Esterases and uses thereof

Patent metadata

Field	Value
Publication number	US-12098398-B2
Application number	US-202217887544-A
Country	US
Kind code	B2
Filing date	Aug 15, 2022
Priority date	Jul 12, 2016
Publication date	Sep 24, 2024
Grant date	Sep 24, 2024

Abstract

The present invention relates to novel esterase, more particularly to esterase variants having improved thermostability compared to the esterase of SEQ ID N o 1 and the uses thereof for degrading polyester containing material, such as plastic products. The esterases of the invention are particularly suited to degrade polyethylene terephthalate, and material containing polyethylene terephthalate.

First claim

We claim:

1. An esterase variant which (i) has a polyester degrading activity, (ii) has at least 75% identity to the full length amino acid sequence set forth in SEQ ID NO: 1, and (iii) has at least one amino acid substitution as compared to SEQ ID NO: 1, wherein the position of the amino acid substitution is numbered by reference to the amino acid sequence set forth in SEQ ID NO: 1 and one of said at least one amino acid substitution is at position D203.

2. The esterase variant of claim 1 , wherein the amino acid substitution at position D203 is D203E, D203K, or D203R.

3. The esterase variant of claim 1 , which further comprises: a) at least one amino acid substitution at position S248; or b) further comprises at least one amino acid substitution at position E173.

4. The esterase variant of claim 1 , which exhibits increased thermostability as compared to the esterase of SEQ ID NO: 1.

5. The esterase variant of claim 1 , which has at least the amino acid substitution selected from D203E/K/R and exhibits increased thermostability as compared to the esterase of SEQ ID NO: 1.

6. A nucleic acid encoding an esterase as defined in claim 1 .

7. An expression cassette or vector comprising the nucleic acid of claim 6 .

8. A host cell comprising the nucleic acid of claim 6 .

9. A method of producing an esterase comprising:
(a) culturing the host cell according to claim 8 under conditions suitable to express the nucleic acid encoding the esterase; and
(b) recovering said esterase from the cell culture.

10. A composition comprising an esterase according to claim 1 and one or several excipients or additives.

11. A method of degrading a plastic product containing at least one polyester comprising (a) contacting the plastic product with an esterase according to claim 1 , thereby degrading the plastic product.

12. The method of claim 11 , further comprising (b) recovering monomers and/or oligomers resulting from the degradation of the at least one polyester.

13. The method of claim 11 , wherein the plastic product comprises at least one polyester selected from polyethylene terephthalate (PET), polytrimethylene terephthalate (PTT), polybutylen terephthalate (PBT), polyethylene isosorbide terephthalate (PEIT), polylactic acid (PLA), polyhydroxyalkanoate (PHA), polybutylene succinate (PBS), polybutylene succinate adipate (PBSA), polybutylene adipate terephthalate (PBAT), polyethylene furanoate (PEF), Polycaprolactone (PCL), poly(ethylene adipate) (PEA), polyethylene naphthalate (PEN) and blends/mixtures of these materials.

14. A polyester containing material comprising an esterase variant according to claim 1 and/or a host cell expressing said esterase variant and at least one polyester selected from polyethylene terephthalate (PET), polytrimethylene terephthalate (PTT), polybutylen terephthalate (PBT), polyethylene isosorbide terephthalate (PEIT), polylactic acid (PLA), polyhydroxyalkanoate (PHA), polybutylene succinate (PBS), polybutylene succinate adipate (PBSA), polybutylene adipate terephthalate (PBAT), polyethylene furanoate (PEF), Polycaprolactone (PCL), poly(ethylene adipate) (PEA), polyethylene naphthalate (PEN) and blends/mixtures of these materials.

15. A plastic compound comprising at least one polyester selected from polyethylene terephthalate (PET), polytrimethylene terephthalate (PTT), polybutylen terephthalate (PBT), polyethylene isosorbide terephthalate (PEIT), polylactic acid (PLA), polyhydroxyalkanoate (PHA), polybutylene succinate (PBS), polybutylene succinate adipate (PBSA), polybutylene adipate terephthalate (PBAT), polyethylene furanoate (PEF), Polycaprolactone (PCL), poly(ethylene adipate) (PEA), polyethylene naphthalate (PEN) and blends/mixtures of these materials and an esterase variant according to claim 1 and/or a host cell expressing said esterase variant.

16. A method of degrading a plastic product containing at least one polyester selected from polyethylene terephthalate (PET) and polybutylene adipate terephthalate (PBAT), comprising (a) contacting the plastic product with an esterase according to claim 1 , thereby degrading the plastic product.

17. A polyester containing material comprising at least one polyester selected from polyethylene terephthalate (PET) and polybutylene adipate terephthalate (PBAT), and an esterase variant according to claim 1 and/or a host cell expressing said esterase variant.

18. The esterase variant of claim 2 , which has at least 90% identity to the full length amino acid sequence set forth in SEQ ID NO: 1.

19. The esterase variant of claim 2 , which has at least 95% identity to the full length amino acid sequence set forth in SEQ ID NO: 1.

20. The esterase variant of claim 5 , which has at least 90% identity to the full length amino acid sequence set forth in SEQ ID NO: 1.

21. The esterase variant of claim 5 , which has at least 95% identity to the full length amino acid sequence set forth in SEQ ID NO: 1.

Assignees

• Carbios

Inventors

• Topham Christopher
• Texier Hélène
• Tournier Vincent
• Desrousseaux Marie-Laure
• Duquesne Sophie
• Andre Isabelle
• Barbe Sophie
• Marty Alain

Classifications

• C12Y301/01

  Carboxylic ester hydrolases (3.1.1) · CPC title

• C08J11/105

  by treatment with enzymes · CPC title

• C12Y301/01074

  Cutinase (3.1.1.74) · CPC title

• Y02W30/62

  Plastics recycling; Rubber recycling · CPC title

• C12N9/18 Primary

  Carboxylic ester hydrolases {(3.1.1)} · CPC title