expected success because Fukui and Clemente both teach recombinant microorganisms expressing PHA synthases.
Claim 15 is rejected under 35 U.S.C. 103 as being unpatentable over Fukui, Riedel, Maruyama, and Clemente as applied to claims 1-3, 8, 10, 19-21, 23-24, and 26-27 above, and further in view of Sato (Regulation of 3-hydroxyhexanoate composition in PHBH synthesized by recombinant Cupriavidus nectar H16 from plant oil by using butyrate as a co-substrate; 2015 – previously cited).
The teachings of Fukui, Riedel, Maruyama, and Clemente, herein referred to as modified-Fukui-Riedel-Maruyama-Clemente, are discussed above as it pertains to producing a polyhydroxyalkanoate copolymer mixture.
However, modified-Fukui-Riedel-Maruyama-Clemente does not teach: wherein the microorganism is a transformed microorganism that has been transformed to supply a greater amount of 3-hydroxyhexanoyl-CoA to an intracellular polyhydroxyalkanoate synthase than a wild strain of the microorganism (claim 15).
Sato’s general disclosure relates to “A (R)-3-hydroxyhexanoate (3HH) composition-regulating technology for poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) production using recombinant Cupriavidus necator H16 with butyrate as a co-substrate” (see, e.g., Sato, abstract). Moreover, Sato teaches a new (R)-3-hydroxyhexanoyl-CoA synthesis pathway (see, e.g., Sato, abstract).
Regarding claim 15 pertaining to the transformed microorganism and 3-hydroxyhexanoyl-CoA, Sato teaches the transformation of C. necator to enhance the synthetic pathway of (R)-3-hydroxyhexanoyl-CoA using a mutant PHA synthase from Aeromonas caviae (see, e.g., Sato, abstract). Sato teaches that this strategy can enhance biosynthesis of 3-ketohexanoyl-CoA as a precursor of (R)-3-hydroxyhexanoyl-CoA (see, e.g., Sato, Introduction, pg. 246); therefore, one of ordinary skill in the art would appreciate that increasing the precursor to (R)-3-hydroxyhexanoyl-CoA would result in increased (R)-3-hydroxyhexanoyl-CoA synthesis (see, e.g., Sato, Figure 1(b), pg. 247).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce modified-Fukui-Riedel-Maruyama-Clemente’s microorganism, wherein the microorganism has enhanced (R)-3-hydroxyhexanoyl-CoA synthesis, as taught by Sato. One would have been motivated to do so because Sato teaches that increasing the synthesis of (R)-3-hydroxyhexanoyl-CoA would result in an increase in the 3HH monomer fraction of poly (3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) (see, e.g., Sato, Introduction, pg. 247). Furthermore, modified-Fukui-Riedel-Maruyama-Clemente teaches “a microbial strain in which a broad substrate-specific PHA synthase (for example, the gene of A. caviae-derived PHA synthase or its mutant enzyme) capable of using 3HHx-CoA having 6 carbons as a substrate has been introduced into a C. necator strain in which PHA synthase inherently present in the native strain has been deleted by mutation or gene destruction, or a microbial strain in which the PHA synthase gene inherently present in the native C. necator strain has been replaced with the gene of a broad substrate-specific PHA synthase or its mutant enzyme, characterized in that it has an ability of synthesizing P(3HB-co-3HHx) which the wild type C. necator strain cannot biosynthesize due to the effect of the broad substrate-specific PHA synthase” (see, e.g., Fukui, [0057]); therefore, 3HHx-CoA is a precursor for HHx production. Therefore, based on the teachings of modified-Fukui-Riedel-Maruyama-Clemente and Sato, it would have been obvious to produce a transformed microorganism with increased 3-hydroxyhexanoyl-CoA synthesis because this would allow for increased 3HH production when producing a polyhydroxyalkanoate copolymer mixture. One would have expected success because modified-Fukui-Riedel-Maruyama-Clemente and Sato both teach production of copolymer mixtures.
Claims 16-17 are rejected under 35 U.S.C. 103 as being unpatentable over Fukui, Riedel, Maruyama, Clemente, and Sato as applied to claims 1-3, 8, 10, 15, 19-21, 23-24, and 26-27 above, and further in view of Arikawa (WO 2019/142845; Date of Publication: July 25, 2019 – cited in the IDS filed on 10/07/2022 – previously cited).
The teachings of Fukui, Riedel, Maruyama, Clemente, and Sato, herein referred to as modified-Fukui-Riedel-Maruyama-Clemente-Sato, are discussed above as it pertains to producing a polyhydroxyalkanoate copolymer mixture.
However, modified-Fukui-Riedel-Maruyama-Clemente-Sato does not teach: wherein the transformed microorganism has been transformed to inhibit degradation of an intermediate metabolite having six carbon atoms in β-oxidation of an oil or fatty acid (claim 16); or wherein the transformed microorganism has been transformed to inhibit expression of a gene encoding β-ketothiolase enzyme having thiolysis activity for β-ketohexanoyl-CoA which is β-ketoacyl-CoA having six carbon atoms (claim 17).
Arikawa’s general disclosure relates to “a transformed micro-organism for producing a PHA copolymer comprising a 3HH monomer unit at a higher composition rate” (see, e.g., Arikawa, abstract). Moreover, Arikawa discloses “a transformed micro-organism for producing a PHA copolymer comprising a 3HH monomer unit at a higher composition rate, specifically, a micro-organism having a PHA synthase gene capable of synthesizing the PHA copolymer comprising the 3HH monomer unit and a gene that encodes a protein having R-specific enoyl-CoA hydratase activity, wherein the transformed micro-organism is characterized in that the expression of a gene that encodes at least one type of β- ketothiolase enzyme exerts thiolysis on a C6 β-ketoacyl-CoA (i.e., β-keto hexanoyl-CoA) is suppressed, and the enzyme activity is eliminated or reduced” (see, e.g., Arikawa, abstract). Furthermore, Arikawa discloses that suppression of a C6 intermediate metabolite results in the compositional ratio of 3HH monomer units in the copolymerization PHA of the final synthesized product to be increased (see, e.g., Arikawa, English translation, pg. 4).
Regarding claim 16 pertaining to the transformed microorganism, Arikawa teaches a transformed microorganism wherein decomposition of the C6 intermediate metabolite in β-oxidation is suppressed (see, e.g., Arikawa, English translation, pg. 4).
Regarding claim 17 pertaining to the transformed microorganism, Arikawa teaches “repressing the expression of at least one or at least two genes encoding a β-ketothiolase enzyme having thiolysis activity for a C6 β-ketoacyl-CoA (i.e., β-ketohexanoyl-CoA)” (see, e.g., Arikawa, English translation, pg. 4).
It would have been first obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to transform modified-Fukui-Riedel-Maruyama-Clemente-Sato’s microorganism to suppress decomposition of the C6 intermediate metabolite, as taught by Arikawa. One would have been motivated to do so because Arikawa teaches that suppression of the C6 intermediate metabolite results in the compositional ratio of 3HH monomer units in the copolymerization PHA of the final synthesized product to be increased (see, e.g., Arikawa, English translation, pg. 4). Furthermore, modified-Fukui-Riedel-Maruyama-Clemente-Sato teaches that increased 3HH levels makes the polymer more soluble (see, e.g., Riedel, “Discussion”, pg. 468), and the amount of HHx alters the flexibility of the polymer (see, e.g., Fukui, [0004]). Therefore, based on the teachings of modified-Fukui-Riedel-Maruyama-Clemente-Sato and Arikawa, it would have been obvious to produce a transformed microorganism that has suppressed decomposition of the C6 intermediate metabolite to produce a PHA containing a higher composition of 3HH monomer units.
It would have been secondly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to transform modified-Fukui-Riedel-Maruyama-Clemente-Sato’s microorganism to repress expression of genes encoding a β-ketothiolase enzyme having thiolysis activity for a C6 β-ketoacyl-CoA (i.e., β-ketohexanoyl-CoA), as taught by Arikawa. One would have been motivated to do so because Arikawa teaches that repression of genes encoding the β-ketothiolase enzyme results in production of “a copolymerized PHA containing 3HH monomer units at a higher composition ratio” (see, e.g., Arikawa, English translation, pg. 2). Furthermore, modified-Fukui-Riedel-Maruyama-Clemente-Sato teaches that increased 3HH levels makes the polymer more soluble (see, e.g., Riedel, “Discussion”, pg. 468), and the amount of HHx alters the flexibility of the polymer (see, e.g., Fukui, [0004]). Therefore, based on the teachings of modified-Fukui-Riedel-Maruyama-Clemente-Sato and Arikawa, it would have been obvious to produce a transformed microorganism that has repression of genes encoding the β-ketothiolase enzyme to produce a PHA containing a higher composition of 3HH monomer units. One would have expected success because modified-Fukui-Riedel-Maruyama-Clemente-Sato and Arikawa both teach PHA synthesis.
Claim 18 is rejected under 35 U.S.C. 103 as being unpatentable over Fukui, Riedel, Maruyama, Clemente, Sato, and Arikawa as applied to claims 1-3, 8, 10, 15-17, 19-21, 23-24, and 26-27 above, and further in view of Asrar (US Patent No. 6,091,002; Date of Publication: July 18, 2000 – previously cited).
The teachings of Fukui, Riedel, Maruyama, Clemente, Sato, and Arikawa, herein referred to as modified-Fukui-Riedel-Maruyama-Clemente-Sato-Arikawa, are discussed above as it pertains to producing a polyhydroxyalkanoate copolymer mixture.
However, modified-Fukui-Riedel-Maruyama-Clemente-Sato-Arikawa does not teach: wherein the β-ketothiolase enzyme has an amino acid sequence having a sequence identity of 90 to 100% with the amino acid sequence of SEQ ID NO: 9 or 10 (claim 18).
Asrar’s general disclosure relates to genetically engineered plants and bacteria “for the biosynthesis of polyhydroxyalkanoate homopolymers and copolymers” (see, e.g., Asrar, abstract). Moreover, Asrar discloses that a β-ketothiolase is capable of producing copolymers containing monomers ranging in size from C4 to C6, which allows for the formation of PHAs (see, e.g., Asrar, [4], [1369]).
Regarding claim 18 pertaining to SEQ ID NO: 9, Asrar teaches a β-ketothiolase protein (see, e.g., Asrar, Example 8) that has 100% sequence identity to instant SEQ ID NO: 9 (see, e.g., Office Action Appendix).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce modified-Fukui-Riedel-Maruyama-Clemente-Sato-Arikawa’s microorganism, wherein the microorganism represses a β-ketothiolase protein, as taught by Asrar. One would have been motivated to do so because Asrar teaches that a β-ketothiolase is capable of producing copolymers containing monomers ranging in size from C4 to C6, which allows for the formation of PHAs (see, e.g., Asrar, [4], [1369]). Furthermore, modified-Fukui-Riedel-Maruyama-Clemente-Sato-Arikawa teaches that repression of genes encoding the β-ketothiolase enzyme results in production of “a copolymerized PHA containing 3HH monomer units at a higher composition ratio” (see, e.g., Arikawa, English translation, pg. 2). Therefore, based on the teachings of modified-Fukui-Riedel-Maruyama-Clemente-Sato-Arikawa and Asrar, it would have been obvious to produce a microorganism that represses β-ketothiolase enzyme corresponding to instant SEQ ID NO: 9 because it would result in PHA production with high HHx ratio. One would have expected success because modified-Fukui-Riedel-Maruyama-Clemente-Sato-Arikawa and Asrar both teach PHA synthesis
Claim 22 is rejected under 35 U.S.C. 103 as being unpatentable over Fukui, Riedel, Maruyama, and Clemente as applied to claims 1-3, 8, 10, 19-21, 23-24, and 26-27 above, and further in view of Murakami (WO 2009/145164; Date of Publication: December 3, 2009 – cited in the IDS filed on 06/24/2024 – previously cited).
The teachings of Fukui, Riedel, Maruyama, and Clemente, herein referred to as modified-Fukui-Riedel-Maruyama-Clemente, are discussed above as it pertains to producing a polyhydroxyalkanoate copolymer mixture.
However, modified-Fukui-Riedel-Maruyama-Clemente does not teach: wherein the middle-chain fatty acid is hexanoic acid (claim 22).
Murakami’s general disclosure relates to a microorganism that produces a PHA copolymer comprising 3-hydroxybutyric acid and 3-hydroxyvaleric acid (see, e.g., Murakami, English translation, pg. 2). Furthermore, Murakami discloses “a microorganism in which the expression of the bktB gene is enhanced, the catalytic activity of the enzyme encoded by the gene is improved, and a polyhydroxyalkanoate synthase gene and a crotonyl-CoA reductase gene are introduced. By culturing the microorganism, it is possible to efficiently produce P (3HB-co-3HH), which is a PHA having excellent flexibility and a wide application range, using an inexpensive carbon source” (see, e.g., Murakami, abstract). Moreover, Murakami discloses that use of hexanoic acid as a carbon source allows for the production of 3HH-containing PHA (see, e.g., Murakami, English translation, pg. 5).
Regarding claim 22 pertaining to hexanoic acid, Murakami teaches that hexanoic acid can be used as a carbon source (see, e.g., Murakami, English Translation, pg. 5).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to culture modified-Fukui-Riedel-Maruyama-Clemente’s microorganism, wherein the microorganism uses hexanoic acid as a carbon source, as taught by Murakami. One would have been motivated to do so because Murakami teaches that use of hexanoic acid as a carbon source allows for the production of 3HH-containing PHA (see, e.g., Murakami, English translation, pg. 5). Furthermore, modified-Fukui-Riedel-Maruyama-Clemente teaches the production of PHA with high 3HHx content makes the polymer more soluble (see, e.g., Riedel, “Discussion”, pg. 468), meanwhile the lower HHx content forms a gel-like phase (see, e.g., Riedel, abstract). Furthermore, modified-Fukui-Riedel-Maruyama-Clemente teaches that the amount of HHx affects polymer flexibility (see, e.g., [0003]-[0004]). Therefore, based on the teachings of modified-Fukui-Riedel-Maruyama-Clemente and Murakami, it would have been obvious to supply hexanoic acid to a microorganism to allow for production of 3HH-containing PHA. One would have expected success because modified-Fukui-Riedel-Maruyama-Clemente and Murakami both teach PHA synthesis
Examiner’s Response to Arguments
Applicant’s amendments and arguments filed on 11/10/2025 have been fully considered but they are not persuasive and deemed insufficient to overcome the prior arts of record.
Regarding Applicant’s arguments regarding the amendment to claim 1, which incorporates the limitations of claim 10 and which was not rejected (remarks, pages 7-9), this argument is not persuasive because, as discussed above, the transitional phrase “having” is synonymous to “comprising”, making the SEQ ID NO: 6 limitation open-ended and does not exclude additional, unrecited elements (see, e.g., MPEP 2111.03(I)). Based on this, Clemente was used as prior art to teach instant SEQ ID NO: 6, as taught above. Therefore, claims 1 and 10 are not free of the art.
Conclusion
Claims 1-3, 8, 10, 15-24, and 26-27 are rejected.
No claims are allowed.
Correspondence Information
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/NATALIE IANNUZO/Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653