Fucose-binding protein, method for producing same, and use of same

Abstract

The present invention addresses the problem of providing a fucose-binding protein which is improved in the productivity when expressed in a host such as Escherichia coli, the binding affinity for a fucose-containing sugar chain such as a sugar chain containing a structure composed of Fuc[alpha]1-2Gal[beta]1-3GlcNAc and / or Fuc[alpha]1-2Gal[beta]1-3GalNAc, and / or the stability to heat. The problem can be solved by, in the amino acid sequence for a fucose-binding protein BC2LCN which is represented by SEQ ID NO: 1, deleting multiple amino acid residues located on the C-terminal side and, if necessary, substituting a glycine residue located at position-36 in SEQ ID NO: 1 by a cysteine residue, substituting a glutamine residue located at position-39 in SEQ ID NO: 1 by a leucine residue or a methionine residue, substituting a glutamine residue located at position-65 in SEQ ID NO: 1 by a leucine residue, substituting a cysteine residue located at position-72 in SEQ ID NO: 1 by a glycine residue or an alanine residue, substituting a glutamic acid residue located at position-81 in SEQ ID NO: 1 by a cysteine residue, a glutamine residue, a histidine residue or a methionine residue and substituting a glycine residue that is specified as a residue located at position-36 in SEQ ID NO: 1 by a cysteine residue.

Description

technical field [0001] The present invention relates to a fucose-binding protein, its production method, and its use. In particular, the present invention may relate to productivity when expressed in hosts such as Escherichia coli , and fucose-containing products such as sugar chains containing a structure composed of Fucα1-2Galβ1-3GlcNAc and / or Fucα1-2Galβ1-3GalNAc. A fucose-binding protein having improved sugar chain binding affinity and / or thermal stability. Background technique [0002] BC2LCN derived from the N-terminal domain of the BC2L-C lectin produced by Gram-negative bacteria (Burkholderia cenocepacia) has binding affinity for sugar chains containing fucose residues. Proteins, for example, are known to be excluding H-type 1-type sugar chains (Fucα1-2Galβ1-3GlcNAc) and H-type In addition to high binding affinity to type 3 sugar chains (Fucα1-2Galβ1-3GalNAc), it also has a high binding affinity for Lewis (Lewis) Y-type sugar chains (Fucα1-2Galβ1-4(Fucα1-3)GlcNAc),...

AI Technical Summary

Problems solved by technology

However, so far, there have been no reports of BC2LCN that has improved thermal stability and binding affinity to sugar chains through amino acid substitutions at specific positions.

In addition, in order to realize the industrial application of BC2LCN, from the viewpoint of stable supply and large-scale supply, a high-productivity method such as production using microorganisms such as Escherichia coli is preferable, but there is no BC2LCN with improved productivity so far. Related reports

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