US 12,480,111 B2
Penicillin-G acylases
Rama Voladri, Pleasanton, CA (US); Christopher Michael Micklitsch, Philadelphia, PA (US); Oscar Alvizo, Fremont, CA (US); Jovana Nazor, Milpitas, CA (US); Da Duan, Foster City, CA (US); Judy Victoria Antonio Viduya, Greenbrae, CA (US); Stephan Jenne, Foster City, CA (US); Chihui An, Scotch Plains, NJ (US); Keith Allen Canada, Freehold, NJ (US); Paul N. Devine, Tinton Falls, NJ (US); Iman Farasat, Rahway, NJ (US); Anna Fryszkowska, New York, NY (US); Katrina W. Lexa, Santa Rosa, CA (US); and Robert Kevin Orr, Cranford, NJ (US)
Assigned to Codexis, Inc., Redwood City, CA (US)
Filed by Codexis, Inc., Redwood City, CA (US)
Filed on Mar. 23, 2023, as Appl. No. 18/189,091.
Application 18/189,091 is a continuation of application No. 16/616,041, granted, now 11,643,642, previously published as PCT/US2018/034073, filed on May 23, 2018.
Claims priority of provisional application 62/527,199, filed on Jun. 30, 2017.
Claims priority of provisional application 62/525,404, filed on Jun. 27, 2017.
Prior Publication US 2023/0272363 A1, Aug. 31, 2023
Int. Cl. C12N 9/84 (2006.01)
CPC C12N 9/84 (2013.01) [C12Y 305/01011 (2013.01)] 13 Claims
 
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.