	US 12,188,055 B2
	Engineered glucosyltransferases
Yougen Li, Glen Mills, PA (US); Ellen D. Semke, Newark, DE (US); Qiong Cheng, Wilmington, DE (US); and Jian Ping Lai, Wallingford, PA (US)
Assigned to NUTRITION & BIOSCIENCES USA 4, INC., Wilmington, DE (US)
Filed by NUTRITION & BIOSCIENCES USA 4, INC., Rochester, NY (US)
Filed on Jan. 20, 2023, as Appl. No. 18/157,512.
Application 18/157,512 is a continuation of application No. 17/118,733, filed on Dec. 11, 2020, granted, now 11,560,551.
Application 17/118,733 is a continuation of application No. 16/127,293, filed on Sep. 11, 2018, granted, now 10,865,393.
Claims priority of provisional application 62/557,840, filed on Sep. 13, 2017.
Prior Publication US 2023/0392128 A1, Dec. 7, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/10 (2006.01); C08B 37/00 (2006.01); C12N 15/56 (2006.01); C12P 19/04 (2006.01); C12P 19/18 (2006.01); G16B 25/00 (2019.01)

CPC C12N 9/1048 (2013.01) [C08B 37/0009 (2013.01); C12N 9/1051 (2013.01); C12P 19/04 (2013.01); C12P 19/18 (2013.01); C12Y 204/01005 (2013.01); C12Y 204/01 (2013.01); G16B 25/00 (2019.02)]

20 Claims

1. A polynucleotide comprising a nucleotide sequence encoding a non-native glucosyltransferase,

wherein the non-native glucosyltransferase comprises at least two amino acid substitutions at positions corresponding with amino acid residues Gln-588, Phe-607, or Arg-741 of SEQ ID NO:62,

wherein the non-native glucosyltransferase synthesizes insoluble alpha-glucan comprising 1,3-linkages,

wherein the non-native glucosyltransferase has:

(i) an insoluble alpha-glucan yield that is higher than the insoluble alpha-glucan yield of a second glucosyltransferase that only differs from the non-native glucosyltransferase at the substitution positions, and/or a leucrose yield that is lower than the leucrose yield of the second glucosyltransferase;

wherein the non-native glucosyltransferase comprises an amino acid sequence that has at least 90% sequence identity to the amino acid sequence of residues 55-960 of SEQ ID NO:4 or the amino acid sequence of residues 54-957 of SEQ ID NO: 65.