Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114.
Applicant's submission filed on 11/25/2025 has been entered.
DETAILED ACTION
Claims 2-3, 6-7 and 10-20 are canceled. Claims 1, 4-5, and 8-9 are pending and under consideration in this office action.
Priority
The instant claims are entitled to an effective filing date of 12/08/2020.
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.
(Maintained) Claims 1, 4-5 and 8-9 are rejected under 35 U.S.C. 103 as being unpatentable over Gaddy (U.S. Patent No. 7,285,402 B2, as provided in the IDS filed 12/02/2021), in view of Phillips (Applied biochemistry and biotechnology, 1993, 39, 559-571) and Phillips (Applied biochemistry and biotechnology, 1994, 45, 145-157; hereafter Phillips1994).
Regarding claim 1, Gaddy teaches a continuous method for producing ethanol from the anaerobic bacterial fermentation of a gaseous substrate comprising carbon monoxide, the method comprises: culturing in a fermentation bioreactor anaerobic, acetogenic C. ljungdahlii bacteria in a continuously fed liquid nutrient medium to provide a fermentation broth and supplying to said fermentation bioreactor said gaseous substrate comprising carbon monoxide, such as carbon monoxide and hydrogen. See claims 1 and 6 of Gaddy. In example 6, Gaddy teaches fermenting C. ljungdahlii in a continuous stirred tank reactor (CSTR) (fermenter) with a feed gas containing CO. See column 22 lines 62-66. Gaddy teaches feeding a liquid medium containing excess salts, vitamins and trace metals to the reactor. See column 23 lines 1-2. The vitamin solution contains 50.5 mg/L pantothenic acid (i.e. vitamin B5), 20.6 mg/L d-biotin (i.e. vitamin B7), and 50.6 mg/L thiamine HCl (i.e. vitamin B1). See table 1. Therefore, Gaddy implies that the vitamin solution contains vitamin B5 in an amount that is 2.45 times that of vitamin B7 and 0.998 times that of vitamin B1 (i.e. nearly equivalent). In analyzing parameters for ethanol production, Gaddy discloses that the ratio of the pantothenate (vitamin B5) fed to the cell production is 97 µg pantothenate/g-cell produced. Gaddy suggests that this level is sufficiently high to assure that pantothenate is not limiting. See column 23 lines 13-16. Under these conditions, the cell concentration is 2.7 g/L and the ethanol productivity is 29 g/L·day, which equates to an ethanol productivity rate of 10.7 g/day/gram cells. See column 23 lines 7-12 and table 2. To clarify the calculation of the ethanol productivity rate, 29 g/L·day ÷2.7g/L cell concentration is 10.7 g/day/gram cells.
In summation, Gaddy teaches a fermentation process comprising: providing a CO-containing gaseous substrate to a fermenter that includes a fermentation broth; providing vitamin B1, B5, and B7 to the fermentation broth, wherein a feed rate of vitamin B5 is 97 µg/g-cell produced, which is within the instantly claimed range of 25 to about 150 ug/g cell produced. Gaddy also teaches fermenting the CO-containing gaseous substrate with C. ljungdahlii, an acetogenic bacteria, wherein the process provides a specific ethanol productivity rate of 10.7 g/day/gram cells, which is more than 8g/day/gram cells.
Gaddy does not teach providing an amount of vitamin B5 at a feed rate that is at least 4 times a feed rate of vitamin B7 (biotin), and providing an amount of vitamin B5 at a feed rate that is at least 3.5 times a feed rate of vitamin B1 (thiamine).
Phillips indicates that reducing the concentration of B-vitamins in a Clostridium ljungdahlii fermentation medium can increase the ethanol-to acetate product ratio. See the second paragraph on page 563. Furthermore, Philips discloses that the relative amounts of ethanol and acetate can be controlled by nutritional factors. See the second paragraph on page 570.
Phillips1994 teaches growing Clostridium ljungdahlii on H2 or CO for the production of ethanol. See the abstract and figure 6. Phillips1994 teaches a CSTR (i.e. continuous stirred tank reactor) medium that includes 0.155 mg/L Ca-D-pantothenate (vitamin B5), 0.040 mg/L biotin (vitamin B7) and 0.01 mg/L thiamine-HCl (vitamin B1). See table 3. Therefore, Phillips1994 implies that the CSTR medium includes vitamin B5 in an amount that is 3.875 times the amount of vitamin B7 and 15.5 times the amount of vitamin B1. Moreover, Phillips1994 teaches a basal defined medium comprising 0.083 mg/L Ca-D-pantothenate, 0.033 mg/L biotin, and 0.083 mg/L thiamine-HCl. See table 3. Thus, Phillips1994 implies that basal defined medium includes a vitamin B5 in an amount that is 2.515 times the amount of vitamin B7 and equivalent to the amount of vitamin B1.
It would have been obvious to a person of ordinary skill in the art prior to the effective filing date of the instantly claimed invention to adjust the vitamin B7 and vitamin B1 feed rates of Gaddy in view of Phillips and the B-vitamin proportions in the CSTR medium of Phillips1994. Doing so is mere optimization through routine experimentation. One would be motivated to optimize the vitamin feed rates of Gaddy because Phillips suggests that reducing the concentration of B-vitamins can increase the ethanol to acetate ratio. There would be a reasonable expectation of success because Gaddy demonstrates feeding, to a CSTR reactor, a vitamin solution containing vitamin B5 in an amount that is 2.45 times that of vitamin B7 and 0.998 times that of vitamin B1 (nearly equivalent); Phillips1994 teaches a basal defined medium with similar B-vitamin proportions compared to the vitamin solution of Gaddy because the basal defined medium of Phillips1994 contains vitamin B5 in an amount that is 2.515 times that of vitamin B7 and equivalent to that of vitamin B1; yet, Phillips1994 also teaches a CSTR medium where vitamin B5 is an amount that is 3.875 times that of vitamin B7 and 15.5 times that of vitamin B1. Therefore, one could reasonably adjust the B-vitamin proportions in the vitamin solution of Gaddy in view of the CSTR medium of Phillips1994. MPEP 2144.05(II) states that “[w]here 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." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955).
Regarding claim 4, Gaddy teaches acetogenic bacteria including Clostridium ljungdahlii. See column 6 lines 31 and 38-39.
Regarding claim 5, Gaddy teaches feeding a gaseous substrate that contains 57% H2 and 36% CO to the CSTR fermentor, which is a 1.5 molar ratio of H2 to CO. See column 22 lines 65-66. Thus, Gaddy teaches a H2/CO molar ratio that is about 0.2 or more.
Regarding claim 8, Gaddy teaches an ethanol production medium that does not contain yeast extract. See table 1 in column 35. Gaddy also teaches using a rich medium containing 1 g/L yeast extract as an alternative to a nutrient medium containing vitamin B5 or calcium pantothenate. See column 17 line 66-66 and column 26 line 65.
Phillips suggests that the reduction of the B-vitamin concentration and elimination of yeast extract diminishes growth potential slightly but causes a significant increase in the ethanol-to-acetate product ratio. See the second paragraph on page 563.
It would have been obvious prior to the effective filing date of the instantly claimed invention to use the ethanol production medium of Gaddy and in the process to arrive at a fermentation broth with 0.01 g/L or less yeast extract. One would be motivated to do so because Phillips suggests that eliminating yeast extract may contribute to an increase in the ethanol-to-acetate product ratio. There would have been a reasonable expectation of success because Gaddy suggested that yeast extract was not necessary.
Regarding claim 9, Gaddy teaches bacterial growth media without sugar. See column 6 line 61.
It would have been obvious prior to the effective filing date of the instantly claimed invention to optimize the amount of carbohydrates in the sugarless fermentation media of Gaddy, and in the process to have arrived a fermentation broth with 0.01 g/L or less carbohydrates as instantly claimed. There would be a reasonable expectation of success because Gaddy suggested that the sugarless medium contained vitamins and minerals sufficient to permit growth. See column 6 lines 58-61.
Response to Arguments
Applicant's arguments filed 11/25/2025 have been fully considered but they are unpersuasive.
§103 rejection of claims 1, 4-5 and 8-9 over Gaddy in view of Phillips and Phillips1994:
Applicant argues that the inventive concept of the claimed process lies in economically increasing specific ethanol productivity with unexpected and surprisingly optimized B vitamin combinations to improve industry competitiveness (see paragraph [0006] of the specification). Increasing specific ethanol productivity increases production, and optimizing vitamin combinations reduces operation costs. See the first full paragraph and argument number 1 on page 4 of the remarks.
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To the extent that this argument represents an attempt to overcome the prior art rejections by reliance on unexpected results, it is unpersuasive. Paragraph [0006] in its entirety states the following:
Therefore, paragraph [0006] does not provide any evidence for unexpected and surprisingly optimized B vitamin combinations, as argued. MPEP 716.02(b)-(e) indicates that unexpected results must be established by factual evidence. Since Applicant has not provided any factual evidence, the argument is unpersuasive.
Applicant argues that the claimed process describes a feed rate, not a concentration. Applicant references the declaration of Senaratne for the feed rate calculation as being vitamin feed rate (µg/g cells produced)= vitamin flow rate (µg/hour)÷cell production rate (g/hour). Applicant asserts that cell density is not equivalent to cell production rate. The cell production rate is measured based on cell purge from the reactor consistent with industry practice. Applicant asserts that claim 1 requires the feed rate of B5 to be 25 to 150 µg per gram of cells produced. The feed rate of B5 is four times that of B7, so the B7 feed rate must be within 6.25 to 37.7 µg/g cells produced. The B5 feed rate is 3.5 times that of B1, so the B1 feed rate must be within 7.14 to 42.85 µg/g cells produced. See argument number 2 spanning pages 4-5 of the remarks.
This argument is not persuasive because the rejection above does not conflate concentrations and feed rates, and the argument is not commensurate in scope with the instant claims. The declaration of Senaratne is addressed separately below. Gaddy teaches a continuous method that includes continuous nutrient feed, substrate feed, cell production in the bioreactor, cell removal or purge from the bioreactor and product removal. A continuous process results in a steady state within the bioreactor, which means that all of the measurable variables (i.e. feed rates, substrate and nutrient concentrations maintained in the bioreactor, cell concentration in the bioreactor and cell removal from the bioreactor, product removal from the bioreactor as well as conditional variables) are constant over time. See column 5 lines 54-67 of Gaddy. Gaddy teaches feeding 97 µg pantothenate/g-cell produced. See column 23 lines 13-16. Absent evidence to the contrary, the “g-cell produced” of Gaddy is measured in a way consistent with industry practice. Gaddy teaches feeding vitamins to the reactor including pantothenic acid (i.e. vitamin B5), 20.6 mg/L d-biotin (i.e. vitamin B7), and 50.6 mg/L thiamine HCl (i.e. vitamin B1). See column 23 lines 1-2 and table 1 of Gaddy. Gaddy implies that the concentrations may be constant over time. Claim 1 requires the active method step of “providing vitamins B1, B5, and B7 to the fermentation broth”. The claims do not limit the time frame of that active method step in anyway. Therefore, the argument that the instantly claimed feed rates are required to be based on an hourly rate is not commensurate in scope with the instant claims, i.e. the argument that vitamin feed rate (µg/g cells produced)= vitamin flow rate (µg/hour)÷cell production rate (g/hour). Moreover, for clarity of the record, the rejection of claim 1 above references the vitamin B concentrations taught by Phillips1994, because of their implied vitamin B proportions.
Applicant argues that the experimental data in the application demonstrate unexpected improvements in specific ethanol productivity as a result of the optimized B vitamin feed rates. Experiment 1 and 2 show that increasing the feed rates of all three vitamins generally increase specific ethanol productivity. Experiment 3 shows that increasing only B5 feed rate from about 20 µg/g to about 108 µg/g cells produced while maintaining B1 and B7 below 20 µg/g results in approximately 42% increase in specific ethanol productivity (SEP). This demonstrates the unexpected result that increasing all three vitamins is not necessary to achieve higher productivity. Experiment 4 shows that increasing B1 and B7 while holding B5 below 30 µg/g does not increase SEP. Experiment 5 shows that increasing B5 from about 48 µg/g to about 82 µg/g, while maintaining B1 and B7 below 30 µg/g and even reducing them below 20 µg/g increases SEP by about 24%. This further confirms that B5 feed rate is a critical driver of ethanol productivity and that higher B1/B7 rates are unnecessary or even disadvantageous. Collectively, the experiments reveal an unexpected and non-linear relationship between vitamin feed rates and SEP. See page 5 of the remarks.
This argument is not persuasive because it is not commensurate in scope with the instant claims. Claim 1 encompasses one or more acetogenic bacteria that are selected from a genus of Clostridium. Experiments 1-4 teach Clostridium ljungdahlii and experiment 5 teaches Clostridium authoethanogenum. Experiments 1 and 2 are not commensurate in scope with the instant claims because the vitamin B5 feed rates disclosed in the experiments are not at least 3.5 times the feed rates of vitamin B1 as the vitamin B1 feed rates are all greater than that of the vitamin B5 feed rates. Regardless, every run disclosed in experiments 1 and 2 show an SEP of more than 8 g/day/gram cells [00059]-[0061]. Applicant argues that experiment 3 demonstrates unexpected results because it shows that increasing all three vitamins is not necessary to achieve higher productivity. However, in the argument Applicant references two runs in which the feed rates of all three B5, B7 and B1 feed are increased. To clarify, experiment 3 run (1) includes the following feed rates in the units of µg/g cells produced: vitamin B5 at 19.33, vitamin B7 at 17.76 and vitamin B1 at 17.36. The SEP of the run is 7.95 g/day/ g cells. Experiment 3 run (5) includes: vitamin B5 at 108.13, vitamin B7 at 19.87 and vitamin B1 at 14.95. The SEP of the run is 11.25 g/day/ g cells. See [0063]. Although the observed SEP of run (5) is higher than that observed for run (1), the results do not clearly show that it is unnecessary to increase all three vitamins, as argued by Applicant. Experiment 4 is not commensurate in scope with the instant claims because the vitamin B5 feed rates used in the experiment are not at least 4 times that of vitamin B7 and 3.5 times that of B1, and because SEP rates are below 8 g/day/grams cells or more. Moreover, Applicant argues that the results of experiment 5 confirm that the B5 feed rate is a critical driver of ethanol productivity. However, the fact that applicant has recognized another advantage which would flow naturally from following the suggestion of the prior art cannot be the basis for patentability when the differences would otherwise be obvious. Ex parte Obiaya, 227 USPQ 58, 60 (Bd. Pat. App. & Inter. 1985). In the instant case, Gaddy states that “[t]he proper pantothenate feed to the growth stage is a variable that must be optimized”. See column 31 lines 18-19. Thus, the arguments and evidence set forth in the remarks do not clearly establish unexpected results compared to the closest prior art of Gaddy.
Applicant argues that Gaddy does not teach or suggest the claimed vitamin feed rate or specific ratios between vitamins B5, B1 and B7. Applicant asserts that the 1575 to 3150 µg/g-cells produced Ca-d-pantothenate feed rate of Gaddy (table 2; column 20 lines 8-10) is much higher than the claimed range of 25-150 µg/g. Applicant asserts that Gaddy provides no teaching or suggestion of the specific feed rate ratios required. See the first four paragraphs on page 6 of the remarks.
This argument is not persuasive because MPEP 2123 states that “[a] reference may be relied upon for all that it would have reasonably suggested to one having ordinary skill the art, including nonpreferred embodiments”. Merck & Co. v. Biocraft Laboratories, 874 F.2d 804, 10 USPQ2d 1843 (Fed. Cir.), cert. denied, 493 U.S. 975 (1989). See also Upsher-Smith Labs. v. Pamlab, LLC, 412 F.3d 1319, 1323, 75 USPQ2d 1213, 1215 (Fed. Cir. 2005). In the instant case, Applicant is referencing example 3 of Gaddy, which does use a higher vitamin B5 range. However, example 3 of Gaddy is not relied upon in the rejection above. Rather, example 6 of Gaddy is relied upon because the example uses a pantothenate feed rate of 97 µg/g-cells produced, which is within the instantly claimed 25-150 µg/g-cells produced range. See column 23 lines 13-16 and table 2 of Gaddy.
Applicant argues that Phillips employs vitamin feed rates that are significantly higher than those required by the instant claims. Applicant asserts that Phillips teaches that reduced vitamin concentrations increased the ethanol-to-acetate ratio, consistent with the notion that excessive vitamin levels inhibit optimal ethanol productivity. Even after such reduction, Applicant asserts that the feed rates remain well above the claimed rages. See the bottom of page 6 and the top of page 7 of the remarks.
This argument is not persuasive because one cannot show non-obviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). Phillips is not relied upon for teaching the vitamin feed rates. Rather, Gaddy is relied upon for directly teaching the instantly claimed “µg/g cells produced unit”, and Phillips is relied upon for providing motivation to optimize the vitamin concentrations in the vitamin solution of Gaddy for ethanol production.
Applicant argues that Phillips1994 fails to provide anything on medium flow rate, cell concentration, purge rate and cell production rate. The only continuous process of Phillips1994 is the seed culture for the batch experiments. Applicant argues that the CSTR medium composition of Phillips1994 is for the seed inoculation continuous process. Applicant asserts that it is impossible to calculate how much vitamin B1, B5 and B7 is used during the continuous process of Phillips1994. As such, Phillips1994 does not provide any meaningful information on vitamin consumption and optimization. See the last paragraph on page 7 and the paragraph bridging pages 7-8 of the remarks.
This argument is not persuasive because Phillips1994 is not relied upon for teaching the instantly claimed vitamin feed rate; and the instant claims do not require a medium flow rate, cell concentration, purge rate or cell production rate. Applicant indicates that Phillips (1994) teaches away from the instant invention because Phillips (1994) teaches a batch culture and only a seed inoculation continuous process. However, the instant claims do not explicitly require the fermentation process to be a continuous process. Moreover, the test for obvious is not whether the features of a secondary reference may be bodily incorporated into the structure of the primary reference; nor is it that the claimed invention must be expressly suggested in any one or all of the references. Rather, the test is what the combined teachings of the references would have suggested to those of ordinary skill in the art. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981). In the instant case, the teachings of Phillips and Phillips (1994) suggest optimizing the vitamin ratios in the vitamin solution of Gaddy.
37 CFR 1.132 Declaration
The examiner acknowledges receipt of the Declaration under 37 CFR 1.132 by Dr. Ryan Senaratne filed on 11/25/2025.
The Declaration is insufficient to overcome the §103 rejection of claims 1, 4-5 and 8-9 based upon Gaddy in view of Phillips and Phillips (1994) as set forth in the last office action because: the arguments are not persuasive for reasons discussed above. The arguments spanning pages 2-6 of the Declaration are identical to the arguments set forth in the remarks spanning pages 4-8. The Declaration does not provide additional evidence of unexpected results. Rather, the Declaration, on page 3, references the same experiments of the instant disclosure that are discussed above.
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 USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The 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/process/file/efs/guidance/eTD-info-I.jsp.
(Maintain) Claims 1, 4-5, and 8-9 are rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1-11 of U.S. Patent No. 10,100,336 B2 to Scott et al. (hereinafter Scott ‘336) in view of Gaddy (US 7,285,402 B2), Phillips (Applied biochemistry and biotechnology, 39, 1993, 559-571) and Phillips (1994) (Applied biochemistry and biotechnology, 1994 45, 145-157).
Claim 1 of Scott ‘336 recites a fermentation process comprising fermenting syngas in a fermentation medium with acetogenic bacteria, the process effective for providing a specific STY of at least about 1 gram of ethanol/(L∙day∙gram cells),
wherein the fermentation medium includes
at least 112 to 125 mg of nitrogen per gram of cells,
at least 10.5 to 15 mg of phosphorous per gram of cells, and
at least 26 to 36 mg of potassium per gram of cells,
wherein the fermentation medium has less than 0.025 ppm boron, less than 0.0025 ppm manganese, less than 0.001 ppm molybdenum, and less than 0.01 ppm copper and wherein the fermentation medium has less than 0.01 g/L carbohydrates and less than 0.01 g/L yeast extract.
Claim 7 of Scott ‘336 recites the fermentation process of claim 1 wherein the syngas has a CO/CO2 ratio of at least about 0.75.
Claim 8 of Scott ‘336 recites the fermentation process of claim 1 wherein the acetogenic bacteria is selected from the group consisting of… Clostridium aceticum, Clostridium acetobutylicum, Clostridium autoethanogenum DSM 19630 of DSMZ Germany, Clostridium autoethanogenum DSM 10061 of DSMZ Germany, Clostridium autoethanogenum DSM 23693 of DSMZ Germany, Clostridium autoethanogenum DSM 24138 of DSMZ Germany, Clostridium carboxidivorans P7 ATCC PTA-7827, Clostridium coskatii ATCC PTA-10522, Clostridium drakei, Clostridium ljungdahlii PETC ATCC 49587, Clostridium ljungdahlii ERI2 ATCC 55380, Clostridium ljungdahlii C-01 ATCC 55988, Clostridium ljungdahlii O-52 ATCC 55889, Clostridium magnum, Clostridium pasteurianum DSM 525 of DSMZ Germany, Clostridium ragsdali P11 ATCC BAA-622, Clostridium scatologenes, Clostridium thermoaceticum, Clostridium ultunense, and mixtures thereof.
The patent claims of Scott ‘336 lack providing vitamin B1, B5, and B7 to a fermentation broth, wherein a feed rate of vitamin B5 is 25 to 150 ug/g cell produced, wherein an amount of vitamin B is provided at a feed rate of at least 4 times as feed rate of vitamin B7, and the amount of vitamin B5 is provided at a feed rate that is at least 3.5 times a feed rate of vitamin B1; and wherein the process provides a specific ethanol productivity rate of 8 g/day/gram cells or more (relevant to instant claim 1). The patent claims of Scott ‘336 lack CO-containing gaseous substrate has a H2/CO molar ratio of about 0.2 or more (relevant to instant claim 5).
However, Gaddy teaches providing a liquid medium containing vitamin B1, B5 and B7 to a fermenter containing broth, wherein the vitamin B5 feed rate is 97 µg/g-cell produced, and wherein the ethanol productivity is 10.7 g/day/gram cells produced. See Gaddy column 22 line 61 to column 23 line 15 for the fermentation process. Phillips indicates that reducing the concentration of b-vitamins in a Clostridium ljungdahlii fermentation medium can increase the ethanol-to acetate product ratio. See the second paragraph on page 563. Phillips (1994) teaches a CSTR (i.e. continuous stirred tank reactor) medium that includes 0.155 mg/L Ca-D-pantothenate, 0.040 mg/L biotin and 0.01 mg/L thiamine-HCl. See table 3. Therefore, Phillips (1994) implies that the CSTR medium includes vitamin B5 in an amount that is 3.875 times the amount of vitamin B7 and 15.5 times the amount of vitamin B1 (relevant to instant claim 1). Gaddy teaches feeding a gaseous substrate containing H2 and CO in a 1.5 molar ratio to a fermenter, see column 22, lines 65-66 (relevant to instant claim 5).
It would have been obvious to a person of ordinary skill in the art, prior to the effective filing date of the instantly claimed invention to modify the fermentation medium of Scott ‘336 by combining the medium with vitamins B1, B5 and B7 of Gaddy, by further optimizing the vitamin B feed rates of Gaddy based on the teachings of Phillips and Phillips (1994); and by finally optimizing molar ratio of H2 and CO in the syngas, as taught by Gaddy, in order to ferment acetogenic bacteria for ethanol production. There would be a reasonable expectation of success because Scott ‘336, Gaddy, Phillips and Phillips (1994) are all in the same field of producing ethanol by fermenting acetogenic bacteria.
(Maintain) Claims 1, 4-5, and 8-9 are rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1-5 of U.S. Patent No. 9,193,947 B2 to Senaratne et al. (hereinafter Senaratne ‘947) in view of Gaddy (US 7,285,402 B2), Phillips (Applied biochemistry and biotechnology, 39, 1993, 559-571) and Phillips (1994) (Applied biochemistry and biotechnology, 1994 45, 145-157).
Claim 1 of Senaratne ‘947 recites a process for culturing acetogenic bacteria on syngas, the process comprising: culturing the acetogenic bacteria on a first substrate to provide a cell density of at least about 0.005 g/L, wherein the first substrate includes a carbon source selected from the group consisting of yeast extract, carbohydrates, alcohol, amino acids, peptone, peptides, protein, fatty acids, lipid and mixtures thereof; sporulating the acetogenic bacteria by replacing at least a portion of the first substrate with syngas having at least about 10 mole % CO and replacing at least a portion of a first medium with a production medium to convert at least a portion of the bacteria to spores, wherein the sporulating provides a spore number to cell number ratio of about 0.05 or more; and germinating the spores in the production medium at a pH of about 4.1 to about 5 with syngas having at least about 10 mole % CO to provide a specific CO uptake of at least about 0.25 mmole/min/gram of cells and a space time yield (STY) of about 1 g or more ethanol/(L·day gram cells).
Claim 2 of Senaratne ‘947 recites the process of claim 1 wherein the syngas has a CO/CO2 molar ratio of at least about 0.75.
Claim 3 of Senaratne ‘947 recites the process of claim 1 wherein the acetogenic bacterium is selected from the group consisting of… Clostridium aceticum, Clostridium acetobutylicum, Clostridium acetobutylicum P262 (DSM 19630 of DSMZ Germany), Clostridium autoethanogenum (DSM 19630 of DSMZ Germany), Clostridium autoethanogenum (DSM 10061 of DSMZ Germany), Clostridium autoethanogenum (DSM 23693 of DSMZ Germany), Clostridium autoethanogenum (DSM 24138 of DSMZ Germany), Clostridium carboxidivorans P7 (ATCC PTA-7827), Clostridium coskatii (ATCC PTA-10522), Clostridium drakei, Clostridium ljungdahlii PETC (ATCC 49587), Clostridium ljungdahlii ERI2 (ATCC 55380), Clostridium ljungdahlii C-01 (ATCC 55988), Clostridium ljungdahlii O-52 (ATCC 55889), Clostridium magnum, Clostridium pasteurianum (DSM 525 of DSMZ Germany), Clostridium ragsdali P11 (ATCC BAA-622), Clostridium scatologenes, Clostridium thermoaceticum, Clostridium ultunense.
The patent claims of Senaratne ‘947 lack providing vitamin B1, B5, and B7 to a fermentation broth, wherein a feed rate of vitamin B5 is 25 to 150 ug/g cell produced, wherein an amount of vitamin B is provided at a feed rate of at least 4 times a feed rate of vitamin B7, and the amount of vitamin B5 is provided at a feed rate that is at least 3.5 times a feed rate of vitamin B1; and wherein the process provides a specific ethanol productivity rate of 8 g/day/gram cells or more (relevant to instant claim 1). The patent claims Senaratne ‘947 lack CO-containing gaseous substrate has a H2/CO molar ratio of about 0.2 or more (relevant to instant claim 5).
However, Gaddy teaches providing a liquid medium containing vitamin B1, B5 and B7 to a fermenter containing broth, wherein the vitamin B5 feed rate is 97 µg/g-cell produced, and wherein the ethanol productivity is 10.7 g/day/gram cells produced. See Gaddy column 22 line 61 to column 23 line 15 for the fermentation process. Phillips indicates that reducing the concentration of b-vitamins in a Clostridium ljungdahlii fermentation medium can increase the ethanol-to acetate product ratio. See the second paragraph on page 563. Phillips (1994) teaches a CSTR (i.e. continuous stirred tank reactor) medium that includes 0.155 mg/L Ca-D-pantothenate, 0.040 mg/L biotin and 0.01 mg/L thiamine-HCl. See table 3. Therefore, Phillips (1994) implies that the CSTR medium includes vitamin B5 in an amount that is 3.875 times the amount of vitamin B7 and 15.5 times the amount of vitamin B1 (relevant to instant claim 1). Gaddy teaches feeding a gaseous substrate that contains H2 and CO in a 1.5 molar ratio into a fermenter, see column 22, lines 65-66 (relevant to instant claim 5).
It would have been obvious to a person of ordinary skill in the art, prior to the effective filing date to modify the production medium of Senaratne ‘947 by combining the medium with vitamins B1, B5 and B7 of Gaddy, by further optimizing the vitamin B feed rates of Gaddy in view of Phillips and Phillips (1994); and by finally optimizing molar ratio of H2 and CO in the syngas, as taught by Gaddy, in order to ferment acetogenic bacteria for ethanol production. There would be a reasonable expectation of success because Senaratne ‘947, Gaddy, Phillips and Phillips (1994) are all in the same field of producing ethanol by fermenting acetogenic bacteria.
(Maintain) Claims 1, 4-5, and 8-9 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-10 of U.S. Patent No. 10,131,872 B2 to Senaratne et al. (hereinafter Senaratne ‘872) in view of Gaddy (US 7,285,402 B2), Phillips (Applied biochemistry and biotechnology, 39, 1993, 559-571) and Phillips (1994) (Applied biochemistry and biotechnology, 1994 45, 145-157).
Claim 1 of Senaratne ‘872 recites a process for fermenting a carbon monoxide (CO)-containing gaseous substrate comprising: growing an inoculum of acetogenic bacteria cells on a first medium that includes a first substrate to provide a cell density of about 0.02 g/L or more, wherein the first medium has a pH of 5.5 to 7.0 and the first substrate includes a carbon source that is selected from the group consisting of yeast extract, carbohydrates, alcohol, amino acids, peptone, peptides, protein, fatty acids, lipid and mixtures thereof, wherein the inoculum of acetogenic bacteria has no measurable consumption of a (CO)-containing gaseous substrate; sporulating at least a portion of the inoculum of bacteria by reducing an amount of the first substrate to provide bacterial spores; germinating the bacterial spores by replacing at least a portion of the first medium with a production medium having a pH of 4.1 to 5 and by replacing at least a portion of the first substrate with a CO-containing gaseous substrate, wherein the CO is a sole carbon source in the production medium; and supplying the CO-containing gaseous substrate to maintain a specific CO uptake of about 0.25 mmol per g cells or more; wherein the process of fermenting CO provides a specific STY (space time yield) of about 1 g of ethanol/(L day gram cells) or more.
Claim 2 of Senaratne ‘872 recites the process of claim 1 wherein the CO containing gaseous substrate is syngas.
Claim 3 of Senaratne ‘872 recites the process of claim 2 wherein the syngas has a CO/CO2 molar ratio of at least about 0.75.
Claim 4 of Senaratne ‘872 recites the process of claim 1 wherein the acetogenic bacteria is selected from the group consisting of…Clostridium aceticurn, Clostridium acetobutylicum, Clostridium acetobutylicum P262 (DSM 19630 of DSMZ Germany), Clostridium autoethanogenum (DSM 19630 of DSMZ Germany), Clostridium autoethanogenum (DSM 10061 of DSMZ Germany), Clostridium autoethanogenum (DSM 23693 of DSMZ Germany), Clostridium autoethanogenum (DSM 24138 of DSMZ Germany), Clostridium carboxidivorans P7 (ATCC PTA-7827), Clostridium coskatii (ATCC PTA-10522), Clostridium drakei, Clostridium ljungdahlii PETC (ATCC 49587), Clostridium ljungdahlii ERI2 (ATCC 55380), Clostridium ljungdahlii C-01 (ATCC 55988), Clostridium ljungdahlii O-52 (ATCC 55889), Clostridium magnum, Clostridium pasteurianum (DSM 525 of DSMZ Germany), Clostridium ragsdali P11 (ATCC BAA-622), Clostridium scatologenes, Clostridium thermoaceticum, Clostridium ultunense.
The patent claims of Senaratne ‘872 lack providing vitamin B1, B5, and B7 to a fermentation broth, wherein a feed rate of vitamin B5 is 25 to 150 ug/g cell produced, wherein an amount of vitamin B is provided at a feed rate of at least 4 times as feed rate of vitamin B7, and the amount of vitamin B5 is provided at a feed rate that is at least 3.5 times a feed rate of vitamin B1; and wherein the process provides a specific ethanol productivity rate of 8 g/day/gram cells or more (relevant to instant claim 1). The patent claims Senaratne ‘872 lack CO-containing gaseous substrate has a H2/CO molar ratio of about 0.2 or more (relevant to instant claim 5). The patent claims of Senaratne ‘872 lack a fermentation broth that has 0.01g/L or less yeast extract (relevant to instant claim 8); and 0.01 g/L or less carbohydrates (relevant to instant claim 9).
However, Gaddy teaches providing a liquid medium containing vitamin B1, B5 and B7 to a fermenter containing broth, wherein the vitamin B5 feed rate is 97 µg/g-cell produced, and wherein the ethanol productivity is 10.7 g/day/gram cells produced. See Gaddy column 22 line 61 to column 23 line 15 for the fermentation process. Phillips indicates that reducing the concentration of b-vitamins in a Clostridium ljungdahlii fermentation medium can increase the ethanol-to acetate product ratio. See the second paragraph on page 563. Phillips (1994) teaches a CSTR (i.e. continuous stirred tank reactor) medium that includes 0.155 mg/L Ca-D-pantothenate, 0.040 mg/L biotin and 0.01 mg/L thiamine-HCl. See table 3. Therefore, Phillips (1994) implies that the CSTR medium includes vitamin B5 in an amount that is 3.875 times the amount of vitamin B7 and 15.5 times the amount of vitamin B1 (relevant to instant claim 1). Gaddy teaches feeding a gaseous substrate containing H2 and CO in a 1.5 molar ratio to a fermenter, see column 22, lines 65-66 (relevant to instant claim 5). Gaddy teaches an ethanol production medium that does not contain yeast extract (relevant to instant claim 8). See table 1 in column 35. Gaddy teaches bacterial growth media without sugar (relevant to instant claim 9). See column 6 line 61.
It would have been obvious to a person of ordinary skill in the art, prior to the effective filing date to modify the fermentation process of Senaratne ‘872 by combining the medium with vitamins B1, B5 and B7 of Gaddy, by further optimizing the vitamin B feed rates of Gaddy based on the teachings of Phillips and Phillips (1994); and by finally optimizing molar ratio of H2 and CO in the syngas, as taught by Gaddy, in order to ferment acetogenic bacteria for ethanol production. There would be a reasonable expectation of success because Senaratne ‘872, Gaddy, Phillips and Phillips (1994) are all in the same field of producing ethanol by fermenting acetogenic bacteria.
(Maintain) Claims 1, 4-5, and 8-9 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-9 of U.S. Patent No. 10,337,074 B2 to Senaratne et al. (hereinafter Senaratne‘337) in view of Gaddy (US 7,285,402 B2), Phillips (Applied biochemistry and biotechnology, 39, 1993, 559-571) and Phillips (1994) (Applied biochemistry and biotechnology, 1994 45, 145-157).
Claim 9 of Senaratne ‘074 recites a method of producing, one or more alcohols from a gaseous substrate, the method comprising: adding gaseous substrate comprising carbon monoxide (CO) and hydrogen (H2) into an aqueous medium in a bioreactor, wherein the aqueous medium includes one or more anaerobic acetogenic microorganisms; agitating the aqueous medium at an agitator speed of 10 to 1000 rpm; measuring conversion of CO; measuring conversion of H2; and increasing a flow rate of gaseous substrate into the bioreactor by a flow factor (proposed quantity of gaseous feed divided by current quantity of gaseous feed) of 1.05 to 2.0 when CO conversion in the bioreactor is 25 to 95%, H2 conversion in, the bioreactor is 25 to 95% and a difference between CO conversion and H2 conversion in the bioreactor is 0% to 25%, wherein the agitator speed is increased when a second CO conversion is 0 to 25% and H2 conversion is 0 to 25%; wherein if a concentration of acetic acid in the aqueous medium is less than about 1.5 grams per liter after reaching a cell density of at least 50% more than an initial cell density, then about 1 ml of a vitamin solution is added per liter of aqueous medium, wherein the vitamin solution includes about 0.08 to about 1 μM biotin, about 0.12 to about 1.5 μM thiamin HCL, and about 0.15 to about 2 μM calcium d-pantothenate.
The claims of Senaratne ‘074 lack providing vitamin B1, B5, and B7 to a fermentation broth, wherein a feed rate of vitamin B5 is 25 to 150 ug/g cell produced, wherein an amount of vitamin B is provided at a feed rate of at least 4 times as feed rate of vitamin B7, and the amount of vitamin B5 is provided at a feed rate that is at least 3.5 times a feed rate of vitamin B1; and wherein the process provides a specific ethanol productivity rate of 8 g/day/gram cells or more (relevant to instant claim 1). The patent claims Senaratne ‘074 lack Clostridium (relevant to instant claim 3) selected from Clostridium ljungdahlii, Clostridium autoethanogun, Clostridium carboxidivorans, Clostridium drakei, Clostridium coskatiii, Clostridium ragsdalei, and mixture thereof (relevant to instant claim 4). The patent claims Senaratne ‘074 lack CO-containing gaseous substrate has a H2/CO molar ratio of about 0.2 or more (relevant to instant claim 5). The patent claims of Senaratne ‘074 lack a fermentation broth that has 0.01g/L or less yeast extract (relevant to instant claim 8); and 0.01 g/L or less carbohydrates (relevant to instant claim 9).
However, Gaddy teaches providing a liquid medium containing vitamin B1, B5 and B7 to a fermenter containing broth, wherein the vitamin B5 feed rate is 97 µg/g-cell produced, and wherein the ethanol productivity is 10.7 g/day/gram cells produced. See Gaddy column 22 line 61 to column 23 line 15 for the fermentation process. Phillips indicates that reducing the concentration of b-vitamins in a Clostridium ljungdahlii fermentation medium can increase the ethanol-to acetate product ratio. See the second paragraph on page 563. Phillips (1994) teaches a CSTR (i.e. continuous stirred tank reactor) medium that includes 0.155 mg/L Ca-D-pantothenate, 0.040 mg/L biotin and 0.01 mg/L thiamine-HCl. See table 3. Therefore, Phillips (1994) implies that the CSTR medium includes vitamin B5 in an amount that is 3.875 times the amount of vitamin B7 and 15.5 times the amount of vitamin B1 (relevant to instant claim 1). Gaddy teaches feeding a gaseous substrate containing H2 and CO in a 1.5 molar ratio to a fermentor, see column 22, lines 65-66 (relevant to instant claim 5). Gaddy teaches an ethanol production medium that does not contain yeast extract (relevant to instant claim 8). See table 1 in column 35. Gaddy teaches bacterial growth media without sugar (relevant to instant claim 9). See column 6 line 61.
It would have been obvious to a person of ordinary skill in the art, prior to the effective filing date, to have modified the alcohol production method of Senaratne ‘074 by fermenting Clostridium ljungdahlii with vitamins B1, B5 and B7 in accordance with the teachings of Gaddy, and by further optimizing the vitamin B feed rates of Gaddy based on the teachings of Phillips and Phillips (1994); and by finally optimizing molar ratio of H2 and CO in the syngas, as taught by Gaddy, in order to ferment acetogenic bacteria for ethanol production. There would be a reasonable expectation of success because Senaratne ‘074, Gaddy, Phillips and Phillips (1994) are all in the same field of producing ethanol by fermenting acetogenic bacteria.
(Maintain) Claims 1, 4-5, and 8-9 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-6 of U.S. Patent No. 10,494,651 to Senaratne et al. (hereinafter Senaratne ‘651) in view of Gaddy (US 7,285,402 B2), Phillips (Applied biochemistry and biotechnology, 39, 1993, 559-571) and Phillips (1994) (Applied biochemistry and biotechnology, 1994 45, 145-157).
Claim 1 of ‘651 to Senaratne recites a process for fermenting a CO-containing gaseous substrate to produce ethanol comprising: (a) providing a CO-containing gaseous substrate to a fermentation medium; (b) contacting the resultant CO-containing fermentation medium of step (a) with one or more acetogenic bacteria to form a mixture; (c) fermenting the mixture of step (b); (d) measuring the specific feed rate of the CO-containing gaseous substrate and the conductivity of the mixture of step c); (e) providing the CO-containing gaseous substrate to the mixture to maintain a conductivity (y) to specific gas feed rate (x) of the CO-containing gaseous substrate according to a formula where y=−6.0327x+12.901 where the conductivity is about 30 mS/cm or less, until reaching a target cell density of the one or more acetogenic bacteria, wherein x is about 0.2 to about 07 mmole/minute/gram of cells of the acetogenic bacteria; maintaining a cell density of the one or more acetogenic bacteria above a target cell density; and obtaining ethanol; wherein the process is effective for maintaining a space time yield (STY) of about 10 g ethanol/(L-day) or more.
Claim 6 of ‘651 to Senaratne recites the fermentation process of claim 1 wherein the one or more acetogenic bacteria is selected from the group consisting of…Clostridium aceticum, Clostridium acetobutylicum, Clostridium autoethanogenum, Clostridium coskatii (ATCC PTA-10522), Clostridium drakei, Clostridium ljungdahlii PETC (ATCC 49587), Clostridium ljungdahlii ERI2 (ATCC 55380), Clostridium ljungdahlii C-01 (ATCC 55988), Clostridium ljungdahlii O-52 (ATCC 55889), Clostridium magnum, Clostridium pasteurianum (DSM 525 of DSMZ Germany), Clostridium ragsdali Pit (ATCC BAA-622), Clostridium scatologenes, Clostridium thermoaceticum, Clostridium ultunense.
The claims of Senaratne ‘651 lack providing vitamin B1, B5, and B7 to a fermentation broth, wherein a feed rate of vitamin B5 is 25 to 150 ug/g cell produced, wherein an amount of vitamin B is provided at a feed rate of at least 4 times as feed rate of vitamin B7, and the amount of vitamin B5 is provided at a feed rate that is at least 3.5 times a feed rate of vitamin B1; and wherein the process provides a specific ethanol productivity rate of 8 g/day/gram cells or more (relevant to instant claim 1). The patent claims Senaratne ‘651 lack CO-containing gaseous substrate has a H2/CO molar ratio of about 0.2 or more (relevant to instant claim 5). The patent claims of Senaratne ‘651 lack a fermentation broth that has 0.01g/L or less yeast extract (relevant to instant claim 8); and 0.01 g/L or less carbohydrates (relevant to instant claim 9).
However, Gaddy teaches providing a liquid medium containing vitamin B1, B5 and B7 to a fermenter containing broth, wherein the vitamin B5 feed rate is 97 µg/g-cell produced, and wherein the ethanol productivity is 10.7 g/day/gram cells produced. See Gaddy column 22 line 61 to column 23 line 15 for the fermentation process. Phillips indicates that reducing the concentration of b-vitamins in a Clostridium ljungdahlii fermentation medium can increase the ethanol-to acetate product ratio. See the second paragraph on page 563. Phillips (1994) teaches a CSTR (i.e. continuous stirred tank reactor) medium that includes 0.155 mg/L Ca-D-pantothenate, 0.040 mg/L biotin and 0.01 mg/L thiamine-HCl. See table 3. Therefore, Phillips (1994) implies that the CSTR medium includes vitamin B5 in an amount that is 3.875 times the amount of vitamin B7 and 15.5 times the amount of vitamin B1 (relevant to instant claim 1). Gaddy teaches feeding a gaseous substrate containing H2 and CO in a 1.5 molar ratio to a fermentor, see column 22, lines 65-66 (relevant to instant claim 5). Gaddy teaches an ethanol production medium that does not contain yeast extract. See table 1 in column 35 (relevant to instant claim 8). Gaddy teaches bacterial growth media without sugar See column 6 line 61 (relevant to instant claim 9).
It would have been obvious to a person of ordinary skill in the art, prior to the effective filing date to modify the fermentation mixture of Senaratne ‘651 by combining the medium with vitamins B1, B5 and B7 of Gaddy, by further optimizing the vitamin B feed rates of Gaddy based on the teachings of Phillips and Phillips (1994); and by finally optimizing molar ratio of H2 and CO in the syngas, as taught by Gaddy, in order to ferment acetogenic bacteria for ethanol production. There would be a reasonable expectation of success because Senaratne ‘651, Gaddy, Phillips and Phillips (1994) are all in the same field of producing ethanol by fermenting acetogenic bacteria.
(Maintain) Claims 1, 4-5, and