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 .
Restriction/Election
In response to the communication received on April 3, 2026, from Mahreen Chaudhry Hoda, the election with traverse of Group I, claims 1-4,6-7,9,11-12,14,16-19,21,23-26,28-33,35,37-39,42-44 and 47-51, is acknowledged. Applicant contends that there would be no serious search and examination burden associated with examining Groups I and II together because the searches for each of these groups would be co-extensive. Group II is directed to a method that requires the microorganism of Group 1 (Remarks, page 2).
As described in the Office action mailed on 02/04/2026, inventions I and II are related as product(s) and process of using the product(s). In the instant case the claimed microorganisms of Group I can be cultured without carboxylic acid, i. e., using different method steps. Further in contrast, the claimed microorganisms of Group I can be used in genomics, physiological and biochemical studies, which are processes unrelated to the instantly claimed method of Group II. In addition, the invention of Group II is directed to isolating products of interest, which certainly requires searches and considerations that are not required by the invention(s) of Group I.
The restriction is still deemed proper and is therefore made FINAL.
Priority
This application is a continuation of PCT International Application No. PCT/US22/036856, which designated the United States, and was filed on Jul. 12, 2022, published in English, which claims priority to U.S. Provisional Patent Application No. 63/220,927, filed Jul. 12, 2021. Accordingly, the effective filing date is 07/12/2021.
Information Disclosure Statement
Initialed and dated copy of Applicants’ information disclosure statement (IDS) filed on 04/03/2026 is attached to the instant Office action. The submission is in compliance with the provisions of 37 C.F.R. § 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
The listing of references throughout the specification (e.g., at pages 28-35; 42-43; 45; 51; 56-62; 64; 68; 71; 74) is not a proper information disclosure statement. 37 CFR § 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the references have been cited by the examiner on form PTO-892, they have not been considered.
Status of Claims
Claims 1-4, 6-7, 9, 11-12, 14, 16-19, 21, 23-26, 28-33, 35, 37-39, 42-44, and 47-53 are pending.
Claims 52-53 are withdrawn from consideration for being directed to non-elected invention(s).
Claims 1-4, 6-7, 9, 11-12, 14, 16-19, 21, 23-26, 28-33, 35, 37-39, 42-44, and 47-51 are examined in this Office action.
Claim Objections
The claims are objected to for the following informalities.
In claim 50, in the phrase describing compound “CO2”, the number “2” should be subscripted.
Appropriate corrections is required.
Claim Rejections - 35 USC § 112
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 the second paragraph of 35 U.S.C. 112:
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.
Indefiniteness
Claims 18 and 50 are 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 that the inventor or a joint inventor, or for pre-AIA the applicant, regards as the invention.
Claim 18 does not separate the relationship between the elements such that “enzymes to convert a 1-carbon substrate to formyl-CoA and a 2-hydroxyacyl-CoA lyase or oxalyl-CoA decarboxylase condensing said 2-hydroxyaldehyde with said formyl-CoA to form a 2,3-dihydroxyacyl-CoA 1 carbon longer than said 2-hydroxyaldehyde” lack clarity for what exactly the claim encompasses.
Claim 50 recites the limitation of a carbon feedstock selected from the group consisting of glucose, xylose, arabinose, glycerol, methane, CO2, methanol, formate, formaldehyde, and “similar substances”. It is unclear what these “similar substances” encompass.
In the interest of compact prosecution, the claims are nevertheless examined.
The following is a quotation of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), first paragraph:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Scope of Enablement
Claims 1-4, 6-7, 9, 11-12, 14, 16-19, 21, 23-26, 28-33, 35, 37-39, 42-44, and 47-51 are rejected under 35 U.S.C. § 112(a), first paragraph, because the specification, while being enabling for genetically modified bacteria capable of converting carboxylic acid to a product, does not reasonably provide enablement for producing all possible genetically modified microorganisms capable of converting carboxylic acid to all possible products. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with these claims. All dependent claims are included in these rejections unless they contain a limitation that overcomes the deficiencies of the parent claim from which they depend.
The Specification provides a plurality of largely prophetic examples for practicing the claimed invention(s), including lists of enzymes, exemplary engineered vectors, and limited number of actually performed experiments using transformed E. coli (Examples 1-28).
For example, while it is known in the art that the diversity of microbes presently living on earth is known to be high, the true extent of microbial diversity is largely unknown. Bacteria, archaea, eukaryotes, and viruses comprise the four major categories of microorganisms. The microbial population represents the richest biodiversity resources on the planet. According to a recent estimate, there are over one trillion different species of microorganisms on the planet, of which 99.999 percent have not yet been identified. These bacteria create communities with astounding diversity in their hosts or habitats (Sangwan et al. 2023, Methods for Measurement of Microbial Diversity, In: Biotechnological Interventions Augmenting Livestock Health and Production, Springer, pp 171-192). These factors do not appear to have been adequately addressed in the instant application.
Given the absence of guidance in the specification, and given the relatively high level of unpredictability in the art, one of skill in the art cannot predict the effect of introducing the myriad of listed enzymes into all possible expression vectors and subsequently microorganisms, and their causal relationship(s) to converting carboxylic acid to all possible products (or even more narrowly the recited products in some of the dependent claims), undue trial and error experimentation would have been required for one skilled in the art to use the claimed invention.
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, 6-7, 9, 11-12, 14, 16-19, 21, 23-26, 28-33, 35, 37-39, 42-44, and 47-51 are rejected under 35 U.S.C. § 103(a) as being unpatentable over in view of GONZALEZ (Gonzalez et al., PCT International Patent Application Publication No. WO 2016/069929 A1, published 6 May 2016; see IDS filed 04/03/2026) in view of LI (Li et al., 2020, Next-generation metabolic engineering of non-conventional microbial cell factories for carboxylic acid platform chemicals, Biotechnology Advances 43: 107605, https://doi.org/10.1016/j.biotechadv.2020.107605, pp 1-23), LIU (Liu and Jarboe, 2012, Metabolic engineering of biocatalysts for carboxylic acids production, 2012, Computational and Structural Biotechnology Journal 3 (4), e201210011, http://dx.doi.org/10.5936/csbj.201210011, pp 1-9), and VILA-SANTA (Vila-Santa et al., 2021, Prospecting Biochemical Pathways to Implement Microbe-Based Production of the New-to-Nature Platform Chemical Levulinic Acid, ACS Synthetic Biology 10: 724-736).
The claims are drawn to a genetically modified microorganism converting a carboxylic acid to a product, which comprises: a) a first set of nucleic acids encoding enzymes to activate said carboxylic acid to the corresponding acyl-CoA intermediate; b) a second set of nucleic acids encoding enzymes to convert said acyl-CoA intermediate to a product; and c) a third set of nucleic acids encoding enzymes to generate reducing equivalents and ATP from an externally supplied energy source; wherein said first set of metabolic enzymes comprises an acyl-CoA synthetase, or an acyl-CoA transferase, or a carboxylate kinase and a phosphotransacylase, or a carboxylic acid reductase and an acyl-CoA reductase, or an aldehyde dehydrogenase and an acyl-CoA reductase converting said carboxylic acid to the corresponding acyl-CoA intermediate; wherein said second set of metabolic enzymes comprises an aldehyde forming acyl-CoA reductase converting said acyl-CoA to an aldehyde; or wherein said second set of metabolic enzymes comprises an aldehyde forming acyl-CoA reductase and alcohol dehydrogenase, or an alcohol forming acyl-CoA reductase converting said acyl-CoA to an alcohol; which microorganism is bacteria or yeast.
GONZALEZ teaches the biosynthesis of products from 1-carbon compounds (entire document; see Title, Abstract, for example). Desirable products include liquid fuels, basic chemicals, consumer chemicals, and specialty chemicals ([0004]; Figure 2). GONZALES describes an engineered microbe that contains a designed platform for the conversion of one-carbon substrates to chemical products. The designed platform embodies a new metabolic architecture that consolidates carbon fixation, central metabolism, and product synthesis into a single pathway. This is made possible by the key finding that 2-hydroxyacyl-CoA lyase, an enzyme in the α-oxidation pathway, is capable of catalyzing the C-C bond formation between formyl-CoA and aldehydes of different chain lengths, allowing for the elongation of the carbon backbone of said aldehyde by one-carbon units. These novel microbes present an opportunity for the production of chemicals from single-carbon feedstocks such as carbon dioxide, carbon monoxide, formate, formaldehyde, methanol or methane (Abstract).
GONZALEZ teaches and claims a genetically modified microbe (i.e., a genetically modified microorganism) converting a carboxylic acid to a product (conversion of single-carbon molecules such as formate (carboxylic acid) to a product; claim 1 of GONZALEZ), comprising: a. a first set of nucleic acids encoding enzymes to activate said carboxylic acid to the corresponding acyl-CoA intermediate (a polypeptide capable of catalyzing a reaction of formate to formyl-CoA); b. a second set of nucleic acids encoding enzymes to convert said acyl-CoA intermediate to a product (polypeptides catalyzing a conversion of formyl-CoA through elongation and termination steps into a desired carboxylic acid, alcohol, or aldehyde product; claim 1 ); and c. a third set of nucleic acids encoding enzymes to generate reducing equivalents and ATP from an externally supplied energy source (methanol (externally supplied energy source) allows for a high growth rate via methanol oxidation producing ATP for growth in E. coli metabolism, implying the presence of nucleic acids encoding enzymes needed for this metabolism; paragraphs [00169]).
GONZALEZ teaches an elongation module shown in FIG. 4A-B and summarized in Table 1B, which allows for the iterative elongation of the carbon backbone. GONZALEZ specifically teaches that 2-hydroxyacyl-CoA can be converted to its corresponding aldehyde via an alternative arrangement of reductions and dehydrations to those described in FIG. 4B. Lyase is first reduced to a 2-hydroxyaldehyde by an acyl-CoA reductase, examples of which are provided above. The 2-hydroxyaldehyde is then reduced further to a 1,2-diol by a 1,2-diol oxidoreductase. E. coli fucO has been shown to encode one such 1,2-diol oxidoreductase. Finally, the 1,2-diol can be converted to the aldehyde by a diol dehydratase [0090-0095].
For example, regarding claim 1, GONZALEZ teaches and claims (claim 1 of GONZALEZ) a genetically modified microbe comprising a synthetic pathway (i.e., a genetically modified microorganism) converting single-carbon molecules such as formate (carboxylic acid) to a product (i.e., converting a carboxylic acid to a product), comprising a first set of nucleic acids encoding enzymes to activate said carboxylic acid to the corresponding acyl-CoA intermediate (i.e., a polypeptide capable of catalyzing a reaction of formate to formyl-CoA), a second set of nucleic acids encoding enzymes to convert said acyl-CoA intermediate to a product (i.e., polypeptides catalyzing a conversion of formyl-CoA through elongation and termination steps into a desired carboxylic acid, alcohol, or aldehyde product), and a third set of nucleic acids encoding enzymes to generate reducing equivalents and ATP from an externally supplied energy source (i.e., methanol, externally supplied energy source), which allows for a high growth rate via methanol oxidation, by producing ATP for growth in E.coli metabolism (implying the presence of nucleic acids encoding enzymes needed for this metabolism [paragraph 00169]; see also infra).
Regarding claim 2, GONZALEZ teaches and claims (claim 6 of GONZALEZ) the microorganism of claim 1, and further teaches that the first set of metabolic enzymes comprises an acyl-CoA synthetase (i.e., acetyl-CoA synthetase), or an acyl-CoA transferase, or a carboxylate kinase and a phosphotransacylase, or a carboxylic acid reductase and an acyl-CoA reductase, or an aldehyde dehydrogenase and an acyl-CoA reductase converting said carboxylic acid to the corresponding acyl-CoA intermediate (i.e., a polypeptide capable of catalyzing a reaction of formate to formyl-CoA, claim 1 of GONZALEZ).
Regarding claim 3, GONZALEZ teaches the microorganism of claim 1, and further teaches that the second set of metabolic enzymes comprises an aldehyde forming acyl-CoA reductase (i.e., an acyl-CoA reductase; paragraph [0094]) converting said acyl-CoA to an aldehyde (i.e., polypeptides catalyzing a conversion of formyl-CoA through elongation and termination steps into a desired carboxylic acid, alcohol, or aldehyde product).
Regarding claim 4. GONZALEZ teaches the microorganism of claim 3, and further teaches and claims that the carboxylic acid is a α-hydroxy acid, and said aldehyde is a α -hydroxy aldehyde (paragraph [00106]; claim 13 of GONZALEZ).
Regarding claim 18, GONZALEZ teaches the microorganism of claim 4 and further teaches a cyclical method of single carbon addition wherein an aldehyde and a formyl-CoA are ligated, with additional steps to convert the produced 2-hydroxyacyl-CoA to an aldehyde that is a single carbon longer than the starting aldehyde. For example, GONZAEZ teaches that the elongation module consists of the ligation of an aldehyde and formyl-CoA, followed by steps that convert the produced 2-hydroxyacyl- CoA back to an aldehyde 1 carbon longer than the starting aldehyde paragraph [00142]) with a 2-hydroxyacyl-CoA lyase (2-hydroxyacyl-CoA lyase; claim 9 of GONZALEZ).
Regarding claim 19, GONZALEZ teaches the method of instant claim 18, and further teaches wherein said second set of metabolic enzymes further comprises a thioesterase, or an acyl-CoA transferase, or a phosphotransacylase and a carboxylate kinase (thioesterase, or an acyl-CoA:acetyl-CoA transferase, or a phosphotransacylase and a carboxylate kinase; claim 20 of GONZALEZ) converting said 2,3-dihydroxyacyl-CoA to a 2,3-dihydroxyacid.
Regarding claim 42-44 and 48, GONZALEZ teaches the microorganism of claim 1, and further teaches that the externally supplied energy source is a reduced 1-carbon substrate (i.e., methanol; single carbon, externally supplied energy source), which allows for a high growth rate via methanol oxidation, by producing ATP for growth in E.coli metabolism (implying the presence of nucleic acids encoding enzymes needed for this metabolism [paragraph 00169]). GONZALEZ teaches that the third set of metabolic enzymes comprises methanol dehydrogenase converting methanol to formaldehyde (methanol to formaldehyde is a methanol dehydrogenase; claim 3), a formaldehyde dehydrogenase converting formaldehyde to formate (acylating aldehyde dehydrogenase (formaldehyde dehydrogenase} converts formaldehyde to formyl-CoA; Table 1A), and a formate dehydrogenase converting formate to CO2 (GONZALEZ shows formate dehydrogenase converting CO2 to formate but it is a reversible enzyme and converts formate to CO2; Fig. 3).
Regarding claim 47, GONZALEZ teaches the microorganism of claims 42-44 and further teaches that the third set of metabolic enzymes further comprises an acylating formaldehyde dehydrogenase converting formaldehyde to formyl-CoA (formaldehyde to formyl-CoA using acylating aldehyde dehydrogenase; Fig. 3), a phosphate formyl transferase converting formyl-CoA to formyl-phosphate (a phosphate formyl transferase is a reversible enzyme therefore it can be used as in GONZALEZ to produce formyl-CoA from formyl-phosphate as well as the reverse to form formyl-phosphate from formyl-CoA, which is also evidenced by the instant application, in Fig. 1; Fig. 3; paragraph [0088]), and a formate kinase converting formyl-phosphate to formate (formate kinase is used in GONZALEZ to form formyl-phosphate from formate, wherein formate kinase is a reversible enzyme such that the opposite reaction is also possible, which is also evidenced in the instant application as such in Fig. 5; claim 7; Table 1A).
GONZALEZ does not explicitly teach every single step and element of all of the instant claims. However, such compositions and methods practiced with them would have been prima facie obvious to a person having ordinary skilled in the art at the time the application was filed for the following reasons.
LI teaches the next-generation metabolic engineering of non-conventional microbial cell factories for carboxylic acid platform chemicals, and teaches that carboxylic acids contain carboxyl groups that can undergo a wide range of chemical transformation. Therefore, they serve as key platform chemicals for the production of high value-added industrial products. The rapid development in metabolic engineering of microbes, specifically Escherichia coli and Saccharomyces cerevisiae, provide a new and promising alternative route for producing carboxylic acids as platform chemicals (entire document; see Title, Abstract, for example).
LI teaches that, in the past decade, rapid progress in metabolic engineering of microbes has enabled sustainable production of a wide array of bioproducts such as biopharmaceuticals, biofuels and biomaterials (page 1, right-hand col., last paragraph). In Table 1 on page 2, LI provides examples of carboxylic acids that have been commercialized.
LI teaches the metabolic engineering for producing short-chain fatty acids (section 3.1; Fig. 1 and accompanying text). In Table 2 on pages 6-8, LI provides representative examples of carboxylic acid platform chemicals produced through systems metabolic engineering. LI teaches the metabolic engineering for producing hydroxy carboxylic acids (section 3.2.)
In Figure 2 on page 10, LI provides a schematic diagram summarizing metabolic pathways and key enzymes for production of hydroxy carboxylic acids in microbes. LI teaches the metabolic engineering for producing dicarboxylic fatty acids (section 3.3.; Figure 3 and accompanying text).
LIU teaches the metabolic engineering of biocatalysts for carboxylic acids production, and teaches the fermentation of renewable feedstocks by microbes to produce sustainable fuels and chemicals, which has the potential to replace petrochemical-based production. For example, carboxylic acids produced by microbial fermentation can be used as platform chemicals to generate primary building blocks of industrial chemicals by either enzymatic or chemical catalysis. Metabolic engineering in the form of overexpression of key pathway genes, as well as deletion of competing pathways, has proved quite effective for improving carboxylic acid production (entire document; see Title, Abstract, for example).
LIU teaches the metabolic engineering by genetic manipulations (section I). In Table 1 and Figure 2, LIU teaches the metabolic pathways for production of lactate, malate and succinate in E. coli.
LIU teaches omics analysis, which can provide the global information from disturbed metabolism and find the potential target genes for problem solving (section 2).
LIU also teaches the engineering of tolerance to product toxicity (section 3).
LIU teaches the combination of directed evolution and genetic engineering. While genetic manipulation and metabolic evolution are both useful on their own, these tools become especially powerful when used together (section 4).
VILA-SANTA teaches the prospecting biochemical pathways to implement microbe-based production of the new-to-nature platform chemical levulinic acid (LA), one of the carboxylic acids consistently identified as having high potential as a platform molecule with application in the production of polycarbonates and plastics, renewable jet fuels, epoxy resins, herbicides, pharmaceuticals, or flavoring agents. Resorting to a combined approach involving complementary computational tools and extensive manual curation, VILA-SANTA provides a thorough prospect of candidate biosynthetic pathways that can be assembled for the production of levulinic acid in Escherichia coli (i.e., bacterium) or in Saccharomyces cerevisiae (i.e., yeast). Not only does the described approach offer a platform for the future implementation of the microbial production of levulinic acid, but also it provides an organized research strategy that can be used as a framework for the implementation of other new-to-nature biosynthetic pathways for the production of value-added chemicals, thus fostering the emerging field of synthetic industrial microbiotechnology (entire document; see Title, Abstract, for example).
VILA-SANTA teaches that recent advances in the field of synthetic biology have rendered possible the production of a panoply of molecules not naturally produced by microbes resorting to the assembly of “new-to-nature” pathways. Paradigmatic examples of these synthetic pathways include the implementation of odd-chain fatty alcohols production in Saccharomyces cerevisiae or the production of 2,4-dihydroxybutyric acid and chiral organoboranes in Escherichia coli (page 725, left-hand col., last paragraph).
VILA-SANTA teaches that S. cerevisiae and E. coli were selected as hosts due to their central role as experimental models and microbial chassis in industrial biotechnology, specifically for the production of carboxylic acids. Altogether, the study provides not only an integrated view of the best performing pathways to produce LA from renewable sources but also a framework for analyzing pathway design strategies that can be implemented for the microbe-based production of value-added compounds that fall outside the natural microbial metabolic repertoire (page 726, left-hand col., second paragraph).
In Figure 3, VILA-SANTA depicts a comparison of the precursors for LA synthesis identified by the three different approaches used in this study. In this figure are shown the
results obtained by the different searches that were performed to identify possible substrates for LA biosynthesis including search for structural similar substrates, the use of ReactPred and the use of MINE. In Table 1 on page 728, VILA-SANTA describes predicted new-to-nature pathways for LA biosynthesis using the candidate precursors, provides a list of precursors, pathway descriptions, predicted reaction rules, and EC numbers of the enzymes assigned. See also Figure 4 for selected LA biosynthetic pathways. See also section 3 on page 731 for the analysis of in vivo implementation of the candidate pathways.
In Figure 6, VILA-SANTA provides an o Outline of the steps taken for the selection of the five assembled pathways. This schematic representation details the different steps that were taken to assemble and rank the five possible LA biosynthetic pathways that were examined, including also the strategies that had to be used for the filtering of the results obtained in the different steps. See also the Materials and Methods section, as well as the associated content/supplemental materials.
Accordingly, before the effective filing date of the claimed invention, it would have been prima facie obvious to a person of ordinary skill in the art to combine and to modify the teachings of GONZALEZ, LI, LIU, and VILA-SANTA, to provide for new-to-nature carbon and energy conversion pathways that facilitate the synthesis of fuels and chemicals from carboxylic acid compounds and intermediates driven by genetically altered microorganisms; thus arriving at the Applicant’s invention, with a reasonable expectation of success, and without any surprising results. Obviously, one of ordinary skill in the art would have been motivated to do so for the purpose of engineering microbes that contain a designed platform for the conversion of one-carbon substrates to chemical products (as taught by GONZALEZ, LI, LIU, and VILA-SANTA). As evidenced above, the instant application parallels the combined teachings of GONZALEZ, LI, LIU, and VILA-SANTA.
It would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to have modified the microorganism as disclosed by GONZALEZ and the other references to incorporate enzymes generating ATP from NADH as the electron transport chain does this in vivo indirectly to form ATP from NADH.
It would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to have modified the microorganism as disclosed by GONZALEZ and the other references to show the reversible reaction of formate to CO2, which the formate dehydrogenase disclosed in GONZALEZ is capable of.
It would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to have shown the reactions from a phosphate formyl transferase converting formyl-CoA to formyl-phosphate and a formate kinase converting formyl-phosphate to formate, as both of these enzymes are reversible.
It would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to have modified the microorganism as taught by GONZALEZ and the other references to have condensed a 2-hydroxyaldehyde with a formyl-CoA to form a 2,3-dihyroxyacyl-CoA as the addition of the formyl-CoA to the previous aldehyde would make a molecule that is a single carbon longer that said 2-hydroxyaldyde, while retaining the previous hydroxyl now at the 3 position thereby changing the molecule to 2,3-dihydroxyacyl-CoA to complete the cycle of single carbon addition.
It would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to have modified utilized the same enzymes as disclosed by GONZALEZ and the other references to convert said 2,3-dihydroxyacyl- CoA to a 2,3-dihydroxyacid as the function of these enzymes remain the same and would catalyze the conversion of an acyl-CoA to a carboxylic acid.
Even though GONZALEZ, LI, LIU, and VILA-SANTA may not explicitly teach every single recited step and elements of all of the instant claims, these are considered design choice and mere routine optimization of the taught compositions and methods, which would be obvious to a person having ordinary skill in the art. They are limitations that are taught and/or suggested by the cited references and by the state of the art of biosynthesis of products from 1-carbon compounds, as they are well known as desirable for the biological production of multi-carbon compounds directly from single carbon compounds without the need for central metabolism, by using engineered microorganisms that can make various chemicals for industrial use.
Summary
No claim is allowed.
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BRATISLAV STANKOVIC, JD, PhD
Supervisory Patent Examiner
Art Units 1661 & 1662
/BRATISLAV STANKOVIC/SPE, Art Units 1661 & 1662
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