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 .
Claim Status
Claims 1-4 and 6-10 are pending (claim set as filed on 08/10/2023). Claims 1-4 and 6-10 are currently under examination and were examined on their merits.
Priority
This application filed on 08/10/2023 claims priority to PCT application no. PCT/EP2022/053368, filed on 02/11/2022, and claims foreign priority to application no. EP 21156553.6, filed on 02/11/2021. Acknowledgment is made of applicant’s claim for foreign priority under 35 U.S.C. 119 (a)-(d). Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
The Information Disclosure Statement (IDS) filed on 08/10/2023 has been received and considered.
Specification
The abstract of the disclosure is objected to because the sentence “[t]he exogenous nucleic acid sequences encode a permease for lactose import and the GMD, WcaG and FucT are chromosomally integrated” (lines 4-6) lacks clarity since proteins, in this case GMD, WcaG and FucT (see line 5), cannot be chromosomally integrated.
A corrected abstract of the disclosure is required and must be presented on a separate sheet, apart from any other text. See MPEP § 608.01(b).
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.
Claims 8 and 10 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 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 8 recites “wherein the permease for lactose import is selected from a group including lactose permease (LacY)), GDP-D-mannose-4,6-dehydratase (GMD), GDP-L-fucose synthase (WcaG), and fucosyltransferase (FucT).”.
The Examiner notes that the instant specification does not disclose wherein GDP-D-mannose-4,6-dehydratase (GMD), GDP-L-fucose synthase (WcaG), and fucosyltransferase (FucT) have permease function, but that GDP-D-mannose-4,6-dehydratase (GMD) converts GDP-mannose to GDP-4-dehydro-6-deoxy-D-mannose (see specification, paragraphs [0213]-[0215] and Fig. 1), GDP-L-fucose synthase (WcaG) converts GDP-4-dehydro-6-deoxy-D-mannose to GDP-L-fucose (see specification, paragraphs [0217]-[0219] and Fig. 1), that fucosyltransferase (FucT) transfers fucose to an acceptor molecule, such as lactose, which, in case of lactose as acceptor molecule, leads to the production of fucosyllactose (see specification, paragraphs [0204]-[0205] and Fig. 1). One of ordinary skill in the art would not be able to determine the metes and bounds of claim 8 and thus, could not clearly determine how to avoid infringement of the claim. In the interest of compact prosecution, the Examiner interprets claim 8 to the broadest embodiment claimed.
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.
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 factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
Determining the scope and contents of the prior art.
Ascertaining the differences between the prior art and the claims at issue.
Resolving the level of ordinary skill in the pertinent art.
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.
Claims 1-4 and 7-8 are rejected under 35 U.S.C. 103 as being unpatentable over Seo et al. (US 2020/0048640 A1, published on 02/13/2020), hereinafter ‘Seo’, in view of Roenneke et al. (“Production of the compatible solute α‑d‑glucosylglycerol by metabolically engineered Corynebacterium glutamicum”, published on 06/16/2018, Microb Cell Fact, Vol. 17, article number 94, pages 1-14), hereinafter ‘Roenneke’.
Seo’s general disclosure relates to a method of producing 2’-fucosyllactose including culturing in a medium supplemented with lactose a recombinant Corynebacterium glutamicum” (see entire document, including abstract).
Regarding claim 1, pertaining to ‘genetically modified or engineered’, it is noted that the instant specification describes that “"genetically modified" describes bacteria, whose genetic material has been modified in comparison to a naturally occurring wild type strain, for example by deleting or removing genetic elements of the wild type strain and/or by inserting additional genetic rnaterial, in particular DNA sequences, for example in form of nonintegrating DNA plasmids or as DNA sequences that chromosomally integrate into the bacterial genome (specification, paragraph [0198]).
Pertaining to “glycosyltransferases involved in corynebacterial cell wall
biosynthesis”, the phrase ‘involved in corynebacterial cell wall biosynthesis’ is not clearly defined within the claim or the specification. The claimed glycosyltransferases could be directly or indirectly involved in corynebacterial cell wall biosynthesis, and as such, any glycosyltransferase in corynebacterium could be involved in corynebacterial cell wall biosynthesis.
Pertaining to a genetically modified or engineered corynebacterium for
production of fucosyllactose, Seo teaches a genetically modified corynebacterium for production of fucosyllactose (“recombinant Corynebacterium glutamicum (C. glutamicum) which is transformed to express a-L2-fucosyltransferase, GDP-D-mannose-4,6-dehydratase (Gmd), GDP-L-fucose synthase (GDP-4-keto-6-deoxymannose-3,5-epimerase-4-reductase, WcaG) and lactose permease (LacY), and to overexpress phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC), and a method for producing fucosyllactose using the same. (paragraphs [0002], [0027]; see Fig. 1).
Regarding claim 3, pertaining to the corynebacterium, Seo teaches wherein the corynebacterium is corynebacterium glutamicum (“a recombinant Corynebacterium glutamicum”; see abstract).
Regarding claims 4 and 8, please note the 112b rejection of claim 8 above.
Pertaining to ‘exogenous nucleic acid’, the instant specification states that “”exogenous nucleic acid” or "exogenous genetic element" relates to any nucleic acid or nucleic acid sequence introduced into the bacterial cell, which is not a component of the cells "original" or "natural" genome.”, and that “[e]xogenous nucleic acids may be integrated or non-integrated” (specification, paragraph [0239]).
Pertaining to a permease for lactose import, Seo teaches wherein the corynebacterium has been modified to express a permease for lactose import from exogenous nucleic acid sequences (“recombinant Corynebacterium glutamicum (C. glutamicum) which is transformed to express … lactose permease (LacY)”, “lactose permease (Lac Y) is an enzyme involved in transporting lactose present outside the strain to the inside thereof”, “genes encoding … and lactose permease (Lac Y) are preferably derived from Escherichia coli”, “effects of incorporation of lacZ-removed lac operon (lac YA) on production of 2'-fucosyllactose in Corynebacterium glutamicum”; paragraphs [0002], [0017], [0019], and [0028]; see Fig. 1) (instant claim 4), wherein the permease for lactose import is lactose permease (LacY) (““recombinant Corynebacterium glutamicum (C. glutamicum) which is transformed to express … lactose permease (LacY)”, “; paragraph [0002], see Fig. 1) (instant claim 8).
Regarding claim 7, pertaining to wherein the corynebacterium exports fucosyllactose, it is noted that the specification states that “export.of FL relates to any kind of process, active or passive, that leads to FL being present outside the bacteria in the culture medium.” (specification, paragraph [0070]).
Pertaining to wherein the corynebacterium exports fucosyllactose, Seo teaches wherein the corynebacterium exports fucosyllactose (see Fig. 1; note the arrow indicating export of 2-FL (2'-fucosyllactose) to the outside of the cell).
Additionally, Seo teaches that, “regarding the method for producing 2'-fucosyllactose according to the present invention, the medium preferably further includes glucose.”, and that “[b]y adding glucose to the medium, growth of strain can be facilitated and 2'-fucosyllactose can thus be produced at a higher productivity.” (paragraphs [0024]).
Seo does not teach wherein the corynebacterium is defective for functional expression of one or more glycosyltransferases involved in corynebacterial cell wall biosynthesis (instant claim 1), wherein the one or more glycosyltransferases is selected from the group consisting of cgp_3313 (MrcB; GT51), cgp_0336 (PonA; GT51); cgp_3166 (GT4); cgp_2400 (GT4); cgp_1876 (GT4); cgp_1268 (GlgA; GT4); cgp_0554 (GT4); cgp_3191 (GlfT; GT2); cgp_1672 (PpmC; GT2); cgp_1180 (GT2); cgp_0848 (WbbL; GT2); cgp_0730 (GT2); cgp_0396 (GT2); cgp_0394 (GT2); cgp_0246 (GT2); cgp_0163 (GT2); cgp_2393 (GT87,GT87); cgp_2390 (GT87); cgp_2389 (GT87);
cgp_2385 (GT87); and cgp_3164 (instant claim 2).
Roenneke’s general disclosure relates to “genetic engineering of C. glutamicum for the production of αGG.” (see entire document, including page 2, right column, paragraph 3; note, “α-d-Glucosylglycerol (αGG)”, see abstract).
Regarding claims 1 and 2, pertaining to a genetically modified Corynebacterium, Roenneke teaches a corynebacterium defective for functional expression of a glycosyltransferases involved in corynebacterial cell wall biosynthesis (“C. glutamicum strains … IMglgA … C. glutamicum WT with inactivation of glgA (cg1268)”, “Inactivation of the chromosomal glgA gene (orf cg1268) in C. glutamicum WT”; page 4, right column, paragraph 2; see Table 1 on page 3; note that the instant specification describes GlgA as a glycosyltransferase involved in corynebacterial cell wall biosynthesis (see specification, paragraph [0118]) (instant claim 1), wherein the glycosyltransferase is GlgA (“C. glutamicum WT with inactivation of glgA (cg1268)”; see Table 1 on page 3) (instant claim 2). The Examiner notes that inactivation of glgA inherently teaches that GlgA cannot be functionally expressed.
Additionally, Roenneke teaches that “[C. glutamicum] transiently accumulates in the course of cultivations with sugars as substrate large amounts of glycogen.”(page 2, right column, paragraph 3), and that “C. glutamicum IMglgA did not accumulate glycogen during cultivation in minimal medium with either 1% (w/v) glucose
or 1% (w/v) sucrose as substrate (page 6, right column, paragraph 1 - page 7, left column, paragraph 1). Roenneke further discloses wherein “[r]edirection of carbon flux towards αGG synthesis by elimination of the competing pathways for glycogen and trehalose synthesis as well … is an efficient strategy to further optimize production of αGG”(see abstract).
While Seo does not teach wherein the corynebacterium is defective for functional expression of one or more glycosyltransferases involved in corynebacterial cell wall biosynthesis (instant claim 1), wherein the one or more glycosyltransferases is selected from the group consisting of cgp_3313 (MrcB; GT51), cgp_0336 (PonA; GT51); cgp_3166 (GT4); cgp_2400 (GT4); cgp_1876 (GT4); cgp_1268 (GlgA; GT4); cgp_0554 (GT4); cgp_3191 (GlfT; GT2); cgp_1672 (PpmC; GT2); cgp_1180 (GT2); cgp_0848 (WbbL; GT2); cgp_0730 (GT2); cgp_0396 (GT2); cgp_0394 (GT2); cgp_0246 (GT2); cgp_0163 (GT2); cgp_2393 (GT87,GT87); cgp_2390 (GT87); cgp_2389 (GT87);
cgp_2385 (GT87); and cgp_3164 (instant claim 2), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined Seo’s teachings with Roenneke’s teachings on shifting carbon flux by inactivating glgA, in order to have created a genetically modified corynebacterium for production of fucosyllactose, wherein the corynebacterium is defective for functional expression of a glycosyltransferase involved in corynebacterial cell wall biosynthesis, wherein the glycosyltransferase is cgp_1268 (GlgA). One would have been motivated to do so, in order to develop a superior genetically modified corynebacterium for increased production of fucosyllactose by shifting the carbon flux from glycogen synthesis towards GDP-L-fucose production. A skilled artisan would have reasonably expected success in combining Seo’s and Roenneke’s teachings since both references are directed to Corynebacterium glutamicum (see above).
Claims 1, 3-4, and 7-8 are rejected under 35 U.S.C. 103 as being unpatentable over Seo et al. (US 2020/0048640 A1, published on 02/13/2020), hereinafter ‘Seo’, in view of Wang et al. (“Eliminating the capsule-like layer to promote glucose uptake for hyaluronan production by engineered Corynebacterium glutamicum”, published on 06/19/2020, Nature Communications, Vol. 11, Article number: 3120, pages 1-10), hereinafter ‘Wang’.
Seo’s general disclosure relates to a method of producing 2’-fucosyllactose including culturing in a medium supplemented with lactose a recombinant Corynebacterium glutamicum” (see entire document, including abstract).
Regarding claim 1, pertaining to ‘genetically modified or engineered’, it is noted that the instant specification describes that “"genetically modified" describes bacteria, whose genetic material has been modified in comparison to a naturally occurring wild type strain, for example by deleting or removing genetic elements of the wild type strain and/or by inserting additional genetic rnaterial, in particular DNA sequences, for example in form of nonintegrating DNA plasmids or as DNA sequences that chromosomally integrate into the bacterial genome (specification, paragraph [0198]).
Pertaining to “glycosyltransferases involved in corynebacterial cell wall
biosynthesis”, the phrase ‘involved in corynebacterial cell wall biosynthesis’ is not clearly defined within the claim or the specification. The claimed glycosyltransferases could be directly or indirectly involved in corynebacterial cell wall biosynthesis, and as such, any glycosyltransferase in corynebacterium could be involved in corynebacterial cell wall biosynthesis.
Pertaining to a genetically modified or engineered corynebacterium for
production of fucosyllactose, Seo teaches a genetically modified corynebacterium for production of fucosyllactose (“recombinant Corynebacterium glutamicum (C. glutamicum) which is transformed to express a-L2-fucosyltransferase, GDP-D-mannose-4,6-dehydratase (Gmd), GDP-L-fucose synthase (GDP-4-keto-6-deoxymannose-3,5-epimerase-4-reductase, WcaG) and lactose permease (LacY), and to overexpress phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC), and a method for producing fucosyllactose using the same. (paragraphs [0002], [0027]; see Fig. 1).
Regarding claim 3, pertaining to the corynebacterium, Seo teaches wherein the corynebacterium is corynebacterium glutamicum (“a recombinant Corynebacterium glutamicum”; see abstract).
Regarding claims 4 and 8, please note the 112b rejection of claim 8.
Pertaining to ‘exogenous nucleic acid’, the instant specification states that “”exogenous nucleic acid” or "exogenous genetic element" relates to any nucleic acid or nucleic acid sequence introduced into the bacterial cell, which is not a component of the cells "original" or "natural" genome.”, and that “[e]xogenous nucleic acids may be integrated or non-integrated” (specification, paragraph [0239]).
Pertaining to a permease for lactose import, Seo teaches wherein the corynebacterium has been modified to express a permease for lactose import from exogenous nucleic acid sequences (“recombinant Corynebacterium glutamicum (C. glutamicum) which is transformed to express … lactose permease (LacY)”, “lactose permease (Lac Y) is an enzyme involved in transporting lactose present outside the strain to the inside thereof”, “genes encoding … and lactose permease (Lac Y) are preferably derived from Escherichia coli”, “effects of incorporation of lacZ-removed lac operon (lac YA) on production of 2'-fucosyllactose in Corynebacterium glutamicum”; paragraphs [0002], [0017], [0019], and [0028]; see Fig. 1) (instant claim 4), wherein the permease for lactose import is lactose permease (LacY) (““recombinant Corynebacterium glutamicum (C. glutamicum) which is transformed to express … lactose permease (LacY)”, “; paragraph [0002], see Fig. 1) (instant claim 8).
Regarding claim 7, pertaining to wherein the corynebacterium exports fucosyllactose, it is noted that the specification states that “export.of FL relates to any kind of process, active or passive, that leads to FL being present outside the bacteria in the culture medium.” (specification, paragraph [0070]).
Pertaining to wherein the corynebacterium exports fucosyllactose, Seo teaches wherein the corynebacterium exports fucosyllactose (see Fig. 1; note, the arrow indicates export of 2-FL (2'-fucosyllactose) to the outside of the cell).
Additionally, Seo teaches that, “regarding the method for producing 2'-fucosyllactose according to the present invention, the medium preferably further includes glucose”, and that “[b]y adding glucose to the medium, growth of strain can be facilitated and 2'-fucosyllactose can thus be produced at a higher productivity.” (paragraphs [0024]).
Seo does not teach wherein the corynebacterium is defective for functional expression of one or more glycosyltransferases involved in corynebacterial cell wall biosynthesis (instant claim 1).
Wang’s general disclosure relates to engineering of “C. glutamicum for high-yield HA production by selecting the most productive HA synthase, overexpressing enzymes of the intermediate pathways to convert glucose into the HA building blocks UDP-GlcA and UDPGlcNAc and decreasing endogenous extracellular polysaccharide biosynthesis” (see entire document, including page 2, left column, paragraph 4; note, HA, hyaluronic acid; page 2, left column, paragraph 1).
Regarding claim 1, pertaining to a genetically modified Corynebacterium, Wang teaches a corynebacterium defective for functional expression of one or more glycosyltransferases involved in corynebacterial cell wall biosynthesis (“we speculated that cg0420 may participate in biosynthesis of C. glutamicum extracellular polysaccharides. Prior to cg0420 deletion, we found three other putative glycosyltransferase genes within the same genomic region (cg0424, cg0419, and cg0438) (Fig. 2b and Supplementary Fig. 5). ... However, we were able to delete
cg0424 to generate strain Delcg0424. Basing on Delcg0424, we further deleted cg0420 and constructed strain Delcg0420,0424.”, “Sugar constituent analysis of the Delcg0424 extracellular polysaccharides showed that the concentration of mannose decreased by 32% while the concentration of arabinose was not affected (Fig. 2d). The concentrations of mannose and arabinose released from strain Delcg0420,0424 extracellular polysaccharides hydrolysis decreased by 24% and 47% respectively, comparing to that of the strain Delcg0424 (Fig. 2d)”; see page 3, left column, paragraph 1 - page 3, right column paragraph 1; see Fig. 2d).
Additionally, Wang teaches that “loss of cg0424 and cg0420 reduced cell surface
polysaccharides such as mannan and arabinomannan (Fig. 2d) and enhanced HA production (Table 1) without impairing cell growth (Fig. 2c)” (page 7, right column, paragraph 2; see Fig. 2d), wherein HA is hyaluronic acid (“Hyaluronan (hyaluronic acid or HA)”; page 2, left column, paragraph 1; see Table 1). Wang further discloses: “Hyaluronan is widely used in cosmetics and pharmaceutics. Development of robust and safe cell factories and cultivation approaches to efficiently produce hyaluronan is of many interests.” (see abstract).
While Seo does not teach wherein the corynebacterium is defective for functional expression of one or more glycosyltransferases involved in corynebacterial cell wall biosynthesis (instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined Seo’s teachings with Wang’s teachings on increased production of a product of interest in the presence of inactivated glycosyltransferases, in order to have created a genetically modified corynebacterium for production of fucosyllactose, wherein the corynebacterium is defective for functional expression of one or more glycosyltransferases involved in corynebacterial cell wall biosynthesis (instant claim 1). One would have been motivated to do so to develop a superior genetically modified corynebacterium with increased production of fucosyllactose. A skilled artisan would have reasonably expected success in combining Seo’s and Wang’s teachings since both references are directed to Corynebacterium glutamicum (see above).
Claims 1, 4, and 6 are rejected under 35 U.S.C. 103 as being unpatentable over Seo et al. (US 2020/0048640 A1, published on 02/13/2020), hereinafter ‘Seo’, in view of Wang et al. (“Eliminating the capsule-like layer to promote glucose uptake for hyaluronan production by engineered Corynebacterium glutamicum”, published on 06/19/2020, Nature Communications, Vol. 11, Article number: 3120, pages 1-10), hereinafter ‘Wang’, and Li et al. (“One-pot synthesis of GDP-L-fucose by a four-enzyme cascade expressed in Lactococcus lactis”, published on 10/16/2017, Journal of Biotechnology, Vol. 264, pages 1-7), hereinafter ‘Li’.
Seo’s and Wang’s teachings have been set forth above.
Regarding claim 6, pertaining to ManB and ManC, Seo teaches wherein the genetically modified corynebacterium according to claim 4 additionally comprises nucleic acids for overexpression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) (“Corynebacterium glutamicum possesses genes encoding phosphomannomutase (ManB) and GTP-mannose-1-phosphate
guanylyltransferase (ManC), and can thus express the same. Therefore, there may be no need to incorporate genes encoding the enzymes, but the enzymes should be overexpressed for mass-production. For this reason, the present invention requires transformation of Corynebacterium glutamicum in order to overexpress the two enzymes.”; paragraph [0018]).
Modified Seo does not teach wherein the corynebacterium additionally comprises exogenous nucleic acids for expression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) (instant claim 6).
Li’s general disclosure relates to the use of “Lactococcus lactis subsp. cremoris NZ9000 to express 4 enzymes, ManB, ManC, Gmd, and WcaG”, and to producing “GDP-L-fucose by using one-pot synthesis method with mannose-6-phosphate as substrate and the enzymes as biocatalyst” (see entire document, including abstract).
Regarding claim 6, pertaining to ‘exogenous nucleic acid’, the instant specification states that “”exogenous nucleic acid” or "exogenous genetic element" relates to any nucleic acid or nucleic acid sequence introduced into the bacterial cell, which is not a component of the cells "original" or "natural" genome.”, and that “[e]xogenous nucleic acids may be integrated or non-integrated” (specification, paragraph [0239]).
Pertaining to exogenous sequences for the expression of ManB and ManC, Li teaches E. coli derived phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) (“ManB, phosphomannomutase; ManC, mannose 1-phosphate guanylyltransferase”, “4 genes (manB, manC, gmd, and wcaG) cloned from Escherichia coli were transformed into L. lactis strains”; see abstract and Fig. 1; it is noted that the instant specification states that GTP-mannose-1-phosphate guanylyltransferase is also called mannose 1-phosphate guanylyltransferase (specification, paragraph [0226]).
Additionally, Li teaches wherein ManB and ManC are involved in the production of GDP-L-fucose (“When mannose-6-phosphate was used as the substrate, the consecutive reactions with ManB, ManC, Gmd, and WcaG resulted in the successful production of GDP-L-fucose”; see abstract).
While modified Seo does not teach wherein the corynebacterium additionally comprises exogenous nucleic acids for expression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) (instant claim 6), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention, to have combined modified Seo’s corynebacterium with Li’s teachings on E. coli derived ManB and ManC, in order to create a genetically modified corynebacterium, wherein the corynebacterium comprises exogenous E. coli derived nucleic acids for expression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC). One would have been motivated to do so to develop a superior genetically modified corynebacterium for improved production of fucosyllactose. A skilled artisan would have reasonably expected success in combining modifed Seo’s and Li’s teachings since both references are directed to using ManB and ManC for the production of GDP-L-fucose (see above).
Claims 1, 4, 8, and 10 are rejected under 35 U.S.C. 103 as being unpatentable over Seo et al. (US 2020/0048640 A1, published on 02/13/2020), hereinafter ‘Seo’, in view of Wang et al. (“Eliminating the capsule-like layer to promote glucose uptake for hyaluronan production by engineered Corynebacterium glutamicum”, published on 06/19/2020, Nature Communications, Vol. 11, Article number: 3120, pages 1-10), hereinafter ‘Wang’, and Sasaki et al. (“Simultaneous utilization of D-cellobiose, D-glucose, and D-xylose by recombinant Corynebacterium glutamicum under oxygen-deprived conditions”, published on 09/23/2008, Appl Microbiol Biotechnol, Vol. 81, pages 691–699), hereinafter ‘Sasaki’.
Seo’s and Wang’s teachings have been set forth above.
Modified Seo does not teach wherein the exogenous nucleic acid sequences for expression of LacY, GMD, WcaG, or FucT are chromosomally integrated (instant claim 10).
Sasaki’s general disclosure relates to Corynebacterium glutamicum R, that was “metabolically engineered to broaden its sugar utilization range to D-xylose and D-cellobiose contained in lignocellulose hydrolysates” (see entire document, including abstract).
Regarding claim 10, please note the 112b rejection above.
Pertaining to chromosomal integration, Sasaki teaches wherein metabolic genes “were integrated into the chromosome of C. glutamicum without a selection marker because foreign genes on a chromosome are more stable than those integrated via plasmids.” (page 692, left column, paragraph 3).
While modified Seo does not teach wherein the exogenous nucleic acid sequences for expression of LacY, GMD, WcaG, or FucT are chromosomally integrated (instant claim 10), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined modifed Seo’s corynebacterium with Sasaki’s teachings on chromosomally integrating metabolic genes, to have created a genetically modified corynebacterium, wherein the exogenous nucleic acid sequences for expression of LacY are chromosomally integrated. One would have been motivated to do so to ensure the stability of the exogenous nucleic acids for expression of LacY in the corynebacterium (see Sasaki above). A skilled artisan would have reasonably expected success in combining modified Seo’s and Sasaki’s teachings, since both are directed to Corynebacterium glutamicum (see above).
Claims 1, 4, 6, and 9 are rejected under 35 U.S.C. 103 as being unpatentable over Seo et al. (US 2020/0048640 A1, published on 02/13/2020), hereinafter ‘Seo’, in view of Wang et al. (“Eliminating the capsule-like layer to promote glucose uptake for hyaluronan production by engineered Corynebacterium glutamicum”, published on 06/19/2020, Nature Communications, Vol. 11, Article number: 3120, pages 1-10), hereinafter ‘Wang’, Li et al. (“One-pot synthesis of GDP-L-fucose by a four-enzyme cascade expressed in Lactococcus lactis”, published on 10/16/2017, Journal of Biotechnology, Vol. 264, pages 1-7), hereinafter ‘Li’, and Sasaki et al. (“Simultaneous utilization of D-cellobiose, D-glucose, and D-xylose by recombinant Corynebacterium glutamicum under oxygen-deprived conditions”, published on 09/23/2008, Appl Microbiol Biotechnol, Vol. 81, pages 691–699), hereinafter ‘Sasaki’.
Seo’s, Wang’s and Li’s teachings have been set forth above.
Modified Seo does not teach wherein the exogenous nucleic acids for expression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) are chromosomally integrated (instant claim 9).
Sasaki’s general disclosure relates to Corynebacterium glutamicum R, that was “metabolically engineered to broaden its sugar utilization range to D-xylose and D-cellobiose contained in lignocellulose hydrolysates” (see entire document, including abstract).
Regarding claim 9, pertaining to chromosomal integration, Sasaki teaches wherein metabolic genes “were integrated into the chromosome of C. glutamicum without a selection marker because foreign genes on a chromosome are more stable than those integrated via plasmids.” (page 692, left column, paragraph 3).
While modified Seo does not teach wherein the exogenous nucleic acids for expression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) are chromosomally integrated (instant claim 9), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined modified Seo’s corynebacterium with Sasaki’s teachings on chromosomally integrating metabolic genes, to have created a genetically modified corynebacterium, wherein the exogenous nucleic acids for expression of phosphomannomutase (ManB) and GTP-mannose-1-phosphate guanylyltransferase (ManC) are chromosomally integrated. One would have been motivated to do so to ensure the stability of the exogenous nucleic acids for expression of ManB and ManC in the corynebacterium (see Sasaki above). A skilled artisan would have reasonably expected success in combining modified Seo’s and Sasaki’s teachings, since both are directed to Corynebacterium glutamicum (see above).
Conclusion
No claims are allowed.
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/SANDRA ZINGARELLI/Examiner, Art Unit 1653
/SHARMILA G LANDAU/ Supervisory Patent Examiner, Art Unit 1653