Variant penicillin-G acylases

Patent metadata

Field	Value
Publication number	US-11180747-B2
Application number	US-202016995630-A
Country	US
Kind code	B2
Filing date	Aug 17, 2020
Priority date	May 7, 2015
Publication date	Nov 23, 2021
Grant date	Nov 23, 2021

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.

First claim

What is claimed is:

1. An engineered penicillin G acylase capable of removing the 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.

2. The engineered penicillin G acylase of claim 1 , wherein said penicillin G acylase comprises at least one substitution or one specific combination of substitutions as provided in Table 5.1, Table 6.2, and/or Table 6.3.

3. The engineered penicillin G acylase of claim 1 , wherein said engineered penicillin G acylase comprises SEQ ID NO: 6.

4. The engineered penicillin G acylase of claim 1 , wherein said engineered penicillin G acylase is encoded by a polynucleotide sequence selected from SEQ ID NOS: 5.

Assignees

• Codexis Inc

Inventors

• Banerjee Goutami
• Yang Jie
• Zhang Xiyun
• Milczek Erika M
• Mayo Melissa Ann
• Jenne Stephan

Classifications

• C12Y305/01011

  Penicillin amidase (3.5.1.11), i.e. penicillin-amidohydrolase · CPC title

• A61P5/50

  for increasing or potentiating the activity of insulin · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• C12P21/00

  Preparation of peptides or proteins (single cell protein C12N1/00) · CPC title

• C12N9/84 Primary

  Penicillin amidase {(3.5.1.11)} · CPC title