Method for production of 4-cyano benzoic acid or salts thereof

Patent metadata

Field	Value
Publication number	US-2022017931-A1
Application number	US-202017424824-A
Country	US
Kind code	A1
Filing date	Jan 20, 2020
Priority date	Jan 22, 2019
Publication date	Jan 20, 2022
Grant date	—

Abstract

Described herein are methods for the production of 4-cyano benzoic acid or salts thereof from terephthalonitrile using nitrilase as catalyst. Also described herein are compositions including 4-cyano benzoic acid.

First claim

1 . An isolated nitrilase capable of catalysing a reaction from terephthalonitrile to ammonium 4-cyano benzoic acid in an aqueous medium comprising water, nitrilase and terephthalonitrile and/or ammonium 4-cyano benzoic acid, wherein the concentration of ammonium 4-cyano benzoic acid in the aqueous medium after incubation is at least 5% (w/w) and the concentration of terephthalonitrile is below 1.0% (w/w).

2 . The isolated nitrilase of claim 1 wherein after incubation the aqueous medium comprises below 0.5% (w/w) terephthalic acid.

3 . The isolated nitrilase of claim 1 comprising a sequence selected from the group consisting of
a. An amino acid molecule of SEQ ID NO: 2, 4, 6 or 8,
b. An amino acid molecule having at least 55% identity to the amino acid molecule of SEQ ID NO: 2, 4, 6 or 8 or a functional fragment thereof,
c. An amino acid molecule encoded by a nucleic acid molecule of SEQ ID NO: 1, 3, 5 or 7 or a functional fragment thereof,
d. An amino acid molecule encoded by a nucleic acid molecule having at least 70% identity to SEQ ID NO: 1, 3, 5 or 7 or a functional fragment thereof, and
e. An amino acid molecule encoded by a nucleic acid molecule hybridizing under stringent conditions to a fragment of at least 250 bases complementary to SEQ ID NO: 1, 3, 5 or 7 or a functional fragment thereof,
wherein the amino acid molecule as defined in b., d. and e. catalyzes the reaction from terephthalonitrile to ammonium 4-cyano benzoic acid in an aqueous medium.

4 . An isolated nitrilase sequence selected from the group consisting of
a. An amino acid molecule of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 20 or 22, and b. An amino acid molecule having at least 55% identity to the amino acid molecule of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 20 or 22 or a functional fragment thereof, and c. An amino acid molecule encoded by a nucleic acid molecule of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, and d. An amino acid molecule encoded by a nucleic acid molecule having at least 70% identity to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, and e. An amino acid molecule encoded by a nucleic acid molecule hybridizing under stringent conditions to a fragment of at least 250 bases complementary to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, wherein the amino acid molecule as defined in b., c., d. and e. catalyzes a reaction from terephthalonitrile to ammonium 4-cyano benzoic acid in an aqueous medium.

5 . A process for producing 4-cyano benzoic acid or salt thereof comprising the steps of
i. Providing an aqueous medium comprising water, one or more nitrilase and terephthalonitrile, ii. Incubating the aqueous medium and iii. Optionally isolating the 4-cyano benzoic acid or salt thereof from the reaction mixture, wherein the one or more nitrilase is capable of catalysing the reaction from terephthalonitrile to ammonium 4-cyano benzoic acid in an aqueous medium comprising water, nitrilase and terephthalonitrile and/or ammonium 4-cyano benzoic acid, wherein the concentration of ammonium 4-cyano benzoic acid in the aqueous medium after incubation is at least 5% (w/w) and the concentration of terephthalonitrile is below 1.0% (w/w).

6 . The process of claim 5 wherein after incubation the aqueous medium comprises below 0.5% (w/w) terephthalic acid.

7 . The process of claim 5 wherein the nitrilase comprises a sequence selected from the group consisting of
a. An amino acid molecule of SEQ ID NO: 2, 4, 6 or 8,
b. An amino acid molecule having at least 55% identity to the amino acid molecule of SEQ ID NO: 2, 4, 6 or 8 or a functional fragment thereof,
c. An amino acid molecule encoded by a nucleic acid molecule of SEQ ID NO: 1, 3, 5 or 7 or a functional fragment thereof,
d. An amino acid molecule encoded by a nucleic acid molecule having at least 70% identity to SEQ ID NO: 1, 3, 5 or 7 or a functional fragment thereof, and
e. An amino acid molecule encoded by a nucleic acid molecule hybridizing under stringent conditions to a fragment of at least 250 bases complementary to SEQ ID NO: 1, 3, 5 or 7 or a functional fragment thereof,
wherein the amino acid molecule as defined in b., d. and e. catalyzes the reaction from terephthalonitrile to ammonium 4-cyano benzoic acid in an aqueous medium.

8 . A process for producing 4-cyano benzoic acid or salt thereof comprising the steps of
i. Providing an aqueous medium comprising water, one or more nitrilase and terephthalonitrile, ii. Incubating the aqueous medium and iii. Optionally isolating the 4-cyano benzoic acid or salt thereof from the reaction mixture, wherein the one or more nitrilase is selected from the group consisting of a. an amino acid molecule of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 20 or 22 or a functional fragment thereof, b. an amino acid molecule having at least 55% identity to the amino acid molecule of SEQ ID NO: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 20 or 22 or a functional fragment thereof, c. an amino acid molecule encoded by a nucleic acid molecule of SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, d. an amino acid molecule encoded by a nucleic acid molecule having at least 70% identity to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, and e. an amino acid molecule encoded by a nucleic acid molecule hybridizing under stringent conditions to a fragment of at least 250 bases complementary to SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, wherein the amino acid molecule as defined in ii., iii., iv. and v. has an activity of converting terephthalonitrile to ammonium 4-cyano benzoic acid in an aqueous medium.

9 . The process of claim 8 wherein the aqueous medium further comprises a divalent cation.

10 . The process of claim 9 wherein the divalent cation is Mg 2+ , Mn 2+ , Ca 2+ , Fe 2+ , Zn 2+ or Co 2+ .

11 . The process of claim 8 wherein the terephthalonitrile is added to the aqueous medium before incubation in a concentration of between 1% and 30% w/w.

12 . The process of claim 8 wherein a pH-value of the aqueous medium is adjusted to below 5 by adding acid to the aqueous medium during or after incubation.

13 . The process of claim 8 wherein a product is isolated by filtration or centrifugation after incubation.

14 . The process of claim 8 wherein the aqueous medium is incubated for at least 2 h.

15 . The process of claim 8 wherein the aqueous medium is incubated between 15 and 50° C.

16 . The process of claim 8 wherein the nitrilase is produced by fermentation.

17 . A recombinant construct comprising a nitrilase wherein the nitrilase is selected from the group consisting of
a. an amino acid molecule of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 20 or 22 or a functional fragment thereof, b. an amino acid molecule having at least 55% identity to the amino acid molecule of SEQ ID NO: SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 20 or 22 or a functional fragment thereof, c. an amino acid molecule encoded by a nucleic acid molecule of SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, d. an amino acid molecule encoded by a nucleic acid molecule having at least 70% identity to SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 19 or 21 or a functional fragment thereof, and e. an amino acid molecule encoded by a nucleic acid molecule hybridizing under stringent conditions to a fragment of at least 250 bases complementary to SEQ ID NO 1, 3, 5, 7, 9, 11, 13, 15, 19 o

Assignees

• Basf Se

Inventors

• Ghislieri Diego
• Willrodt Christian
• Koradin Christopher
• Seemayer Stefan
• Schachtschabel Doreen
• Baldenius Kai-Uwe
• Goetz Roland

Classifications

• C12P13/002 Primary

  Nitriles (-CN) · CPC title

• C12Y305/05001

  Nitrilase (3.5.5.1) · CPC title

• C12Y305/05007

  Aliphatic nitrilase (3.5.5.7) · CPC title

• C12P7/40

  containing a carboxyl group {including Peroxycarboxylic acids} · CPC title

• C12N9/78

  acting on carbon to nitrogen bonds other than peptide bonds (3.5) · CPC title