Oxidation and amination of primary alcohols

Patent metadata

Field	Value
Publication number	US-9580732-B2
Application number	US-201214233505-A
Country	US
Kind code	B2
Filing date	Jul 17, 2012
Priority date	Jul 20, 2011
Publication date	Feb 28, 2017
Grant date	Feb 28, 2017

Abstract

A method including the steps a) providing a primary alcohol of the formula HO—(CH 2 ) x —R 1 , where R 1 is —OH, —SH, —NH 2 or —COOR 2 ; x is at least 3; and R 2 is H, alkyl or aryl, b) oxidizing the primary alcohol by contacting it with an NAD(P) + -dependent alcohol dehydrogenase, and c) contacting the oxidation product of step a) with a transaminase, where the NAD(P) + -alcohol dehydrogenase and/or the transaminase is a recombinant or isolated enzyme. A whole cell catalyst for carrying out the method. The use of such a whole cell catalyst for oxidizing a primary alcohol.

First claim

The invention claimed is:

1. A method for producing an amine or a diamine, comprising:
(a) contacting a primary alcohol of the formula
HO—(CH)—R 1 ,

with an NAD(P) + -dependent alcohol dehydrogenase to oxidize said primary alcohol to form an aldehyde or ketone oxidation product, wherein R 1 is selected from the group consisting of —OH, —SH, —NH 2 and —COOR 2 , x is at least 3 and R 2 is selected from the group consisting of H, alkyl and aryl; and

(b) contacting said aldehyde or ketone oxidation product with a transaminase to transfer α-amino groups from an amino acid to an α-ketocarboxylic acid to produce said amine or said diamine,
wherein said NAD(P) + -dependent alcohol dehydrogenase, said transaminase, or both, are a recombinant or isolated enzyme.

2. The method of claim 1 , further comprising
hydroxylating an alkane of the formula
H—(CH 2 ) x —R 1

by contacting said alkane with a monooxygenase to produce said primary alcohol of the formula
HO—(CH)—R 1 , wherein R 1 has the same meaning as defined above.

3. The method of claim 1 , wherein said NAD(P) + -dependent alcohol dehydrogenase is an NAD(P) + -dependent alcohol dehydrogenase having zinc as a cofactor.

4. The method of claim 3 , wherein said NAD(P) + -dependent alcohol dehydrogenase is a Bacillus stearothermophilus NAD(P) + -dependent alcohol dehydrogenase.

5. The method of claim 2 , wherein monooxygenase is selected from the group consisting of Pseudomonas putida AlkBGT, Candida tropicales cytochrome P450, and Cicer arietinum cytochrome P450.

6. The method of claim 1 , wherein said transaminase is selected from the group consisting of:
(a) a Chromobacterium violaceum ATCC 12472 transaminase, wherein in said transaminase an amino acid corresponding to the amino acid residue at position 23 of the amino acid sequence of SEQ ID NO: 1 is isoleucine, valine, phenylalanine, methionine, or leucine; and
(b) a Chromobacterium violaceum ATCC 12472 transaminase, wherein in said transaminase an amino acid corresponding to the amino acid residue at position 29 of the amino acid sequence of SEQ ID NO: 1 is an amino acid other than threonine.

7. The method of claim 1 , wherein steps (a), (b), or both, are carried out in the presence of an isolated or recombinant alanine dehydrogenase and an inorganic nitrogen source, wherein said alanine dehydrogenase catalyzes the conversion of L-alanine, with consumption of water and NAD + , to form pyruvate, ammonia and NADH.

8. The method of claim 1 , wherein at least one of said NAD(P) + -dependent alcohol dehydrogenase and said transaminase are a recombinant enzyme and are in the form of a whole cell catalyst comprising said enzyme.

9. The method of claim 8 , wherein both of said NAD(P) + -dependent alcohol dehydrogenase and said transaminase are in the form of a whole cell catalyst.

10. The method of claim 1 , wherein said step (a) further comprises the presence of an organic cosolvent having a log P of greater than −1.38.

11. The method of claim 10 , wherein said organic cosolvent is selected from unsaturated fatty acids.

12. The method of claim 10 , wherein said organic cosolvent is a compound of the formula
R 1 —O—(CH 2 ) x —O—R 4 ,

wherein R 3 and R 4 are each independently of one another, selected from the consisting of methyl, ethyl, propyl and butyl, and x is 1 to 4.

13. A method for producing an amine or a diamine, comprising
(a) providing a whole cell catalyst comprising a recombinant NAD(P) + -dependent alcohol dehydrogenase, a recombinant transaminase, optionally a recombinant monooxygenase, and optionally a recombinant alanine dehydrogenase
(b) contacting a primary alcohol of the formula
HO—(CH 2 ) x —R 1

with said whole cell catalyst, wherein said recombinant NAD(P) + -dependent alcohol dehydrogenase present in said whole cell catalyst oxidizes said primary alcohol to form an aldehyde or ketone oxidation product, said recombinant transaminase present in said whole cell catalyst transfers α-amino groups from an amino acid to an α-ketocarboxylic acid to produce said amine or said diamine, said optional alanine dehydrogenase catalyzes the conversion of L-alanine, with consumption of water and NAD + , to form pyruvate, ammonia and NADH, said optional monooxygenase hydroxylates an alkane of the formula H—(CH 2 ) x —R 1 to produce said primary alcohol wherein R 1 is selected from the group consisting of —OH, —SH, —NH 2 , and —COOR 2 , x is at least 3 and R 2 is selected from the group consisting of H, alkyl and aryl.

14. The method of claim 13 , wherein step (b) is performed in the presence of an organic cosolvent having a log P of greater than −1.38.

15. The method of claim 14 , wherein said organic cosolvent is selected from unsaturated fatty acids.

16. The method of claim 10 , wherein said organic cosolvent has a log P of 0 to 1.2.

17. The method of claim 11 , wherein said organic cosolvent is oleic acid.

18. The method of claim 2 , wherein said monooxygenase is a recombinant or isolated enzyme.

19. The method of claim 8 , wherein the whole cell catalyst further comprises a recombinant or isolated monooxygenase and alanine dehydrogenase, wherein said monooxygenase hydroxylates an alkane of the formula H—(CH 2 ) x —R 1 to produce said primary alcohol and said alanine dehydrogenase catalyzes the conversion of L-alanine, with consumption of water and NAD + , to form pyruvate, ammonia and NADH.

Assignees

• Pötter Markus
• Haas Thomas
• Pfeffer Jan Christoph
• Skerra Arne
• Kroutil Wolfgang
• Lerchner Alexandra
• Sattler Johann H
• Schaffer Steffen
• Tauber Katharina Christin
• Evonik Degussa Gmbh

Inventors

• Pötter Markus
• Haas Thomas
• Pfeffer Jan Christoph
• Skerra Arne
• Kroutil Wolfgang
• Lerchner Alexandra
• Sattler Johann H
• Schaffer Steffen
• Tauber Katharina Christin

Classifications

• C12P13/001 Primary

  Amines; Imines · CPC title

• Y02P20/582

  Recycling of unreacted starting or intermediate materials · CPC title