Udp-glycosyltransferases

Abstract

The present disclosure relates a polypeptide having UGT activity, which polypeptide comprises an amino acid sequence which, when aligned with a polypeptide having UGT activity comprising the sequence set out in SEQ ID NO: 2, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 35, 189, 280, 284, 285, 334 or 373, said positions being defined with reference to SEQ ID NO: 2 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having UGT activity. A polypeptide according to the disclosure may be used in a recombinant cell for the production of steviol or a steviol glycoside.

1 . A polypeptide having UGT activity, which polypeptide comprises an amino acid sequence which, when aligned with a polypeptide having UGT activity comprising the sequence set out in SEQ ID NO: 2, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions
 35, 189, 280, 284, 285, 334 or 373   said positions being defined with reference to SEQ ID NO: 2 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having UGT activity.   
     
     
         2 . A polypeptide according to  claim 1 , wherein the modified property is modified UGT activity. 
     
     
         3 . A polypeptide according to  claim 1 , wherein the UGT activity is UGT3 activity. 
     
     
         4 . A polypeptide according to  claim 1 , wherein the reference polypeptide comprises the UGT of SEQ ID NO: 2. 
     
     
         5 . A polypeptide according to  claim 1 , wherein:
 (i) a valine is present at position 35;   (ii) a alanine is present at position 189;   (iii) an asparagine is present at position 280;   (iv) an asparagine is present at position 284;   (v) a glycine is present at position 285   (vi) an asparagine is present at position 285   (vii) a serine is present at position 285   (viii) an alanine is present at position 334; and/or   (ix) an alanine is present at position 373.   said positions being defined with reference to SEQ ID NO: 2.   
     
     
         6 . A polypeptide according to  claim 1 , comprising an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% sequence identity with SEQ ID NO: 2. 
     
     
         7 . A polypeptide having UGT activity comprising an amino acid sequence having at least about 95% sequence identity, at least 96%, at least 97%, at least 98% or at least 99% sequence identity to any one of SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18 or 20.