US 12,480,104 B2
Engineered sucrose phosphorylase variant enzymes
Jonathan Vroom, South San Francisco, CA (US); Santhosh Sivaramakrishnan, Redwood City, CA (US); and Nandhitha Subramanian, Cambridge (GB)
Assigned to Codexis, Inc., Redwood City, CA (US)
Appl. No. 17/617,533
Filed by Codexis, Inc., Redwood City, CA (US)
PCT Filed Jun. 30, 2020, PCT No. PCT/US2020/040250
§ 371(c)(1), (2) Date Dec. 8, 2021,
PCT Pub. No. WO2021/003132, PCT Pub. Date Jan. 7, 2021.
Claims priority of provisional application 62/869,670, filed on Jul. 2, 2019.
Prior Publication US 2022/0243183 A1, Aug. 4, 2022
Int. Cl. C12N 9/00 (2006.01); C12N 1/20 (2006.01); C12N 9/10 (2006.01); C12N 15/72 (2006.01)
CPC C12N 9/1051 (2013.01) [C12N 1/20 (2013.01); C12N 15/72 (2013.01); C12Y 204/01007 (2013.01); C12N 2800/101 (2013.01)] 21 Claims
 
1. An engineered sucrose phosphorylase comprising a polypeptide sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to SEQ ID NO: 2, wherein the polypeptide sequence of said engineered sucrose phosphorylase comprises at least one substitution at position 397 and wherein the amino acid positions of said polypeptide sequence are numbered with reference to SEQ ID NO: 2, and wherein the engineered sucrose phosphorylase exhibits improved activity on a substrate selected from the group consisting of sucrose, sucrose-related disaccharides and/or inorganic phosphate as compared to wild-type Alloscardovia omnicolens sucrose phosphorylase set forth in SEQ ID NO: 2.