IP Library › Granted Patent US 10,760,037
Granted Patent B2
US 10,760,037 · App. 15/741,519 · Granted Sep 1, 2020

Sulfite compositions

Inventors: Marco Malten (Copenhagen NV, DK); Thomas Agersten Poulsen (Ballerup, DK); Lise Munch Mikkelsen (Roedovre, DK); Dorte Marie Koefoed Klitgaard (Bagsvaerd, DK); Vibeke Skovgaard Nielsen (Nordhavn, DK); Katarina Larson (Malmo, SE)
Assignee: NOVOZYMES A/S
C11D3/38627C11D3/122C11D3/38681C12N9/20C12Y301/01
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Quick Facts
Patent No.
US 10,760,037
App. No.
15/741,519
Granted
Sep 1, 2020
Kind
B2
Abstract

The present invention relates to a composition comprising: (a) a source of sulfite; and (b) a lipase variant of a parent lipase, wherein said variant has improved stability towards sulfite as compared to the parent lipase.

Claims (183)

1. A composition comprising: (a) a source of sulfite; and (b) a lipase variant of a parent lipase,

wherein the variant has lipase activity;

wherein the variant has improved stability in presence of the source of sulfite as compared to the parent lipase;

wherein the parent lipase has the amino acid sequence of SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; SEQ ID NO: 8; SEQ ID NO: 10; or SEQ ID NO: 12; and

wherein the variant is a polypeptide having at least 80%, but less than 100% sequence identity to SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; SEQ ID NO: 8; SEQ ID NO: 10; or SEQ ID NO: 12; and wherein the lipase variant comprises a set of amino acid substitutions selected from the group consisting of:

N33K F51V E96V K98E E210K L259F;

N33K F51V S85K K98E;

N33K F51V K98E E210V;

P250V;

N33K F51V E87T K98E E210K I252W L259F;

N33K F51V L93K K98E;

S83T I86P G91A N92D E96L L97P K98Q;

N33K F51V N92K K98E;

N33K A40E F51V K98E;

N33K F51V E87V K98E;

Q15R N33K F51V K98E;

N33K F51V E87L K98E;

N33K F51V K98E L264P;

N33K A40Q F51V K98E;

N33K F51V K98E L264W;

E210R;

N33K F51V E56K L69R K98E I196V;

Q4K N33K F51V K98E;

N33K F51V G91T K98E;

N33K A38L F51V K98E;

V2K N33K F51V E56K L69R K98E V230R;

N33K F51V G91V K98E;

N33K F51V E56K L69R E96T K98E;

N33K F51V K98E T189D;

N33K F51V K98E R125Q;

N33K A40I F51V K98E;

N33K F51V K98E K223T;

R27D A38G E96D A111D K163G S254D T256P;

N33K F51V K98E R133Q;

N33K F51V L69C K98E;

N33K F51V G91A K98E;

N33K F51V G91F K98E;

N33K F51V K98E S216P;

N33K A40G F51V K98E;

N33K F51V K98E N251G;

N33K F51V G91Y K98E;

N33K F51V K98E V154L;

G23S N33K F51V K98E;

N33K F51V K98E R108K;

E1H N33K F51V K98E;

N33K F51V K98E S170T;

G23T N33K F51V K98E;

N33K F51V K98E T267V;

N33K F51V K98E H135Y;

N33K F51V K98E L269D;

F51V E87K K98E E210Q L227G;

N33K F51V K98E T267L;

N33K F51V K98E R139M;

R27D N33K F51V K98E;

N33K F51V K98E N162G;

N33K E210R;

N33K F51V L69R E96V K98E E210K F211W;

N33K F51V K98E N251G;

R27D N33K F51V K98E;

G23H N33K F51V K98E;

N33K F51V E96T K98E E210K;

N33K E87K;

N33K F51V E96D K98E E210K;

N33K F51V K98E L264E;

N33K F51V L69R E96V K98E E210K F211W;

N33K F51V E56K K98E K163P V176L E210K Y220F L227G;

F51V E210K;

V2K N33K F51V L227G;

N33K F51V K98I;

N33K F51V K98E E210K F211W L259F;

F51V E56K L69R G91A N92D E96L K98Q;

N33K F51V E56K L69R K98E I196V;

F51V E96T K98E E210K;

N33K F51V K98E E210K F211W I252W;

F51V E87K K98I;

N33K F51V K98E N101D K163P E210K;

F51V N101D E210K;

F51V E96D K98E E210K;

N33K F51V K98E V176L E210K L227G;

N33K F51V E96T K98E E210K F211W;

N33K F51V E56K L69R E96D K98E;

N33K F51V E56K L69R K98E T267L;

N33K F51V K98E N101D;

N33K F51V E56K L69R K98E A180D;

N33K F51V E56K L69R E96D K98E; and

N33K F51V E56K L69R K98E D165N.

2. The composition according to claim 1 , wherein the source of sulfite is selected from: sulfites, including sodium sulfite, potassium sulfite, or calcium sulfite; bisulfite, including potassium bisulfite, sodium bisulfite, or calcium bisulfite; metabisulfite, including potassium metabisulfite, sodium metabisulfite, or calcium metabisulfite; or any combination thereof.

3. The composition according to claim 2 , wherein an amount of sulfite (as SO 3 2− ) or metabisulfite (as S 2 O 5 2− ) is from 0.1 wt % to 3 wt %.

4. The composition according to claim 1 , wherein the variant further comprises one or more alterations at positions corresponding to any of positions: 33, 58, 60, 83, 94, 150, 163, 225, 231, 233, 249, 263, 265, and 266 of the parent lipase.

5. The composition according to claim 1 further comprising a surfactant.

6. A lipase variant of a parent lipase,

wherein the variant has lipase activity,

wherein the variant has improved stability in presence of a source of sulfite as compared to the parent lipase,

wherein the variant has at least 80% but less than 100% sequence identity to SEQ ID NO: 2; SEQ ID NO: 4; SEQ ID NO: 6; SEQ ID NO: 8; SEQ ID NO: 10; or SEQ ID NO: 12, and

wherein the variant comprises

a set of amino acid substitutions selected from the group consisting of:

N33K F51V E96V K98E E210K L259F;

N33K F51V S85K K98E;

N33K F51V K98E E210V;

P250V;

N33K F51V E87T K98E E210K I252W L259F;

N33K F51V L93K K98E;

S83T I86P G91A N92D E96L L97P K98Q;

N33K F51V N92K K98E;

N33K A40E F51V K98E;

N33K F51V E87V K98E;

Q15R N33K F51V K98E;

N33K F51V E87L K98E;

N33K F51V K98E L264P;

N33K A40Q F51V K98E;

N33K F51V K98E L264W;

E210R;

N33K F51V E56K L69R K98E I196V;

Q4K N33K F51V K98E;

N33K F51V G91T K98E;

N33K A38L F51V K98E;

V2K N33K F51V E56K L69R K98E V230R;

N33K F51V G91V K98E;

N33K F51V E56K L69R E96T K98E;

N33K F51V K98E T189D;

N33K F51V K98E R125Q;

N33K A40I F51V K98E;

N33K F51V K98E K223T;

R27D A38G E96D A111D K163G S254D T256P;

N33K F51V K98E R133Q;

N33K F51V L69C K98E;

N33K F51V G91A K98E;

N33K F51V G91F K98E;

N33K F51V K98E S216P;

N33K A40G F51V K98E;

N33K F51V K98E N251G;

N33K F51V G91Y K98E;

N33K F51V K98E V154L;

G23S N33K F51V K98E;

N33K F51V K98E R108K;

E1H N33K F51V K98E;

N33K F51V K98E S170T;

G23T N33K F51V K98E;

N33K F51V K98E T267V;

N33K F51V K98E H135Y;

N33K F51V K98E L269D;

F51V E87K K98E E210Q L227G;

N33K F51V K98E T267L;

N33K F51V K98E R139M;

R27D N33K F51V K98E;

N33K F51V K98E N162G; End of first example

N33K E210R;

N33K F51V L69R E96V K98E E210K F211W;

N33K F51V K98E N251G;

R27D N33K F51V K98E;

G23H N33K F51V K98E;

N33K F51V E96T K98E E210K;

N33K E87K;

N33K F51V E96D K98E E210K;

N33K F51V K98E L264E;

N33K F51V L69R E96V K98E E210K F211W;

N33K F51V E56K K98E K163P V176L E210K Y220F L227G;

F51V E210K;

V2K N33K F51V L227G;

N33K F51V K98I; End of second example

N33K F51V K98E E210K F211W L259F;

F51V E56K L69R G91A N92D E96L K98Q;

N33K F51V E56K L69R K98E I196V;

F51V E96T K98E E210K;

N33K F51V K98E E210K F211W I252W;

F51V E87K K98I;

N33K F51V K98E N101D K163P E210K;

F51V N101D E210K;

F51V E96D K98E E210K;

N33K F51V K98E V176L E210K L227G;

N33K F51V E96T K98E E210K F211W;

N33K F51V E56K L69R E96D K98E;

N33K F51V E56K L69R K98E T267L;

N33K F51V K98E N101D;

N33K F51V E56K L69R K98E A180D;

N33K F51V E56K L69R E96D K98E; and

N33K F51V E56K L69R K98E D165N.

7. The variant according to claim 6 , wherein the variant further comprises one or more amino acid substitutions at a position corresponding to any of positions 33, 58, 60, 83, 94, 150, 163, 225, 231, 233, 249, 263, 265 and 266 of the parent lipase.

8. A polynucleotide encoding the variant according to claim 6 .

9. A nucleic acid construct comprising the polynucleotide according to claim 8 .

10. An expression vector comprising the polynucleotide according to claim 8 .

11. A host cell comprising the polynucleotide according to claim 8 .

12. A method of producing a lipase variant, comprising: (a) cultivating the host cell according to claim 11 under conditions suitable for expression of the variant; and (b) recovering the variant.

Priority Claims (1)
EP 15175251 · Jul 3, 2015 · regional
Continuity (1)
Related Publication 20190071619A1 · Mar 7, 2019