US 11,999,976 B2
Engineered ketoreductase polypeptides and uses thereof
Haibin Chen, Ningbo (CN); Chuanyang Shang, Ningbo (CN); Baoqin Cai, Ningbo (CN); Marco Bocola, Ningbo (CN); Shumin Shi, Ningbo (CN); Ruimei Hong, Ningbo (CN); Lei Sun, Ningbo (CN); and Yong Koy Bong, Ningbo (CN)
Assigned to Enzymaster (Ningbo) Bio-Engineering Co., Ltd, Ningbo (CN)
Appl. No. 17/290,916
Filed by Enzymaster (Ningbo) Bio-Engineering Co., Ltd, Ningbo (CN)
PCT Filed Dec. 11, 2019, PCT No. PCT/CN2019/124709
§ 371(c)(1), (2) Date May 3, 2021,
PCT Pub. No. WO2020/119740, PCT Pub. Date Jun. 18, 2020.
Claims priority of application No. 201811537509.4 (CN), filed on Dec. 15, 2018.
Prior Publication US 2021/0371829 A1, Dec. 2, 2021
Int. Cl. C12N 9/04 (2006.01); C12N 11/02 (2006.01); C12P 7/22 (2006.01); C12P 7/62 (2022.01); C12P 41/00 (2006.01)
CPC C12N 9/0006 (2013.01) [C12N 11/02 (2013.01); C12P 7/22 (2013.01); C12P 7/62 (2013.01); C12P 41/002 (2013.01); C12Y 101/01184 (2013.01)] 19 Claims
 
1. An engineered ketoreductase polypeptide which is, under suitable reaction conditions, capable of catalyzing the conversion of 4-hydroxy-2-butanone to (R) -(-)-1,3-butanediol, wherein the amino acid sequence of said polypeptide comprises one or more of substitutions selected from the group consisting of substitutions corresponding to substitutions X46T, X185C, and X228A in the polypeptide of SEQ ID NO 2, wherein the numbering refers to SEQ ID NO:2, wherein said amino acid sequence is selected from the group consisting of SEQ ID NO: 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 66, 68, 70, 72, 74, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 126, 128, 130, 132, 134, 136, 138, 140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, and 332.