Jovana Nazor, Milpitas, CA (US); Vesna Mitchell, Santa Clara, CA (US); David Elgart, San Mateo, CA (US); Katrina W. Lexa, Santa Rosa, CA (US); Nikki Dellas, San Carlos, CA (US); Robert Kevin Orr, Cranford, NJ (US); Oscar Alvizo, Fremont, CA (US); Ravi David Garcia, Los Gatos, CA (US); Judy Victoria Antonio Viduya, Greenbrae, CA (US); and Courtney Dianne Moffett, San Francisco, CA (US)
Assigned to Codexis, Inc., Redwood City, CA (US)
Filed by Codexis, Inc., Redwood City, CA (US)
Filed on Nov. 30, 2021, as Appl. No. 17/537,941.
Application 16/902,001 is a division of application No. 15/588,378, filed on May 5, 2017, granted, now 10,724,025, issued on Jul. 28, 2020.
Application 17/537,941 is a continuation of application No. 16/902,001, filed on Jun. 15, 2020, granted, now 11,214,786.
Claims priority of provisional application 62/332,103, filed on May 5, 2016.
Prior Publication US 2022/0154164 A1, May 19, 2022
1. A method for producing phenyl acetate mono-protected or di-protected insulin, comprising: i) providing an engineered penicillin G acylase having a polypeptide sequence that is at least 95% identical to SEQ ID NO: 1220, wherein said penicillin G acylase comprises a leucine residue at position X2 as compared to the sequence of SEQ ID NO: 1220 and free insulin; and ii) exposing said engineered penicillin G acylase to said insulin, under conditions such that said engineered penicillin G acylase acylates up to two positions selected from the group of positions consisting of the A1 position of insulin, the B1 position of insulin, and the B29 position of insulin, thereby producing mono-protected or di-protected insulin.