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 .
Applicants’ Amendment
1) Acknowledgment is made of Applicants’ amendment filed 01/29/26 in response to the non-final Office Action mailed 10/31/25.
Status of Claims
2) Claims 1-10 have been amended via the amendment filed 01/29/2026.
New claims 11-18 have been added via the amendment filed 01/29/2026.
Claims 1-18 are pending and are under examination.
Prior Citation of Title 35 Sections
3) The text of those sections of Title 35 U.S. Code not included in this action can be found in a prior Office Action.
Prior Citation of References
4) The references cited or used as prior art in support of one or more rejections in the instant Office Action and not included on an attached form PTO-892 or form PTO-1449 have been previously cited and made of record.
Objection(s) Withdrawn
5) The objection to the specification set forth in paragraph 14(a) of the Office Action mailed 10/31/25 is withdrawn in light of Applicants’ amendments to the specification.
6) The objection to claim 10 set forth in paragraph 14(b) of the Office Action mailed 10/31/25 is withdrawn in light of Applicants’ amendments to the claim.
7) The objection to claims 1, 7 and 9 set forth in paragraph 14(c) of the Office Action mailed 10/31/25 is withdrawn in light of Applicants’ amendments to the claims.
Objection(s) Maintained
8) The objection to claim 8 set forth in paragraph 14(c) of the Office Action mailed 10/31/25 is maintained. The issue raised therein remains.
Rejection(s) Withdrawn
9) The rejection of claims 1 and 6-9 set forth on paragraph 8(a) of the Office Action mailed 10/31/25 under 35 U.S.C § 112(b) or 35 U.S.C § 112 (pre-AIA ), second paragraph, as being indefinite is withdrawn in light of Applicants’ claim amendments.
10) The rejection of claims 3, 5 and 10 set forth on paragraph 8(b) of the Office Action mailed 10/31/25 under 35 U.S.C § 112(b) or 35 U.S.C § 112 (pre-AIA ), second paragraph, as being indefinite is withdrawn in light of Applicants’ claim amendments.
11) The rejection of claim 1 set forth on paragraph 8(c) of the Office Action mailed 10/31/25 under 35 U.S.C § 112(b) or 35 U.S.C § 112 (pre-AIA ), second paragraph, as being indefinite is withdrawn in light of Applicants’ claim amendment.
12) The rejection of claim 10 set forth on paragraph 8(d) of the Office Action mailed 10/31/25 under 35 U.S.C § 112(b) or 35 U.S.C § 112 (pre-AIA ), second paragraph, as being indefinite is withdrawn in light of Applicants’ claim amendments.
13) The rejection of claims 2-10 set forth on paragraph 8(e) of the Office Action mailed 10/31/25 under 35 U.S.C § 112(b) or 35 U.S.C § 112 (pre-AIA ), second paragraph, as being indefinite is withdrawn in light of Applicants’ claim amendments.
14) The rejection of claims 9-10 set forth on paragraph 8(f) of the Office Action mailed 10/31/25 under 35 U.S.C § 112(b) or 35 U.S.C § 112 (pre-AIA ), second paragraph, as being indefinite is withdrawn in light of Applicants’ amendments to the base claim.
15) The rejection of claims 1-9 set forth on paragraph 10 of the Office Action mailed 10/31/25 under 35 U.S.C § 102(a)(1) as being anticipated by WO 2016172282 A1 (WO ‘282, of record) is withdrawn in light of Applicants’ claim amendments. Applicants’ arguments are moot in light of the withdrawal of the rejection.
16) The rejection of claims 1-9 set forth on paragraph 11 of the Office Action mailed 10/31/25 under 35 U.S.C § 102(a)(1) or 102(a)(2) as being anticipated by WO 2019/058000 A1 (of record) is withdrawn in light of Applicants’ claim amendments. Applicants’ arguments are moot in light of the withdrawal of the rejection.
17) The rejection of claim 10 set forth on paragraph 13 of the Office Action mailed 10/31/25 under 35 U.S.C § 103 as being unpatentable over WO 2019/058000 A1 (of record) as applied to claims 9 and 1 above and further in view of WO 2020141168 A1 (of record) (WO ‘168) is withdrawn in light of Applicants’ claim amendments. Applicants’ arguments are moot in light of the withdrawal of the rejection.
Rejection(s) under 35 U.S.C § 112(b) or (Pre-AIA ) Second Paragraph
18) 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.
19) Claims 1-18 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 inventor or a joint inventor or for the pre-AIA the Applicants regard as the invention.
(a) Claim 1, as amended, is ambiguous and indefinite in the limitations “reducing ...... endogenous lipase activity corresponding to Yarrowia lipase activity 8”. First, it is unclear what is encompassed in “lipase activity 8”, what precise structure is responsible for this lipase activity, and how does this “lipase activity 8” differ from any other lipase activity. Second, the term “reducing” is a relative term which is not specifically defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the claim. What precise level of reduction is encompassed, and reduction compared to what, is unclear. One of ordinary skill in the art cannot understand in an unambiguous way that which is being claimed and the scope of the claim.
(b) Analogous rejection applies to new claims 15 and 16 with regard to the limitations “reduction ...... of endogenous lipase activity corresponding to Yarrowia lipase activity 3” and “reduction ...... of endogenous lipase activity corresponding to Yarrowia lipase activity 4” respectively and to claim 8, as amended, with regard to the limitations “reduction ...... of endogenous lipase activity corresponding to Yarrowia lipase activity 2”. One of ordinary skill cannot understand in an unambiguous way that which is being claimed.
(c) Claim 10, as amended, and new claim 14 are indefinite for having improper antecedence in the limitation “the acetylating enzyme”. Claim 10 depends directly from claim 9 and indirectly from claim 1, and new claim 14 depends indirectly from claim 1. Neither claim 9 nor claim 1 includes the recitation of any “acetylating enzyme”.
(d) New claim 17 is ambiguous and indefinite in the limitations “the heterologous genes encoding acetylating enzymes catalyzing the conversion of retinol to retinyl acetate ....”. New claim 17 depends directly from claim 9 and indirectly from claim 1, which do not recite any heterologous genes “encoding acetylating enzymes” catalyzing the conversion of retinol to retinyl acetate.
(e) Claims 9-18, which depend directly or indirectly from claim 1, are also rejected as being indefinite, because of the ambiguity or indefiniteness identified above in the base claim.
Claim(s) Interpretation
20) The claim limitation “a two-phase fermentation process” is interpreted as per the following definition on page 1 of the as-filed specification [Emphasis added]:
The present invention is related to fermentative production of retinoids, including retinol or retinyl acetate, comprising cultivation of a retinoid-producing host cell, such as fungal host cell, particularly oleaginous host such as e.g. Yarrowia, in a two-phase culture system in the presence of vegetable oil as second phase.
Further, lines 16-18 of page 2 of the as-filed specification state that the host cell is cultivated in the presence of a plant-based second phase, e.g. vegetable oil, such as e.g. corn oil, as second phase solvent. The vegetable corn oil is identified as an oleic acid in the last paragraph of page 2 of the as-filed specification. Accordingly, an oleic acid such as corn oil when present or added after or following the initial phase of a culturing or fermentation process is interpreted as necessarily serving as a lipophilic solvent. Corn oil when present after or following the initial phase of a culturing or fermentation process serves necessarily and inherently as a second phase lipolytic solvent.
Rejection(s) under 35 U.S.C § 102
21) The following is a quotation of the appropriate paragraphs of 35 U.S.C § 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(2) the claimed invention was described in a patent issued under section 151, or in application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
22) Claims 1-18 are rejected under 35 U.S.C § 102(a)(2) as being anticipated by WO 2021136689 A1 (Applicants’ IDS) as evidenced by WO 2016/172282 A1 (Applicants’ IDS) and WO 2008042338 A2 (Applicants’ IDS).
WO 2021136689 A1 disclosed a process of extracting retinoids in a fermentation process comprising cultivation of an oleaginous yeast such as Yarrowia lipolytica in the presence of oils containing free fatty acids such as oleic acid or triglyceride oils such as vegetable corn oil. The oleaginous Yarrowia lipolytica yeast is modified to reduce or abolish the activity of Yarrowia LIP8 (SEQ ID NO: 5), or Yarrowia LIP2 (SEQ ID NO: 1), or Yarrowia LIP3 (SEQ ID NO: 3), or Yarrowia LIP4 (SEQ ID NO: 15). Surprisingly, the modification of the oleaginous Yarrowia lipolytica yeast such as the blockage of certain enzymes involved in pre-digestion of vegetable oil into glycerol and fatty acids lead to an increase in retinyl acetate formation, i.e., increase in the percentage of retinyl acetate based on total retinoids by at least about 30%. The modified oleaginous Yarrowia lipolytica yeast comprising the genetic modification(s), i.e., reduction or abolishment of endogenous lipases, including but not limited to modification in the activity of an endogenous lipase gene with at least 90% or 95% identity to SEQ ID NO: 5 that corresponds to LIP8 obtainable from Yarrowia lipolytica. The modified strain was grown in the presence of oleic acid or vegetable corn oil. The process of fermentation(s) of the modified strain was identical to the conditions previously described in Example 6 of WO 2016/172282 A1, but with a different oil, i.e., oleic acid. The fermentation process for the production of beta-ionone taught in Example 6 of the evidentiary reference of WO 2016172282 A1, which is expressly identified by the prior art WO 2021136689 A1 in the first full paragraph of page 23 therein, was a 2-phase fermentation process, wherein glucose was used as a carbon source in the initial fermentation batch phase, and corn oil was used after or following the initial phase, i.e., in the second phase. See the 2nd half of section [0160] of the evidentiary reference of WO 2016172282 A1. That the vegetable corn oil used in the second phase of the prior art method served intrinsically as an extracting solvent is inherent from the disclosure of WO 2021136689 A1 in light of what was well known in the art. For instance, the evidentiary reference of WO 2008042338 A2, expressly incorporated by reference in the second full paragraph of page 14 of WO 2021136689 A1, describes the vegetable corn oil as an extracting solvent used in a method of extracting a carotenoid, retinolic compound, retinal, retinoic acid, or retinol after cultivating a genetically modified Y. lipolytica. See claims 675, 674, 669 and 664-667 of WO 2008042338 A2. The method disclosed by the prior art of WO 2021136689 A1 resulted in increased percentage of retinyl acetate from the conversion of retinol such as at least about 70% retinyl acetate based on total retinoids present in the Yarrowia lipolytica. WO 2021136689 A1 disclosed the construction of the Lip8-deleted (i.e., endogenous lipase genes-deleted or lipase activity-abolished) strain expressing a highly active heterologous transferase of the Lachancea mirantina fungus, i.e., LmATF1, SEQ ID NO: 13 disclosed in WO 2019058001 A1 (incorporated therein by reference) carrying the amino acid substitutions S480Q_G409A_V407I_H69A_I484L, i.e., the instantly recited SEQ ID NO: 12 having the amino acid substitutions at positions 480, 409, 407, 69 and 484, which strain showed zero percent increase or abundance of retinyl acetate. See the entire reference including pages 2-7; the first full sentence under Example 4; Tables 1, 2, 3A and 6; 1st full paragraph of page 23; and claims 9-14 and 1-10 of WO 2021136689 A1; and the sequence alignment reports below.
Lipase 8:
BJP31651
ID BJP31651 standard; protein; 371 AA.
AC BJP31651
DT 19-AUG-2021 (first entry)
DE Yarrowia lipolytica lipase lip8, SEQ 5.
KW EC 3.1.1; Lipase; enzyme identification; fermentation;
KW genetically engineered microorganism; lip8 protein; retinoid; retinol;
KW retinyl acetate; screening.
OS Yarrowia lipolytica.
PN WO 2021136689-A1.
PD 08-JUL-2021.
PF 18-DEC-2020; 2020WO-EP087018.
PR 30-DEC-2019; 2019CH-00001716.
PR 27-JUL-2020; 2020EP-00187829.
PA (STAM ) DSM IP ASSETS BV.
PI Mcmahon J, Kooi EI, Wu L, De Jong RM, Vyas VK, Houston PL.
DR WPI; 2021-770980/059.
DR N-PSDB; BJP31652.
PT Retinoid-producing host cell is capable of retinyl acetate formation,
PT particularly retinyl acetate-producing host cell such as fungal host
PT cells, where oleaginous yeast cell is Yarrowia, which comprises genetic
PT modification(s).
PS Claim 4; SEQ ID NO 5; 35pp; English.
CC The present invention relates to a novel retinoid-producing host cell
CC capable of retinyl acetate formation, particularly retinyl acetate-
CC producing host cell, such as fungal host cells, preferably oleaginous
CC yeast cell (e.g., Yarrowia). The host cell comprises genetic
CC modifications, such as reduction or abolishment of endogenous enzymes of
CC SEQ ID NO:1-15 (see BJP31647-BJP31661, only odd numbers) involved in pre-
CC digestion of vegetable oil into glycerol and fatty acids, where the
CC endogenous enzymes belonging to EC class 3.1.1.-, or enzymes with
CC esterase or lipase activity. The invention also provides: a method for
CC reducing or abolishing percentage of retinoids other than retinyl acetate
CC in a retinoid mix generated in a fermentation process; a method for
CC producing a product such as retinol, retinyl acetate, vitamin A, and a
CC mix comprising retinol, retinyl acetate and vitamin A; and a method for
CC identifying suitable endogenous hydrolases to get modified in order to
CC increase percentage of retinyl acetate in fermentation of retinyl acetate
CC producing host cell grown on vegetable oil as carbon source. The retinoid
CC -producing host cell is stable intermediate in vitamin A production and
CC industrially intractable, improves product-specificity and productivity,
CC and increases retinyl acetate formation.
SQ Sequence 371 AA
Query Match 100%; Score 1994; Length 371; Best Local Similarity 100%;
Matches 371; Conservative 0; Mismatches 0; Indels 0; Gaps 0.
Qy 1 MVSLSARIKDFFSVLLLGAATITPSTQTAGVSQGFYDFARDFAHLSNIAYCVNAPITPLN 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MVSLSARIKDFFSVLLLGAATITPSTQTAGVSQGFYDFARDFAHLSNIAYCVNAPITPLN 60
Qy 61 PDFTCGNSCKHFPEIELVKTFGGNFFKTSITGYLAVDHVKKEKYVVFRGTFSLADAITDM 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 PDFTCGNSCKHFPEIELVKTFGGNFFKTSITGYLAVDHVKKEKYVVFRGTFSLADAITDM 120
Qy 121 QFQLSPFLVDVPALNTFSANDTTAEAQTHCEGCKIHDGFSKAFTETWGNIGEDLQKHLDA 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 QFQLSPFLVDVPALNTFSANDTTAEAQTHCEGCKIHDGFSKAFTETWGNIGEDLQKHLDA 180
Qy 181 NPDYQLYVTGHSLGAAMALLGATSIKLKGYDPILINYGQPRVGNKPFAEFINKLWFGEGN 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 NPDYQLYVTGHSLGAAMALLGATSIKLKGYDPILINYGQPRVGNKPFAEFINKLWFGEGN 240
Qy 241 GLEITPERKLYRMTHWNDIFVGLPNWEGYTHSNGEVYINNRFINPPLKDVISCAGGENSK 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GLEITPERKLYRMTHWNDIFVGLPNWEGYTHSNGEVYINNRFINPPLKDVISCAGGENSK 300
Qy 301 CYRSSFSLLSQINLLQNHLAYIDYIGYCALNIGRRELADQEHYTGPYYYGHRSEEDFKKL 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 CYRSSFSLLSQINLLQNHLAYIDYIGYCALNIGRRELADQEHYTGPYYYGHRSEEDFKKL 360
Qy 361 GLELSTPQVEN 371
|||||||||||
Db 361 GLELSTPQVEN 371
Lipase 3:
BJP31649
ID BJP31649 standard; protein; 498 AA.
AC BJP31649
DT 19-AUG-2021 (first entry)
DE Yarrowia lipolytica lipase lip3, SEQ 3.
KW EC 3.1.1; Lipase; enzyme identification; fermentation;
KW genetically engineered microorganism; lip3 protein; retinoid; retinol;
KW retinyl acetate; screening.
OS Yarrowia lipolytica.
PN WO 2021136689-A1.
PD 08-JUL-2021.
PF 18-DEC-2020; 2020WO-EP087018.
PR 30-DEC-2019; 2019CH-00001716.
PR 27-JUL-2020; 2020EP-00187829.
PA (STAM ) DSM IP ASSETS BV.
PI Mcmahon J, Kooi EI, Wu L, De Jong RM, Vyas VK, Houston PL
DR WPI; 2021-770980/059.
DR N-PSDB; BJP31650.
PT Retinoid-producing host cell is capable of retinyl acetate formation,
PT particularly retinyl acetate-producing host cell such as fungal host
PT cells, where oleaginous yeast cell is Yarrowia, which comprises genetic
PT modification(s).
PS Claim 4; SEQ ID NO 3; 35pp; English.
CC The present invention relates to a novel retinoid-producing host cell
CC capable of retinyl acetate formation, particularly retinyl acetate-
CC producing host cell, such as fungal host cells, preferably oleaginous
CC yeast cell (e.g., Yarrowia). The host cell comprises genetic
CC modifications, such as reduction or abolishment of endogenous enzymes of
CC SEQ ID NO:1-15 (see BJP31647-BJP31661, only odd numbers) involved in pre-
CC digestion of vegetable oil into glycerol and fatty acids, where the
CC endogenous enzymes belonging to EC class 3.1.1.-, or enzymes with
CC esterase or lipase activity. The invention also provides: a method for
CC reducing or abolishing percentage of retinoids other than retinyl acetate
CC in a retinoid mix generated in a fermentation process; a method for
CC producing a product such as retinol, retinyl acetate, vitamin A, and a
CC mix comprising retinol, retinyl acetate and vitamin A; and a method for
CC identifying suitable endogenous hydrolases to get modified in order to
CC increase percentage of retinyl acetate in fermentation of retinyl acetate
CC producing host cell grown on vegetable oil as carbon source. The retinoid
CC -producing host cell is stable intermediate in vitamin A production and
CC industrially intractable, improves product-specificity and productivity,
CC and increases retinyl acetate formation.
SQ Sequence 498 AA
Query Match 100.0%; Score 2664; Length 498; Best Local Similarity 100%;
Matches 498; Conservative 0; Mismatches 0; Indels 0; Gaps 0.
Qy 1 MPLELPSLNASIVGNTVQNGAVEQFLNIRYADIPGKFEKPVLKNDWNGAEIDATKVGPVC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MPLELPSLNASIVGNTVQNGAVEQFLNIRYADIPGKFEKPVLKNDWNGAEIDATKVGPVC 60
Qy 61 PQPRTPFNFFSVPDDLWEKVNVDTYQDGLLCDNLIVTRPKGVSANARLPTVVWIHGGSNI 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 PQPRTPFNFFSVPDDLWEKVNVDTYQDGLLCDNLIVTRPKGVSANARLPTVVWIHGGSNI 120
Qy 121 EGSIYNLIYEPQFLVAESVRVGKPIVHVCIEYRLGLAGFLTKNGKGNWGTWDQYTGCQWV 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 EGSIYNLIYEPQFLVAESVRVGKPIVHVCIEYRLGLAGFLTKNGKGNWGTWDQYTGCQWV 180
Qy 181 NRHIQDFGGDPLNVTLTGESAGSVAVHNMLIKDSMNGRKLFRNAVMMSGTLETITPQPPK 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 NRHIQDFGGDPLNVTLTGESAGSVAVHNMLIKDSMNGRKLFRNAVMMSGTLETITPQPPK 240
Qy 241 WHARLEEKVAKVTGKEVADLASLSDKELLDAQIKLNVAVCMTCDDGDFFEPGWKQHLTPD 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 WHARLEEKVAKVTGKEVADLASLSDKELLDAQIKLNVAVCMTCDDGDFFEPGWKQHLTPD 300
Qy 301 WLDKLIISDCKDEGMLYFLPVNAQDDEELLAKVAKSPVGKEISELYGIKEGGDIKSACLD 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 WLDKLIISDCKDEGMLYFLPVNAQDDEELLAKVAKSPVGKEISELYGIKEGGDIKSACLD 360
Qy 361 LKTDATFNYFNHLLFKKMEEARNNGSTSRVYRLAVDEPNPHNPDQRAHHAVDVLYMFNST 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 LKTDATFNYFNHLLFKKMEEARNNGSTSRVYRLAVDEPNPHNPDQRAHHAVDVLYMFNST 420
Qy 421 KFNEHGDKLSRLFQSHFLRLAYGLEPWDHRNFGVYRNGGYQQLPLSELNKVRPVERYEAL 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 KFNEHGDKLSRLFQSHFLRLAYGLEPWDHRNFGVYRNGGYQQLPLSELNKVRPVERYEAL 480
Qy 481 SKMDFGQVGRLSNALSRL 498
||||||||||||||||||
Db 481 SKMDFGQVGRLSNALSRL 498
Lipase 4:
BJP31661
ID BJP31661 standard; protein; 406 AA.
AC BJP31661
DT 19-AUG-2021 (first entry)
DE Yarrowia lipolytica lipase lip4, SEQ 15.
KW EC 3.1.1; Lipase; enzyme identification; fermentation;
KW genetically engineered microorganism; lip4 protein; retinoid; retinol;
KW retinyl acetate; screening.
OS Yarrowia lipolytica.
PN WO 2021136689-A1.
PD 08-JUL-2021.
PF 18-DEC-2020; 2020WO-EP087018.
PR 30-DEC-2019; 2019CH-00001716.
PR 27-JUL-2020; 2020EP-00187829.
PA (STAM ) DSM IP ASSETS BV.
PI Mcmahon J, Kooi EI, Wu L, De Jong RM, Vyas VK, Houston PL.
DR WPI; 2021-770980/059.
DR N-PSDB; BJP31662.
PT Retinoid-producing host cell is capable of retinyl acetate formation,
PT particularly retinyl acetate-producing host cell such as fungal host
PT cells, where oleaginous yeast cell is Yarrowia, which comprises genetic
PT modification(s).
PS Claim 4; SEQ ID NO 15; 35pp; English.
CC The present invention relates to a novel retinoid-producing host cell
CC capable of retinyl acetate formation, particularly retinyl acetate-
CC producing host cell, such as fungal host cells, preferably oleaginous
CC yeast cell (e.g., Yarrowia). The host cell comprises genetic
CC modifications, such as reduction or abolishment of endogenous enzymes of
CC SEQ ID NO:1-15(see BJP31647-BJP31661, only odd numbers) involved in pre-
CC digestion of vegetable oil into glycerol and fatty acids, where the
CC endogenous enzymes belonging to EC class 3.1.1.-, or enzymes with
CC esterase or lipase activity. The invention also provides: a method for
CC reducing or abolishing % of retinoids other than retinyl acetate
CC in a retinoid mix generated in a fermentation process; a method for
CC producing a product such as retinol, retinyl acetate, vitamin A, and a
CC mix comprising retinol, retinyl acetate and vitamin A; and a method for
CC identifying suitable endogenous hydrolases to get modified in order to
CC increase % of retinyl acetate in fermentation of retinyl acetate
CC producing host cell grown on vegetable oil as carbon source. The retinoid
CC -producing host cell is stable intermediate in vitamin A production and
CC industrially intractable, improves product-specificity and productivity,
CC and increases retinyl acetate formation.
SQ Sequence 406 AA
Query Match 100.0%; Score 2197; Length 406; Best Local Similarity 100%;
Matches 406; Conservative 0; Mismatches 0; Indels 0; Gaps 0.
Qy 1 MAGFNFTFGQVISYLASMLYGQVDATSSSTRIQATQDLYDFTAKFSRLSNIAYCINAPFT 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MAGFNFTFGQVISYLASMLYGQVDATSSSTRIQATQDLYDFTAKFSRLSNIAYCINAPFT 60
Qy 61 PLRTDFTCGESCRYFPDLQLDSVFGGNFSSASTTGYIAYDHKKKEKYIVFRGTFSIPDII 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 PLRTDFTCGESCRYFPDLQLDSVFGGNFSSASTTGYIAYDHKKKEKYIVFRGTFSIPDII 120
Qy 121 TDIQFQTAPWLTSLPTHLIPTKEDFEHKQAILKHYAAENKGLSNLEERQDVVHEDPSLVP 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TDIQFQTAPWLTSLPTHLIPTKEDFEHKQAILKHYAAENKGLSNLEERQDVVHEDPSLVP 180
Qy 181 KKMDKCENCQIHDGFAKGFNETIEHAGPQIEKFLGNNTDYKMFVVGHSLGAAQAQLFATQ 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 KKMDKCENCQIHDGFAKGFNETIEHAGPQIEKFLGNNTDYKMFVVGHSLGAAQAQLFATQ 240
Qy 241 FKLLGFDPYMINFGQPRLGNPEFAAYINQLWFNDTGLVVNDARRFYRVTHWNDIVVGVPD 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 FKLLGFDPYMINFGQPRLGNPEFAAYINQLWFNDTGLVVNDARRFYRVTHWNDIVVGVPD 300
Qy 301 WLNYTHSIGEVFIDEESVYPKLDKVVVCEGGENPLCHRGTFNLWSRINFLQNHLAYIFYI 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 WLNYTHSIGEVFIDEESVYPKLDKVVVCEGGENPLCHRGTFNLWSRINFLQNHLAYIFYI 360
Qy 361 GLCAFNIGRRDVLNMPQYQGNFSYQHNIDPNYNYDTKVPTRISKSN 406
||||||||||||||||||||||||||||||||||||||||||||||
Db 361 GLCAFNIGRRDVLNMPQYQGNFSYQHNIDPNYNYDTKVPTRISKSN 406
Lachancea mirantina acetyl transferase 1 (LmATF1)
BGE36662
ID BGE36662 standard; protein; 559 AA.
AC BGE36662
DT 16-MAY-2019 (first entry)
DE Lachancea mirantina acetyl transferase 1 (LmATF1), SEQ:13.
KW ATF1 protein; EC 2.3.1.84; acetyl transferase 1;
KW acetyl-CoA:alcohol O-acetyltransferase; alcohol O-acetyltransferase;
KW genetically engineered microorganism; retinol; retinyl acetate.
OS Lachancea mirantina.
PN WO 2019058001-A1.
PD 28-MAR-2019.
PF 25-SEP-2018; 2018WO-EP076034.
PR 25-SEP-2017; 2017US-0562672P.
PR 20-APR-2018; 2018EP-00168564.
PA (STAM) DSM IP ASSETS BV.
PI Balch N, Blomquist P, Doten R, Houston P, Lam E, Mcmahon J
PI Trueheart J, Viarouge C, De Jong RM.
DR WPI; 2019-288905/25.
DR N-PSDB; BGE36663, BGE36664.
PT New carotenoid producing host cell useful for producing retinyl ester mix
PT or vitamin A, comprising enzyme with acetylating activity.
PS Claim 7; SEQ ID NO 13; 27pp; English.
CC The present invention relates to a novel carotenoid-producing host cell
CC comprising an enzyme with acetylating activity, where the acetylating
CC activity includes retinol acetylating activity, preferably acetyl
CC transferase (EC 2.3.1.84) activity, more preferably acetyl transferase 1
CC activity. The invention further discloses: (1) a process for producing
CC retinyl ester mix comprising retinyl acetate through enzymatic activity
CC of acetyl transferase preferably activity of acetyl transferase 1, which
CC involves contacting retinol with the acetyl transferase, preferably
CC acetyl transferase 1; and (2) a process for producing vitamin A,
CC involves: (a) introducing a nucleic acid molecule encoding acetyl
CC transferase into a suitable carotenoid-producing host cell (b) enzymatic
CC conversion of retinol comprising trans-and cis-retinol into a retinyl
CC acetate mix comprising cis-and trans-retinyl acetate; and (c) converting
CC retinyl acetate into vitamin A under suitable culture conditions. The
CC novel carotenoid-producing host cell of the invention is useful for
CC producing retinyl esters, particular retinyl acetate, which is an
CC important building block for the production of vitamin A. The present
CC sequence represents a Lachancea mirantina acetyl transferase 1 (LmATF1),
CC which is useful for constructing a plasmid in order to produce a strain
CC carrying heterologous ATF1 gene.
SQ Sequence 559 AA
Query Match 100%; Score 2950; Length 559; Best Local Similarity 100%;
Matches 559; Conservative 0; Mismatches 0; Indels 0; Gaps 0.
Qy 1 MGDLDARGTSAHPELSERPSIMPSMSDIQDPSGDDKATPRGSAAGLPQLELAGHARRLGH 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MGDLDARGTSAHPELSERPSIMPSMSDIQDPSGDDKATPRGSAAGLPQLELAGHARRLGH 60
Qy 61 LENFFAVQHRQQIYSSFAVFCEFDTACSLAQLASAVRNVCLSNPLLLHTVEPKHPDIAGF 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 LENFFAVQHRQQIYSSFAVFCEFDTACSLAQLASAVRNVCLSNPLLLHTVEPKHPDIAGF 120
Qy 121 YHSDEYLSRPWPQHDYMRVLREVHVADVVMNGQKEHAHVVRDAVDVFQAHGNQVTSELLE 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 YHSDEYLSRPWPQHDYMRVLREVHVADVVMNGQKEHAHVVRDAVDVFQAHGNQVTSELLE 180
Qy 181 LMTQIEIPHASQTRPSWRLLCFPHGEANRWRTFAFVSNHCSSDGLSGLNFFRDLQKELAH 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 LMTQIEIPHASQTRPSWRLLCFPHGEANRWRTFAFVSNHCSSDGLSGLNFFRDLQKELAH 240
Qy 241 GPTSGAPGAPGASGVIFDYAQDAATLPKLPPPIDQKLDYRPSKKALLGLLAGKFVREKLG 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GPTSGAPGAPGASGVIFDYAQDAATLPKLPPPIDQKLDYRPSKKALLGLLAGKFVREKLG 300
Qy 301 YVSAAPPTTPTSDLAHPEGHQYYCYLVNVPTSSVAHIKTQVRENVPHKCTLTPFLQACWL 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 YVSAAPPTTPTSDLAHPEGHQYYCYLVNVPTSSVAHIKTQVRENVPHKCTLTPFLQACWL 360
Qy 361 VSLFKYGRVFSGSWLERYTDVLVAMNTRQLLPEDLELQRQYRYGSNVGGVRYNYPIAPLD 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 VSLFKYGRVFSGSWLERYTDVLVAMNTRQLLPEDLELQRQYRYGSNVGGVRYNYPIAPLD 420
Qy 421 VRDNDQKFWSLVESYRLALSDARDKNDYLYALGALMLPEIYEKKNVDAVVNDTILNQRRS 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 VRDNDQKFWSLVESYRLALSDARDKNDYLYALGALMLPEIYEKKNVDAVVNDTILNQRRS 480
Qy 481 GTLISNVGYVRDEQPTAFAIKNHVFSQGVGANRNAFVLNICATDQGGLNIAISIAKGTLA 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 GTLISNVGYVRDEQPTAFAIKNHVFSQGVGANRNAFVLNICATDQGGLNIAISIAKGTLA 540
Qy 541 SRQEGQELCDIFKSTLLRF 559
|||||||||||||||||||
Db 541 SRQEGQELCDIFKSTLLRF 559
Claims 1-18 are anticipated by WO 2021136689 A1. The reference of WO 2016/172282 A1 or WO 2008042338 A2 is not used as a secondary reference in combination with WO 2021136689 A1, but rather is used to show that every element of the claimed subject matter is disclosed by WO 2021136689 A1 with the unrecited limitation(s) being inherent as evidenced by the state of the art. See In re Samour 197 USPQ (CCPA 1978).
Claim Objection(s) - Suggestion(s)
23) New claim 17 is incorrect in reciting the plural limitations “the heterologous genes encoding acetylating enzymes catalyzing the conversion of retinol to retinyl acetate” in associated with the non-plural limitation “is a fungal enzyme” [Emphasis added]. Appropriate correction is needed.
Conclusion
24) No claims are allowed.
In line 2 of claim 3, for proper antecedence, it is suggested that Applicants replace the limitation “retinoids” with the limitation --the retinoids--.
25) Applicants’ amendment necessitated the new ground(s) of rejection presented in this Office action. THIS ACTION IS MADE FINAL. Applicants are reminded of the extension of time policy as set forth in 37 C.F.R 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 extension fee pursuant to 37 C.F.R 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.
Correspondence
26) Any inquiry concerning this communication or earlier communications from the Examiner should be directed to S. Devi, Ph.D., whose telephone number is (571) 272-0854. A message may be left on the Examiner’s voice mail system. The Examiner is on a flexible work schedule, however she can normally be reached Monday to Friday from 8.00 a.m. to 4.00 p.m. (EST). If attempts to reach the Examiner by telephone are unsuccessful, the Examiner’s supervisor Jeffrey Stucker, can be reached at (571) 272-0911. The fax phone number for the organization where this application or proceeding is assigned (571) 273-8300.
27) Information regarding the status of an application may be obtained from Patent Center. Status information for unpublished applications is available through Patent Center or Private PAIR to authorized users only. Should you have questions about access to Patent Center or the Private PAIR system, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). 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.
/S. DEVI/
S. Devi, Ph.D.Primary Examiner
Art Unit 1645
May, 2026
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