Penicillin-G acylases

US12480111B2 · US · B2

Patent metadata
FieldValue
Publication numberUS-12480111-B2
Application numberUS-202318189091-A
CountryUS
Kind codeB2
Filing dateMar 23, 2023
Priority dateJun 27, 2017
Publication dateNov 25, 2025
Grant dateNov 25, 2025

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  1. Title

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  2. Abstract

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  3. Assignees and inventors

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  4. Key dates

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  5. First independent claim

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  6. CPC / IPC classifications

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  7. Citations and related patents

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Abstract

Official abstract text for this publication.

The present invention provides engineered penicillin G acylase (PGA) enzymes, polynucleotides encoding the enzymes, compositions comprising the enzymes, and methods of using the engineered PGA enzymes.

First claim

Opening claim text (preview).

We claim: 1 . An engineered polynucleotide encoding an engineered penicillin G acylase variant having at least 95% sequence identity to SEQ ID NO:6, and substitutions at positions 254, 255, and 370 and at least one additional substitution selected from 54C, 62G, 115A/P, 125L/T, 127S/V, 185V, 253K/V, 256Q, 257I, 257V, 260A/P, 268S/V, 322P, 325G, 348C, 348Q, 369L, 369P, 369V, 369W, 372A/H/L, 373F/M, 377P 378H, 384A, 384F/513Q/536M, 384G/L, 388T, 389L, 391P/S, 435R, 461A, 517L/P, 530C/Y, 554A/E/P/V, 556G, 557G/S, 559P/S, 560I, 600T/623V, 623A/G/R/W, 624A, 626G, 627G/H, 705G/P, 706G, 707S, 723A/G, 740L, 748G, and 752E, and wherein said engineered penicillin G acylase variant is capable of deacylating a di or tetra-protected insulin dimer, wherein said positions are numbered with reference to SEQ ID NO:6. 2 . The engineered polynucleotide encoding an engineered penicillin G acylase variant of claim 1 comprising the substitutions 254W, 255G, and a substitution set selected from 103V/370F/444S/706G/766G, 103V/369W/370F/442I/444S/536M/556G/766G, 103V/369W/370F/444S, 103V/369W/370F/444S/556G/706G/766G, 103V/369W/370F/444S/765P/766G, 103V/369W/370F/765P/766G, 257V/362V/370I/384A/451R, 257V/362V/370I/384L/451R/723L, 362V/370I/451R/705D, 369P/370F, 369P/370F/444S/706G/766G, 369P/370F/556G/766G, 369V/370F/388T/444S/556G/766G, 369V/370F/444S, 369V/370F/444S/556G/766G, 369V/370F/556G, 369V/370F/556G/765P, 369V/370F/556G/766G, 369V/370F/766G, 369W/370F/444S/556G, 369W/370F/444S/556G/612A/766G, 369W/370F/444S/556G/706G/765P, 369W/370F/444S/706G/765P/766G, 370I/372A/373M/384L/513Q/560G, 370I/372A/384L/451R/705D, 370I/372A/384L/560G/705D, 370I/384A/451R/560G/705D/723L, 370I/384L/451R/705D/723L, 370I/451R/560G/705D/723L, and 370I/451R/705D/723L. 3 . The engineered polynucleotide encoding an engineered penicillin G acylase variant of claim 2 comprising the substitutions 370I, 384L, 513Q, 560G and a substitution set is selected from 103V/254W/255G/372S/373F/557G, 253H/254W/255G/322T/369W/372A/373M/623G, 253H/254Q/255G/322T/369W/372A/373M/623G, 253H/254Q/255G/369W/372A/373M/391A/623G/723A, 253H/254Q/255G/369W/372A/373M/619R/623G/723A, 253H/254Q/255G/369W/372A/373M/623G/723A, 253H/254Q/255G/372A/373L/623G/723A, 253H/254S/255V/369W/372A/373M/623S/723A, 253H/254S/255G/369W/372A/373M, 253H/254W/255G/322T/369W/372A/373W/723A, 253H/254W/255G/369W/372A/373M/623G/723A, 253H/254W/255G/372A/373L/623S, 253S/254S/255V/322T/369W/372A/373M/619R/723A, 254W/255G/260S/372S/373F/556G, 254W/255G/260S/372S/373F/556G/557V/559S, 254W/255G/322T/369W/372A/373M, 254W/255G/322T/369W/372A/373W/723A, 254W/255G/322T/369W/372A/373M/623G/723A, and 254W/255G/369W/372A/373F/556G. 4 . The engineered polynucleotide encoding an engineered penicillin G acylase variant of claim 1 , wherein said engineered penicillin G acylase variant comprises a histidine tag. 5 . The engineered polynucleotide encoding an engineered penicillin G acylase variant of claim 4 , wherein said histidine tag is present at the C-terminus of said engineered penicillin G acylase variant. 6 . The engineered polynucleotide encoding an engineered penicillin G acylase variant of claim 1 , wherein said engineered penicillin G acylase variant comprises a polypeptide sequence set forth in variant numbers 1-308. 7 . The engineered polynucleotide encoding an engineered penicillin G acylase variant of claim 1 , wherein said engineered penicillin G acylase variant comprises a polypeptide sequence selected from the even-numbered sequences between SEQ ID NO:4 and SEQ ID NO:90. 8 . The engineered polynucleotide sequence of claim 1 , said wherein said sequence comprises a polynucleotide sequence that is at least 95% identical to a sequence selected from the odd-numbered sequences between SEQ ID NO:5 and SEQ ID NO:89. 9 . A vector comprising the engineered polynucleotide sequence of claim 1 . 10 . The vector of claim 9 , further comprising at least one control sequence. 11 . A host cell comprising the vector of claim 10 . 12 . A method for producing an engineered penicillin G acylase variant, comprising culturing said host cell of claim 11 under conditions that said engineered penicillin G acylase variant is produced by said host cell. 13 . The method of claim 12 , further comprising the step of recovering said engineered penicillin G acylase variant produced by said host cell.

Assignees

Inventors

Classifications

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

  • C12N9/84Primary

    Penicillin amidase {(3.5.1.11)} · CPC title

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Frequently asked questions

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What does patent US12480111B2 cover?
The present invention provides engineered penicillin G acylase (PGA) enzymes, polynucleotides encoding the enzymes, compositions comprising the enzymes, and methods of using the engineered PGA enzymes.
Who is the assignee on this patent?
Codexis Inc
What technology area does this patent fall under?
Primary CPC classification C12N9/84. Mapped technology areas include Chemistry & Metallurgy.
When was this patent published?
Publication date Tue Nov 25 2025 00:00:00 GMT+0000 (Coordinated Universal Time) (B2). Legal status and post-grant events are not shown on this page.
What related patents are in patentsdb?
We list 7 related publications on this page (citations in our corpus or others sharing the same primary CPC).