Biocatalysts and methods for the synthesis of armodafinil

Patent metadata

Field	Value
Publication number	US-9365835-B2
Application number	US-201614997277-A
Country	US
Kind code	B2
Filing date	Jan 15, 2016
Priority date	Dec 8, 2010
Publication date	Jun 14, 2016
Grant date	Jun 14, 2016

Abstract

The present invention relates to non-naturally occurring polypeptides useful for preparing armodafinil, polynucleotides encoding the polypeptides, and methods of using the polypeptides. The non-naturally occurring polypeptides of the present invention are effective in carrying out biocatalytic conversion of the (i) 2-(benzhydrylsulfinyl)acetamide to (−)-2-[(R)-(diphenylmethyl)sulfinyl]acetamide (armodafinil), or (ii) benzhydryl-thioacetic acid to (R)-2-(benzhydrylsulfinyl)acetic acid, which is a pivotal intermediate in the synthesis of armodafinil, in enantiomeric excess.

First claim

What is claimed is:

1. A non-naturally occurring polynucleotide encoding a polypeptide having cyclohexanone monooxygenase (CHMO) activity wherein the amino acid sequence of the polypeptide has at least 95% sequence identity to SEQ ID NO: 136.

2. The polynucleotide of claim 1 , wherein said encoded polypeptide is capable of converting the acid substrate (1b) to compound (2b) (R-enantiomer) or its opposite enantiomer compound (S-enantiomer) with at least 2-fold improved activity relative to the wild-type polypeptide of SEQ ID NO: 2,

3. The polynucleotide of claim 2 , wherein said encoded polypeptide is capable of converting the acid substrate of compound (1b) to the R-enantiomer compound (2b) in at least 50% enantiomeric excess.

4. The polynucleotide of claim 1 , wherein said encoded polypeptide is capable of converting compound (1a) to compound (2a)

in at least 75%, at least 85%, at least 90%, at least 95%, at least 98%, or at least 99% enantiomeric excess under suitable reaction conditions.

5. The polynucleotide of claim 4 , wherein said encoded polypeptide is capable of converting compound (1a) to compound (2a) with an activity increased at least 2-fold, at least 4-fold, at least 10-fold, at least 25-fold, at least 40-fold, or at least 60-fold relative to the activity of the polypeptide of SEQ ID NO: 2 under suitable conditions.

6. The polynucleotide of claim 1 , wherein said encoded polypeptide is capable of converting compound (1b) to compound (2b)

in enantiomeric excess under suitable reaction conditions.

7. The polynucleotide of claim 6 , wherein said encoded polypeptide is capable of converting compound (1b) to compound (2b) in at least 75%, at least 85%, at least 90%, at least 95%, at least 98% or at least 99% enantiomeric excess under suitable reaction conditions.

8. The polynucleotide of claim 2 , wherein said encoded polypeptide is capable of converting compound (1b) to compound (2b) with an activity increased at least 2-fold, at least 4-fold, at least 10-fold, at least 25-fold, at least 40-fold, or at least 60-fold relative to the activity of the polypeptide of SEQ ID NO: 38 under suitable reaction conditions.

9. The polynucleotide of claim 2 , wherein said encoded polypeptide is capable of at least 90% or greater conversion of compound (1b) to compound (2b) in 24 h with a substrate loading of about 50 g/L.

10. The polynucleotide of claim 1 , wherein the polynucleotide encodes a polypeptide which comprises the amino acid sequence of SEQ ID NO: 90, 96, 98, 102, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140 or 142.

Assignees

• Codexis Inc

Inventors

• Ang Ee Lui
• Alvizo Oscar
• Behrouzian Behnaz
• Clay Michael D
• Collier Steven J
• Eberhard Ellen D
• Fan Fu
• Song Shiwei
• Smith Derek J
• Widegren Magnus
• Wilson Robert
• Xu Junye
• Zhu Jun

Classifications

• C12P13/02

  Amides, e.g. chloramphenicol {or polyamides; Imides or polyimides; Urethanes, i.e. compounds comprising N-C=O structural element or polyurethanes (peptides C12P21/00 or C07K)} · CPC title

• C12N9/0073 Primary

  with NADH or NADPH as one donor, and incorporation of one atom of oxygen 1.14.13 · CPC title

• C12Y114/13022

  Cyclohexanone monooxygenase (1.14.13.22) · CPC title

• C12P11/00

  Preparation of sulfur-containing organic compounds · CPC title

• Y02P20/52

  using catalysts, e.g. selective catalysts · CPC title