DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 02/23/2026 has been entered.
Priority
This application is a 371 of PCT/IN2021/050558 filed 06/09/2021. Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged.
Acknowledgment is made of applicant’s claim for foreign priority under 35 U.S.C. 119 (a)-(d) based on IN20201 1024273 filed 06/09/2020. Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Status of the Claims
Claims 1, 2, 5 and 8-10 are pending. Claims 1, 5 and 8 are amended. Claims 3, 4, 6 and 7 are cancelled. Claims 9 and 10 were withdrawn.
Claims 1, 2, 5 and 8 (claim set filed 01/26/2026) are examined on the merits herein.
Withdrawal of Rejections
The response and amendment filed on 01/26/2026 are acknowledged. All of the amendment and arguments have been thoroughly reviewed and considered.
For the purposes of clarity of the record, the reasons for the Examiner's withdrawal and/or maintaining if applicable, of the substantive or essential claim rejections are detailed directly below and/or in the Examiner's response to arguments section.
The previous claim 1 objection has been withdrawn necessitated by amendment of claim 1.
New Rejections
Claim Rejections - 35 USC § 112
The following is a quotation of the first paragraph 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 the first paragraph of pre-AIA 35 U.S.C. 112:
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.
Claims 1, 2, 5 and 8 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for synthesis of 40 g/lt of poly-gamma-glutamic acid (PGA) by incubating a raw material broth comprising tomato waste with Bacillus paralicheniformis MCC 0196 and isolation of PGA, does not reasonably provide enablement for synthesis of more than 40 g/lt and up-to 300 g/lt of PGA. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims.
Claims were analyzed based on Wands factors (MPEP 2164.01(a)):
(B) The nature of the invention: Claim 1 is directed to a process of synthesis of PGA by incubating a raw material comprising tomato waste with Bacillus paralicheniformis MCC 0196 and isolation of PGA by centrifugation of broth and treatment with ice-cold methanol and the process provides yield of 40-300 g/lt.
The breadth of the claims: claim 1 is broad. Although claim 1 specifies the Bacillus strain
used, its amount and conditions of fermentation, i.e. time, temperature and pH and requires tomato waste as a raw material, however the amount of tomato waste and its preparation are not limited and additional components are not excluded from the broth due to transitional phrase “comprising”. Besides, isolation of PGA is described broadly.
(C) The state of the prior art; (E) The level of predictability in the art: The prior art of Hsueh (Hsueh et al. Int. J. Molec. Sci., 2017, 18, 2644, 1-20 on record in IDS) teaches synthesis of poly-gamma-glutamic acid (PGA) by bacteria including Bacillus species, mechanisms of synthesis and gene regulation, applications of PGA and genetic modifications of bacterial strains to produce high level of PGA (Abstract). Hsueh describes conditions of fermentation providing variable yields with the yields covering the claimed limitation. For instance, B. licheniformis NCIM 2324 was shown to provide maximal yield of 98.64 g/l (p. 14, 1st paragraph). Tables 2 and 3 provide fermentation parameters and components of the fermentation medium including natural sources such as starch, yeast extract and molasses. However, Hsueh does not teach tomato waste as raw material. Hijosa-Valsero (Hijosa-Valsero et al. BioEnergy Research, 2019, 12, 1000-1011) teaches application of tomato waste for fermentation for production of ethanol by bacterial strains (Abstract) and describes presence of saccharides as carbon source, proteins as nitrogen source as well as potassium, magnesium, calcium and iron necessary for fermentation (p. 1001, right column, 1st paragraph). However, Hijosa-Valsero does not describe synthesis of PGA. Therefore, the prior art cannot predict the yield of PGA produced with a raw material comprising tomato waste.
(D) The level of one of ordinary skill: Working in this art are highly skilled.
(F) The amount of direction provided by the inventor; (G) The existence of working examples; (H) The quantity of experimentation needed to make or use the invention based on the content of the disclosure: The specification provides three working examples of PGA synthesis with tomato waste as a raw material, i.e. Examples 10-12 from which fermentation in Example 10 was performed in the presence of salts, “ammonium nitrate 0.6%; K2HPO4 1%; MgSO4 0.5%; CaCl2 0.2%; FeCl3 0.03% (p. 13, Example 10). The yields achieved were: 40 g/lt in Examples 10 and 12 and 39 g/lt in Example 11. The higher yields of production were achieved with sucrose as carbon source in the fermentation medium: 198 g/L were obtained in Example 17 for PGA production in a liter fermenter in the presence of 200-2300 g/L of sucrose (p. 16) and 284 g/L yield was reached during non-sterile fermentation (p. 17) in the presence of 300-600 g/L of sucrose. However, these fermentations did not comprise tomato waste as a raw material. Therefore, one of ordinary skill in the art would have to undergo undue experimentation to achieve the claimed yield of 40-300 g/lt when using raw material comprising tomato waste.
Based on the unpredictability taught by the prior art and absence of working examples and directions provided by inventors, one of ordinary skill in the art would have to undergo undue experimentation to practice the full scope of the invention. Therefore, claim 1 is rejected under 35 U.S.C. 112(a) for failing to disclose sufficient supporting information to enable a person of skill in the art to synthesize more than 40 g/lt and up-to 300 g/lt of PGA by incubating a raw material broth comprising tomato waste with Bacillus paralicheniformis MCC 0196 and isolation of PGA.
Claims 2 and 5, dependent on claim 1, do not resolve the issue mentioned above and are rejected.
Claim 8, dependent on claim 1, in addition to not resolving issue mentioned for claim 1 above, recites concentration of salts not supported by working example. However, since the specification provides examples showing that in the absence of the recited salts the fermentation of tomato waste results in the same amount of PGA, i.e. 40 g/lt, (Example 10) compared to that in the presence of salts (Example 11), claim 8 is interpreted as enabling production of 40 g/lt but not higher than that. Therefore, claim 8 is rejected under 35 U.S.C. 112(a) for failing to disclose sufficient supporting information to enable a person of skill in the art to synthesize more than 40 g/lt and up-to 300 g/lt of PGA by incubating a raw material broth comprising tomato waste with Bacillus paralicheniformis MCC 0196 and isolation of PGA.
Maintained/Modified Rejections
The following rejections are maintained and/or modified taking into consideration amendment
to claims filed on 01/26/2026.
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.
Claims 1 and 5 are rejected under 35 U.S.C. 103 as being unpatentable over Hsueh (Hsueh et al. Int. J. Molec. Sci., 2017, 18, 2644, 1-20 on record in IDS) in view of Fang (Fang et al. J. Cleaner Production, 2020, 255, 120248, 1-11), Kreyenschulte (Kreyenschulte et al. Crit. Review Biotechnol., 2014, 34, 1-15) and Hijosa-Valsero (Hijosa-Valsero et al. BioEnergy Research, 2019, 12, 1000-1011).
Regarding claim 1, Hsueh teaches synthesis of poly-gamma-glutamic acid (PGA) by bacteria including Bacillus species, mechanisms of synthesis and gene regulation, applications of PGA and genetic modifications of bacterial strains to produce high level of PGA (Abstract). Hsueh describes that PGA is produced mainly by gram-positive bacteria belonging to the genus Bacillus, including B. subtilis, and B. licheniformis (p. 2, last paragraph). Hsueh mentions the molecular weight of PGA between 10 and 1000 kDa (p. 1, 1st paragraph). Hsueh describes genes involved in PDA synthesis and performs phylogenetic analysis based on these genes. Hsueh discloses the conservation of the following genes: “conservation of genes decreased in the following order: pgsC > pgdS > pgsE > pgsA > pgsB” and mentions that: “pgsC is important for γ-PGA synthesis, and thus, it is difficult to modify or replace it, but pgsB, pgsA, and pgsE. can vary or be replaced without affecting γ -PGA synthesis” (p. 7, last paragraph). Hsueh describes that B. paralicheniformis has 4 of these genes (pgsC, pgsA, pgsB, pgdS) indicating that B. paralicheniformis strains can synthesize PGA (p. 9, Figure 8). Hsueh describes multiple recombinant strains of Bacillus genus generated for increased PGA production (p. 12-13, Table 3) with the yields covering the claimed limitation for yield. For instance, B. licheniformis NCIM 2324 was shown to provide maximal yield of 98.64 g/l and B. subtilis NX-2 – 107.7 g/l (Table 3). Tables 2 and 3 provide ingredients of the fermentation medium such as glucose or sucrose as carbon source, glutamic acid, citric acid and ammonium sulfate, ammonium chloride as nitrogen sources and natural sources such as starch, yeast extract and molasses and various salt ingredients. Hsueh describes fermentation parameters in Table 3, including temperature and pH, corresponding to claimed limitations.
Hsueh does not teach a raw material broth comprising tomato waste for PGA synthesis, does not teach Bacillus paralicheniformis MCC 0196 strain, does not teach time of fermentation, amount of bacteria and does not teach obtaining PGA from the broth by centrifugation and ice cold methanol treatment.
Fang teaches production of PGA by Bacillus amyloliquefaciens JX-6 using agricultural waste as substrates (Abstract). Fang describes using corn stalk and soybean meal as solid substrates supplemented with monosodium glutamate. The fermentation is performed at 33-37°C (p. 3, left column, 2nd paragraph). Fang shows the time course of PGA production under sterilized and non-sterilized conditions on Figure 2 (p. 4). As can be seen the highest production, up-to 150 g/kg, can be reached at 36-48 hours of fermentation that covers claim limitation for fermentation time. Fang determined the amount of viable bacteria during fermentation. Figure 6 (p. 8) shows that the amount of viable bacteria during fermentation under sterilized condition is higher than 1x109 CFU/g for most of fermentation time reaching 12-14x109 CFU/g at 72h. That covers claim 1 limitation for the amount of bacteria. Fang teaches centrifugation of the fermentation mixture at 12,000 x g for 20 min to separate cells and collect supernatant and precipitation of PGA with cold ethanol (p. 3, left column, 4th paragraph). Feng compares the level of produced PGA during fermentation of Bacillus strains using agro-industrial residues in Table 3 (p. 10) and shows highest level reached in the described study, i.e. 112.82 g/kg.
Kreyenschulte teaches different alcohols for PGA precipitation. Kreyenschulte discloses that alcohol induced precipitation is the most common method of PGA recovery and that cold methanol, ethanol, 2-propanol were reported to be used (p. 7, left column, 2nd paragraph). Kreyenschulte describes that: “The addition of alcohol reduces the water activity leading to a precipitation of the polymer molecules, which can then be separated from the supernatant…” (p. 7, left column, 2nd paragraph). Therefore, ethanol in Fang teaching can be substituted with methanol as taught by Kreyenschulte.
Hijosa-Valsero teaches application of tomato waste to bioethanol production by twelve different yeast and bacterial strains (Abstract). Hijosa-Valsero describes composition of tomato pomace, a solid waste generated during processing of tomatoes to obtain tomato juice, paste, sauce, puree or ketchup (p. 1001, right column, 1st paragraph). Hijosa-Valsero discloses presence of carbon source such as starch, cellulose, simple sugars and nitrogen source, such as proteins in tomato waste: “The chemical composition of tomato pomace is variable depending on the sample analyzed, but it is normally in the range of 10–18% starch, 27–32% cellulose, 5–18% hemicellulose, 11–26% simple sugars, 7.6% pectin, 31% lignin, 12– 23% protein, 5–20% fat and 4–6% ash” (p. 1001, right column, 1st paragraph). Hijosa-Valsero mentions presence in tomato waste of potassium, magnesium, calcium and iron: “Tomato waste contains important amounts of Ca, K (~ 7–11 g/kg), Mg, Na and P (~ 2–3 g/kg), and lower amounts of Fe, Mn and Cu (15–30 mg/kg)” (p. 1001, right column, 1st paragraph). Therefore, tomato waste contains all the necessary ingredients for bacterial fermentation and described by Hsueh for PGA production and hence can be used for synthesis of PGA by fermentation.
First, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to combine teachings of Hsueh and Fang and use conditions for fermentation during synthesis of PGA from Feng teaching, including fermentation time and amount of bacteria used, and method of recovery of PGA by alcohol (ethanol) precipitation from Feng teaching and use Bacillus strains, including strain of Bacillus paralicheniformis, as described in Hsueh teaching. One would have been motivated to do so since Hsueh teaches multiple Bacillus species producing PGA and strains genetically modified for enhanced production and Fang describes fermentation providing 112.82 g/kg yield of PGA. A skilled artisan would have reasonably expected success in the combination because Hsueh and Feng teach production of PGA by Bacillus strains.
Second, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Kreyenschulte teaching and use cold methanol for precipitation of PGA produced based on Hsueh and Feng teachings. One would have been motivated to do so with reasonably expected success since Kreyenschulte teaches functional equivalence of ethanol and methanol for PGA precipitation.
Third, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use tomato waste as a raw material as described by Hijosa-Valsero for the synthesis of PGA based on Hsueh, Fang and Kreyenschulte teachings. One would have been motivated to do so since tomato waste contains the necessary ingredients for synthesis of PGA as taught by Hsueh, Fang teaches production of PGA using agricultural waste and Hijosa-Valsero showed application of tomato waste for bioethanol production. A skilled artisan would have reasonably expected success in that because Hsueh and Fang provide method of production of PGA by Bacillus strains and Hijosa-Valsero provides source of fermentation medium containing necessary nutrients.
Last, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention that instant Bacillus paralicheniformis MCC 0196 is the same or an obvious variant of Bacillus paralicheniformis strains of Hsueh teaching that can perform the same function of PGA synthesis. One would have been motivated to use Bacillus paralicheniformis strains for PGA synthesis with reasonably expected success since Hsueh showed presence of several genes necessary for PGA synthesis in the genome of Bacillus paralicheniformis strains. Therefore, absent evidence to the contrary, if the strain is not the same, the use of an obvious variant of Bacillus paralicheniformis with the same function and used for the same purpose is obvious.
Thus, Hsueh, Fang, Kreyenschulte and Hijosa-Valsero teachings render claim 1 obvious.
Regarding claims 5, Hsueh teaches fermentation broth ingredients and sources of potassium (KH2PO4), magnesium (MgSO4), calcium (CaCl2) and iron (FeCl3) (Table 2). Hijosa-Valsero mentions presence in tomato waste of potassium, magnesium, calcium and iron (p. 1001, right column, 1st paragraph). Thus, Hsueh, Feng, Kreyenschulte and Hijosa-Valsero teachings render claim 5 obvious.
Claim 2 is rejected under 35 U.S.C. 103 as being unpatentable over Hsueh (Hsueh et al. Int. J. Molec. Sci., 2017, 18, 2644, 1-20 on record in IDS) in view of Fang (Fang et al. J. Cleaner Production, 2020, 255, 120248, 1-11), Kreyenschulte (Kreyenschulte et al. Crit. Review Biotechnol., 2014, 34, 1-15) and Hijosa-Valsero (Hijosa-Valsero et al. BioEnergy Research, 2019, 12, 1000-1011) as applied to claim 1 above, and further in view of Lopez (Lopez and Alippi J. Microbiol. Methods, 2019, 165, 105690, 1-11).
Teachings of Hsueh, Fang, Kreyenschulte and Hijosa-Valsero have been set forth above.
Hsueh, Fang, Kreyenschulte and Hijosa-Valsero do not teach B. paralicheniformis isolated from honey.
Lopez teaches multiple aerobic spore-forming bacteria isolated from honey with most of them belonging to Bacillus genus (p. 1, 2nd paragraph). Lopez mentions that some bacterial groups are closely related and discloses a method for rapid differentiation of isolates based on PCR-amplified 16S rRNA gene and restriction analysis (Abstract). Lopez describes differentiation between 80 isolates belonging to 26 species reported in honey and other apiarian sources (Abstract).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Lopez teaching and use Bacillus strains from honey in the process of PGA synthesis based on Hsueh, Fang, Kreyenschulte and Hijosa-Valsero teachings. One would have been motivated to do so since Hsueh teaches multiple Bacillus species producing PGA and Lopez provides method for identification of Bacillus species from honey and reports differentiation of 80 isolates from honey and other apiarian sources. A skilled artisan would have reasonably expected success in the combination because Hsueh, Feng, and Lopaz teach Bacillus species and Hsueh and Feng teach production of PGA by Bacillus strains and Lopaz provides honey as the source for novel Bacillus strains. Thus, Hsueh, Fang, Kreyenschulte, Hijosa-Valsero and Lopez teachings render claim 2 obvious.
Claim 8 is rejected under 35 U.S.C. 103 as being unpatentable over Hsueh (Hsueh et al. Int. J. Molec. Sci., 2017, 18, 2644, 1-20 on record in IDS) in view of Fang (Fang et al. J. Cleaner Production, 2020, 255, 120248, 1-11), Kreyenschulte (Kreyenschulte et al. Crit. Review Biotechnol., 2014, 34, 1-15) and Hijosa-Valsero (Hijosa-Valsero et al. BioEnergy Research, 2019, 12, 1000-1011) as applied to claim 1 above and further in view of Ju (Ju et al. Biotechnol. Lett., 2014, 36, 2319-2324).
Teachings of Hsueh, Fang, Kreyenschulte and Hijosa-Valsero have been set forth above.
Regarding claim 8, Hsueh teaches different conditions for PGA production (Table 3), including pH 7.5 and temperature of fermentation as low as 30°C which is close to claimed temperature of 28°C and Hsueh teaches the recited salt ingredients although not mentioning their concentrations (Table 3). Hijosa-Valsero describes presence of potassium, magnesium, calcium and iron in tomato waste (p. 1001, right column, 1st paragraph). Feng teaches significant production level reached at 36 hours of fermentation at sterilized and non-sterilized conditions (p. 8, Figure 6). However, Hsueh, Fang, Kreyenschulte and Hijosa-Valsero do not teach the recited concentrations of salts in the fermentation.
Regarding claim 8, Ju teaches enhanced production of PGA by Bacillus subtilis MJ80 strain isolated from soil samples (Abstract). Ju describes optimization of fermentation conditions and reaching productivity of 75.5 g/l of PGA in 3 days (Abstract). Ju discloses the optimized fermentation media to contain 20 g/l starch and 80 g/l glycerol as carbon sources; 70 g/l glutamic acid; 7 g/l urea as nitrogen source and 12 g/l citric acid (p. 2322 right column, last paragraph). Ju describes that the optimized composition of the fermentation media contains 0.5% NH4Cl, 0.05% K2PO4, 0.025% MgSO4x7H2O, 0.015% CaCl2 and 0.004% FeCl3 (p. 2322 right column, last paragraph) that corresponds to 5 g/l NH4Cl, 0.5g/l K2PO4, 0.25g/l MgSO4x7H2O, 0.15 g/l CaCl2 and 0.04 g/l FeCl3. Thus, Ju teaching contains the claimed salt ingredients concentrations of which are close to claimed parameters. It is noted that "where the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation" (See MPEP 2144.05 II).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to follow Ju and Hsueh teachings and add salt ingredients at specified or optimized concentrations to the fermentation broth comprising tomato waste for PGA production based on Hsueh, Fang, Kreyenschulte and Hijosa-Valsero teachings. One would have been motivated to do so since Hsueh discloses presence of salt ingredients during production of PGA by various Bacillus strains and Ju describes optimized conditions including salt concentrations for fermentation providing 75.5 g/l yield of PGA. A skilled artisan would have reasonably expected success in the combination because Hsueh, Feng and Ju teach production of PGA by Bacillus strains.
It would have been obvious to one of ordinary skill in the art that parameters of fermentation and concentrations of fermentation broth ingredients can be optimized. One would have been motivated to optimize the concentrations of ingredients and temperature of fermentation to achieve higher yield of PGA. A skilled artisan would have reasonably expected success in this optimization because selection of the conditions of fermentation is routine and conventional. Thus, Hsueh, Fang, Kreyenschulte, Hijosa-Valsero and Ju teachings render claim 8 obvious.
Response to Arguments
Applicant's arguments filed 1/26/2026 have been fully considered but they are not persuasive.
Applicant argues (addressing p. 4-7 of the Remarks) that claim 1 presents unexpected results exemplified in Example 11 and summarized in Table 1 showing that “incubating 6x108 to 6x109 CFU/ml of Bacillus paralicheniformis MCC 0196 in a tomato waste produces a remarkably high yield of PGA”. Applicant continues that Table 1 provides the highest γ-PGA production obtained to date in a batch fermentation process (40 g/L) that is much higher as compared to yield obtained with other waste products of Table 1. Applicant further argues that the person having ordinary skill in the art would have had no basis to believe that using tomato waste in the synthesis of PGA would result in much higher yields of PGA as compared to other natural or synthetic nutrient media. These arguments are not persuasive because:
The primary prior art of Hsueh teaches the yields γ-PGA production by different Bacillus strains and using fermentation medium with natural or synthetic ingredients that exceed 40 g/l (p. 12- 13, Table 3). For instance, for batch fermentation B. licheniformis NCIM 2324 was shown to provide maximal yield of 98.64 g/l in the presence of glutamic acid and citric acid (p. 14, 1st paragraph) and B. subtilis NX-2 provides 107.7 g/l of γ-PGA with fermentation of natural substrates (Table 3). Fang compares γ-PGA production yields for fermentation using agricultural waste and provides examples of yields exceeding 100 g/kg such as 112.82 g/kg when using corn stack and soybean meal waste (Table 3, p. 10).
Hijosa-Valsero teaches that tomato waste contains nutrients providing carbon source such as starch, cellulose, simple sugars and nitrogen source, such as proteins in tomato waste (p. 1001, right column, 1st paragraph). Additionally, Hijosa-Valsero mentions presence in tomato waste of potassium, magnesium, calcium and iron (p. 1001, right column, 1st paragraph). Besides nutrients mentioned by Hijosa-Valsero, tomatoes contain glutamic acid and citric acid as evidenced by Missio (Missio et al. Scientia Agricola, 2015, 72, 314-321). Missio teaches chemical changes in sugar-acid profile of tomatoes during storage and describes presence of glutamic acid and citric acid in tomatoes (Abstract, Figure 3). Therefore, tomato waste contains all the necessary nutrients as taught by Hsueh (Table 3), i.e. carbon source, nitrogen source, glutamate, trace elements, and Hijosa-Valsero showed application of tomato waste for bioproduction (Abstract) providing motivation to use tomato waste as raw material for production of PGA by Bacillus strains expecting to reach the same or higher yield than reported by Hsueh.
Therefore, the 40 g/l yield of PGA produced using tomato waste as raw material is not an unexpected result.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to LIOUBOV G KOROTCHKINA whose telephone number is (571)270-0911. The examiner can normally be reached Monday-Friday: 8:00-5:30.
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/L.G.K./Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653
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