DETAILED ACTION
Applicant’s amendment submitted on 7/1/2024 is acknowledged. Claims 1-8, 10-12, and 14-24 are pending in the instant application. Claims 22-24 are newly added. Claims 1-5, 7, 12, and 16-17, are currently amended. Claims 8, 10, and 21 are withdrawn pursuant to 37 CFR 1.142(b) as being drawn to a non-elected invention.
The instant action is Non-Final because it presents new rejections that were not necessitated by the amendment.
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 .
Priority
The instant application is a U.S. National Phase of PCT/CN2020/137779 filed on 12/18/2020 and claims foreign priority to CN202010460035.9 filed on 5/27/2020, and a receipt of a certified copy of that document is acknowledged. However, no English translation of the foreign priority document has been provided to perfect priority, therefore the effective filing date of the instant claims is considered to be the filing date of the international application, 12/18/2020. See 37 CFR 1.55.
Election/Restrictions
The restriction requirement was made final in the Non-Final Rejection mailed on 5/15/2024 and claims 8, 10, and 21 remain withdrawn. Applicant’s arguments submitted 7/1/2024 are based on the currently amended claims and not on the claims as originally filed and restricted.
Response to Amendment
Applicants’ amendment to the claims overcomes the 112(b) rejections previously set forth in the Non-Final Rejection mailed on 5/15/2024. Accordingly, the 112(b) rejections are withdrawn.
Claims 16 and 17 are no longer substantial duplicates in view of Applicants’ amendment.
Claim Objections
Claims 2 and 23 are objected to because of the following informalities:
Claim 2 recites “(7) enhancing activity of AHAS and/or ilvD” in the last line of the claim. The recitation of the abbreviation AHAS should be preceded by the fully written out phrase in its first instance in the claims. Additionally, claim 2 should be amended to indicate an ilvD gene for consistency with previous recitations of genes in the claim.
Claim 23 recites “by substituting the pflB gene of the microorganism itself with the ilvD gene”; “by substituting the frd gene of the microorganism itself with the leuDH gene”; and “by substituting the mgsA gene of the microorganism itself with a ilvC gene” which are improper for the following reason. The conventional manner of reciting a substitution is as follows: “substituting y for x;” where x is the variable being replaced by y. The term “substituting” should be amended to “replacing”.
Appropriate correction is required.
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.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
New Rejection Necessitated by Amendment: Claim 23 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 23 recites the limitation "the leuDH gene" in lines 3-4. There is insufficient antecedent basis for this limitation in the claim. Claim 23 depends from claim 2, which depends from claim 1. Neither claim 1 nor claim 2 recite a leuDH gene, so it is unclear to what “the leuDH gene,” as recited in claim 23, refers to in claim 23. One of ordinary skill in the art would not be able to determine the metes and bounds of the claim limitation, and thus, claim 23 is indefinite.
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 (i.e., changing from AIA to pre-AIA ) 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, 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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
New Rejection (claim 4 was omitted in the previous 103 rejection): Claims 1-2, 4-7, 12, 14-15, and 24 are rejected under 35 U.S.C. 103 as being unpatentable over Xie et al. (US 11,802,300 B2; corresponds to CN110607268A of record in IDS filed 10/19/2021) in view of Hasegawa et al. (Metabolic Engineering 2020, Vol 59: pp. 24-35; of record) and as evidenced by Part_BBa K3866001.
Regarding claim 1, Xie teaches a “genetically engineered strain [of Escherichia coli] with high yield of L-valine” (Abstract) wherein “a gene bcd [under a trc promoter] encoding leucine dehydrogenase of Bacillus subtilis replaces a gene ilvE” (see Claim 1) among other genetic modifications. Using the bcd instead of the ilvE leads to NADH instead of NADPH being used as the coenzyme during L-valine synthesis (see Col. 4, lines 29-42). The modifications of the strain focuses on removal of L-valine feedback inhibition, increasing the supply of reducing power NADPH, and enhancement of L-valine output (see Col. 1, lines 47-63). With regard to activating an Entner-Doudoroff metabolic pathway, the act of transferring the amino acid dehydrogenase gene into an E. coli in itself may inherently activate the ED pathway as claimed.
Xie does not explicitly teach activating an ED metabolic pathway in the microorganism.
Hasegawa teaches the production of isobutanol in Corynebacterium glutamicum wherein the productivity of the microorganism is enhanced via the Entner-Doudoroff pathway (Title). Figure 1 (shown below) teaches the metabolic pathway from glucose to isobutanol including the overexpressed genes (underlined), disrupted genes (crossed out), and genes involved in the ED pathway (black arrows). As seen in the figure, L-valine and isobutanol share a late metabolite precursor, 2-ketoisovalerate, suggesting the pathways to reach this molecule would be advantageous for the production of both L-valine and isobutanol. Hasegawa teaches a way to enhance glycolytic flux by introducing the Entner-Doudoroff pathway (EDP) wherein the zwf, edd, and eda were enhanced, reading on claim 15 (page 32, last paragraph; page 33, first paragraph). The enhancement of the EDP generates a higher ratio of reducing equivalents that permit higher production levels of the L-valine and isobutanol precursor, 2-ketoisovalerate. Although the figure fails to show the enhancement of the pgl gene, the instant specification refers to pgl as “a lactonase encoding gene” (page 4, last paragraph). The figure 1 legend refers to devB as 6-phosphogluconolactase and it clearly lies within the EDP.
Therefore, it would have been obvious to one of ordinary skill in the art, before the effective filing date of the claimed invention, to activate the Entner-Doudoroff Pathway in an L-valine producing microorganism using overexpression of the zwf, lactonase (such as pgl), edd, and eda genes. One of ordinary skill in the art would have been motivated with a reasonable expectation of success to enhance the EDP as taught by Hasekawa in a microorganism comprising a transferred leucine dehydrogenase gene, as taught by Xie, to increase the availability of the NADPH and the production of the L-valine, yielding predictable results.
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Regarding claim 2, Xie teaches “a lactate dehydrogenase gene ldhA, a pyruvate formate lyase I gene pflB and genes frdA, frdB, frdC, and frdD for four subunits of fumarate reductase are knocked out from the genome of the E. coli W3110” (page 10, column 2, lines 54-57) corresponding to the limitations (2) and (6).
Regarding claims 4 and 5, Xie teaches the bcd gene encoding leucine dehydrogenase (leuDH) is under a trc promoter, reading on claims 4 and 5 (see Col. 3, lines 55-59).
Regarding claim 6, Xie teaches an E. coli W3110 recombinant microorganism (see Abstract, Col. 3, lines 40-63, and Claim 1).
Regarding claim 7, Xie teaches the development of a recombinant microorganism for producing L-valine, but differs from the claim insofar as it does not teach the use of metabolic evolution wherein the L-valine strain undergoes 50-120 generations as defined by the instant specification. However, it would have been obvious to one of ordinary skill in the art prior to the effective filing date to obtain the microorganism through the use of ‘metabolic evolution’. The passage of generations of bacteria is a known technique in the field for optimizing growth and productivity of a bacterial isolate. One of ordinary skill in the art would have been motivated to pass the recombinant L-valine producing organism through 50-120 generations to yield the predictable results of a stable, productive isolate with a reasonable expectation of success.
Regarding claim 12, Xie teaches “an acetohydroxy acid reductoisomerase gene ilvC of E. coli is replaced with an acetohydroxy acid isomeroreductase mutant L67E/R68F/K75E gene ilvCM” (page 10, column 2, lines 60-64). Thus, an acetohydroxy acid reductoisomerase encoding gene is being inserted into the microorganism as per claim 12.
Regarding claim 14, Xie teaches “a gene bcd [under a trc promoter] encoding leucine dehydrogenase of Bacillus subtilis replaces a gene ilvE” (see Claim 1) among other genetic modifications, reading on claim 14.
Regarding claim 24, the trc promoter comprises a RBS artificial regulatory element as evidenced by Part_BBa K3866001, reading on claim 24.
New Rejection Necessitated by Amendment: Claim 11 is rejected under 35 U.S.C. 103 as being unpatentable over Xie et al. (US 11,802,300 B2; corresponds to CN110607268A of record in IDS filed 10/19/2021) in view of Hasegawa et al. (Metabolic Engineering 2020, Vol 59: pp. 24-35; of record) and as evidenced by Part_BBa K3866001, as applied to claims 1-2, 4-7, 12, 14-15, and 24 above, and further in view of Park et al. (PNAS 2007, Vol 104(19): pp. 7797-7802; of record in IDS filed 09/29/2022).
Xie in view of Hasegawa teach the invention of claim 1 as outlined in the rejection above.
Regarding claim 11, neither Xie nor Hasegawa teach knocking out a 6-phosphoglucokinase gene pfkA.
Park teaches the “[m]etabolic engineering of Escherichia coli for the production of L-valine” (Title) wherein “pfkA/B knockout seems to increase the availability of NADPH, an important cofactor for L-valine production, by pushing more carbon flux towards the pentose phosphate pathway” (page 7800, column 2, paragraph 1).
Therefore, it would have been obvious to one of ordinary skill in the art, prior to the effective filing date, to knockout the pfkA gene in a recombinant L-valine producing microorganism. As the limiting factor of L-valine is often the availability of NADPH, one of ordinary skill in the art would be motivated to combine the teachings of Xie and Park to create a recombinant L-valine-producing microorganism that has an NADH-dependent amino acid dehydrogenase and the pfkA gene knocked out. With increased levels of NAPH available, a higher titer of L-valine could be produced from the microorganism.
New Rejection Necessitated by Amendment: Claims 16-20 and 22 are rejected under 35 U.S.C. 103 as being unpatentable over Xie et al. (US 11,802,300 B2; corresponds to CN110607268A of record in IDS filed 10/19/2021) in view of Hasegawa et al. (Metabolic Engineering 2020, Vol 59: pp. 24-35; of record) and as evidenced by Part_BBa K3866001, as applied to claims 1-2, 4-7, 12, 14-15, and 24 above, and further in view of Savrasova et al. (Heliyon 2019, Vol 5: pp. 1-25; of record in IDS filed 10/19/2021).
Xie teaches the limitations of items (2) and (6) from claim 2 wherein by knocking out lactate dehydrogenase gene ldhA, pyruvate formate lyase I gene pflB and four fumarate reductase subunit genes frdA, frdB, frdC and frdD, the accumulation of byproducts (lactate, formate and succinate) under anaerobic conditions is reduced (page 11, column 4, lines 24-28), but differs from the claims insofar as it does not teach the enhancement of the AHAS nor ilvD.
Savrasova teaches “[t]he valine biosynthetic pathway in E. coli consists of four reactions catalyzed by enzymes: acetohydroxy acid synthase, which catalyzes the first common step in BCAA biosynthesis (isoenzymes AHAS I, II, III, encoded by ilvBN, ilvGM, and ilvIH, respectively); ketol-acid reductoisomerase (KARI), encoded by ilvC; dihydroxy acid dehydratase (DHAD), encoded by ilvD; and BCAA aminotransferase (BCAT, hereinafter AT), encoded by ilvE” wherein “[t]he key enzyme among the four is AHAS” (page 4, paragraph 2). Last, Savrasova teaches of “E. coli valine-producing strains containing feedback-resistant AHAS I encoded by the ilvBNN17K genes as part of the artificial operon […] in the chromosome,” reading on claim 17 (sentence spanning pages 4-5).
Regarding claims 16-20 and 22, it would have been obvious to one of ordinary skill in the art prior to the effective filing date to combine the teachings of Xie with the teachings of Savrasova. One of ordinary skill in the art would have been motivated with a reasonable expectation of success to knock out either or both the lactate dehydrogenase gene and/or the fumarate reductase genes to limit the accumulation of undesirable metabolic byproducts as taught by Xie while also enhancing the activity of the AHAS (ilvBN) gene to limit the amount of feedback inhibition received and to ensure an increased production of L-valine as taught by Savrasova, which would yield predictable results.
New Rejection Necessitated by Amendment: Claims 3 and 23 are rejected under 35 U.S.C. 103 as being unpatentable over Xie et al. (US 11,802,300 B2; corresponds to CN110607268A of record in IDS filed 10/19/2021) in view of Hasegawa et al. (Metabolic Engineering 2020, Vol 59: pp. 24-35; of record) and as evidenced by Part_BBa K3866001, as applied to claims 1-2, 4-7, 12, 14-15, and 24 above, and further in view of Shi et al. (Metabolic Engineering 2013, Vol.16, p.1-10).
Xie teaches knocking out a pflB gene, a frd gene, and an ilvC gene in E. coli W3110 (see Claim 1).
Regarding claim 23, Xie does not teach substituting an ilvD gene for the pflB gene, substituting a leuDH gene for the frd gene, or substituting the ilvC gene for mgsA.
Shi teaches methods for improving isobutanol production in E. coli (see Abstract). As seen in the figure above in the rejection of claim 1, L-valine and isobutanol share a late metabolite precursor, 2-ketoisovalerate, suggesting the pathways to reach this molecule would be advantageous for the production of both L-valine and isobutanol. Shi teaches integration of ilvD at pflB and integrating ilvC at mgsA (see p.2, right column, 4th paragraph and p.3, left column, 1st paragraph).
Therefore, it would have been obvious to one of ordinary skill in the art, before the effective filing date of the claimed invention, to have integrated ilvD at pflB and ilvC at mgsA, as taught by Shi, in the E. coli for producing L-valine as taught by Xie and modified by Hasegawa above, to arrive at the claimed invention. One of ordinary skill in the art would have been motivated to apply Shi’s knock-out/knock-in of pflB and mgsA for ilvD and ilvC, respectively, to the E. coli of Xie as modified by Hasegawa because it is a standard technical practice in the relevant field and one of ordinary skill would recognize the benefits of improving the production of isobutanol as it relates to the production of L-valine, as discussed in the rejection of claim 1 above.
With regards to claim 3, Shi teaches performing their methods in E. coli ATCC 8739 as an initial strain (see p.4, left column, last paragraph and Fig. 2). It would have been obvious to substitute The ATCC 8739 strain, as used by Shi, for the W3110 strain, as used by Xie, to arrive at the claimed invention. One of ordinary skill in the art would have been substituting known E. coli strains used in the relevant technologies, yielding predictable results.
Nonstatutory Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
New Rejection Necessitated by Amendment: Claims 1-3, 6-7, 12, 14-16, 18-20, 22, and 24 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-4, 6-7, 11, 13, 15-20, and 22-23 of copending Application No. 17/603,006 in view of Hasegawa et al. (Metabolic Engineering 2020, Vol 59: pp. 24-35; of record).
This is a provisional nonstatutory double patenting rejection.
Both instant claim 1 and co-pending claim 1 recite a construction method of a recombinant microorganism for producing L-valine comprising (lines 1-2): transferring an amino acid dehydrogenase gene into a microorganism (lines 2-3), the microorganism is Escherichia coli; the amino acid dehydrogenase gene is NADH-dependent (lines 5-6). Claim 1 of the ‘006 application is directed towards a similar method of inserting an amino acid dehydrogenase into a microorganism, but rather than activating the Entner-Doudoroff pathway, the method as claimed activates the activity of a transhydrogenase and a NAD kinase in the microorganism. However, the same genes are knocked out or enhanced in the recombinant microorganisms of the two applications (claim 2). Last, although the instant claim 1 does not specifically require the activation of the transhydrogenase and/or NAD kinase as claimed in the ‘006 application, using the specification of the patent as a dictionary (see MPEP § 804(II)(B)(1)) to define the scope of the claimed method, it is clear that the claimed method can be used to overcome the reducing power imbalance during the anaerobic fermentation (page 4, middle paragraph). Co-pending claim 1 does not recite activating an Entner-Doudoroff pathway as instantly claimed. Hasegawa teaches the production of isobutanol in Corynebacterium glutamicum wherein the productivity of the microorganism is enhanced via the Entner-Doudoroff pathway (Title). Figure 1 (shown below) teaches the metabolic pathway from glucose to isobutanol including the overexpressed genes (underlined), disrupted genes (crossed out), and genes involved in the ED pathway (black arrows). As seen in the figure, L-valine and isobutanol share a late metabolite precursor, 2-ketoisovalerate, suggesting the pathways to reach this molecule would be advantageous for the production of both L-valine and isobutanol. Hasegawa teaches a way to enhance glycolytic flux by introducing the Entner-Doudoroff pathway (EDP) wherein the zwf, edd, and eda were enhanced, reading on claim 15 (page 32, last paragraph; page 33, first paragraph). The enhancement of the EDP generates a higher ratio of reducing equivalents that permit higher production levels of the L-valine and isobutanol precursor, 2-ketoisovalerate. Although the figure fails to show the enhancement of the pgl gene, the instant specification refers to pgl as “a lactonase encoding gene” (page 4, last paragraph). The figure 1 legend refers to devB as 6-phosphogluconolactase and it clearly lies within the EDP.
Therefore, it would have been obvious to one of ordinary skill in the art, before the effective filing date of the claimed invention, to activate the Entner-Doudoroff Pathway in an L-valine producing microorganism using overexpression of the zwf, lactonase (such as pgl), edd, and eda genes. One of ordinary skill in the art would have been motivated with a reasonable expectation of success to enhance the EDP as taught by Hasekawa in a microorganism comprising a transferred leucine dehydrogenase gene, as taught by Xie, to increase the availability of the NADPH and the production of the L-valine, yielding predictable results.
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Instant claims 2, 3, 6, 7, 12, 14, 16, 18, 19, 20, and 24 correspond to co-pending claims 2, 3, 6, 7, 11, 13, 15, 16, 17, 18, and 23, respectively. Instant claim 22 corresponds to co-pending claims 19, 20, and 22.
Response to Arguments
Applicant requests that the double patenting rejection over co-pending application 17/603,006 be held in abeyance (Remarks p.9, 1st----2nd passages). A request to hold a rejection in abeyance is not a proper response to a rejection. Rather, a request to hold a matter in abeyance may only be made in response to an OBJECTION or REQUIREMENTS AS TO FORM (see 37 CFR 1.111(b) and MPEP § 714.02). Thus, the double patenting rejections of record are still applicable as no response to these rejections has been filed by Applicant at this time. Claims 1-3, 6-7, 12, 14-16, 18-20, 22, and 24 are provisionally rejected on the ground of nonstatutory double patenting.
Applicant’s arguments, see p.10, last passage, filed 7/1/2024, with respect to claims 1-3, 5-6, and 12-14 have been fully considered and are persuasive. The 35 U.S.C. 102(a)(1) rejection of claims 1-3, 5-6, and 12-14 has been withdrawn.
Applicant's arguments filed 7/1/2024 have been fully considered but they are not persuasive.
In Applicant’s Remarks, see p.11, last passage and p.12, 1st-2nd passages, Applicant argues that the present application concerns a one-step anaerobic fermentation while the prior art of Xie concerns a two-stage aerobic-anaerobic fermentation, and thus the microbial metabolic conditions are different. This is not found persuasive as this argument is not commensurate in scope with the claimed invention which does not recite any fermentation conditions. Additionally, the rejection above teaches each and every limitation of the claimed invention.
In Applicant’s Remarks, see p.12, 3rd-last passages, Applicant argues the prior art of Xie concerns the TCA cycle, which is among the prior arts to be abandoned. Applicant further explains glucose metabolism under anaerobic conditions and the dependency on cofactors. Applicant further submits the claimed invention improves the redox balance by transferring an amino acid dehydrogenase gene into the microorganism and activating an Entner-Doudoroff metabolic pathway in the microorganism; the amino acid dehydrogenase gene is NADH-dependent. This is not found persuasive. The fermentation conditions are not recited in the claimed invention and thus Applicant’s arguments are not commensurate in scope with the claimed invention. Further, the prior art of Xie in view of Hasegawa as used in the rejection set forth above teach each and every limitation of the claimed invention.
In Applicant’s Remarks, see p.13, 4th-5th passages, Applicant argues that park teaches knocking out pfkA/B for reasons other than those of Applicant’s. This is not found persuasive because a rationale for a teaching, suggestion, or modification may differ from that of Applicant’s. See MPEP § 2144(IV).
In Applicant’s Remarks, see p.13, 6th-last passages, Applicant argues that Hasegawa is directed to activation of the Entner-Doudoroff pathway (EDP) in Corynebacterium glutamicum and not Escherichia coli, as presently claimed. Applicant further argues a person skilled in the art cannot predict that activating the EDP in E. coli can effectively improve the yield of L-valine and solve the problem of reducing power imbalance during the anaerobic fermentation from the teachings of Hasegawa relating to C. glutamicum. Applicant further argues Hasegawa fails to disclose “transferring an NADH-dependent amino acid dehydrogenase gene into a microorganism” and “activating an Entner-Doudoroff metabolic pathway in Escherichia coli” to solve the problem of reducing power imbalance during anaerobic fermentation. This is not found persuasive. Hasegawa teaches the production of isobutanol in Corynebacterium glutamicum wherein the productivity of the microorganism is enhanced via the Entner-Doudoroff pathway (Title). Figure 1 (shown below) teaches the metabolic pathway from glucose to isobutanol including the overexpressed genes (underlined), disrupted genes (crossed out), and genes involved in the ED pathway (black arrows). As seen in the figure, L-valine and isobutanol share a late metabolite precursor, 2-ketoisovalerate, suggesting the pathways to reach this molecule would be advantageous for the production of both L-valine and isobutanol. Hasegawa teaches a way to enhance glycolytic flux by introducing the Entner-Doudoroff pathway (EDP) wherein the zwf, edd, and eda were enhanced. The enhancement of the EDP generates a higher ratio of reducing equivalents that permit higher production levels of the L-valine and isobutanol precursor, 2-ketoisovalerate. Thus, one of skill in the art would recognize the benefit of activating the EDP in an L-valine producing microorganism. With respect to Applicant’s arguments concerning what Hasegawa fails to teach, these limitations are not relied upon to be taught by Hasegawa in the rejection set forth above, and are taught by the prior art of Xie.
In Applicant’s Remarks, see passage bridging p.14-15, Applicant further argues the cited prior art do not teach the fermentation conditions of the claimed invention. This is not found persuasive because presently no fermentation conditions are recited in the claimed invention. Thus, Applicant’s arguments are not commensurate in scope with the claimed invention.
In Applicant’s Remarks, see p.15, 3rd-last passages and p.16, 1st-4th passages, Applicant argues the present application achieves unexpected effects and cites to Examples 13-20 for evidence of achieving unexpected increased amounts of L-valine. This is not found persuasive. Each of the examples requires anaerobic fermentation conditions of which are not recited in the claimed invention, and therefore, the argument is not commensurate in scope with the claimed invention. Further, there is no requirement for producing L-valine in the claimed invention which is directed to methods of constructing an E. coli for producing L-valine. Therefore, Applicant’s arguments are not persuasive.
Conclusion
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/LOUISE W HUMPHREY/Supervisory Patent Examiner, Art Unit 1657
/J.P.S./Examiner, Art Unit 1657