Biological processes for the production of aryl sulfates
Inventors
Jendresen, Christian Bille • Nielsen, Alex Toftgaard
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11390894-B2
Publication Date
2022-07-19
Expiration Date
Abstract
The present invention generally relates to the field of biotechnology as it applies to the production of aryl sulfates using polypeptides or recombinant cells comprising said polypeptides. More particularly, the present invention pertains to polypeptides having aryl sulfotransferase activity, recombinant host cells expressing same and processes for the production of aryl sulfates employing these polypeptides or recombinant host cells.
Core Innovation
The invention relates to a process for producing a sulfated phenolic compound by contacting a medium comprising a phenolic compound or a precursor of a phenolic compound with a recombinant host cell. The recombinant host cell comprises a heterologous polypeptide having aryl sulfotransferase activity, and the host cell is a bacterium or yeast. The sulfated phenolic compound is represented by the general formula (I), while the precursor of a phenolic compound is a compound of general Formula (p-I).
The aryl sulfotransferase polypeptide is selected from polypeptides comprising the amino acid sequences set forth in SEQ ID NO: 1-13, or from polypeptides having at least about 95% sequence identity to those sequences and having aryl sulfotransferase activity. Structural options for the phenolic compound and phenolic precursor are defined by substituent selections for R1-R6, including hydroxyl groups, and by R7 and R8 selected from specified groups.
The document further describes host engineering to enhance sulfation productivity by improving sulfur-related metabolism, including providing PAPS supply and PAP removal through sulfur metabolism enzymes such as ATP sulfurylase components (cysD/cysN), APS kinase (cysC), and PAP phosphatase (cysQ). It also describes in vivo generation of the phenolic precursor using tyrosine ammonia lyase polypeptides (SEQ ID NO: 14-23) followed by sulfation by the heterologous aryl sulfotransferase.
Claims Coverage
The independent claim covers 2 inventive features. It centers on use of a recombinant bacterium or yeast containing a heterologous aryl sulfotransferase, and contacting a medium comprising a phenolic compound or precursor defined by the stated general formulas to produce a sulfated phenolic compound.
Recombinant host cell with heterologous aryl sulfotransferase activity
A recombinant host cell that is a bacterium or yeast comprises a heterologous polypeptide having aryl sulfotransferase activity selected from SEQ ID NO: 1-13 or from polypeptides having at least about 95% sequence identity to those sequences and having aryl sulfotransferase activity.
Sulfated phenolic compound from a phenolic compound or precursor of Formula (p-I)
A process contacts a medium comprising a phenolic compound represented by general formula (I) or a precursor of phenolic compound represented by general Formula (p-I), with substituent selections for R1-R6 and R7-R8 as specified.
The claimed coverage centers on producing sulfated phenolic compounds by using a recombinant bacterium or yeast expressing a heterologous aryl sulfotransferase and by providing phenolic compounds or precursors defined by the stated formulas, with dependent claims adding host engineering and structural limitations.
Stated Advantages
Improved yields or conversion of sulfated product when host sulfur-related enzymes are co-expressed, as described in the examples.
Reduced toxicity of the sulfated product (zosteric acid) compared with p-coumaric acid, as described in the examples.
Documented Applications
Producing sulfated phenolic compounds, including zosteric acid, by contacting a medium containing a phenolic compound or phenolic precursor with a recombinant bacterium or yeast expressing a heterologous aryl sulfotransferase.
In vivo generation of a phenolic precursor using tyrosine ammonia lyase polypeptides followed by sulfation by heterologous aryl sulfotransferase within the recombinant host.
Using host engineering to enhance PAPS supply and PAP removal through sulfur metabolism enzymes to increase sulfation productivity.
Producing sulfated phenolic compounds in yeast, including S. cerevisiae.
Interested in licensing this patent?