DETAILED ACTION
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
The amendment filed May 12, 2026, has been received and entered.
Claims 2, 4, 6-10, 13, 15-17, and 23-38 are cancelled.
Claim 40 is new.
Claims 1, 3, 5, 11, 12, 14, 18-22, 39, and 40 are pending and examined on the merits.
Notice Re: Prior Art Available Under Pre-AIA and AIA
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.
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.
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.
Claim 40 is rejected under 35 U.S.C. 103 as being unpatentable over Kalum (US 2007/0292938. Previously cited) in view of Ason (JP 2013053083. Machine Translation cited below. Previously cited).
Kalum discloses a method for extracting one or more components from yeast cells, wherein the one or more components to be extracted comprises a protein (claims 1 and 8 of Kalum; paragraph [0007]). The method comprises treating yeast cells with a phospholipase (claim 1 of Kalum; paragraph [0003]). The phospholipase activity may be provided by enzymes having other activities as well, such as a lipase with phospholipase activity (paragraph [0024]). The yeast can be Saccharomyces cerevisiae (paragraph [0005]). Therefore, Kalum meets limitations of the claimed invention by disclosing contacting Saccharomyces cerevisiae with the enzyme lipase (lipase with phospholipase activity).
In extracting proteins and separating the one or more components (including protein) from the treated yeast cells (claims 1 and 8 of Kalum) wherein the yeast is S. cerevisiae, then the method of Kalum is directed to a ‘method for optimizing a protein content extracted from Saccharomyces cerevisiae’ as claimed, wherein the steps result in ‘optimizing the protein content extracted from S. cerevisiae’ as claimed.
Additionally, Kalum discloses that the skilled person will know how to adjust parameters such as pH to achieve the desired results when discussing suitable conditions under which to perform the treatment of phospholipase (paragraph [0072]). In one embodiment of the invention, treatment with phospholipase is conducted at a pH between 2 and 10, such as between 3 and 9 (paragraph [0072]).
Kalum differs from the claimed invention in that Kalum does not expressly disclose that their step of treating S. cerevisiae with a lipase (a lipase with phospholipase activity) at a pH between 2 and 10, such as between 3 and 9, further includes contacting the S. cerevisiae with β-glucanase. Kalum further differs from the claimed invention in that Kalum does not disclose that after that step, there is a step of purifying a composition comprising a protein content ranging between 60% and 80% by weight of said composition.
Kalum discloses that in one or more embodiments, the one or more components to be extracted is a yeast extract (paragraph [0015]). Also, the value of yeast extracts depends to a high degree on the amount of protein in the extract, there is thus a desire to achieve the highest possible protein yield (paragraph [0015]).
Ason discloses a method for producing yeast protein comprising treating yeast cells with a cell wall lytic enzyme (page 3, second paragraph; claim 4 of Ason). The yeast can be Saccharomyces cerevisiae (page 2, second paragraph of ‘DESCRIPTION-OF-EMBDOIMENT’ section). The cell wall lytic enzyme has almost no protease activity and can be a β-glucanase (page 3, third paragraph). Thereafter, the method comprises heat treatment at 50ºC or higher (preferably 50 to 100ºC), and separation of a fraction containing mainly protein (page 3, fifth paragraph; abstract). The yeast protein obtained by the method can have a protein content of 60% or more, or 80% or more (page 3, seventh paragraph). Additionally, in the examples of Ason, the treatment with β-glucanase (FiltraseBRX, Denateam GEL) was performed at pH 4.5 and pH 6.0 (page 3, last paragraph; page 4, second paragraph; page 3, third paragraph for names of the β-glucanase of the examples).
Before the effective filing date of the claimed invention, it would have been obvious to the person of ordinary skill in the art to further include a β-glucanase with the lipase having phospholipase activity when performing the method of Kalum in order to also treat the S. cerevisiae with a β-glucanase. One of ordinary skill in the art would have been motivated to do treat the S. cerevisiae with both the lipase having a phospholipase activity and a β-glucanase because it would have enabled acquisition of yeast protein having a high protein content, as indicated in Ason; this would have been desirable for the invention of Kalum since Kalum recognizes the value of a yeast product having the highest possible protein yield (paragraph [0015]). There would have been a reasonable expectation of performing the invention of Kalum with the combination of the lipase and a β-glucanase because both Kalum and Ason disclose that S. cerevisiae can successfully be treated with these enzymes for obtaining protein from yeast cells. Also, in performing the method rendered obvious by Kalum in view of Ason, it would have been obvious to the person of ordinary skill in the art to perform the enzyme treatment at a pH suitable for both phospholipase and β-glucanase as taught in Kalum and Ason, specifically pH 4.5 taught in Ason for β-glucanase treatment which falls in the pH ranges disclosed in Kalum of a pH between 2 and 10, such as between 3 and 9. The pH of 4.5 falls withing the claimed pH of ‘ranging between 3.5 and 5.5.’ Therefore, Kalum in view of Ason renders obvious step a) of instant claim 40.
Regarding step b) of instant claim 40, Ason discloses that following the reaction with the cell wall lytic enzyme (e.g. β-glucanase), a heat treatment is performed, and then centrifuged (page 3, fifth paragraph). Then, the cell wall constituents are removed by a machine to obtain a fraction mainly composed of proteins (page 3, fifth paragraph). The fraction containing mainly protein is used as it is are dried to obtain yeast protein (page 3, fifth paragraph). As pointed out above, Ason teaches that the yeast protein obtained by their method can have a protein content of 60% or more, or 80% or more (page 3, seventh paragraph).
In performing the method rendered obvious by Kalum in view of Ason, it would have been obvious to perform the steps taught by Ason for obtaining a fraction containing mainly protein as described in the preceding paragraph. One of ordinary skill in the art would have been motivated to do this in order to obtain a yeast protein having a protein of 60% or more, or 80% or more, which suggest a protein content ranging between 60% and 80% by weight of the yeast protein. The steps taught by Ason after β-glucanase treatment are directed to purifying a composition comprising a protein content ranging between 60% and 80% by weight of said composition. Thus Kalum in view of Ason also renders obvious step b) of instant claim 40. There would have been a reasonable expectation that the protein content of the yeast protein obtained by the method rendered obvious by Kalum in view of Ason is between 60% and 80% by weight of the yeast protein because Ason teaches that their treatment involving β-glucanase resulted in yeast protein of a protein content of 60% or more, or 80% or more (page 3, seventh paragraph).
Therefore, Kalum in view of Ason renders obvious instant claim 40.
Claims 1, 3, 5, 11, 12, 14, and 19-22 are rejected under 35 U.S.C. 103 as being unpatentable over Menin (AU 2019260558 A1. Listed on IDS filed 12/4/25) in view of Siddiqi (Biotechnology and Bioengineering. 1996. 50: 145-150) and Morel (EP 3378334).
Menin discloses a method for obtaining yeast proteins (abstract). The method starts with cell lysis, preferably via thermal plasmolysis, and then subjecting the material to an enzyme of glucanase type and an enzyme of ribonuclease type, followed by separation after which the insoluble fraction represents the product of interest having a content of true proteins of at least 72% (page 2, line 30 through page 3, line 3). See also page 7, lines 11-23 for one embodiment of the invention. When discussing the broader scope of the invention, Menin discloses that a yeast cream is obtained by harvesting a culture of yeast by centrifugation or filtration, which can be washed (page 5, lines 10-14). The cells are then subjected to mechanical or chemical rupture using known methods such as high-pressure homogenization, mechanical grinding, and ultrasound disintegration (page 5, lines 15-18). In one preferred embodiment of the invention, the yeast is subjected to thermal plasmolysis (page 5, lines 18-20). The mechanical rupture (e.g., high-pressure homogenization, mechanical grinding, ultrasound disintegration) and its alternative, thermal plasmolysis, of the yeast cells is directed to breaking the cell wall of yeast material comprising a cell wall, meeting some limitations of step (a) of instant claim 1.
The material obtained from the cell lysis (via mechanical rupture, e.g., high-pressure homogenization, or thermal plasmolysis) is a whole material containing a soluble fraction and insoluble fraction (page 5, lines 25-32). That material is then subjected to one or more enzyme activities with the objective of solubilizing the maximum number of constituents that are not proteins, without attacking the proteins (page 5, line 30 through page 6, line 2). Preferably ribonuclease activity and glucanase activity (EC 3.2.1) are used, which can be implemented sequentially or simultaneously (page 6, lines 2-4). This enzymatic step allows a novel soluble fraction and an insoluble fraction essentially comprising the proteins (page 6, lines 13-16). Therefore, Menin meets some limitations of step (b) of instant claim 1 by disclosing contacting the yeast material with enzymes comprising glucanase. Menin further discloses separation of the soluble fraction and insoluble fraction (page 6, line 17). As pointed out above, the insoluble fraction has a content of true proteins of at least 72% (page 2, line 30 through page 3, line 3). See also page 8, lines 29-30 of Menin, in combination with page 4, lines 4-10, which makes clear that the at least 72% protein is in terms of dry matter. As such, the separation of the soluble fraction and the insoluble fraction is directed to purifying a first fraction (Menin’s soluble fraction) comprising soluble materials and a second fraction (Menin’s insoluble fraction) comprising insoluble proteins, wherein the second fraction comprises a protein content overlapping with the claimed protein content of ‘between 10% and 90% by dry weight’ of the second fraction, meeting some limitations of step (b) of instant claim 1. In obtaining the yeast proteins, then the method of Menin results in optimizing the protein content extracted from the yeast material comprising a cell wall (yeast cells), meeting a limitation of instant claim 1.
Menin differs from the claimed invention in that Menin does not expressly disclose:
The cell lysis (by mechanical rupture, e.g. high pressure homogenization, or thermal plasmolysis) breaks the cell wall of the yeast material (yeast cells) to a particle size between 2 µm and 10 µm;
Their step of treating the lysed yeast cells (obtained after mechanical rupture or thermal plasmolysis) with enzymes comprising contacting the yeast with enzymes comprising lipase and beta-glucanase, and the first fraction (soluble fraction of Menin) comprises soluble proteins, wherein each one of the first fraction (soluble fraction of Menin) and the second fraction (insoluble fraction of Menin) comprises a protein fraction between 10% and 90% by dry weight of said first fraction and said second fraction, respectively.
Regarding difference (1) (The cell lysis breaks the cell walls of the yeast materials into a particle size between 2 µm and 10 µm):
Siddiqi discloses a study which took measurements of size distributions for the breakage of commercial packed bakers’ yeast cells, i.e. Saccharomyces cerevisiae (page 146, left column, first paragraph), as a function of operating pressure and number of passes through a specific high-pressure homogenizer (abstract). In the background of the study, Siddiqi discusses the disruption of cells in high-pressure homogenization for the isolation of intracellular products such as proteins (page 145, left column, first two paragraphs). In their study, >99% of the total mass is >1 µm in size (page 146, right column, second-to-last paragraph). Figure 1 on page 147 shows the particle size distribution of packed baker’s yeast cells homogenized at 500 bar operating pressure and varying passes, including no pass (N = 0). The initial size distribution (i.e., N = 0) had a modal size of approximately 4.5 µm (page 146, right column, last paragraph; see N = 0 in Figure 1). It is evident from Figure 1 that the initial cells ranged in diameter from about 2.2 µm to about 6 µm. The sizes range in Figure 1 was reduced for the various passes through the homogenizer, with sizes ranging from about 1 µm to about 5.5 µm. See also Figure 3 on page 147 showing a median size (d50) of the homogenate size distribution with various number of passes, ranging from as low as about 2.5 µm to as high as about 4.2 µm.
Before the effective filing date of the claimed invention, it would have been obvious to the person of ordinary skill in art to expect that the mechanical rupture, e.g., high-pressure homogenization, or thermal plasmolysis would have broken the yeast cells, in particular Menin’s preferred embodiment is Saccharomyces cerevisiae cells (page 5, lines 1-4), to a particle size that falls in the claimed range of between 2 µm and 10 µm, in particular a range of greater than 1 µm and less than about 6 µm which renders obvious the claimed particle size because of overlap, when performing the method of Menin. One of ordinary skill in the art would have expected this based on Siddiqi which discloses the particle size distribution of Saccharomyces cerevisiae from high-pressure homogenization, and based on the size of Saccharomyces cerevisiae cells as provided by the initial particle size distribution in Siddiqi without any pass through a high-pressure homogenizer – the broken S. cerevisiae cells must be smaller than the maximum size of about 6 µm of intact S. cerevisiae cells.
Regarding difference (2) (The enzymatic treatment of Menin does not comprise contacting the lysed cells (obtained after mechanical rupture, e.g., high pressure homogenization, or thermal plasmolysis) with enzymes comprising lipase and β-glucanase; the soluble fraction of Menin comprises soluble proteins; each one of the first fraction, i.e. the soluble fraction of Menin, and the second fraction, i.e., the insoluble fraction of Menin, comprises a protein content between 10% and 90% by dry weight of said first fraction and said second fraction, respectively):
Menin discloses that the obtained protein extract still containing lipids derived from the membrane can be delipidated with methods known to persons skilled in the art, including via treatment with lipases or phospholipases followed by separation from the solubilized phase (page 3, lines 9-12). See also page 6, line 30 through page 7, line 3, which teaches that optionally, the amount of lipids in the insoluble fraction can be reduced through the action of a lipase or phospholipase, achieving true protein purities of more than 80%.
Morel discloses a method for producing a flavour composition comprising contacting disrupted yeast cells with “a glucanase and/or endoprotease” to obtain a flavour composition (which can be liquid) comprising a soluble fraction, wherein the process is able to efficiently solubilize the disrupted yeast cell walls (paragraph [0019]). The disrupted yeast cells are obtained with standard techniques known in the art, such a physical process with milling or grinding (paragraph [0020]). In a preferred embodiment, the glucanase is a laminaripentaose-producing β-1,3-glucanase (paragraph [0030]). Laminaripentaose-producing β-1,3-glucanase is efficiently used for degrading the glucans which are present in the yeast cell walls into smaller glucan fragments, such as laminaripentaoses, and degradation of the glucans into glucan fragments having a length of 1 to 50 kDa is found to efficiently solubilize the yeast cell walls (paragraph [0030]). In a preferred embodiment, the amount of protein or protein fragments in the flavour composition is within the range of 0.5 to 40%, more preferably within the range of 1 to 40%, more preferably within the range of 5 to 35%, most preferably within the range of 10, 15, 20, or 25 to 35% (w/w, on carrier free dry matter) (paragraph [0043]). In a preferred embodiment, the proteins are water soluble or substantially water soluble (paragraph [0055]).
Before the effective filing date of the claimed method, it would have been obvious to the person of ordinary skill in the art to further include lipase and/or phospholipase (directed to a lipase, meeting the limitation of instant claim 14) in the enzymatic treatment step of the method rendered obvious by Menin in view of Siddiqi. One of ordinary skill in the art would have been motivated to do this in order to remove constituents that are not proteins, which would have been sought based on the teachings of Menin for the enzyme treatment (page 5, line 30 through page 6, line 2). Since Menin itself recognizes using a lipase or a phospholipase to reduce the amount of lipids in their insoluble fraction to increase the protein content, then one of ordinary skill in the art would have been motivated to include lipase and/or phospholipase with the glucanase in the enzyme treatment step of the method rendered obvious by Menin in view of Siddiqi.
Moreover, before the effective filing date of the claimed method, it would have been obvious to the person of ordinary skill in the art to substitute the glucanase of the method rendered obvious by Menin and Siddiqi with laminaripentaose-producing β-1,3-glucanase (directed to β-glucanase) for the predictable result of treating the lysed yeast cells (lysed S. cerevisiae cells) with a glucanase activity that allows wall polysaccharides to be solubilized to soluble oligosaccharides as taught in Menin (page 6, lines 8-10). It would have been an obvious matter of simple substitution of one known glucanase for another. Additionally, one of ordinary skill in the art would have been motivated to make this substitution because Morel successfully used laminaripentaose-producing β-1,3-glucanase for treating disrupted yeast cells to obtain a desirable protein-containing product, specifically a soluble fraction, wherein this enzyme efficiently degrades the glucans which are present in the yeast cell walls into smaller glucan fragments, and degradation of the glucans into glucan fragments having a length of 1 to 50 kDa is found to efficiently solubilize yeast cell walls. There would have been a reasonable expectation of practicing the method rendered obvious by Menin in view of Siddiqi with laminaripentaose-producing β-1,3-glucanase since Menin teaches using glucanase in general for practicing their invention, which the laminaripentaose-producing β-1,3-glucanase is directed to.
In performing the method rendered obvious by Menin in view of Siddiqi and Morel, which includes contacting yeast material (broken to a particle size rendering obvious the claimed limitation) with enzymes comprising lipase (and/or phospholipase) and β-glucanase, then it would have been obvious to the person of ordinary skill in the art that the soluble fraction necessarily comprises soluble proteins and is directed to the claimed ‘first fraction’ and each of the soluble fraction (claimed first fraction) and the insoluble fraction (claimed second fraction comprising insoluble proteins) comprises a protein content that meets the claimed limitation of being ‘between 10% to 90% by dry weight’ of said first fraction and said second fraction, respectively. One of ordinary skill in the art would have expected this because Menin found that with lipase treatment, the protein content is increased to 80% (by dry weight, which falls in the claimed range) in the insoluble fraction, and Morel found that treating disrupted yeast cells with laminaripentaose-producing β-1,3-glucanase is used for obtaining a soluble fraction comprising soluble proteins in an amount of most preferably within the range of 10, 15, 20, or 25 to 35% (w/w, on carrier free dry matter). Moreover, since Menin in view of Siddiqi and Morel renders obvious the claimed steps of breaking the cell wall of yeast material to a particle size between 2 µm and 10 µm, and contacting the yeast material with enzyme comprising lipase (and/or phospholipase) and β-glucanase, then it follows that the resulting separated soluble and insoluble fractions necessarily have the same properties as claimed.
Therefore, Menin in view of Siddiqi and Morel render obvious instant claims 1, 3 (high-pressure homogenization), 5 (Saccharomyces cerevisiae as discussed above, see page 5, lines 1-4 of Menin), and 14.
Regarding instant claim 11, Menin teaches that most of the protein profile of the obtained yeast protein extract is distributed around 40-45 kDa (page 10, lines 7-9). The yeast protein extract is the soluble fraction of Menin. Therefore, in the invention rendered obvious by Menin, Siddiqi, and Morel, it would have been obvious that at least one protein of the insoluble proteins is characterized by a molecular weight around 40-45 kDa which falls in the claimed range of ‘between 1 kDa and 250 kDa.’ Therefore, instant claim 11 is rendered obvious.
Regarding instant claim 12, Menin discloses that the solubility of a protein extract of their invention is less than 3.5% in water (page 10, line 3). This converts to 35 g/L aqueous solubility. The protein extract of Menin is the soluble fraction. Therefore, in the invention rendered obvious by Menin, Siddiqi, and Morel, it would have been obvious that a protein of the insoluble proteins is characterized by an aqueous solubility of less than 35 g/L which overlaps with the claimed range of ‘less than 300 g/L.’ Thus, instant claim 12 is rendered obvious.
Regarding instant claim 19, the references differ from the claimed invention in that they do not expressly disclose repeating and applying pressure between 150 bars and 1500 bars. However, Siddiqi used high-pressure homogenization (limitation of parent instant claim 3) such that S. cerevisiae cells were homogenized at 500 bar under multiple passes (Figure 1 on page 147). For the method rendered obvious by Menin in view of Siddiqi and Morel, it would have been obvious to the person of ordinary skill in the art to perform high-pressure homogenization at 500 bar under multiple passes because it would have successfully ruptured the S. cerevisiae cells based on Siddiqi. Since 500 bars falls in the claimed range of ‘between 150 bars and 1500 bars’ and multiple passes in the homogenizer is directed to repeating applying that pressure, then instant claim 19 is rendered obvious.
Regarding instant claim 20, Menin teaches that initially, a culture of the yeast is carried out, the biomass is harvested by centrifuge or filtration, and then can be washed (page 5, lines 10-14). This is directed to a step comprising washing. Therefore, instant claim 20 (washing) is rendered obvious.
Regarding instant claim 21, Morel discloses obtaining a liquid fraction (abstract). The liquid fraction is the soluble fraction comprising soluble proteins. Therefore, in performing the method rendered obvious by Menin in view of Siddiqi and Morel, it would have been obvious to the person of ordinary skill in the art that the soluble fraction (directed to the first fraction) is in the form of a solution (as it comprises the water-soluble components, including water-soluble protein). Thus, instant claim 21 (solution) is rendered obvious.
Regarding instant claim 22, Menin discloses that the product obtained (their insoluble fraction) can be freeze-dried or dried with a method known to skilled person, such as spray drying and vacuum drying (page 7, lines 8-10). In drying the insoluble fraction (directed to the claimed second fraction) in the method rendered obvious by Menin in view of Siddiqi and Morel, then proteins, including any single protein, is concentrated since water is removed. Therefore, instant claim 22 (concentrating) is rendered obvious.
Claim 18 is rejected under 35 U.S.C. 103 as being unpatentable over Menin, Siddiqi, and Morel as applied to claims 1, 3, 5, 11, 12, 14, and 19-22 above, and further in view of Kalum (US 2007/0292938. Previously cited).
As discussed above, Menin in view of Siddiqi and Morel renders obvious claims 1, 3, 5, 11, 12, 14, and 19-22. The references differ from claim 18 in that they do not expressly disclose that the enzymatic reaction is performed at a pH between 3.5 and 5.5.
As discussed above, Morel teaches using a glucanase, preferably laminaripentaose-producing β-1,3-glucanase, for obtaining a soluble fraction comprising soluble proteins from disrupted yeast cells. In a preferred embodiment, Morel teaches that the set of contacting the disrupted yeast cell walls with a glucanase is carried out at a pH within the range of 3-10, more preferably within the range of 4 to 8 or from 4 to 7, such as from 4.5 to 6.0 or from 5.0 to 6.0 (paragraph [0023]).
Kalum discloses a method for extracting one or more components from yeast cells, wherein the one or more components to be extracted comprises a protein (claims 1 and 8 of Kalum; paragraph [0007]). The method comprises treating yeast cells with a phospholipase (claim 1 of Kalum; paragraph [0003]). Additionally, Kalum discloses that the skilled person will know how to adjust parameters such as pH to achieve the desired results when discussing suitable conditions under which to perform the treatment of phospholipase (paragraph [0072]). In one embodiment of the invention, treatment with phospholipase is conducted at a pH between 2 and 10, such as between 3 and 9 (paragraph [0072]).
Before the effective filing date of the claimed invention, it would have been obvious to the person of ordinary skill in the art to perform the enzymatic treatment at a pH suitable for both the phospholipase (as rendered obvious above, directed to a lipase) and the β-glucanase (laminaripentaose-producing β-1,3-glucanase) when performing the method rendered obvious by Menin in view of Siddiqi and Morel as the pH is a condition for reaction of the enzymes. The full pH range suitable for phospholipase of between 2 and 10, including between 3 and 9, as taught in Kalum includes the full pH range disclosed in Morel for glucanase (preferably laminaripentaose-producing β-1,3-glucanase). The narrower pH range of Morel overlaps the claimed range, with significant overlap of the preferred pH ranges of Morel with the claimed range. Therefore, instant claim 18 is rendered obvious.
Claim 39 is rejected under 35 U.S.C. 103 as being unpatentable over Menin, Siddiqi, and Morel as applied to claims 1, 3, 5, 11, 12, 14, and 19-22 above, and further in view of Ason (JP 2013053083. Machine Translation cited below. Previously cited).
As discussed above, Menin in view of Siddiqi and Morel renders obvious claims 1, 3, 5, 11, 12, 14, and 19-22. The references differ from claim 39 in that they do not expressly disclose that the enzymes further comprise any one of the recited enzymes or any combination thereof.
Ason discloses a method for producing yeast protein comprising treating yeast cells with a cell wall lytic enzyme (page 3, second paragraph; claim 4 of Ason). The yeast can be Saccharomyces cerevisiae (page 2, second paragraph of ‘DESCRIPTION-OF-EMBDOIMENT’ section). The cell wall lytic enzyme has almost no protease activity and can be a β-glucanase (page 3, third paragraph). Also, Ason discloses that the cell wall lytic enzyme used in their method includes glucanase and mannanase (page 3, third paragraph).
Before the effective filing date of the claimed invention, it would have been obvious to the person of ordinary skill in the art to further include mannanase with the lipase (and/or phospholipase, directed to a lipase) and the β-glucanase (laminaripentaose-producing β-1,3-glucanase) when performing the method rendered obvious by Menin in view of Siddiqi and Morel. One of ordinary skill in the art would have been motivated to do this because mannanase is recognized as a cell wall lytic enzyme suitable for producing yeast protein from yeast cells, thereby assisting with obtaining yeast proteins as sought by Menin. It would have been an obvious matter of combining enzymes recognized for the same purpose of obtaining yeast proteins. See MPEP 2144.06(I). Thus, instant claim 39 is rendered obvious.
Claim 40 is rejected under 35 U.S.C. 103 as being unpatentable over Menin (AU 2019260558 A1. Listed on IDS filed 12/4/25) in view of Morel (EP 3378334) and Kalum (US 2007/0292938. Previously cited).
Menin discloses a method for obtaining yeast proteins (abstract). The method starts with cell lysis, preferably via thermal plasmolysis, and then subjecting the material to an enzyme of glucanase type and an enzyme of ribonuclease type, followed by separation after which the insoluble fraction represents the product of interest having a content of true proteins of at least 72% (page 2, line 30 through page 3, line 3). See also page 7, lines 11-23 for one embodiment of the invention.
The material obtained from the cell lysis (via mechanical rupture, e.g., high-pressure homogenization, or thermal plasmolysis; see page 5, lines 15-29) is a whole material containing a soluble fraction and insoluble fraction (page 5, lines 25-32). That material is then subjected to one or more enzyme activities with the objective of solubilizing the maximum number of constituents that are not proteins, without attacking the proteins (page 5, line 30 through page 6, line 2). Preferably ribonuclease activity and glucanase activity (EC 3.2.1) are used, which can be implemented sequentially or simultaneously (page 6, lines 2-4). This enzymatic step allows a novel soluble fraction and an insoluble fraction essentially comprising the proteins (page 6, lines 13-16). Preferred yeasts include Saccharomyces cerevisiae (page 5, lines 1-4). Therefore, Menin meets some limitations of step (a) of instant claim 40 by disclosing contacting Saccharomyces cerevisiae with enzymes comprising glucanase.
Menin further discloses separation of the soluble fraction and insoluble fraction (page 6, line 17). As pointed out above, the insoluble fraction has a content of true proteins of at least 72% (page 2, line 30 through page 3, line 3). As such, the separation of the soluble fraction and the insoluble fraction is directed to purifying a composition (Menin’s insoluble fraction) comprising a protein content overlapping with the claimed protein content of ‘between 60% and 80% by weight’ of the composition, meeting limitations of step (b) of instant claim 40. In obtaining the yeast proteins, then the method of Menin results in optimizing the protein content extracted from S. cerevisiae, meeting a limitation of instant claim 40.
Menin differs from the claimed invention in that Menin does not expressly disclose that the enzymatic treatment is contacting the S. cerevisiae with enzymes comprising lipase and beta-glucanase under pH ranging between 3.5 and 5.5.
Menin discloses that the obtained protein extract still containing lipids derived from the membrane can be delipidated with methods known to persons skilled in the art, including via treatment with lipases or phospholipases followed by separation from the solubilized phase (page 3, lines 9-12). See also page 6, line 30 through page 7, line 3, which teaches that optionally, the amount of lipids in the insoluble fraction can be reduced through the action of a lipase or phospholipase, achieving true protein purities of more than 80%.
Morel discloses a method for producing a flavour composition comprising contacting disrupted yeast cells with “a glucanase and/or endoprotease” to obtain a flavour composition (which can be liquid) comprising a soluble fraction, wherein the process is able to efficiently solubilize the disrupted yeast cell walls (paragraph [0019]). The disrupted yeast cells are obtained with standard techniques known in the art, such a physical process with milling or grinding (paragraph [0020]). In a preferred embodiment, the glucanase is a laminaripentaose-producing β-1,3-glucanase (paragraph [0030]). Laminaripentaose-producing β-1,3-glucanase is efficiently used for degrading the glucans which are present in the yeast cell walls into smaller glucan fragments, such as laminaripentaoses, and degradation of the glucans into glucan fragments having a length of 1 to 50 kDa is found to efficiently solubilize the yeast cell walls (paragraph [0030]). In a preferred embodiment, the amount of protein or protein fragments in the flavour composition is within the range of 0.5 to 40%, more preferably within the range of 1 to 40%, more preferably within the range of 5 to 35%, most preferably within the range of 10, 15, 20, or 25 to 35% (w/w, on carrier free dry matter) (paragraph [0043]). In a preferred embodiment, the proteins are water soluble or substantially water soluble (paragraph [0055]).
Before the effective filing date of the claimed method, it would have been obvious to the person of ordinary skill in the art to further include lipase and/or phospholipase (directed to a lipase) in the enzymatic treatment step of the method of Menin. One of ordinary skill in the art would have been motivated to do this in order to remove constituents that are not proteins, which would have been sought based on the teachings of Menin for the enzyme treatment (page 5, line 30 through page 6, line 2). Since Menin itself recognizes using a lipase or a phospholipase to reduce the amount of lipids in their insoluble fraction to increase the protein content, then one of ordinary skill in the art would have been motivated to include lipase and/or phospholipase with the glucanase in the enzyme treatment step of the method of Menin.
Moreover, before the effective filing date of the claimed method, it would have been obvious to the person of ordinary skill in the art to substitute the glucanase of the method rendered obvious by Menin with laminaripentaose-producing β-1,3-glucanase (directed to β-glucanase) for the predictable result of treating the lysed yeast cells (lysed S. cerevisiae cells) with a glucanase activity that allows wall polysaccharides to be solubilized to soluble oligosaccharides as taught in Menin (page 6, lines 8-10). It would have been an obvious matter of simple substitution of one known glucanase for another. Additionally, one of ordinary skill in the art would have been motivated to make this substitution because Morel successfully used laminaripentaose-producing β-1,3-glucanase for treating disrupted yeast cells to obtain a desirable protein-containing product, specifically a soluble fraction, wherein this enzyme efficiently degrades the glucans which are present in the yeast cell walls into smaller glucan fragments, and degradation of the glucans into glucan fragments having a length of 1 to 50 kDa is found to efficiently solubilize yeast cell walls. There would have been a reasonable expectation of practicing the method rendered obvious by Menin with laminaripentaose-producing β-1,3-glucanase since Menin teaches using glucanase in general for practicing their invention, which the laminaripentaose-producing β-1,3-glucanase is directed to.
In performing the method rendered obvious by Menin in view of Morel, which includes contacting S. cerevisiae with enzymes comprising lipase (and/or phospholipase) and β-glucanase, then the insoluble fraction (claimed composition that is purified) comprises a protein content that meets the claimed limitation of being ‘between 60% to 80% by weight.’ One of ordinary skill in the art would have expected this because Menin found that with lipase treatment, the protein content is increased to 80% (by dry weight, which falls in the claimed range) in the insoluble fraction. Moreover, since Menin in view of Morel renders obvious the claimed steps, then it follows that the resulting separated insoluble fraction (the claimed ‘composition’ that is purified) necessarily have the same properties as claimed.
Regarding the claimed pH:
As discussed above, Morel teaches using a glucanase, preferably laminaripentaose-producing β-1,3-glucanase, for obtaining a soluble fraction comprising soluble proteins from disrupted yeast cells. In a preferred embodiment, Morel teaches that the set of contacting the disrupted yeast cell walls with a glucanase is carried out at a pH within the range of 3-10, more preferably within the range of 4 to 8 or from 4 to 7, such as from 4.5 to 6.0 or from 5.0 to 6.0 (paragraph [0023]).
Kalum discloses a method for extracting one or more components from yeast cells, wherein the one or more components to be extracted comprises a protein (claims 1 and 8 of Kalum; paragraph [0007]). The method comprises treating yeast cells with a phospholipase (claim 1 of Kalum; paragraph [0003]). Additionally, Kalum discloses that the skilled person will know how to adjust parameters such as pH to achieve the desired results when discussing suitable conditions under which to perform the treatment of phospholipase (paragraph [0072]). In one embodiment of the invention, treatment with phospholipase is conducted at a pH between 2 and 10, such as between 3 and 9 (paragraph [0072]).
Before the effective filing date of the claimed invention, it would have been obvious to the person of ordinary skill in the art to perform the enzymatic treatment at a pH suitable for both the phospholipase (as rendered obvious above, directed to a lipase) and the β-glucanase (laminaripentaose-producing β-1,3-glucanase) when performing the method rendered obvious by Menin in view of Morel as the pH is a condition for reaction of the enzymes. The full pH range suitable for phospholipase of between 2 and 10, including between 3 and 9, as taught in Kalum includes the full pH range disclosed in Morel for glucanase (preferably laminaripentaose-producing β-1,3-glucanase). The narrower pH range of Morel overlaps the claimed range, with significant overlap of the preferred pH ranges of Morel with the claimed range. Therefore, instant claim 40 is rendered obvious.
Response to Arguments
Applicant’s arguments, filed May 12, 2026, with respect to the objection to the specification, the objections to claims 1, 3, 5, 11, 12, 14, 18-22, and 39, the rejection under 35 U.S.C. 112(b) of claims 1, 3, 5, 11, 12, 14, 18-22, and 39, the rejection under 35 U.S.C. 112(a) of claims 1, 3, 5, 11, 12, 14, 18-22, and 39, and the rejection under 35 U.S.C. 103 of claims 1, 3, 5, 11, 12, 14, 18, 20-22, and 39 as being unpatentable over Kalum in view of Ason, have been fully considered and are persuasive. In particular, the objection to the specification has been overcome by the amendment to the specification. The objections to the claims have been overcome by the amendments to claims 1, 3, 5, and 39. The rejections under 35 U.S.C. 112(b) have been overcome by the amendments to claims 1, 5, 11, 12, and 39. The rejection under 35 U.S.C. 112(a) has been overcome by the amendment to claim 1. The rejection under 35 U.S.C. 103 has been overcome by the amendment to claim 1 because Kalum and Ason do not expressly disclose breaking the cell wall of the yeast material to a particle size between 2 µm and 10 µm, prior to contacting the yeast material with enzymes comprising lipase and β-glucanase. Therefore, these objections and rejections have been withdrawn.
However, upon further consideration, a new ground(s) of rejection is made in view of the previously cited references Kalum and Ason in order to render obvious new claim 40, and the newly cited references Menin, Siddiqi, and Morel in order to render obvious claim 1 and its dependent claims as well as new claim 40.
Conclusion
No claims are allowed.
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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/SUSAN E. FERNANDEZ/Examiner, Art Unit 1651
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