US 11,999,976 B2 · Engineered ketoreductase polypeptides and uses thereof

Abstract

Provided are amino acid sequences of ketoreductase polypeptides that are useful for asymmetrically synthesizing chiral alcohol compounds, its preparation process as well as reaction process under industrial-relevant conditions. Also provided are polynucleotide sequences encoding engineered ketoreductase polypeptides, engineered host cells capable of expressing engineered ketoreductase polypeptides, and methods of producing chiral alcohol compounds using engineered ketoreductase polypeptides. Compared to other enzymes, the engineered ketoreductase polypeptides provided by the invention have better catalytic activity and thermal stability, allowing purification of the enzyme solution by heat treatment, which is advantageous for the production of enzymes and the industrial application of enzymatic reactions. The use of the engineered polypeptides of the present invention greatly simplifies the production process of chiral alcohol compounds, reduces the cost of production and the impact on the environment, and has good industrial application prospects.

Claims19

Dependent claims

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.

Dependencies are inferred from the claim text.

Engineered ketoreductase polypeptides and uses thereof

Life of this patent

Engineered ketoreductase polypeptides and uses thereof

Life of this patent

Abstract

Claims19