Carbohydrate binding module with affinity for insoluble xylan
AssigneeThe Board of Trustees of the University of Illinois
InventorsShosuke YOSHIDA, Roderick I. Mackie, Isaac K. O. Cann
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Abstract
The present disclosure relates to isolated polynucleotides with two polynucleotide sequences linked within one open reading frame, in which the first polynucleotide sequence encodes a peptide that binds to a carbohydrate. The present disclosure also relates to vectors and genetically modified host cells containing such isolated polynucleotides and polypeptides encoded by such isolated polynucleotides. The present disclosure further relates to methods of increasing the ability of a recombinant protein to bind to a carbohydrate and methods of identifying a protein having an ability to bind to a carbohydrate.
Claims22
An isolated polynucleotide comprising a first polynucleotide sequence that encodes SEQ ID NO:
- 1 wherein said first polynucleotide is linked within one open reading frame to a second polynucleotide sequence to form a linked polynucleotide, wherein SEQ ID NO:
- 1 binds to a carbohydrate and wherein the linked polynucleotide does not encode a naturally occurring polypeptide.
The isolated polynucleotide of claim 9, wherein the polypeptide comprises an enzyme.
An isolated polypeptide comprising SEQ ID NO:
- 1 conjugated to an atom or molecule.
A method of increasing the ability of a recombinant protein to bind to a carbohydrate, comprising:
- linking a first isolated polynucleotide encoding SEQ ID NO:
- 1 to a second isolated polynucleotide encoding a polypeptide, a peptide, or a protein tag to form a linked polynucleotide, wherein the linked polynucleotide encodes a recombinant protein having an increased ability to bind to a carbohydrate compared to the polypeptide, peptide, or protein tag alone.
A method of increasing the ability of a recombinant protein to bind to a carbohydrate, comprising:
- linking a first isolated polynucleotide encoding SEQ ID NO:
- 1 to a second isolated polynucleotide encoding an amino acid sequence selected from a library of amino acid sequences to form a linked polynucleotide, wherein the linked polynucleotide encodes a recombinant protein having an increased ability to bind to a carbohydrate compared to the amino acid sequence alone.
A method of identifying a protein having an ability to bind a carbohydrate, comprising
- providing a labeled polynucleotide, wherein the polynucleotide encodes SEQ ID NO:
- 1;
- hybridizing the labeled polynucleotide to a homologous sequence in a nucleotide library;
- and
- isolating the sequence bound by the labeled polynucleotide, wherein the sequence encodes a protein having an ability to bind to a carbohydrate.
Dependencies are inferred from the claim text.