IP Library › Granted Patent US 12,378,541
Granted Patent B2
US 12,378,541 · App. 17/504,852 · Granted Aug 5, 2025

Penicillin-G acylases

Inventors: Goutami Banerjee (Hayward, CA); Jie Yang (Foster City, CA); Xiyun Zhang (Fremont, CA); Erika M. Milczek (New York, NY); Melissa Ann Mayo (Foster City, CA); Stephan Jenne (Foster City, CA)
Assignee: Codexis, Inc.
C12N9/84C12P21/00C12Y305/01011
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Quick Facts
Patent No.
US 12,378,541
App. No.
17/504,852
Filed
Oct 19, 2021
Granted
Aug 5, 2025
Kind
B2
Art Unit
1657
USPC
435/68.1
Abstract

The present disclosure relates to engineered penicillin G acylase (PGA) enzymes having improved properties, polynucleotides encoding such enzymes, compositions including the enzymes, and methods of using the enzymes.

Claims (3)

1. A method for producing free insulin, comprising: i) providing an engineered penicillin G acylase capable of removing A1/B1/B29 tri-phenyl acetate protecting groups from insulin to produce free insulin, wherein said penicillin G acylase is at least 95% identical to SEQ ID NO: 6, and wherein said engineered penicillin G acylase comprises an alanine at position 264, an asparagine at position 484, and a lysine at position 547, wherein said positions are numbered with reference to SEQ ID NO: 6 and insulin comprising A1/B1/B29 tri-phenyl acetate protecting groups; and ii) exposing said engineered penicillin G acylase to said insulin comprising A1/B1/B29 tri-phenyl acetate protecting groups under conditions such that said engineered penicillin G acylase removes the A1/B1/B29 tri-phenyl acetate protecting groups and free insulin is produced.

2. The method of claim 1 , wherein said engineered penicillin G acylase removes the A1/B1/B29 tri-phenyl acetate protecting groups from more than 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% of the insulin comprising A1/B1/B29 tri-phenyl acetate protecting groups to produce free insulin.

3. The method of claim 1 , wherein said penicillin G acylase comprises SEQ ID NO: 6, 10, or 12.

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 (6)
Continuation 16995630 · Aug 17, 2020
Continuation 16524546 · Jul 29, 2019
Continuation 15914258 · Mar 7, 2018
Division 15148056 · May 6, 2016
Provisional Application 62158118 · May 7, 2015
Related Publication 20220042004A1 · Feb 10, 2022
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