IP Library › Granted Patent US 9,624,478
Granted Patent B2
US 9,624,478 · App. 14/799,180 · Granted Apr 18, 2017

Biocatalysts and methods for the synthesis of substituted lactams

Inventors: Fabien Louis Cabirol (Dusseldorf, DE); Haibin Chen (Beijing, CN); Anupam Gohel (Bekasi, ID); Paulina Elena Salim (Singapore, SG); Derek J. Smith (Singapore, SG); Jacob Janey (New York, NY); Birgit Kosjek (Westfield, NJ); Weng Lin Tang (Singapore, SG); Helen Hsieh (Singapore, SG); Son Q. Pham (Singapore, SG)
Assignee: Codexis, Inc.
C12N9/1096C07D211/76C12P13/001C12P13/005C12P17/12C12Y206/01
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Quick Facts
Patent No.
US 9,624,478
App. No.
14/799,180
Granted
Apr 18, 2017
Kind
B2
Abstract

The present disclosure relates to transaminase polypeptides capable of aminating a dicarbonyl substrate, and polynucleotides, vectors, host cells, and methods of making and using the transaminase polypeptides.

Claims (18)

1. An engineered polypeptide comprising an amino acid sequence having at least 80% sequence identity to SEQ ID NO:4 and a substitution at positon X192 selected from X192A, X192G, X192H, X192K, X192N, X192Q, X192R, and X192S.

2. The engineered polypeptide of claim 1 , in which the amino acid sequence further comprises one or more residue differences as compared to SEQ ID NO:4 selected from: X42G; X54P; X54R; X69A; X69G; X69T; X122M; X124F; X124L; X124N; X124R; X126A; X126T; X136Y, X136F; X150S; X152C; X152G; X152I; X152L; X152S; X156A; X156S; X156T; X157L; and X165N.

3. The engineered polypeptide of claim 1 , in which the amino acid sequence further comprises a combination of residue differences selected from:

(a) X124W and X327L;

(b) X209M and X300G;

(c) X122F, X223V and X284A;

(d) XI92A, X215H and X311T;

(e) X62N, X124F, X126A and X136L;

(f) X124W, X126A, X136L, X192A and X284A; and

(g) X124W, X126A, X136L, X152R/X152L/X152I, and X192A.

4. The engineered polypeptide of claim 1 , in which the amino acid sequence further comprises at least one or more residue differences as compared to SEQ ID NO:4 selected from: X54A; X54L; X56D; X56E; X61G; X61W; X62A; X62L; X62N; X62S; X64C; X68I; X122F, X122W; X122Y; X124K; X124M; X124S; X124W; X126C; X126I; X136K; X136L; X136M; X139E; X140K; X143V; X150L; X155I; X155L; X159G; X160L; X176C; X199L; X199V; X209F; X209M; X209Q; X209V; X223S; X223T; X223V; X227I; X282L, X282V; X284A; X284P; X284S; X284T; and X284V.

5. An engineered polynucleotide encoding the polypeptide of claim 1 .

6. A vector comprising the polynucleotide of Claim 5 .

7. A host cell comprising the vector of claim 6 .

8. A method of converting compound 2 to compound 1,

comprising contacting compound 2 with the engineered polypeptide of claim 1 under suitable reaction conditions to produce compound 1.

9. A method of converting compound 2 to compound 1d and compound 1a

comprising contacting compound 2 with the engineered polypeptide of claim 1 under suitable reaction conditions to produce compound 1d and 1a, under suitable reaction conditions wherein said engineered polypeptide comprises at least 1.5 fold the activity of SEQ ID NO:4.

Assignments (1)
SECURITY INTEREST Recorded Feb 15, 2024
From: CODEXIS, INC.
To: INNOVATUS LIFE SCIENCES LENDING FUND I, LP, AS COLLATERAL AGENT
Reel/Frame 066600/0650 →
Continuity (3)
Division 14342713
Provisional Application 61532259 · Sep 8, 2011
Related Publication 20150307857A1 · Oct 29, 2015