DETAILED CORRESPONDENCE
Status of the Application
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claims 1-3, 5-12, 14-19, and 22 are pending in the application.
Applicant’s amendment to the claims, filed February 20, 2026, is acknowledged. This listing of the claims replaces all prior versions and listings of the claims.
Applicant’s remarks filed February 20, 2026 in response to the non-final rejection mailed December 3, 2025 have been fully considered.
Any objection or rejection previously applied to claims 20 and 21 is withdrawn in view of applicant’s amendment to cancel these claims.
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claim Objections
The objection to claim 22 is withdrawn in view of applicant’s amendment to replace “A” with “The.”
Claim 1 is objected to in the recitation of “a strain of Trichoderma reesei or of Trichoderma reesei modified by selective mutation or genetic recombination…growing said strain of Trichoderma reesei or of Trichoderma reesei modified by selective mutation or genetic recombination…wherein the enzymes are produced by a strain CL847 or TR3002” and in the interest of improving claim form, it is suggested that claim 1 be amended to recite (with markings to show changes made) “A process for producing enzymes by a strain of Trichoderma reesei
a step of growing said strain of Trichoderma reesei
(b) a step of producing enzymes, starting from the culture medium obtained in step (a), which is optionally diluted, in the presence of at least one inductive carbon-based substrate;
wherein the pH in step (a) is identical to the pH in step (b) and wherein the pH in step (a) and in step (b) is 4.0 to 4.4;
wherein step (a) and step (b) are performed in the same bioreactor; and
wherein the process is operated in the absence of an antifoam; and
wherein said strain of Trichoderma reesei is
Claim Rejections - 35 USC § 112(d)
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
Claims 10 and 12 are rejected under 35 U.S.C. 112(d) as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends.
This rejection is necessitated by applicant’s amendment to incorporate the limitations of claim 20 into claim 1.
Claim 1 has been amended to recite “wherein the enzymes are produced by a strain CL847 or TR3002.” Given that CL847 and TR3002 are strains of Trichoderma reesei, claim 10 does not further limit claim 1, and given that CL847 and TR3002 are mutant strains of Trichoderma reesei, claim 12 does not further limit claim 1.
Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
Claim Rejections - 35 USC § 103
The rejection of claims 1-3, 5-12, 14, 15, and 18 under 35 U.S.C. 103 as being unpatentable over Edwards et al. (WO 2009/026716 A1; cited on the IDS filed October 20, 2021; hereafter “Edwards”), and
the rejection of claims 16, 17, and 19 under 35 U.S.C. 103 as being unpatentable over Edwards in view of Ben Chaabane et al. (US 2013/0210119 A1; cited on Form PTO-892 mailed November 27, 2024; hereafter “Ben Chaabane”)
are withdrawn in view of applicant’s amendments to claim 1 to incorporate the limitations of claims 20 and 21.
Claims 1-3, 5-12, 14, 15, 18, and 22 are rejected under 35 U.S.C. 103 as being unpatentable over Edwards in view of Herpoël-Gimbert et al. (Biotechnol. Biofuels 1:18, 2008, 12 pages; cited on the attached Form PTO-892; hereafter “Herpoël-Gimbert”).
This rejection was previously applied to claims 20-22 and has been modified to address applicant’s amendment to claim 1.
As amended, the claims are drawn to a process for producing enzymes by a strain of Trichoderma reesei or of Trichoderma reesei modified by selective mutation or genetic recombination, comprising
a step of growing said strain of Trichoderma reesei or of Trichoderma reesei modified by selective mutation or genetic recombination, in the presence of at least one carbon-based growth substrate, in a stirred and aerated bioreactor in batch phase,
(b) a step of producing enzymes, starting from the culture medium obtained in step (a), which is optionally diluted, in the presence of at least one inductive carbon-based substrate;
wherein the pH in step (a) is identical to the pH in step (b) and wherein the pH in step (a) and in step (b) is 4.0 to 4.4;
wherein step (a) and step (b) are performed in the same bioreactor; and
wherein the process is operated in the absence of an antifoam; and
wherein the enzymes are produced by a strain CL847 or TR3002.
Regarding claim 1, Edwards generally teaches a fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures (see Abstract) and Example 2 beginning at p. 15 of Edwards teaches fermentations using hemicellulose-derived carbohydrates (HDC) and cellulase-inducing carbohydrates (CIC).
Regarding the step (a) limitation of “a step of growing said strain of Trichoderma reesei or of Trichoderma reesei modified by selective mutation or genetic recombination, in the presence of at least one carbon-based growth substrate,” Edwards teaches Trichoderma reesei strains RutC30 and P59G (paragraph [0057]) and teaches culturing the Trichoderma in batch mode until glucose in the media was depleted (paragraph [0059].
Regarding the step (a) limitation of “in a stirred and aerated bioreactor in batch phase, at a pH of 3.8 to 4.4” and the claim 1 limitation “wherein the pH in step (a)…is 4.0 to 4.4,” Edwards teaches culturing the Trichoderma in batch mode and teaches the operational parameters during the batch portion of the run included impeller agitation at 500 rpm and air sparging at 8 standard liters per minute (paragraph [0059]). Edwards teaches the culture pH was maintained at 4.0-4.5 during batch growth (paragraph [0059]). Edwards does not expressly teach the pH range of 4.0 to 4.4, however, the recited pH range of 4.0 to 4.4 overlaps with the pH range of 4.0-4.5 taught by Edwards and according to MPEP 2144.05.I, in the case where the claimed ranges “overlap or lie inside ranges disclosed by the prior art” a prima facie case of obviousness exists.
Regarding the step (b) limitation of “a step of producing enzymes, starting from the culture medium obtained in step (a)…in the presence of at least one inductive carbon-based substrate,” Edwards teaches that following the depletion of glucose in the media, the carbon source was added (paragraph [0059]). Edwards teaches the carbon source was either HDC alone or a combination of HDC and cellulase-inducing carbohydrates (CIC) (paragraph [0062]). Edwards teaches that the addition of HDC alone or HDC and CIC resulted in the production of secreted protein (paragraph [0062]). Given that the addition of HDC alone or HDC and CIC resulted in the production of secreted protein, HDC alone or a combination of HDC and CIC are considered to be encompassed by “at least one inductive carbon-based substrate.”
Regarding the claim 1 limitation “wherein the pH…in step (b) is 4.0 to 4.4,” Edwards teaches the culture pH was maintained at 3.5 during cellulase enzyme production (paragraph [0059]).
Regarding the limitation of “wherein step (a) and step (b) are performed in the same bioreactor,” Edwards teaches the batch growth phase and the enzyme production phase were carried out in the same 14L pilot scale fermentation vessel (paragraph [0059]).
Regarding the limitation of “wherein the process is operated in the absence of an antifoam,” the method of Edwards does not include an antifoam.
The differences between claim 1 and the method of Edwards are:
the enzyme production phase of claim 1 (i.e., step (b) of claim 1) is carried out at pH 4.0 to 4.4, while the enzyme production phase of the method of Edwards is carried out at pH 3.5; and
claim 1 recites the enzymes are produced by a strain CL847 or TR3002, while Edwards teaches using a Trichoderma reesei cell for producing cellulases, which is exemplified by Trichoderma reesei strain RutC30.
Regarding difference 1), Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, and 4.4 (paragraph [0034]). In view of the explicit teachings of Edwards, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. One would have been motivated and would have expected success to do this because Edwards teaches the pH of the batch growth phase is 4.0-4.5 and teaches the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, and 4.4, and one of ordinary skill in the art would have recognized that using the same pH for the batch growth phase and the enzyme production phase simplifies the process by avoiding the time and materials (e.g., additional acid) needed to adjust the pH between the batch growth phase and the enzyme production phase.
Regarding difference 2), Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph). Herpoël-Gimbert reports a comparison of the secretomes of strain Rut-C30 and strain CL847, noting that strain CL847 secretes a more diversified set of enzymes than strain Rut-C30 (p. 9, column 1; p. 10, column 1).
In view of the combined teachings of Edwards and Herpoël-Gimbert, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the method of Edwards to use strain CL847 as the Trichoderma reesei cell for producing cellulases. One would have been motivated and would have expected success to do this because Edwards teaches Trichoderma reesei strain RutC30 for producing cellulases while Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases and secretes a more diversified set of enzymes than strain Rut-C30.
Regarding claims 2, 3, and 22, Edwards teaches the culture pH was maintained at pH 4.0-4.5 during batch growth (paragraph [0059]), and Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, and 4.4 (paragraph [0034]), and for the reasons stated above, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. Thus, it would have been obvious to one of ordinary skill in the art before the effective filing date for the pH of the batch growth phase to be 4.0, 4.1, 4.2, 4.3, or 4.4 and for the pH of the enzyme production phase to be the same as the batch growth phase.
Regarding claim 5, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]).
Regarding claim 6, Edwards teaches fed-batch fermentation for cellulase enzyme production (paragraph [0062]).
Regarding claim 7, the recitation of “wherein the culture medium obtained in step (a) is diluted” is interpreted as encompassing the addition of a liquid comprising a carbon source, which would have the effect of diluting the culture medium from the batch mode growth phase. Edwards teaches that following the batch growth phase and for the cellulase production phase, the carbon source was added with peristaltic pumps (paragraph [0059]).
Regarding claim 8, Edwards teaches the culture medium for the batch fermentation comprises 6.00 g/L dry corn steep liquor and 13.00 g/L glucose (p. 16, middle). Dry corn steep liquor and glucose are considered to be carbon sources.
Regarding claims 9 and 18, Edwards teaches peristaltic pumps were used to deliver the carbon source (HDC alone or a combination of HDC and CIC) at a feed rate of 0.4 grams (i.e., 400 milligrams) of carbon/liter culture/hour (paragraph [0059]).
Regarding claims 10, 12, and 22, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph), and Herpoël-Gimbert reports a comparison of the secretomes of strain Rut-C30 and strain CL847, noting that strain CL847 secretes a more diversified set of enzymes than strain Rut-C30 (p. 9, column 1; p. 10, column 1), and for reasons stated above, it would have been obvious to one of ordinary skill in the art before the effective filing date to combine Edwards and Herpoël-Gimbert to modify the method of Edwards to use strain CL847 as the Trichoderma reesei cell for producing cellulases.
Regarding claim 11, Edwards teaches production of cellulases (e.g., Example 2 beginning at p. 15), which are cellulolytic enzymes.
Regarding claim 14, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]). Aqueous ammonia is another name for ammonium hydroxide.
Regarding claim 15, the recitation of “wherein the culture medium obtained in step (a) is not diluted” is interpreted as encompassing no dilution of the culture medium of step (a) immediately before step (b). Edwards does not teach diluting the batch mode growth phase culture medium prior to the cellulase production phase.
Therefore, the invention of claims 1-3, 5-12, 14, 15, 18, and 22 would have been obvious to one of ordinary skill in the art before the effective filing date.
Claims 16, 17, and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Edwards in view of Herpoël-Gimbert as applied to claims 1-3, 5-12, 14, 15, 18, and 22 above, and further in view of Ben Chaabane.
Claim 16 is drawn to the process as claimed in claim 1, wherein step (b) is operated with a stream of an inductive carbon-based substrate of between 30 and 140 mg.g-1.h-1 (i.e., mg per gram of cells per hour).
Claim 17 is drawn to the process as claimed in claim 1, wherein step (b) is operated with a stream of an inductive carbon-based substrate of between 35 and 45 mg.g-1.h-1.
Claim 19 is drawn to the process as claimed in claim 1, wherein step (b) is operated with a stream of an aqueous solution of inductive carbon-based substrate at a concentration of between 200 and 600 g/L.
The relevant teachings of Edwards and Herpoël-Gimbert as applied to claims 1-3, 5-12, 14, 15, 18, and 22 are set forth above.
The combination of Edwards and Herpoël-Gimbert does not teach or suggest the cellulase production phase is operated with a stream of inductive carbon-based substrate of between 30 and 140 mg·g−1·h−1 (claim 16), the cellulase production phase is operated with a stream of inductive carbon-based substrate of between 35 and 45 mg·g−1·h−1 (claim 17), and the cellulase production phase is operated with a stream of inductive carbon-based substrate at a concentration of between 200 and 600 g/L (claim 19).
Similar to Edwards, Ben Chaabane teaches a two-phase process for the production of cellulolytic and/or hemicellulolytic enzymes (paragraph [0022]) by Trichoderma reesei (paragraph [0037]). Ben Chaabane teaches the inducing substrate in the enzyme production phase is a mixture of glucose or cellulosic hydrolysates, lactose and xylose or a solution of hemicellulolytic hydrolysates (paragraph [0022]). Specifically, Ben Chaabane teaches that the inducing substrate in the enzyme production phase is an aqueous solution comprising the inducing substrate constituted by a glucose/ lactose/xylose mixture or hemicellulose hydrolysate solution at a concentration of 450 to 550 g/L with a flow rate of 30 to 45 mg per gram of cells per hour (paragraphs [0040] to [0042]). According to Ben Chaabane, enzyme production methods including these parameters increased protein production ([0090] and [0099]) and achieved a high quality enzyme cocktail (paragraph [0111]). Ben Chaabane does not expressly teach the exact ranges as recited in claims 16, 17, and 19, however, the recited ranges overlap with the ranges taught by Ben Chaabane and according to MPEP 2144.05.I, in the case where the claimed ranges “overlap or lie inside ranges disclosed by the prior art” a prima facie case of obviousness exists.
In view of Edwards, Herpoël-Gimbert, and Ben Chaabane, it would have been obvious to one of ordinary skill in the art before the effective filing date to further modify Edwards to use and deliver the inducing substrate of Ben Chaabane. One would have been motivated and would have expected success to do this because Ben Chaabane teaches that enzyme production methods including these parameters increased protein production and achieved a high quality enzyme cocktail.
Therefore, the invention of claims 16, 17, and 19 would have been obvious to one of ordinary skill in the art before the effective filing date.
RESPONSE TO REMARKS: Applicant argues that Example 2 of Edwards teaches a pH of 3.5 for the enzyme production phase and because the teachings of paragraph [0034] of Edwards are not disclosed in the Examples section, one of ordinary skill in the art would not have consulted paragraph [0034] of Edwards for alternative pH values for the enzyme production phase of Example 2.
Applicant’s arguments are not found persuasive. According to paragraph [0055] of Edwards, “it is to be understood that these examples are for illustrative purposes only and should not be used to limit the scope of the present invention in any manner.” Thus, Edwards expressly teaches that the examples are non-limiting and one of ordinary skill in the art would have recognized that paragraph [0034] of Edwards is teaching culture pH values of the “process of the invention,” which includes the enzyme production phase, and would have recognized that the culture pH values at paragraph [0034] can be applied to the enzyme production phase of Example 2. As stated above, Edwards teaches the pH of the batch growth phase is 4.0-4.5 and teaches the pH of the “process of the invention,” which includes the enzyme production phase, can be pH 4.0, 4.1, 4.2, 4.3, and 4.4. In view of the teachings of Edwards, one of ordinary skill in the art would have recognized that using the same pH for the batch growth phase and the enzyme production phase simplifies the process by avoiding the time and materials (e.g., additional acid) needed to adjust the pH between the batch growth phase and the enzyme production phase.
Applicant argues that since Herpoël-Gimbert teaches a growth phase at pH 4.8, the combined teachings would have rather motivated one of ordinary skill to modify Edwards to use a growth phase pH of 4.8 while leaving the production phase pH at 3.5, which is even further from the claimed invention. Applicant argues that claim 22 requires both “strain CL847” and “wherein the pH in step (a) and in step (b) is 4.0” and one of ordinary skill in the art would have had even less of a reason or chance to arrive at the invention of claim 22.
Applicant’s arguments are not found persuasive. First, it is noted that applicant’s argument appears to conflict with applicant’s earlier argument that one would not have modified the culture pHs of Example 2 of Edwards. Second, it is acknowledged that paragraph [0034] of Edwards teaches a culture pH of 4.8. However, paragraph [0034] of Edwards also teaches additional pH values of 4.0, 4.1, 4.2, 4.3, and 4.4 and since there is a normal desire of scientists or artisans to improve upon what is already generally known (MPEP 2144.05.II.A), it would have been obvious to one of ordinary skill in the art for the pH of the enzyme production phase to be pH 4.0, 4.1, 4.2, 4.3, and 4.4 and to use the same pH for the enzyme production phase that is used for the batch growth phase. Moreover, given the Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases and secretes a more diversified set of enzymes than strain Rut-C30, it would have been obvious to modify the method of Edwards to use strain CL847 rather than Rut-C30.
Applicant argues the declaration under 37 CFR 1.132 filed April 25, 2025 (hereafter “Margeot declaration”) continues to be dismissed while it should be considered as fact for purposes of examination. According to applicant, the example for comparison in the Margeot declaration is closer to the claimed invention than any embodiment disclosed in the prior art and thus, applicant has provided a comparison with the closest prior art.
Applicant’s arguments and evidence of the Margeot declaration are not found persuasive. Contrary to applicant’s position, the Margeot declaration has been fully considered but fails to rebut a prima facie case of obviousness. For example, the claimed subject matter is not compared with the closest prior art as required by MPEP 716.02(e). The Margeot declaration (p. 2, top) discloses a growth phase at pH 4.8 and an enzyme production phase at pH 4.0, 3.5 or 3.0. However, the reference of Edwards, which is the closest prior art, discloses a growth phase at pH 4.0-4.5 and an enzyme production phase at pH 3.5, which, contrary to applicant’s position, is closer to the claimed invention than the example for comparison in the Margeot declaration.
Applicant argues “commensurate in scope” does not require a comparison with every embodiment within the scope of the claims.
Applicant’s arguments are not found persuasive. It is acknowledged that a comparison with every embodiment within the scope of the claims is not required to establish unexpected results. Rather, according to MPEP 716.02(d), “[t]he nonobviousness of a broader claimed range can be supported by evidence based on unexpected results from testing a narrower range if one of ordinary skill in the art would be able to determine a trend in the exemplified data which would allow the artisan to reasonably extend the probative value thereof.” The alleged unexpected results of the Margeot declaration are based on strain CL847, a growth phase at pH 4.8, an enzyme production phase at pH 4.0, and lactose as a carbon source, which are not reasonably commensurate in scope with the claimed invention because it is applicant’s burden to establish unexpected results (MPEP 716.02(b).I) and there is no evidence of record or line of reasoning presented by the applicant that the untested embodiments encompassed by the claims would behave in the same manner as the tested embodiments. See MPEP 716.02(d).I. For these reasons, it is the examiner’s position that the results of the Margeot declaration fail to rebut a prima facie case of obviousness.
Applicant argues the results reported in Figures 1 and 2 clearly show that protein production performance and enzymatic activity are lower at pH 3.5 and also at pH 3.0 than at pH 4.0.
Applicant’s arguments are not found persuasive. Applicant has the burden of explaining the data they proffer as evidence of unexpected results and the evidence relied upon should establish unexpected results (MPEP 716.02(b)). Figures 1 and 2 are photos of bioreactors (see specification at p. 8, lines 30-35) and without further explanation by applicant as to how Figures 1 and 2 are evidence of unexpected results, applicant has failed to satisfy the burden of establishing unexpected results.
Claim Rejections - Double Patenting
The nonstatutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
U.S. Patent No. 9,885,027 B2
The rejection of claims 1-3, 5-12, and 14-19 on the ground of nonstatutory double patenting as being unpatentable over claims 1, 7, 11, and 12 of U.S. Patent No. 9,885,027 B2 (cited on the IDS filed on October 20, 2021) in view of Edwards is withdrawn in view of applicant’s amendments to claim 1 to incorporate the limitations of claims 20 and 21.
Claims 1-3, 5-12, 14-19, and 22 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 7, 11, and 12 of U.S. Patent No. 9,885,027 B2 (cited on the IDS filed on October 20, 2021) in view of Edwards and Herpoël-Gimbert.
This rejection was previously applied to claims 20-22 and has been modified to address applicant’s amendment to claim 1.
Regarding claim 1 of this application, claim 1 of the patent is drawn to a process for the production of an enzymatic cocktail with a cellulolytic microorganism, comprising two phases:
a phase a) comprising growing said microorganism in the presence of at least one carbonaceous growth substrate in a closed reactor, said phase a) being carried out with a concentration of carbonaceous growth substrate in the range of 10 to 90 g/L; and
a phase b) comprising producing the enzymatic cocktail, in which at least one carbonaceous inducer substrate is supplied to the reactor, said carbonaceous inducer substrate being at least one solid residue obtained by enzymatic partial hydrolysis of lignocellulosic materials which have undergone a pre-treatment step, said solid residue having a liquid portion and a solid portion wherein the solid portion constitutes 10% to 40% by weight of the solid residue and said solid residue having no solid residue obtained from ethanolic fermentation of enzymatic hydrolysates from the enzymatic partial hydrolysis of lignocellulosic materials which have undergone a pre-treatment step, said phase b) being carried out with a concentration of carbonaceous inducer substrate in the range of 150 to 400 g/L, wherein the partial hydrolysis is conducted with an initial enzymatic cocktail, wherein, in the partial hydrolysis, only 20% to 70% by weight of the lignocellulosic materials subject to the enzymatic partial hydrolysis are hydrolysed and wherein the partial hydrolysis is conducted with: a dry matter content of more than 20% by weight; and/or with less than 10 mg of the initial enzymatic cocktail per gram of cellulose in the lignocellulosic materials which have undergone a pre-treatment step;
claim 11 of the patent recites the process according to claim 1, in which the cellulolytic microorganism is selected from the group consisting of strains of fungi belonging to the genuses Trichoderma, Aspergillus, Penicillium and Schizophyllum; and
claim 12 of the patent recites the process according to claim 11, in which the cellulolytic microorganism belongs to the species Trichoderma reesei.
The claims of the patent do not recite the following limitations of instant claim 1:
the growth phase is a batch fermentation in a stirred and aerated bioreactor,
the growth and production phases are conducted at a pH of 4.0 to 4.4,
the pH of the growth phase is identical to the pH of the enzyme production phase,
the growth phase and the enzyme production phase are performed in the same bioreactor,
the process is operated in the absence of an antifoam, and
the enzymes are produced by a strain CL847 or TR3002.
Regarding a batch fermentation growth phase in a stirred and aerated bioreactor, the growth phase and the enzyme production phase are performed in the same bioreactor, Edwards generally teaches a fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures with a batch growth phase with operational parameters during the batch growth portion of the run included impeller agitation at 500 rpm and air sparging at 8 standard liters per minute (paragraph [0059]) with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
In view of Edwards, it would have been obvious to one of ordinary skill in the art before the effective filing date to perform the growth phase of the patent using batch mode fermentation at pH 4.0-4.4 with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor, and without antifoam. One would have been motivated to and would have had a reasonable expectation of success to do so because Edwards acknowledges that the growth phase for cellulolytic enzyme production can be conducted using batch mode fermentation with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
Regarding growth and production phases being conducted at a pH of 4.0 to 4.4 and the pH of the growth phase is identical to the pH of the enzyme production phase, Edwards teaches maintaining a culture pH at 4.0-4.5 during batch growth and teaches that the pH of the cellulase production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]) and it would have been obvious to one of ordinary skill in the art to use the same pH for the enzyme production phase that is used for the batch growth phase in the claims of the patent. One would have been motivated to do this because Edwards acknowledges that the pH of a batch growth phase is 4.0-4.5 and teaches the pH of an enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4, and one of ordinary skill in the art would have recognized that using the same pH for the batch growth phase and the cellulase production phase avoids the time and materials needed to adjust the pH between the batch growth phase and the enzyme production phase.
Regarding the process is operated in the absence of an antifoam, given that the claims of the patent do not recite an antifoam and further given that the claims of the patent are presumed to be adequately described and enabled without requiring an antifoam, one of ordinary skill in the art would recognize that the process of the patent does not include or require an antifoam. Also, the method of Edwards, which is similar to the claims of the patent, does not include an antifoam and similar to the claims of the patent. Nowhere does Edwards even mention an antifoam.
Regarding the enzymes being produced by a strain CL847 or TR3002, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph).
In view of the teachings of Herpoël-Gimbert, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the process of the claims of the patent to use strain CL847 as the Trichoderma reesei cell for producing the enzymatic cocktail. One would have been motivated and would have expected success to do this because Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases and secretes a more diversified set of enzymes than strain Rut-C30.
Regarding claims 2 and 3 of this application, Edwards teaches the culture pH was maintained at 4.0-4.5 during batch growth (paragraph [0059]), Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]), and for the reasons set forth above, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. Thus, it would have been obvious to one of ordinary skill in the art before the effective filing date for the pH of the batch growth phase to be 4.0, 4.1, 4.2, 4.3, or 4.4 and for the pH of the enzyme production phase to be the same as the batch growth phase.
Regarding claim 5 of this application, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]).
Regarding claim 6 of this application, Edwards teaches fed-batch fermentation for cellulase production (paragraph [0062]).
Regarding claim 7 of this application, the recitation of “wherein the culture medium obtained in step (a) is diluted” is interpreted as encompassing the addition of a liquid comprising a carbon source, which would have the effect of diluting the culture medium from the batch mode growth phase. Edwards teaches that following the batch growth phase and for the cellulase production phase, the carbon source was added with peristaltic pumps (paragraph [0059]).
Regarding claim 8 of this application, Edwards teaches the culture medium for the batch fermentation comprises 6.00 g/L dry corn steep liquor and 13.00 g/L glucose (p. 16, middle). Dry corn steep liquor and glucose are considered to be carbon sources.
Regarding claims 9 and 18 of this application, Edwards teaches peristaltic pumps were used to deliver the carbon source (HDC alone or a combination of HDC and CIC) at a feed rate of 0.4 grams (i.e., 400 milligrams) of carbon/liter culture/hour (paragraph [0059]).
Regarding claims 10, 12, and 22, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph) and for reasons stated above, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the method of the claims of the patent to use strain CL847 as the cellulolytic microorganism for producing cellulases.
Regarding claim 11 of this application, Edwards teaches production of cellulases (e.g., Example 2 beginning at p. 15), which are cellulolytic enzymes.
Regarding claim 14 of this application, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]). Aqueous ammonia is another name for ammonium hydroxide.
Regarding claim 15 of this application, the recitation of “wherein the culture medium obtained in step (a) is not diluted” is interpreted as encompassing no dilution of the culture medium of step (a) immediately before step (b). The claims of the patent do not recite and Edwards does not teach diluting the batch mode growth phase culture medium prior to the cellulase production phase.
Regarding claims 16 and 17 of this application, claim 7 of the patent recites the specific flow rate at which the carbonaceous inducer substrate used in said phase b) is supplied in the range of 35 to 65 mg of carbonaceous inducer substrate per gram of microorganism per hour.
Regarding claim 19 of this application, claim 1 of the patent recites the concentration of carbonaceous inducer substrate is in the range of 150 to 400 g/L.
Therefore, claims 1-3, 5-12, 14-19, and 22 of this application are unpatentable over claims 1, 7, 11, and 12 of the patent in view of Edwards and Herpoël-Gimbert.
U.S. Patent No. 10,457,925 B2
The rejection of claims 1-3, 5-12, and 14-19 on the ground of nonstatutory double patenting as being unpatentable over claims 1, 5, 6, and 11 of U.S. Patent No. 10,457,925 B2 (cited on the IDS filed on October 20, 2021) in view of Edwards is withdrawn in view of applicant’s amendments to claim 1 to incorporate the limitations of claims 20 and 21.
Claims 1-3, 5-12, 14-19, and 22 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1, 5, 6, and 11 of U.S. Patent No. 10,457,925 B2 (cited on the IDS filed on October 20, 2021) in view of Edwards and Herpoël-Gimbert.
This rejection was previously applied to claims 20-22 and has been modified to address applicant’s amendment to claim 1.
Regarding claim 1 of this application, claim 1 of the patent is drawn to a process for the production of cellulolytic and/or hemicellulolytic enzymes by a cellulolytic and/or hemicellulolytic microorganism, comprising
(a) growing the microorganism in a culture medium in the presence of a source of carbon;
(b) inducing the production of the enzymes by the microorganism in the presence of an inducing substrate, wherein said inducing substrate is a mixture of cellulosic hydrolysates, lactose and a solution of hemicellulosic hydrolysates, the quantities of each of the constituents of the mixture are defined by the following limits:
50% to 65% by weight of cellulosic hydrolysates;
22% to 24% by weight of lactose; and
15% to 25% by weight of a solution of hemicellulosic hydrolysates;
the sum of these three constituents being equal to 100%, and
wherein the microorganism belongs to the species Trichoderma reesei which is deleted for catabolic repression by glucose, wherein the inducing substrate is supplied in a solution having a concentration of 350 to 600 g/L, and wherein both the source of carbon and the inducing substrate comprise sugar, and wherein the cellulosic hydrolysates and the solution of hemicellulosic hydrolysates are obtained from a pretreatment of lignocellulosic biomass, and/or the cellulosic hydrolysates are obtained directly from a process for the transformation of lignocellulosic biomass into ethanol;
claim 5 of the patent recites the process of claim 1, wherein growth step (a) is conducted at a pH of 3.5 to 6 and the temperature is 20° C to 35° C.
The claims of the patent do not recite the following limitations of instant claim 1:
the growth phase is a batch fermentation in a stirred and aerated bioreactor,
the growth and production phases are conducted at a pH of 4.0 to 4.4,
the pH of the growth phase is identical to the pH of the enzyme production phase,
the growth phase and the enzyme production phase are performed in the same bioreactor,
the process is operated in the absence of an antifoam, and
the enzymes are produced by a strain CL847 or TR3002.
Regarding a batch fermentation growth phase in a stirred and aerated bioreactor and the growth phase and the enzyme production phase are performed in the same bioreactor, Edwards generally teaches a fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures with a batch growth phase with operational parameters during the batch growth portion of the run included impeller agitation at 500 rpm and air sparging at 8 standard liters per minute (paragraph [0059]) with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
In view of Edwards, it would have been obvious to one of ordinary skill in the art before the effective filing date to perform the growth phase of the patent using batch mode fermentation at pH 4.0-4.4 with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor. One would have been motivated to and would have had a reasonable expectation of success to do so because Edwards acknowledges that the growth phase for cellulolytic enzyme production can be conducted using batch mode fermentation with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
Regarding growth and production phases being conducted at a pH of 4.0 to 4.4 and the pH of the growth phase is identical to the pH of the enzyme production phase, Edwards teaches maintaining a culture pH at 4.0-4.5 during batch growth and teaches that the pH of the cellulase production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]) and it would have been obvious to one of ordinary skill in the art to use the same pH for the enzyme production phase that is used for the batch growth phase in the claims of the patent. One would have been motivated to do this because Edwards acknowledges that the pH of a batch growth phase is 4.0-4.5 and teaches the pH of an enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4, and one of ordinary skill in the art would have recognized that using the same pH for the batch growth phase and the enzyme production phase avoids the time and materials needed to reduce the pH between the batch growth phase and the enzyme production phase.
Regarding the process is operated in the absence of an antifoam, given that the claims of the patent do not recite an antifoam and further given that the claims of the patent are presumed to be adequately described and enabled without requiring an antifoam, one of ordinary skill in the art would recognize that the process of the patent does not include or require an antifoam. Also, the method of Edwards, which is similar to the claims of the patent, does not include an antifoam and similar to the claims of the patent. Nowhere does Edwards even mention an antifoam.
Regarding the enzymes being produced by a strain CL847 or TR3002, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph).
In view of the teachings of Herpoël-Gimbert, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the process of the claims of the patent to use strain CL847 as the Trichoderma reesei cell for producing the enzymatic cocktail. One would have been motivated and would have expected success to do this because Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases and secretes a more diversified set of enzymes than strain Rut-C30.
Regarding claims 2 and 3 of this application, Edwards teaches the culture pH was maintained at 4.0-4.5 during batch growth (paragraph [0059]), Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]), and for the reasons set forth above, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. Thus, it would have been obvious to one of ordinary skill in the art before the effective filing date for the pH of the batch growth phase to be 4.0, 4.1, 4.2, 4.3, or 4.4 and for the pH of the enzyme production phase to be the same as the batch growth phase.
Regarding claim 5 of this application, Edwards teaches maintaining culture pH by addition of a 10% ammonium hydroxide solution (paragraph [0059]).
Regarding claim 6 of this application, claim 11 of the patent recites wherein the production step is performed in a fed-batch reactor.
Regarding claim 7 of this application, the recitation of “wherein the culture medium obtained in step (a) is diluted” is interpreted as encompassing the addition of a liquid comprising a carbon source, which would have the effect of diluting the culture medium from the batch mode growth phase. Edwards teaches that following the batch growth phase and for the cellulase production phase, the carbon source was added with peristaltic pumps (paragraph [0059]).
Regarding claim 8 of this application, claim 1 of the patent recites the growth phase a) is conducted in the presence of a source of carbon and Edwards teaches the culture medium for the batch growth phase fermentation comprises 6.00 g/L dry corn steep liquor and 13.00 g/L glucose (p. 16, middle). Dry corn steep liquor and glucose are considered to be carbon sources.
Regarding claims 9 and 18 of this application, claim 1 of the patent recites conducting an enzyme production phase b) to produce an enzyme medium in which at least one inducing substrate is introduced, the inducing substrate including cellulosic hydrolysates, lactose, and a solution of hemicellulosic hydrolysates. Edwards teaches peristaltic pumps were used to deliver the carbon source (HDC alone or a combination of HDC and CIC) at a feed rate of 0.4 grams (i.e., 400 milligrams) of carbon/liter culture/hour (paragraph [0059]).
Regarding claims 10, 12, and 22, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph) and for reasons stated above, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the method of the claims of the patent to use strain CL847 as the cellulolytic microorganism for producing cellulases.
Regarding claim 11 of this application, claim 1 of the patent recites a process for producing cellulolytic or hemicellulolytic enzymes.
Regarding claim 14 of this application, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]). Aqueous ammonia is another name for ammonium hydroxide.
Regarding claim 15 of this application, the recitation of “wherein the culture medium obtained in step (a) is not diluted” is interpreted as encompassing no dilution of the culture medium of step (a) immediately before step (b). The claims of the patent do not recite and Edwards does not teach diluting the batch mode growth phase culture medium prior to the cellulase production phase.
Regarding claims 16 and 17 of this application, claim 6 of the patent recites the induction solution is introduced at flow rate of 35 to 45 mg of carbonaceous inducer substrate per gram of microorganism per hour.
Regarding claim 19 of this application, claim 1 of the patent recites the concentration of carbonaceous inducer substrate is in the range of 350 to 600 g/L.
Therefore, claims 1-3, 5-12, 14-19, and 22 of this application are unpatentable over claims 1, 5, 6, and 11 of the patent in view of Edwards and Herpoël-Gimbert.
U.S. Patent No. 10,822,600 B2
The rejection of claims 1-3, 5-12, 14, 15, and 18 on the ground of nonstatutory double patenting as being unpatentable over claims 1 and 5 of U.S. Patent No. 10,822,600 B2 (cited on the IDS filed on October 20, 2021) in view of Edwards, and
the rejection of claims 16, 17, and 19 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1 and 5 of U.S. Patent No. 10,822,600 B2 in view of Edwards as applied to claims 1-3, 5-12, 14, 15, and 18 above, and further in view of Ben Chaabane.
are withdrawn in view of applicant’s amendments to claim 1 to incorporate the limitations of claims 20 and 21.
Claims 1-3, 5-12, 14, 15, 18, and 22 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1 and 5 of U.S. Patent No. 10,822,600 B2 (cited on the IDS filed on October 20, 2021) in view of Edwards and Herpoël-Gimbert.
This rejection was previously applied to claims 20-22 and has been modified to address applicant’s amendment to claim 1.
Regarding claim 1 of this application, claim 1 of the patent is drawn to a process for producing cellulolytic or hemicellulolytic enzymes comprising:
conducting a growth phase a) of a cellulolytic microorganism in a closed reactor, in the presence of at least one carbonaceous growth substrate at a concentration of between 10 and 90 g/l, at a temperature of 25-30° C and a pH of 4-5.5, and
conducting an enzyme production phase b) to produce an enzyme medium in which at least one inducer carbonaceous substrate is introduced at a temperature of 25-27° C and a pH of 4-5,
wherein
said inducer carbonaceous substrate is a pretreated pomace obtained from a pretreatment process of lignocellulosic material, said pomace not having undergone enzymatic hydrolysis and being introduced into the closed reactor in fed-batch or continuous mode,
said pretreated pomace, when suspended at room temperature at 10 wt. % of DM, has an apparent viscosity of less than 1 Pa·s at a shear rate of 10 s−1,
said pomace is introduced at a rate of between 0.3 and 0.8 gram of dry matter (“DM”) per litre of medium and per hour when in continuous mode; or, when in fed-batch mode, the amount of pomace added every f hours, f being between 0.5 h and 48 h, is between 0.3f and 0.8f grams of dry matter per litre of medium.
The claims of the patent do not recite the following limitations of instant claim 1:
the growth phase is a batch fermentation in a stirred and aerated bioreactor,
the growth and production phases are conducted at a pH of 4.0 to 4.4,
the pH of the growth phase is identical to the pH of the enzyme production phase,
the growth phase and the enzyme production phase are performed in the same bioreactor,
the process is operated in the absence of an antifoam, and
the enzymes are produced by a strain CL847 or TR3002.
Regarding a batch fermentation growth phase in a stirred and aerated bioreactor and the growth phase and the enzyme production phase are performed in the same bioreactor, Edwards generally teaches a fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures with a batch growth phase with operational parameters during the batch growth portion of the run included impeller agitation at 500 rpm and air sparging at 8 standard liters per minute (paragraph [0059]) with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
In view of Edwards, it would have been obvious to one of ordinary skill in the art before the effective filing date to perform the growth phase of the patent using batch mode fermentation at pH 4.0-4.5 with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor. One would have been motivated to and would have had a reasonable expectation of success to do so because Edwards acknowledges that the growth phase for cellulolytic enzyme production can be conducted using batch mode fermentation with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
Regarding growth and production phases being conducted at a pH of 4.0 to 4.4 and the pH of the growth phase is identical to the pH of the enzyme production phase, Edwards teaches maintaining a culture pH at 4.0-4.5 during batch growth and teaches that the pH of the cellulase production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]) and it would have been obvious to one of ordinary skill in the art to use the same pH for the enzyme production phase that is used for the batch growth phase in the claims of the patent. One would have been motivated to do this because the claims of the patent recite a growth phase pH that is between 4.0-4.5, Edwards acknowledges that the pH of a batch growth phase is 4.0-4.5 and teaches the pH of an enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4, and one of ordinary skill in the art would have recognized that using the same pH for the batch growth phase and the enzyme production phase avoids the time and materials needed to reduce the pH between the batch growth phase and the enzyme production phase.
Regarding the process is operated in the absence of an antifoam, given that the claims of the patent do not recite an antifoam and further given that the claims of the patent are presumed to be adequately described and enabled without requiring an antifoam, one of ordinary skill in the art would recognize that the process of the patent does not include or require an antifoam. Also, the method of Edwards, which is similar to the claims of the patent, does not include an antifoam and similar to the claims of the patent. Nowhere does Edwards even mention an antifoam.
Regarding the enzymes being produced by a strain CL847 or TR3002, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph).
In view of the teachings of Herpoël-Gimbert, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the process of the claims of the patent to use strain CL847 as the Trichoderma reesei cell for producing the enzymatic cocktail. One would have been motivated and would have expected success to do this because Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases and secretes a more diversified set of enzymes than strain Rut-C30.
Regarding claims 2 and 3 of this application, Edwards teaches the culture pH was maintained at 4.0-4.5 during batch growth (paragraph [0059]), Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]), and for the reasons set forth above, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. Thus, it would have been obvious to one of ordinary skill in the art before the effective filing date for the pH of the batch growth phase to be 4.0, 4.1, 4.2, 4.3, or 4.4 and for the pH of the enzyme production phase to be the same as the batch growth phase.
Regarding claim 5 of this application, Edwards teaches maintaining culture pH by addition of a 10% ammonium hydroxide solution (paragraph [0059]).
Regarding claim 6 of this application, claim 1 of the patent recites the inducer carbonaceous substrate is introduced into the closed reactor in fed-batch or continuous mode.
Regarding claim 7 of this application, the recitation of “wherein the culture medium obtained in step (a) is diluted” is interpreted as encompassing the addition of a liquid comprising a carbon source, which would have the effect of diluting the culture medium from the batch mode growth phase. Edwards teaches that following the batch growth phase and for the cellulase production phase, the carbon source was added with peristaltic pumps (paragraph [0059]).
Regarding claim 8 of this application, claim 1 of the patent recites the growth phase a) is conducted in the presence of at least one carbonaceous growth substrate at a concentration of between 10 and 90 g/l. Given that the range of concentration of carbon-based substrate of claim 1 of this application overlaps with the range of concentration of carbonaceous growth substrate of claim 1 of the patent, a prima facie case of obviousness exists.
Regarding claims 9 and 18 of this application, claim 1 of the patent recites conducting an enzyme production phase b) to produce an enzyme medium in which at least one inducer carbonaceous substrate is introduced and Edwards teaches peristaltic pumps were used to deliver the carbon source (HDC alone or a combination of HDC and CIC) at a feed rate of 0.4 grams (i.e., 400 milligrams) of carbon/liter culture/hour (paragraph [0059]).
Regarding claims 10, 12, and 22, Herpoël-Gimbert teaches Trichoderma reesei strain CL847 is an industrial mutant strain that yields a high production of cellulases (p. 2, column 1, second paragraph) and for reasons stated above, it would have been obvious to one of ordinary skill in the art before the effective filing date to modify the method of the claims of the patent to use strain CL847 as the cellulolytic microorganism for producing cellulases.
Regarding claim 11 of this application, claim 1 of the patent recites a process for producing cellulolytic or hemicellulolytic enzymes.
Regarding claim 14 of this application, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]). Aqueous ammonia is another name for ammonium hydroxide.
Regarding claim 15 of this application, the recitation of “wherein the culture medium obtained in step (a) is not diluted” is interpreted as encompassing no dilution of the culture medium of step (a) immediately before step (b). The claims of the patent do not recite and Edwards does not teach diluting the batch mode growth phase culture medium prior to the cellulase production phase.
Therefore, claims 1-3, 5-12, 14, 15, 18, and 22 of this application are unpatentable over claims 1 and 5 of the patent in view of Edwards and Herpoël-Gimbert.
Claims 16, 17, and 19 are rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1 and 5 of U.S. Patent No. 10,822,600 B2 in view of Edwards and Herpoël-Gimbert as applied to claims 1-3, 5-12, 14, 15, 18, and 22 above, and further in view of Ben Chaabane.
The claims of the patent and the teachings of Edwards and Herpoël-Gimbert as applied to claims 1-3, 5-12, 14, 15, 18, and 22 are set forth above. The claims of the patent do not recite and Edwards and Herpoël-Gimbert do not teach the cellulase production phase is operated with a stream of inductive carbon-based substrate of between 30 and 140 mg·g−1·h−1 (claim 16), the cellulase production phase is operated with a stream of inductive carbon-based substrate of between 35 and 45 mg·g−1·h−1 (claim 17), and the cellulase production phase is operated with a stream of inductive carbon-based substrate at a concentration of between 200 and 600 g/L (claim 19).
Similar to the claims of the patent and the method of Edwards, Ben Chaabane teaches a two-phase process for the production of cellulolytic and/or hemicellulolytic enzymes (paragraph [0022]) by Trichoderma reesei (paragraph [0037]). Ben Chaabane teaches that the inducing substrate in the enzyme production phase is a mixture of glucose or cellulosic hydrolysates, lactose and xylose or a solution of hemicellulolytic hydrolysates (paragraph [0022]). Specifically, Ben Chaabane teaches that the inducing substrate in the enzyme production phase is an aqueous solution comprising the inducing substrate constituted by a glucose/ lactose/xylose mixture or hemicellulose hydrolysate solution at a concentration of 450 to 550 g/L with a flow rate of 30 to 45 mg per gram of cells per hour (paragraphs [0040] to [0042]). According to Ben Chaabane, enzyme production methods including these parameters increased protein production ([0090] and [0099]) and achieved a high quality enzyme cocktail (paragraph [0111]). Ben Chaabane does not expressly teach the exact ranges as recited in claims 16, 17, and 19 of this application, however, the recited ranges overlap with the ranges taught by Ben Chaabane and according to MPEP 2144.05.I, in the case where the claimed ranges “overlap or lie inside ranges disclosed by the prior art” a prima facie case of obviousness exists.
In view of Ben Chaabane, it would have been obvious to one of ordinary skill in the art to use and deliver the inducing substrate of Ben Chaabane in the process of the claims of the patent. One would have been motivated to and would have had a reasonable expectation of success to do this because the claims of the patent, Edwards, and Ben Chaabane are related to a two-phase process for the production of cellulolytic enzymes, and Ben Chaabane teaches that enzyme production methods including these parameters increased protein production and achieved a high quality enzyme cocktail.
Therefore, claims 16, 17, and 19 of this application are unpatentable over claims 1 and 5 of the patent in view of Edwards, Herpoël-Gimbert, and Ben Chaabane.
U.S. application no. 14/239,690
Claims 1-3, 5-12, 14, 15, 18, and 22 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1 of co-pending U.S. application no. 14/239,690 (reference application) in view of Edwards.
Regarding claim 1 of this application, claim 1 of the reference application is drawn to a process for the production of cellulase enzymes in a stirred, aerated bioreactor, comprising:
(a) growing, in batch phase, a filamentous fungus strain in a culture medium comprising at least one water-soluble carbonaceous growth substrate at a concentration in the range of 10 to 30 g/L for 24 hours; and
(b) culturing the filamentous fungus strain of phase (a) in a fed batch growth and production phase, which initially (phase b1) simultaneously produces both biomass and cellulase enzymes, then reaches an equilibrium state (phase b2) wherein substantially only enzymes are produced, in the presence of at least one water-soluble inducer carbonaceous substrate, wherein the culture medium is continuously and without interruption fed with a limiting flow of carbon source in the range 80-90 mg per gram of cellular biomass per hour throughout the entire growth and production phase (b),
wherein the bioreactor has a volumetric oxygen transfer coefficient, kLa, in the range 40 to 100 h-1, and whereby equivalent cellulase productivity is obtained using a bioreactor with an oxygen transfer capacity which is at least two and a half times smaller than is needed for an analogous process comprising the same biomass concentration, in which the growth phase (a) comprises the water- soluble carbonaceous growth substrate added in excess, resulting in higher viscosity in the culture medium, and thus a higher oxygen transfer coefficient, and which analogous process does not comprise a simultaneous biomass and cellulase production phase b1, and
wherein the filamentous fungus strain is a strain of Trichoderma reesei CL847,
and
wherein in change in kLa with time is achieved substantially as represented in Fig. 5.
The claims of the patent do not recite the following limitations of instant claim 1:
the growth phase is a batch fermentation in a stirred and aerated bioreactor,
the growth and production phases are conducted at a pH of 3.8 to 4.4,
the pH of the growth phase is identical to the pH of the enzyme production phase,
the growth phase and the enzyme production phase are performed in the same bioreactor, and
the process is operated in the absence of an antifoam.
Regarding a batch fermentation growth phase in a stirred and aerated bioreactor and the growth phase and the enzyme production phase are performed in the same bioreactor, Edwards generally teaches a fermentation process using hemicellulose-derived carbohydrates for the production of cellulase mixtures with a batch growth phase with operational parameters during the batch growth portion of the run included impeller agitation at 500 rpm and air sparging at 8 standard liters per minute (paragraph [0059]) with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
In view of Edwards, it would have been obvious to one of ordinary skill in the art before the effective filing date to perform the growth phase of the reference application using batch mode fermentation at pH 4.0-4.5 with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor. One would have been motivated to and would have had a reasonable expectation of success to do so because Edwards acknowledges that the growth phase for cellulolytic enzyme production can be conducted using batch mode fermentation with impeller agitation and air sparging, with the batch growth phase and enzyme production phase being carried out in the same bioreactor.
Regarding growth and production phases being conducted at a pH of not more than 4.6 and the pH of the growth phase is identical to the pH of the enzyme production phase, Edwards teaches maintaining a culture pH at 4.0-4.5 during batch growth and teaches that the pH of the cellulase production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]) and it would have been obvious to one of ordinary skill in the art to use the same pH for the enzyme production phase that is used for the batch growth phase in the claims of the reference application. One would have been motivated to do this because Edwards acknowledges that the pH of a batch growth phase is 4.0-4.5 and teaches the pH of an enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 and one of ordinary skill in the art would have recognized that using the same pH for the batch growth phase and the enzyme production phase avoids the time and materials needed to reduce the pH between the batch growth phase and the enzyme production phase.
Regarding the process is operated in the absence of an antifoam, given that the claims of the reference application do not recite an antifoam and further given that the claims of the reference application are presumed to be adequately described and enabled without requiring an antifoam, one of ordinary skill in the art would recognize that the process of the reference application does not include or require an antifoam. Also, the method of Edwards, which is similar to the claims of the reference application, does not include an antifoam. Nowhere does Edwards even mention an antifoam.
Regarding claims 2 and 3 of this application, Edwards teaches the culture pH was maintained at 4.0-4.5 during batch growth (paragraph [0059]), Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, or 4.4 (paragraph [0034]), and for the reasons set forth above, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. Thus, it would have been obvious to one of ordinary skill in the art before the effective filing date for the pH of the batch growth phase to be 4.0, 4.1, 4.2, 4.3, or 4.4 and for the pH of the enzyme production phase to be the same as the batch growth phase in the process of the claims of the reference application.
Regarding claim 5 of this application, Edwards teaches maintaining culture pH by addition of a 10% ammonium hydroxide solution (paragraph [0059]).
Regarding claim 6 of this application, claim 1 of the reference application recites the inducer carbonaceous substrate is introduced in a fed batch growth and production phase.
Regarding claim 7 of this application, the recitation of “wherein the culture medium obtained in step (a) is diluted” is interpreted as encompassing the addition of a liquid comprising a carbon source, which would have the effect of diluting the culture medium from the batch mode growth phase. Edwards teaches that following the batch growth phase and for the cellulase production phase, the carbon source was added with peristaltic pumps (paragraph [0059]).
Regarding claim 8 of this application, claim 1 of the reference application recites the growth phase a) is conducted in the presence of at least one carbonaceous growth substrate at a concentration of between 10 and 30 g/l. Given that the range of concentration of carbon-based substrate of claim 8 of this application overlaps with the range of concentration of carbonaceous growth substrate of claim 1 of the reference application, a prima facie case of obviousness exists.
Regarding claims 9 and 18 of this application, claim 1 of the reference application recites conducting an enzyme production phase b) to produce an enzyme medium in which at least one inducer carbonaceous substrate is introduced and Edwards teaches peristaltic pumps were used to deliver the carbon source (HDC alone or a combination of HDC and CIC) at a feed rate of 0.4 grams (i.e., 400 milligrams) of carbon/liter culture/hour (paragraph [0059]).
Regarding claims 10, 12, and 22 of this application, claim 1 of the reference application recites the filamentous fungus strain is a strain of Trichoderma reesei CL847.
Regarding claim 11 of this application, claim 1 of the reference application recites a process for producing cellulase enzymes.
Regarding claim 14 of this application, Edwards teaches the culture pH was maintained by addition of a 10% ammonium hydroxide solution (paragraph [0059]). Aqueous ammonia is another name for ammonium hydroxide.
Regarding claim 15 of this application, the recitation of “wherein the culture medium obtained in step (a) is not diluted” is interpreted as encompassing no dilution of the culture medium of step (a) immediately before step (b). The claims of the reference application do not recite and Edwards does not teach diluting the batch mode growth phase culture medium prior to the cellulase production phase.
Regarding claims 21 and 22, claim 1 of the reference application recites the filamentous fungus strain is a strain of Trichoderma reesei CL847 and as stated above and reiterated here for clarity, Edwards teaches the culture pH was maintained at pH 4.0-4.5 during batch growth (paragraph [0059]), Edwards acknowledges that the pH of the enzyme production phase can be pH 4.0, 4.1, 4.2, 4.3, and 4.4 (paragraph [0034]), and for the reasons set forth above, it would have been obvious to one of ordinary skill in the art before the effective filing date to use the same pH for the enzyme production phase that is used for the batch growth phase. Thus, it would have been obvious to one of ordinary skill in the art before the effective filing date for the pH of the batch growth phase to be 4.0, 4.1, 4.2, 4.3, or 4.4 and for the pH of the enzyme production phase to be the same as the batch growth phase.
Therefore, claims 1-3, 5-12, 14, 15, 18, and 22 of this application are unpatentable over claim 1 of the reference application in view of Edwards. This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
Claims 16, 17, and 19 are provisionally rejected on the ground of nonstatutory double patenting as being unpatentable over claim 1 of co-pending U.S. application no. 14/239,690 (reference application) in view of Edwards as applied to claims 1-3, 5-12, 14, 15, 18, and 22 above, and further in view of Ben Chaabane.
The claims of the reference application and the teachings of Edwards as applied to claims 1-3, 5-12, 14, 15, 18, and 22 are set forth above. The claims of the reference application do not recite and Edwards does not teach the cellulase production phase is operated with a stream of inductive carbon-based substrate of between 30 and 140 mg·g−1·h−1 (claim 16), the cellulase production phase is operated with a stream of inductive carbon-based substrate of between 35 and 45 mg·g−1·h−1 (claim 17), and the cellulase production phase is operated with a stream of inductive carbon-based substrate at a concentration of between 200 and 600 g/L (claim 19).
Similar to the claims of the reference application and the method of Edwards, Ben Chaabane teaches a two-phase process for the production of cellulolytic and/or hemicellulolytic enzymes (paragraph [0022]) by Trichoderma reesei (paragraph [0037]). Ben Chaabane teaches that the inducing substrate in the enzyme production phase is a mixture of glucose or cellulosic hydrolysates, lactose and xylose or a solution of hemicellulolytic hydrolysates (paragraph [0022]). Specifically, Ben Chaabane teaches that the inducing substrate in the enzyme production phase is an aqueous solution comprising the inducing substrate constituted by a glucose/ lactose/xylose mixture or hemicellulose hydrolysate solution at a concentration of 450 to 550 g/L with a flow rate of 30 to 45 mg per gram of cells per hour (paragraphs [0040] to [0042]). According to Ben Chaabane, enzyme production methods including these parameters increased protein production ([0090] and [0099]) and achieved a high quality enzyme cocktail (paragraph [0111]). Ben Chaabane does not expressly teach the exact ranges as recited in claims 16, 17, and 19 of this application, however, the recited ranges overlap with the ranges taught by Ben Chaabane and according to MPEP 2144.05.I, in the case where the claimed ranges “overlap or lie inside ranges disclosed by the prior art” a prima facie case of obviousness exists.
In view of Ben Chaabane, it would have been obvious to one of ordinary skill in the art to use and deliver the inducing substrate of Ben Chaabane in the process of the claims of the reference application. One would have been motivated to and would have had a reasonable expectation of success to do this because the claims of the reference application, Edwards, and Ben Chaabane are related to a two-phase process for the production of cellulolytic enzymes, and Ben Chaabane teaches that enzyme production methods including these parameters increased protein production and achieved a high quality enzyme cocktail.
Therefore, claims 16, 17, and 19 of this application are unpatentable over claim 1 of the reference application in view of Edwards and Ben Chaabane. This is a provisional nonstatutory double patenting rejection because the patentably indistinct claims have not in fact been patented.
RESPONSE TO REMARKS: Applicant argues the combined data of record overcomes the elements allegedly rendered obvious by Edwards.
Applicant’s argument is not found persuasive. For the reasons set forth above, the combined data of record fails to rebut a prima facie case of obviousness.
Conclusion
Status of the claims:
Claims 1-3, 5-12, 14-19, and 22 are pending.
Claims 1-3, 5-12, 14-19, and 22 are rejected.
No claim is in condition for allowance.
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/David Steadman/Primary Examiner, Art Unit 1656