Variant enzymes

Patent metadata

Field	Value
Publication number	US-2016177281-A1
Application number	US-201414909031-A
Country	US
Kind code	A1
Filing date	Jul 24, 2014
Priority date	Jul 29, 2013
Publication date	Jun 23, 2016
Grant date	—

Abstract

Disclosed are glycosyl hydrolase enzyme variants, particularly variants of certain oxidoreductases of glycosyl hydrolase family 61. Nucleic acids encoding the glycosyl hydrolyase variants, compositions including the glycosyl hydrolase variants, methods of producing the variants, and methods of using the variants are also described.

First claim

1 . (canceled)

2 . A variant of a parent glycosyl hydrolase 61 (GH61) enzyme, wherein said variant has cellulase augmenting activity, and has at least 80% sequence identity to SEQ ID NO:3, wherein said variant comprises at least one amino acid substitution at a position selected from the group consisting of: T287, W13, L288, P19, T20, P29, A37, L39, D47, A48, S68, T107, S121, G122, T127, S164, G166, Q167, A168, N216, I218, I219, V224, T289, Y291, S297, T303, R304, A306, P308, S312, T313, N315, Y317, Y318, N5, I7, V12, Q15, T21, F22, S26, P28, V32, G33, T35, A36, N41, Q50, N51, K58, N59, V80, P89, T108, G116, L120, G123, V132, S135, N147, A161, A181, S183, S185, Q189, G195, T196, H200, A201, T202, I208, P222, T223, S226, S231, G235, L290, G295, G296, G298, Y299, S300, G301, P302, P307, A309, T310, P316, A319, L322, I4, I9, N10, P23, Y24, E25, N27, D38, F43, V44, Y49, P52, I54, T61, N62, K64, V69, K70, A71, I90, V91, N96, N98, D100, T103, E110, V117, L119, D124, P125, G126, V141, D146, A149, I160, H163, A171, N173, P175, I180, S187, L188, S191, L194, D197, P204, V206, L207, N209, Y211, S213, P214, Y217, T230, V232, A233, G292, Q293, Q320, and N323, wherein the position of each amino acid substitution corresponds to SEQ ID N0:3.

3 . The variant of claim 2 , wherein said variant comprises one or more of:
an amino acid substitution at position T287 selected from the group consisting of: H, A, F, G, S, and I; an amino acid substitution at position W13 selected from the group consisting of: G, L, N, R, T, V, F, Q, Y, A, E, I, K, and M; an amino acid substitution at position L288 selected from the group consisting of: E, F, G, I, K, M, N, R, S, T, A, C, D, V, and W; an amino acid substitution at position P19 selected from the group consisting of: F, M, E, H, Q, T, K, and W; an amino acid substitution at position T20 selected from the group consisting of: K, P, G, M, R, Y, A, and N; an amino acid substitution at position P29 selected from the group consisting of: C, E, F, K, T, Y D, G, M, R, and S; an amino acid substitution at position A37 selected from the group consisting of: D, S, E, L, W, F, K, and Q; an amino acid substitution at position L39 selected from the group consisting of: A, D, E, M, T, Y, F, I, N, P, Q, S, and V; an amino acid substitution at position D47 selected from the group consisting of: A, K, T, H, R, P, and W; an amino acid substitution at position A48 selected from the group consisting of: C, E, G, K, L, N, T, Y, Q, W, H, I, and S; an amino acid substitution at position S68 selected from the group consisting of: E, I, L, M, N, T, V, and Y; an amino acid substitution at position T107 selected from the group consisting of: A, D, E, K, M, R, S, and G; an amino acid substitution at position S121 selected from the group consisting of: E, F, N, T, C, L, V, D, G, K, M, and Y; an amino acid substitution at position G122 selected from the group consisting of: E, N, S, V, A, M, T, F, and L; an amino acid substitution at position T127 selected from the group consisting of: A, E, F, I, L, M, V, W, and Y; an amino acid substitution at position S164 selected from the group consisting of: A, D, E, G, K, L, M, Q, R, V, and Y; an amino acid substitution at position G166 selected from the group consisting of: A, D, E, F, H, K, M, N, Q, and S; an amino acid substitution at position Q167 selected from the group consisting of: E, A, D, F, G, K, L, N, R, V, and Y; an amino acid substitution at position A168 selected from the group consisting of: E, M, D, F, I, L, N, P, Q, R, T, and Y; an amino acid substitution at position N216 selected from the group consisting of: D, Q, T, K, and P; an amino acid substitution at position I218 selected from the group consisting of: T, A, D, E, K, M, N, P, S, and V; an amino acid substitution at position I219 selected from the group consisting of: M, A, D, L, N, P, and Q; an amino acid substitution at position V224 selected from the group consisting of: Q, I, A, E, N, P, S, and T; an amino acid substitution at position T289 selected from the group consisting of: A, D, K, L, M, R, and S; an amino acid substitution at position Y291 selected from the group consisting of: A, F, G, I, K, L, M, N, R, S, V, W, D, and E; an amino acid substitution at position S297 selected from the group consisting of: D, E, K, N, Q, R, and T; an amino acid substitution at position T303 selected from the group consisting of: K, M, P, R, S, V, and Y; an amino acid substitution at position R304 selected from the group consisting of: A, C, D, E, N, Q, T, and V; an amino acid substitution at position A306 selected from the group consisting of: E, F, G, I, K, L, M, Q, T, and Y; an amino acid substitution at position P308 selected from the group consisting of: A, G, H, I, K, L, Q, R, S, T, V, W, and Y; an amino acid substitution at position S312 selected from the group consisting of: Q, F, G, I, K, L, M, N, T, V, D, R, and Y; an amino acid substitution at position T313 selected from the group consisting of: S, D, G, L, P, V, E, F, I, K, M, and Y; an amino acid substitution at position N315 selected from the group consisting of: S, H, L, Q, R, V, Y, E, K, and M; an amino acid substitution at position Y317 selected from the group consisting of: Q, S, V, R, E, G, L, M, P, and T; a Y318T amino acid substitution; an amino acid substitution at position N5 selected from the group consisting of: A, D, G, and S; an amino acid substitution at position I7 selected from the group consisting of: L, M, and V; an amino acid substitution at position V12 selected from the group consisting of: M and T; an amino acid substitution at position Q15 selected from the group consisting of: H, K, and D; an amino acid substitution at position T21 selected from the group consisting of: I, K, M, and S; an amino acid substitution at position F22 selected from the group consisting of: K, L, and R; an amino acid substitution at position S26 selected from the group consisting of: M, P, and T; an amino acid substitution at position P28 selected from the group consisting of: M, S, and V; an amino acid substitution at position V32 selected from the group consisting of: A and P; an amino acid substitution at position G33 selected from the group consisting of: Q, A, and S; an amino acid substitution at position T35 selected from the group consisting of: A, D, E, K, N, and S; an amino acid substitution at position A36 selected from the group consisting of: T, I, N, S, and Y; an amino acid substitution at position N41 selected from the group consisting of: D, E, L, and M; an amino acid substitution at position Q50 selected from the group consisting of: A, T, Y, D, E, and K; an amino acid substitution at position N51 selected from the group consisting of: S, E, H, K, L, and T; an amino acid substitution at position K58 selected from the group consisting of: I, R, and V; an amino acid substitution at position N59 selected from the group consisting of: G, E, Q, and S; an amino acid substitution at position V80 selected from the group consisting of: A, E, L, and M; an amino acid substitution at position P89 selected from the group consisting of: T, S, V, and L; an amino acid substitution at position T108 selected from the group consisting of: A, Q, R, and S; an amino acid substitution at position G116 selected from the group consisting of: H, Q, and S; an amino acid substitution at position L120 selected from the group consisting of: V, T, I, and S; an amino acid substitution at position G123 selected from the group consisting of: E, Q, and R; an amino acid substitution at position V132 selected from the group consisting of: D, E, M, and R; an amino acid substitution at position S135 selected from the group consisting of: A, E, G, H, M, and N; an amino acid substitution at position N147 selected from the group consisting of: E, M, S, and T; an amino acid substitution at position A161 selected from the group consisting of: E, G, L, and S; an amino ac

Assignees

• Danisco Us Inc

Inventors

• Foukaraki Maria
• Hommes Ronaldus Wilhelmus
• Kaper Thijs
• Kelemen Bradley R
• Kralj Slavko
• Lantz Suzanne E
• Nikolaev Igor
• Van Der Kley Wilhelmus
• Van Lieshout Johan Franciscus Thomas
• Van Stigt Thans Sander

Classifications

• C12N9/2437 Primary

  Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150) · CPC title

• C12N9/2434

  acting on beta-1,4-glucosidic bonds · CPC title

• C12P19/14

  produced by the action of a carbohydrase {(EC 3.2.x)}, e.g. by alpha-amylase {, e.g. by cellulase, hemicellulase} · CPC title

• C12N9/0083

  Miscellaneous (1.14.99) · CPC title

• C12N9/2402

  hydrolysing O- and S- glycosyl compounds (3.2.1) · CPC title