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 .
Claim Objections
Claims 1-5, 7-8 and 13 are objected to because of the following informalities:
Regarding claim 1, 2, 3, 5, all of these claims contain the word “respectively” but nothing is being claimed or described that would be respective to the list. This extra word should be deleted.
Regarding claim 3, The claim recites each enzyme and then recite each enzyme with the adjective thermostable, such a construction adds a lot of words wherein the subject matter is encompassed by “is thermostable” or “are thermostable”. The conjunction or should also be and as it is meant to refer to all of the enzymes as noted in dependent claims. Amending the claims to streamline the language improves readability.
Regarding claim 4, the claim recited “the” thermostable, this word is not required and appears to be a typo.
Claim 7 refers to poly(diallyldimethylammonium chloride) and then claim 8 uses the common abbreviation PDADMAC, this abbreviation should be added after the first use of the term in claim 7.
Claim 13 refers to “the immobilized cell” which are already present. These repeated words should be deleted.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
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 1-14 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.
Regarding claim 1 and claims dependent thereon, and the limitation “fermenting to obtain fermentation broths comprising an Escherichia coli or Bacillus subtilis”, it is unclear as currently worded what is being fermented. Nothing which might be fermented, such as a inoculated broth, has been claimed and thus it is unclear what is being claims. It is also unclear what is being claimed as it is unclear what the broths are. From dependent claims and the specification it appears that applicant is culturing in separate broths recombinant E. coli or recombining B. subtilis where each broth is expressing one of the five enzymes listed. Following this broths containing the five different strains are mixed to form a fermentation mixture comprising bacteria expressing all of the enzymes desired for mannose production. Amendment to make clear that a series of broths are being inoculated with a series of strains expressing the different enzymes would provide clarity.
Regarding claim 2, the claim recites “characterized in that the method comprises the steps of: fermenting to obtain fermentation broths comprising an Escherichia coli or Bacillus subtilis expressing a-glucan phosphorylase, phosphoglucomutase, phosphoglucose isomerase, mannose 6-phosphate isomerase, or mannose 6-phosphate phosphatase respectively, and mixing the above fermentation broths to obtain a fermentation mixture; adding 1-10% w/v of an inorganic soil to the fermentation mixture, and stirring homogeneously; further adding 0.1-2% w/v of a flocculant to the fermentation mixture to flocculate bacteria, then adding 0.05-3% v/v of a cross-linking agent, and cross-linking for 1-4 hours; filtering under vacuum to obtain a filter cake, extruding the filter cake to granulate into a strip with a rotary granulator, and then breaking into particles having a uniform length with a spherical shot blasting machine; and subjecting the resulting particles to drying by boiling to obtain the immobilized cells for mannose production, wherein the temperature at an air inlet for the drying by boiling is controlled at 60-90°C.” It is unclear as currently worded if applicant is claiming addition steps or defining the steps of the claim from which it depends as the claim language repeats multiple steps. This appears to be an artifact of a previous amendment changing an independent claim to a dependent claim. If these are addition steps, using the words “further comprising” would add clarity if it is clear what further steps are being added. If the previous steps of claim 1 are being further defined use of language like “wherein the inorganic soil is added in an amount of 1-10% w/v” or the like would provided clarity.
Regarding claim 5 and the limitation “the wet bacteria expressing the thermostable a-glucan phosphorylase, the thermostable phosphoglucomutase, the thermostable phosphoglucose isomerase, the thermostable mannose 6-phosphate isomerase, and the thermostable mannose 6-phosphate phosphatase respectively are mixed in a ratio of (0.1-10): (0.1-10) : (0.1-10) : (0.1-10) : (0.1-10), and the bacteria suspension obtained by mixing has an OD600 of 10-150”, The term “wet bacteria” does not have antecedent basis, this term has not been used and it is unclear what it refers to. It also appears in the claim to refer to multiple instances as a ratio is given for different enzymes. It is therefore unclear what is being claimed.
Allowable Subject Matter
The following is a statement of reasons for the indication of allowable subject matter:
The closes prior art is represented by Ma (CN109750011A/IDS submitted).
Ma teaches the in vitro synthesis of mannose from starch using a cocktail mixture of recombinant enzymes expressed and isolated separately. Ma does not teach or suggest to immobilize a mixture of cells where each cell expresses one of the enzymes and use the immobilized mixture to synthesis mannose. The claims further recite the use of spherical shot blasting as a method of forming particles which is uncommon in the field of fermentations. Applicant has further demonstrated that the immobilized cells have a superior property of reuse.
Claims 1-14 would be allowable if rewritten or amended to overcome the rejection(s) under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), 2nd paragraph, set forth in this Office action.
The following claims 1-14 drafted by the examiner and considered to distinguish patentably over the art of record in this application, are presented to applicant for consideration:
1. (Currently Amended) A method for preparing immobilized cells for mannose production, ing[[es ]]the steps of:
inoculating a series of broths wherein each broth contains a recombinant Escherichia coli or Bacillus subtilis expressing α-glucan phosphorylase, phosphoglucomutase, phosphoglucose isomerase, mannose 6-phosphate isomerase, or mannose 6-phosphate phosphatase, to obtain a series of broths expressing each of α -glucan phosphorylase, phosphoglucomutase, phosphoglucose isomerase, mannose 6-phosphate isomerase, and mannose 6-phosphate phosphatase;
[[F]]fermenting the series of broths to obtain fermentation broths comprising [[an]] Escherichia coli or Bacillus subtilis expressing α-glucan phosphorylase, phosphoglucomutase, phosphoglucose isomerase, mannose 6-phosphate isomerase, [[or]] and mannose 6-phosphate phosphatase
adding an inorganic soil to the fermentation mixture, and stirring homogeneously;
further adding a flocculant to the fermentation mixture to flocculate bacteria, and then adding a cross-linking agent for cross-linking;
filtering under vacuum to obtain a filter cake, extruding the filter cake to granulate into a strip with a rotary granulator, and then breaking into particles having a uniform length with a spherical shot blasting machine; and
subjecting the resulting particles to drying by boiling to obtain the immobilized cells for mannose production.
2. (Currently amended) The method according to claim 1, wherein
the inorganic soil is added to the fermentation mixture
the flocculant is added to the fermentation mixture to flocculate the bacteria, then the cross-linking agent is added, and the cross-linking is carried out for 1-4 hours;
3. (Currently amended) The method according to claim 1, wherein the α-glucan phosphorylase, the phosphoglucomutase, the phosphoglucose isomerase, the mannose 6-phosphate isomerase, [[or]]and the mannose 6-phosphate phosphatase [[is]]are thermostable
4. (Currently Amended) The method according to claim 3, characterized in that [[the]] thermostable refers to having an enzymatic activity at 40°C or above.
5. (Currently Amended) The method according to claim 3, characterized in that the
6. (Previously amended) The method according to claim 1, characterized in that the inorganic soil is
7. (Currently amended) The method according to claim 1, characterized in that the flocculant is selected from polyethyleneimine, chitosan, poly(diallyldimethylammonium chloride)(PDADMAC), or polyacrylamide.
8. (Currently Amended) The method of claim 7, characterized in that the flocculant is polyethyleneimine having a molecular weight of 600-70,000 Da, or PDADMAC.
9. (previously amended) The method according to claim 1, characterized in that the cross-linking agent is
10. (previously amended) The method according to claim 1, characterized in that the method further comprises a step of sieving the obtained immobilized cells to obtain morphologically uniform immobilized cells.
11. (Currently amended) A method for producing mannose with an immobilized cell, comprising contacting starch or a starch derivative with the immobilized cells of claim 1 to produce mannose.
12. (Currently Amended) The method according to claim 11, ing[[es]] a step of filtering and recovering the immobilized cells after the reaction is completed.
13. (Currently Amended) The method according to claim 11, wherein the method is performed at 50-300 g/L of starch or starch derivative, in a buffer at pH 5.0-8.0, 10-50 mM inorganic phosphate, and 3-7 mM divalent magnesium ions
14. (Currently Amended) The method according to claim [[11]]13, wherein the buffer is an HEPES buffer, a phosphate buffer, a Tris buffer, or an acetate buffer; and wherein the inorganic phosphate is sodium phosphate or potassium phosphate.
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHARLES Z CONSTANTINE whose telephone number is (571)270-5533. The examiner can normally be reached Mon-Fri 9-5.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Louise Humphrey can be reached at 571-272-5543. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/CHARLES Z CONSTANTINE/Examiner, Art Unit 1657
/LOUISE W HUMPHREY/Supervisory Patent Examiner, Art Unit 1657
Prosecution Insights