DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Election/Restrictions
Applicant's election with traverse of invention Group I, Claims 1-9 and 11-20, drawn to a preparation method for a yeast protein having an antibacterial function, in the reply filed on 06/18/2026 is acknowledged.
In Applicant’s reply, Applicant states that “the Examiner has not explained why each group lacks unity with each other group and has not described the unique special technical feature in each group” (remarks, page 2), that “[t]he Saccharomyces cerevisiae strains used in the present application differ from those in Senthilraja” (remarks, page 3), and that “[t]he method for preparing yeast protein in the present application differs from that in Senthilraja” (remarks, page 3).
The Examiner responds that, as discussed in the Restriction/Election Requirement, Groups I-I I lack unity of invention because even though the inventions of these groups require the technical feature of a yeast protein having an antibacterial function, this technical feature is not a special technical feature as it does not make a contribution over the prior art in view of Senthilraja et al. ("Isolation and Identification of Antimicrobial Protein from Saccharomyces cerevisiae and its Efficacy Against the Human Pathogens", published in 2015, Research Journal of Microbiology, Vol. 10 (1 ), pages 24-32), hereinafter 'Senthilraja'. Regarding the shared technical feature, Senthilraja teaches a yeast protein having an antibacterial function (see entire document, including abstract). Further, the technical feature of a yeast protein having an antibacterial function does not make a contribution over the prior art in view of Xu et al. (CN111513180A, published on 08/11/2020), Hu et al. (CN109207384A, published on 01/15/2019), Shuai et al. (CN103520221A, published on 01/22/2014), and Kollar et al. (“Induction and Acceleration of Yeast Lysis by Addition of Fresh Yeast Autolysate”, published in 1991, Biotechnology Letters, Vol. 13, No. 8, pages 543-546), which in combination disclose the claimed Saccharomyces cerevisiae strains and the instant method for preparing yeast protein having an antibacterial function, as discussed below.
In conclusion, Applicant’s arguments are not found persuasive. The requirement
is still deemed proper and is therefore made FINAL.
Claim Status
Claims 1-20 are pending (claim set as filed on 07/23/2024). Claim 10 is withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Applicant timely traversed the restriction (election) requirement in the reply filed on 06/18/2026.
Claims 1-9 and 11-20 are currently under examination and were examined on their merits.
Priority
This application filed on 07/23/2024 claims priority to PCT application no. PCT/CN2022/142484, filed on 12/27/2022, and claims foreign priority to application no. CN202111631106.8, filed on 12/28/2021. Acknowledgment is made of applicant’s claim for foreign priority under 35 U.S.C. 119 (a)-(d). Receipt is acknowledged of certified copies of papers required by 37 CFR 1.55.
Information Disclosure Statement
The Information Disclosure Statements (IDS) filed on 05/28/2026 and 06/26/2024 have been received and considered.
Claim Rejections - 35 USC § 112(a)
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 1-9 and 11-20 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention.
The microorganisms Saccharomyces cerevisiae FX-2 CCTCC NO: M2016418 and Saccharomyces cerevisiae d8.8 CCTCC NO: M2017148 are recited in the claims and, thus, are essential to the claimed invention. Because the microorganisms are essential to the claimed invention, they must be obtainable by a repeatable method set forth in the specification or otherwise readily available to the public. If the microorganisms are not so obtainable or available, the requirements of 35 U.S.C. § 112 may be satisfied by a deposit of the biological materials.
The specification does not disclose a repeatable process to obtain the microorganisms, and it is not apparent if the biological materials are readily available to the public. It is noted that applicant has deposited the organisms (see specification page 5), but there is no indication in the specification as to public availability.
If the deposit is made under the Budapest Treaty, then an affidavit or declaration by applicant, or a statement by an attorney of record over his or her signature and registration number, stating that the specific strain has been deposited under the Budapest Treaty and that the specific strain will be irrevocably and without restriction or condition released to the public upon the issuance of a patent, would satisfy the deposit requirement made herein.
If the deposit has not been made under the Budapest Treaty, then in order to certify that the deposit meets the criteria set forth in 37 C.F.R. §§ 1.801-1.809, applicant may provide assurance of compliance by an affidavit or declaration, or by a statement by an attorney of record over his or her signature and registration number, showing that:
(a) during the pendency of this application, access to the invention will be afforded to the Commissioner upon request;
(b) all restrictions upon availability to the public will be irrevocably removed upon granting of the patent;
(c) the deposit will be maintained in a public depository for a period of 30 years or 5 years after the last request or for the effective life of the patent, whichever is longer;
(d) a test of the viability of the biological material at the time of deposit will be made (see 37 C.F.R. §1.807); and
(e) the deposit will be replaced if it should ever become inviable.
Applicant’s attention is directed to M.P.E.P. § 2400 in general, and specifically to § 2411.05, as well as to 37 C.F.R. § 1.809(d), wherein it is set forth that “the specification shall contain the accession number for the deposit, the date of the deposit, the name and address of the depository, and a description of the deposited material sufficient to specifically identify it and to permit examination.” It is noted that the instant specification fails to provide any information about whether the deposit was made under the Budapest Treaty or whether the deposited organism is available to the public; the specification should be amended to include this information.
Claim Rejections - 35 USC § 112 (b)
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 6-8 and 14-20 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claims 6, 7, and 14 recite “the process”, and claim 20 recites the “spray drying process” which is indefinite for lacking antecedent basis because ‘process’ is not recited within the claims or in the claims from which claims 6, 7, 14 and 20 depend. One of ordinary skill in the art would not be able to determine the metes and bounds of the claims, and thus, could not clearly determine how to avoid infringement of claims 6, 7, 14, and 20.
In the interest of compact prosecution, claims 6, 7, and 14 are interpreted to the broadest embodiment claimed.
Dependent claim 16 which depends from claim 6, is rejected because it does not clarify the indefinite language in claim 6. Dependent claims 8 and 17-19 which depend from claim 7, are further rejected since they do not clarify the indefinite language in claim 7.
Claim 15 recites “the compound protease is protease EF108” (page 9, line 1), wherein ‘EF108’ is a product name (see specification, page 8, paragraph 4) and EF108 is not further described in the specification. The claim scope is uncertain since the product name cannot be used properly to identify any particular material or product. The product name is used to identify a source of goods, and not the goods themselves. Thus, the product name does not identify or describe the goods associated with the product name. In the present case, the product name ‘EF108’ is used to identify/describe a compound protease and, accordingly, the identification/description is indefinite. One of ordinary skill in the art would not be able to determine the metes and bounds of the claim, and thus, could not clearly determine how to avoid infringement of claim 15.
In the interest of compact prosecution, claim 15 is interpreted to the broadest embodiment claimed.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
Determining the scope and contents of the prior art.
Ascertaining the differences between the prior art and the claims at issue.
Resolving the level of ordinary skill in the pertinent art.
Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-2, 5-9, 11-12, 15-20 are rejected under 35 U.S.C. 103 as being unpatentable over Xu et al. (CN111513180A, published on 08/11/2020), hereinafter ‘Xu’, in view of Hu et al. (CN109207384A, published on 01/15/2019), hereinafter ‘Hu’, in view of Shuai et al. (CN103520221A, published on 01/22/2014), hereinafter ‘Shuai’, and as evidenced by Jackisch (EP1531178A1, published on 05/18/2005), hereinafter ‘Jackisch’.
Xu’s general disclosure relates to a feed-grade high-protein nucleotide-type yeast hydrolysate, its preparation method, and its application (see entire document, including paragraph [0002]).
Regarding claim 1, pertaining to a preparation method, Xu teaches a preparation method for yeast protein (paragraphs [0002], [0018]-[0021]), comprising the following steps:
S1 performing liquid fermentation on a Saccharomyces cerevisiae strain to obtain yeast milk (paragraphs [0019], [0067]), wherein the Saccharomyces cerevisiae strain is Saccharomyces cerevisiae d8.8 CCTCC NO: M2017148 (paragraphs [0017], [0067]). It is noted that the term yeast milk indicates separation of the yeast cells from the fermentation broth, as evidenced by Jackisch (paragraph [0023]). Xu teaches
S2, acidifying the yeast milk to obtain an acidified yeast milk (paragraphs [0028]);
S3, performing autolysis (paragraphs [0020], [0068]-[0069]), wherein a concentration of the autolysis solution is 18% by mass of dry yeast (S3) (paragraphs [0068]-[0069]);
S4, performing enzymolysis on the autolysis solution with a compound protease to obtain an enzymolysis solution (paragraphs [0021], [0030]-[0031]);
S6, evaporating and concentrating the enzyme solution (paragraph [0035]).
It is noted that since Xu’s dried enzyme solution comprises all cell components from the claimed strain, it inherently comprises the claimed yeast protein having an antibacterial function.
Regarding claim 2, pertaining to the fermentation medium in S1, Xu teaches a carbon source, a nitrogen source, and a phosphorus source are added by using a fed-batch method during liquid fermentation (paragraph [0023], [0055]), wherein the carbon source is molasses (paragraph [0024], [0055]), the nitrogen source is ammonia water (paragraphs [0055], [0067]), and the phosphorus source is ammonium dihydrogen phosphate (paragraph [0055]). Since the recited nutrients are provided by fed batch in Xu’s method (paragraph [0023]), Xu’s ammonium dihydrogen phosphate reads on an ammonium dihydrogen phosphate solution.
Regarding claims 5 and 15, please note the 112b rejection of claim 15 above.
Pertaining to the compound protease, Xu teaches wherein the compound protease comprises papain, neutral protease, alkaline protease, and acid protease (paragraph [0082], [0093]).
Regarding claim 6, please note the 112b rejection above. Pertaining to the compound protease, Xu teaches that “[a] compound enzyme preparation containing 6‰ papain, 0.5‰ alkaline protease, 0.5‰ neutral protease, and 0.5‰ acidic protease was added based on the dry matter of the yeast milk,” (paragraph [0082]), which corresponds to a final concentration of the compound protease of 7.5‰ of the weight of dry bases in the autolysis solution.
Regarding claim 9, pertaining to S6, Xu teaches wherein the method comprises:
Heating the yeast protein emulsion to 80°C to perform enzyme inactivation for 2h (paragraph [0059]); and
evaporating and concentrating the yeast protein emulsion that has been subjected to enzyme inactivation, until a solid content reaches 35-45% (paragraph [0059]), so as to obtain a yeast protein product. Since Xu’s dried yeast protein product comprises all cell components of the claimed strain, it inherently comprises the claimed yeast protein having an antibacterial function.
Regarding claim 11, pertaining to the carbon, nitrogen, and phosphorus source, Xu teaches wherein a total sugar content in the molasses is 28.5-31.5% (paragraph 0024]), a nitrogen content in the ammonia water is 16% (paragraph [0067]), and a phosphorus content in ammonium dihydrogen phosphate is 27.0% (paragraph [0056]).
Regarding claim 12, pertaining to the fermentation, Xu teaches wherein a temperature during the liquid fermentation is 35°C, a fermentation time is 12 h, and a pH value of the fermentation was 6.0 (paragraph [0078]).
Regarding claim 16, please note the 112b rejection above. Pertaining to the enzymolyis with compound protease, the enzymolysis temperature is 59°C, a pH is 6.0, and an enzymolysis temperature is 9 h (paragraph [0071]).
Regarding claim 20, please note the 112b rejection above. Pertaining to spray drying, Xu teaches wherein the preparation method further comprises performing spray drying on a concentrated solution obtained in the evaporation and concentration step (paragraphs [0074]-[0075]); and in the spray drying process, an inlet air temperature is controlled at 155 °C-175 °C, an outlet air temperature is controlled at 90 °C-115 °C, an atomization pressure is controlled at 100 bar-160 bar, and a material-receiving temperature is controlled at 25 °C-45 °C (paragraph [0075]). Since Xu’s spray dried preparation comprises all cell components from the claimed strain, it inherently comprises the claimed yeast protein having an antibacterial function.
Xu does not teach wherein the Saccharomyces cerevisiae strain is Saccharomyces cervisiae FX-2 CCTCC NO: M2016418 (S1, instant claim 1).
Xu does not expressly teach mixing the acidified yeast milk (S3, instant claim 1).
Xu does not teach performing second enzymolysis on the first enzymolysis solution with seminase to obtain a second enzymolysis solution; and performing third enzymolysis on the second enzymolysis solution with beta-glucanase and cellulase to obtain a yeast protein emulsion (S5, instant claim 1).
Xu does not teach wherein in the second enzymolysis process, an addition amount of the seminase is 4‰ -8‰ of the weight of dry bases in the first enzymolysis solution (instant claim 7), wherein an enzymolysis temperature of the second enzymolysis process is 55°C-60°C, a pH is 5.0-6.0, and an enzymolysis time is 6 h-12 h (instant claim 17).
Xu does not teach wherein in the third enzymolysis process, an addition amount of the beta-glucanase is 3‰-6‰ of the weight of dry bases in the second enzymolysis solution, and an addition amount of the cellulase is 1‰-3‰ of the weight of the dry bases in the second enzymolysis solution (instant claim 8), wherein an enzymolysis temperature of the third enzymolysis process is 45°C-55°C, a pH is 4.0-5.0, and an enzymolysis time is 8 h-16 h (instant claim 18), and wherein in the third enzymolysis process, a pH value is regulated to 4.0-5.0 by using an organic acid and/or inorganic acid; and the organic acid is one or more of malic acid, citric acid, and lactic acid, the inorganic acid is hydrochloric acid and/or sulfuric acid, and the pH value is regulated by using the organic acid (instant claim 19).
Xu does not teach wherein the phosphorus content in the ammonium dihydrogen phosphate solution is 9%-11% (instant claim 11).
Hu’s general disclosure relates to a modified yeast cell wall and its preparation thereof (see entire document, including paragraph [0002]).
Regarding claim 1, Hu teaches Saccharomyces cerevisiae FX-2 CCTCC NO: M2016418 (paragraph [0041]), and further teaches an enzymolysis step with beta-glucanase and cellulase of autolysed yeast cell wall (paragraph [0037]) to hydrolyse the cell wall (paragraphs [0017], [0059]).
Regarding claims 8 and 18-19, please note the 112b rejection above.
Regarding claim 8, pertaining to enzyme amounts, Hu teaches wherein an addition amount of the beta-glucanase and cellulase is each 0.2-1‰ based on the mass of the yeast cell wall dry matter (see Hu’s claim 8).
Regarding claim 18, pertaining to enzymolysis with beta-glucanase and cellulase, Hu teaches wherein enzymolysis by beta-glucanase is performed at a temperature of 40-67℃ and a pH of 4.5-6.5 for 6-12h (paragraphs [0030] and[0036]), and wherein enzymolysis by cellulase is performed at a temperature of 50-68℃ and a pH of 4.0-5.5 for 3-8h (paragraphs [0029] and [0035]).
Shuai’s general disclosure relates to a process for producing medical dried yeast powder (see entire document, including abstract).
Regarding claim 1, Shuai teaches seminase as a cell wall breaking enzyme to produce a yeast powder (see Shuai’s claim 2).
Regarding claims 7 and 17, please note the 112b rejection above.
Regarding claim 7, pertaining to the enzyme amount, Shuai teaches about 1.6‰ seminase based on the weight of the total yeast suspension (see embodiment 3 on page 3).
Regarding claim 17, pertaining to enzymolysis with seminase, Shuai teaches wherein the seminase enzymolysis is performed at a temperature of 50°C for 7 h (see embodiment 3 on page 3).
Although Xu does not teach wherein the Saccharomyces cerevisiae strain is Saccharomyces cerevisiae strain FX-2 CCTCC NO: M2016418 (S1, instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined Xu’s method with Hu’s Saccharomyces cerevisiae strain FX-2 CCTCC NO: M2016418 to create a method wherein yeast proteins are prepared from Saccharomyces cerevisiae strain FX-2 CCTCC NO: M2016418. One would have been motivated to substitute Xu’s strain d8.8 with Hu’s strain FX-2 in Xu’s method since Hu teaches using strain FX-2 for cell autolysis and enzymolysis (paragraphs [0020]-[0021]).
Although modified Xu does not expressly teach mixing the acidified yeast milk (S3, instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have mixed the acidified yeast milk. One would have been motivated to do so to ensure even distribution of the added acid taught by Xu in the yeast milk (paragraph [0028]).
Although modified Xu does not teach performing a second enzymolysis with seminase on Xu’s enzymolysis solution comprising compound protease, to obtain a second enzymolysis solution, and performing third enzymolysis on the second enzymolysis solution with beta-glucanase and cellulase to obtain a yeast protein emulsion (S5, instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined Xu’s method with Shuai’s teachings on seminase as a cell wall breaking enzyme and with Hu’s teachings on beta-glucanase and cellulase for cell wall hydrolysis, to create a method comprising an enzymolysis step with seminase, and enzymolysis steps with beta-glucanase and cellulase. One would have been motivated to do so to increase degradation of the cell wall and thereby increase the amount of released protein. A skilled artisan would have reasonably expected success in combining Xu’s, Shuai’s and Hu’s teachings, since all references are directed to yeast lysis.
Although modified Xu does not specify combining beta-glucanase and cellulase in a single enzymolysis step, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined beta-glucanase and cellulase in a single enzymolysis step since Hu’s teaches similar enzymolysis temperatures, pH values and times for both enzymes (paragraph [0030], [0036]), [0029] and [0035]). One would have been motivated to do so to simplify the method and to accelerate hydrolysis of the cell wall.
Although modified Xu does not specify the sequence of enzymolysis steps wherein the second enzymolysis with seminase is performed on the first enzymolysis solution comprising compound protease, and the third enzymolysis with beta-glucanase and cellulase is performed on the second enzymolysis solution, the instantly recited sequence of steps would have been within the realm of routine experimentation of a skilled artisan. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to determine the optimal sequence of enzymolysis steps for maximal cell wall breakdown.
Regarding the yeast protein emulsion in S5 and the prepared yeast protein having antibacterial function in S6 in instant claim 1, it is noted that the claimed yeast protein emulsion and the prepared yeast protein having antibacterial function naturally result from performing the claimed method steps S1-S5 and S1-S6, respectively. Since Xu in view of Hu, and Shuai teaches the claimed method steps S1-S6, it is highly likely (i.e.obvious) that modified Xu’s method results in the same or highly similar yeast protein emulsion in S5 and yeast protein having antibacterial function in S6.
Although modified Xu does not teach wherein in the second enzymolysis process, an addition amount of the seminase is 4‰ -8‰ of the weight of dry bases in the first enzymolysis solution (instant claim 7), wherein an enzymolysis temperature of the second enzymolysis process is 55°C-60°C, a pH is 5.0-6.0, and an enzymolysis time is 6 h-12 h (instant claim 17), and further does not teach wherein in the third enzymolysis process, an addition amount of the beta-glucanase is 3‰-6‰ of the weight of dry bases in the second enzymolysis solution, and an addition amount of the cellulase is 1‰-3‰ of the weight of the dry bases in the second enzymolysis solution (instant claim 8), wherein an enzymolysis temperature of the third enzymolysis process is 45°C-55°C, a pH is 4.0-5.0, and an enzymolysis time is 8 h-16 h (instant claim 18), the instantly recited enzyme concentrations, temperatures, pH values and enzymolysis times of the second and third enzymolysis would be within the realm of routine experimentation since enzyme concentrations, temperature, pH values and enzymolysis times are known result-effective variables in enzymolysis. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to determine the optimal enzyme concentrations, temperatures, pH values, and enzymolysis times in the second and third enzymolyis for maximal cell wall degradation.
Although modified Xu does not teach wherein in the third enzymolysis process, a pH value is regulated to 4.0-5.0 by using an organic acid and/or inorganic acid, regulating a pH value using an organic acid and/or inorganic acid is within the skill of an artisan. Although modified Xu does not teach wherein the organic acid is one or more of malic acid, citric acid, and lactic acid, the inorganic acid is hydrochloric acid and/or sulfuric acid, and the pH value is regulated by using the organic acid (instant claim 19), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have used citric acid to regulate the pH value. One would have been motivated to do so to simplify the method by reducing the number of required chemicals since Xu teaches the preferred use of citric acid for acidifying the yeast milk (paragraph [0028]).
Although modified Xu does not teach wherein the phosphorus content in the ammonium dihydrogen phosphate solution is 9%-11% (instant claim 11), the recited phosphorus content would be within the realm of routine experimentation of a skilled artisan, since fermentation is a well known technique in the art and nutrient concentrations including phosphorus are result-effective variables. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to manipulate the phosphorus concentration in the solution to obtain maximal yeast growth during fermentation.
Regarding the recited protein content in instant claim 12, it is noted that the recited protein content greater than or equal to 50% in the obtained yeast milk would naturally result from performing the method according to instant claim 12. Since modified Xu teaches the claimed strains and the claimed method according to claim 12, it is highly likely (i.e. obvious) that modified Xu’s yeast milk has the same or highly similar protein content.
Claims 1, 3, and 4 are rejected under 35 U.S.C. 103 as being unpatentable over Xu et al. (CN111513180A, published on 08/11/2020), hereinafter ‘Xu’, in view of Hu et al. (CN109207384A, published on 01/15/2019), hereinafter ‘Hu’, in view of Kollar et al. (“Induction and Acceleration of Yeast Lysis by Addition of Fresh Yeast Autolysate”, published in 1991, Biotechnology Letters, Vol. 13, No. 8, pages 543-546), hereinafter ‘Kollar’, in view of Shuai et al. (CN103520221A, published on 01/22/2014), hereinafter ‘Shuai’, and as evidenced by Jackisch (EP1531178A1, published on 05/18/2005), hereinafter ‘Jackisch’.
Xu’s general disclosure relates to a feed-grade high-protein nucleotide-type yeast hydrolysate, its preparation method, and its application (see entire document, including paragraph [0002]).
Regarding claim 1, pertaining to a preparation method, Xu teaches a preparation method for yeast protein (paragraphs [0002], [0018]-[0021]), comprising the following steps:
S1 performing liquid fermentation on a Saccharomyces cerevisiae strain to obtain yeast milk (paragraphs [0019], [0067]), wherein the Saccharomyces cerevisiae strain is Saccharomyces cerevisiae d8.8 CCTCC NO: M2017148 (paragraphs [0017], [0067]). It is noted that the term yeast milk indicates separation of the yeast cells from the fermentation broth, as evidenced by Jackisch (paragraph [0023]). Xu teaches
S2, acidifying the yeast milk to obtain an acidified yeast milk (paragraphs [0028]);
Performing autolysis (paragraphs [0020], [0068]-[0069]), wherein a concentration of the autolysis solution is 18% by mass of dry yeast (S3) (paragraphs [0068]-[0069]). Xu’s autolysis of yeast milk reads on the optional autolysis in S2 and autolysis in S3. Since the instant cell wall emulsion in S2 naturally results from performing yeast milk autolysis, Xu’s yeast milk autolysis inherently results in a cell wall emulsion. Xu teaches
S4, performing enzymolysis on the autolysis solution with a compound protease to obtain an enzymolysis solution (paragraphs [0021], [0030]-[0031]);
S6, evaporating and concentrating the enzyme solution (paragraph [0035]).
Since modified Xu’s dried enzyme solution comprises all cell components from the claimed Saccharomyces cerevisiae d8.8, it inherently comprises the claimed antibacterial yeast protein.
Regarding claim 4, Xu teaches wherein the yeast milk is acidified by using citric acid (paragraph [0057]); an addition amount of the citric acid is 2% or 3% of dry matter weight of the yeast milk (paragraphs [0080], [0091]).
Xu does not teach wherein the Saccharomyces cerevisiae strain is Saccharomyces cervisiae FX-2 CCTCC NO: M2016418 (S1, instant claim 1).
Xu does not teach mixing the acidified yeast milk and the cell wall emulsion obtained from autolysis (S3, instant claim 1).
Xu does not teach performing second enzymolysis on the first enzymolysis solution with seminase to obtain a second enzymolysis solution; and performing third enzymolysis on the second enzymolysis solution with beta-glucanase and cellulase to obtain a yeast protein emulsion (S5, instant claim 1).
Xu does not teach wherein in S1, Xu’s yeast milk prepared from Saccharomyces cerevisiae d8.8 CCTCC NO: M2017148 is a first yeast milk, and wherein S1 further comprises performing liquid fermentation on the Saccharomyces cerevisiae FX-2 CCTCC NO: M2016418, and then performing separation to obtain a second yeast milk; in S2, the first yeast milk is acidified to obtain the acidified yeast milk; and the autolysis is performed on the second yeast milk to obtain the cell wall emulsion; and in S3, the acidified yeast milk and the cell wall emulsion are mixed and subjected to autolysis, so as to obtain the autolysis mixed solution (instant claim 3).
Xu does not teach wherein in S2, an addition amount of the citric acid is 1%-4% of the weight of the yeast milk (instant claim 4).
Hu’s general disclosure relates to a modified yeast cell wall and its preparation thereof (see entire document, including paragraph [0002]).
Regarding claim 1, Hu teaches Saccharomyces cerevisiae FX-2 CCTCC NO: M2016418 (paragraph [0041]), and further teaches an enzymolysis step with beta-glucanase and cellulase of autolyzed yeast cell wall (paragraph [0037]) to hydrolyze the cell wall (paragraphs [0017], [0059]).
Kollar’s general disclosure relates to accelerating yeast cell autolysis by adding fresh yeast autolysate (see entire document, including abstract).
Regarding claims 1 and 3, Kollar teaches mixing a yeast cell suspension with fresh yeast autolysate to accelerate cell autolysis (see abstract).
Shuai’s general disclosure relates to a process for producing medical dried yeast powder (see entire document, including abstract).
Regarding claim 1, Shuai teaches seminase as a cell wall breaking enzyme to produce a yeast powder (see Shuai’s claim 2).
Although Xu does not teach wherein the Saccharomyces cerevisiae strain is Saccharomyces cerevisiae strain FX-2 CCTCC NO: M2016418 (S1, instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined Xu’s method with Hu’s Saccharomyces cerevisiae strain FX-2 CCTCC NO: M2016418 to create a method wherein yeast proteins are prepared from Saccharomyces cerevisiae strain FX-2 CCTCC NO: M2016418. One would have been motivated to substitute Xu’s strain d8.8 with Hu’s strain FX-2 in Xu’s method since Hu teaches using strain FX-2 for cell autolysis and enzymolysis (paragraphs [0020]-[0021]).
Although modified Xu does not teach mixing the acidified yeast milk and the cell wall emulsion obtained from autolysis (S3, instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined modified Xu’s method with Kollar’s teaching on accelerating autolysis, to have mixed the acidified yeast milk and the cell wall emulsion obtained from autolysis. One would have been motivated to do so to accelerate autolysis of the yeast milk. A skilled artisan would have reasonably expected success in combining modified Xu’s and Kollar’s teachings since both are directed to yeast autolysis.
Although modified Xu does not teach performing a second enzymolysis with seminase on the Xu’s enzymolysis solution comprising compound protease, to obtain a second enzymolysis solution, and performing third enzymolysis on the second enzymolysis solution with beta-glucanase and cellulase to obtain a yeast protein emulsion (S5, instant claim 1), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined Xu’s method with Shuai’s teachings on seminase as a cell wall breaking enzyme and with Hu’s teachings on beta-glucanase and cellulase for cell wall hydrolysis, to create a method comprising an enzymolysis step with seminase, and enzymolysis steps with beta-glucanase and cellulase. One would have been motivated to do so to increase degradation of the cell wall and thereby increase the amount of released protein. A skilled artisan would have reasonably expected success in combining Xu’s, Hu’s, Shuai’s and Kollar’s teachings, since all references are directed to yeast lysis.
Although modified Xu does not specify combining beta-glucanase and cellulase in a single enzymolysis step, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined beta-glucanase and cellulase in a single enzymolysis step since Hu’s teaches similar enzymolysis temperatures, pH values and times for both enzymes (paragraph [0030], [0036]), [0029] and [0035]). One would have been motivated to do so to simplify the method and to accelerate hydrolysis of the cell wall.
Although modified Xu does not specify the sequence of enzymolysis steps wherein the second enzymolysis with seminase is performed on the first enzymolysis solution comprising compound protease, and the third enzymolysis with beta-glucanase and cellulase is performed on the second enzymolysis solution, the instantly recited sequence of steps would have been within the realm of routine experimentation of a skilled artisan. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to determine the optimal sequence of enzymolysis steps for maximal cell wall breakdown.
Regarding the yeast protein emulsion in S5 and the prepared yeast protein having antibacterial function in S6, it is noted that the claimed yeast protein emulsion and the prepared yeast protein having antibacterial function naturally result from performing the claimed method steps S1-S5 and S1-S6, respectively. Since Xu in view of Hu, Kollar, and Shuai teaches the claimed method steps S1-S6, it is highly likely (i.e.obvious) that modified Xu’s method results in the same or highly similar yeast protein emulsion in S5 and yeast protein having antibacterial function in S6.
Although modified Xu does not teach wherein in S1, Xu’s yeast milk prepared from Saccharomyces cerevisiae d8.8 CCTCC NO: M2017148 is a first yeast milk, and wherein S1 further comprises performing liquid fermentation on the Saccharomyces cerevisiae FX-2 CCTCC NO: M2016418 to obtain a second yeast milk; wherein in S2, the first yeast milk is acidified to obtain the acidified yeast milk; and the autolysis is performed on the second yeast milk to obtain the cell wall emulsion; and in S3, the acidified yeast milk and the cell wall emulsion are mixed and subjected to autolysis, so as to obtain the autolysis mixed solution (instant claim 3), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have further modified Xu’s method wherein a yeast milk from each claimed strain d8.8 and FX-2 is prepared (first and second yeast milk), wherein the first yeast milk is acidified and the second yeast milk is subjected to autolysis, and the resulting autolysate (cell wall emulsion) obtained from the second yeast milk is added to the first yeast milk. One would have been motivated to do so to accelerate the autolysis of the first yeast milk and to increase the amount of released protein in the autolysis mixed solution. Although modified Xu does not specify wherein the first yeast milk is prepared from Saccharomyces cerevisiae d8.8 CCTCC NO: M2017148, and the second yeast milk is obtained from Saccharomyces cerevisiae FX-2 CCTCC NO: M2016418, the instant autolysis conditions would have been within the realm of routine experimentation of a skilled artisan to determine which strain provides a more suitable autolysate as an accelerator for yeast milk autolysis. One would have been motivated to do so to obtain a final autolysate preparation with maximal protein content.
Although modified Xu does not teach wherein in S2, an addition amount of the citric acid is 1%-4% of the weight of the yeast milk (instant claim 4), the recited concentration of citrate would be within the realm of routine experimentation, since autolysis is well-known and the amount of added acid is a result-effective variable. It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have determined the optimal concentration of citrate based on the weight of yeast milk, in order to optimize the autolysis reaction. Regarding the protein content in the cell wall emulsion recited in instant claim 4, it is noted that a protein content in the cell wall emulsion of 15%-30% would naturally result from performing the method according to claim 4. Since modified Xu teaches the claimed strains and method, it is highly likely, i.e. obvious that in S2, a protein content in the obtained cell wall emulsion is 15%-30%.
Claims 1, 4, and 13-14 are rejected under 35 U.S.C. 103 as being unpatentable over Xu et al. (CN111513180A, published on 08/11/2020), hereinafter ‘Xu’, in view of Hu et al. (CN109207384A, published on 01/15/2019), hereinafter ‘Hu’, in view of Kollar et al. (“Induction and Acceleration of Yeast Lysis by Addition of Fresh Yeast Autolysate”, published in 1991, Biotechnology Letters, Vol. 13, No. 8, pages 543-546), hereinafter ‘Kollar’, in view of Shuai et al. (CN103520221A, published on 01/22/2014), hereinafter ‘Shuai’, as evidenced by Jackisch (EP1531178A1, published on 05/18/2005), hereinafter ‘Jackisch’, and in view of Griffon (US 3,975,553; published on 08/17/1976), hereinafter ‘Griffon’.
Xu’s, Hu’s, Kollar’s, and Shuai’s teachings have been set forth above.
Regarding claim 13, pertaining to autolysis, Xu teaches adjusting the solid content before performing autolysis (paragraphs [0057], [0080], and [0091]).
In addition, Xu teaches a protein content of 50-60% in a yeast hydrolysate (paragraphs [0045] and [0047])
Regarding claim 14, please note the 112b rejection above. Pertaining to autolysis, Xu teaches wherein an autolysis temperature is 53°C (paragraph [0091]), and a temperature holding time is 10 h (paragraph [0102]).
Modified Xu does not teach wherein S3 comprises: mixing the acidified yeast milk and the cell wall emulsion to obtain a premixed solution with a protein content greater than or equal to 35%; and adjusting a solid content of the premixed solution, and then performing heating with steam to perform autolysis, so as to obtain the autolysis mixed solution (instant claim 13).
Modified Xu does not teach wherein in the process of performing heating with steam to perform autolysis, a temperature is controlled at 45°C-55 °C, and a temperature-holding time is 8 h-14 h (instant claim 14).
Griffon’s general disclosure relates to deproteinated yeast cell membrane product being produced by autolysis of fresh compressed yeast (see entire document, including abstract).
Regarding claim 13, pertaining to autolysis, Griffon teaches performing heating with steam to perform autolysis (column 4, lines 1-7; see abstract).
Regarding claim 14, pertaining to autolysis, Griffon teaches an autolysis temperature of 55°C (column 3, lines 6-11; column 4, lines 6-7).
Although modified Xu does not teach wherein S3 comprises: mixing the acidified yeast milk and the cell wall emulsion to obtain a premixed solution with a protein content greater than or equal to 35%; and adjusting a solid content of the premixed solution (instant claim 13), the recited protein content and adjusting the solid content in the premixed solution would be within the realm of routine experimentation since autolysis is a well-known technique, and Xu further teaches a protein content of 50-60% in a yeast hydrolysate (paragraphs [0045] and [0047]) and adjusting the solids content before autolysis (paragraphs [0057], [0080], and [0091]). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have determined the optimal protein content in the premixed solution and to have adjusted the solid content in the premixed solution to maximize the amount of released protein during autolysis.
Although modified Xu does not teach heating with steam to perform autolysis (instant claim 13), it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have combined modified Xu’s method with Griffon’s teachings on steam heating, in order to create a method that comprises performing heating with steam to perform autolysis. One would have been motivated to do so to accelerate the autolysis reaction. A skilled artisan would have reasonably expected success in combining modified Xu’s with Griffon’s teachings since both are directed to yeast autolysis.
Although modified Xu does not teach wherein in the process of performing heating with steam to perform autolysis, a temperature is controlled at 45°C-55°C, and a temperature-holding time is 8 h-14 h (instant claim 14), the recited temperatures and holding times would be within the realm of routine experimentation of a skilled artisan since autolysis and the result-effective variables temperature and holding time are well-known in the art, and Xu further teaches an autolysis temperature of 53°C (paragraph [0091]), and a holding time of 10 h (paragraph [0102]). It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to have determined the optimal temperature and holding time to maximize yeast autolysis.
Conclusion
No claims are allowed.
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/SANDRA ZINGARELLI/ Examiner, Art Unit 1653
/SHARMILA G LANDAU/ Supervisory Patent Examiner, Art Unit 1653
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