Engineered Glucosyltransferases for Alpha-Glucan Yield

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Solution Overview

Problem

Current glucosyltransferase enzymes used for producing polysaccharides like alpha-1,3-glucan have limited focus on improving glucan yields, with less attention given to optimizing the production properties of these enzymes.

Innovation Solution

Engineered glucosyltransferases with modified amino acid sequences, specifically with substitutions at positions corresponding to Gln-588, Phe-607, and Arg-741, are developed to enhance alpha-glucan production yields and reduce leucrose production.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional glucosyltransferase enzymes are used for producing alpha-1,3-glucan, then the basic enzymatic function is maintained, but the glucan yield is limited and leucrose byproduct is produced

Engineering Contradiction:
Improvealpha-glucan yieldVSAvoidleucrose byproduct
Core Design Contradiction:
ProductivityVSLoss of substance

Solution Approach 1:

The patent applies parameter changes by modifying specific amino acid residues in the glucosyltransferase enzyme sequence. Three specific positions are targeted: residue 588 (preferably substituted with hydrophobic amino acids like leucine, isoleucine, or valine), residue 607 (preferably substituted with aromatic amino acids like tryptophan or tyrosine), and residue 741 (preferably substituted with small polar amino acids like serine or threonine). These parameter changes in the enzyme's primary structure directly improve glucan yield and reduce leucrose byproduct formation.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies local quality by making specific localized modifications at three critical positions within the enzyme sequence rather than attempting global optimization. Each substitution targets a specific location with predetermined amino acid preferences, creating localized improvements in catalytic efficiency and product selectivity while maintaining the overall enzyme structure and function.

Inventive Principle:
Principle #3Local quality

2Productivity

If amino acid substitutions are made to improve glucan yield, then productivity increases, but enzyme sequence complexity increases

Engineering Contradiction:
Improveglucan production efficiencyVSAvoidenzyme amino acid sequence
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent applies parameter changes by modifying specific amino acid residues in the glucosyltransferase enzyme sequence. Three specific positions are targeted: residue 588 (preferably substituted with hydrophobic amino acids like leucine, isoleucine, or valine), residue 607 (preferably substituted with aromatic amino acids like tryptophan or tyrosine), and residue 741 (preferably substituted with small polar amino acids like serine or threonine). These parameter changes in the enzyme's primary structure directly improve glucan yield and reduce leucrose byproduct formation.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent applies local quality by making specific localized modifications at three critical positions within the enzyme sequence rather than attempting global optimization. Each substitution targets a specific location with predetermined amino acid preferences, creating localized improvements in catalytic efficiency and product selectivity while maintaining the overall enzyme structure and function.

Inventive Principle:
Principle #3Local quality

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

The modified enzymes demonstrate increased alpha-glucan yields and decreased leucrose yields, improving the efficiency and selectivity of glucan synthesis.

Implementation Method 1

the disclosure pertains to glucosyltransferase enzymes with modified amino acid sequences

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

a glucosyltransferase enzyme isolated from Streptococcus salivarius synthesizes alpha-glucan comprising 1,3-linkages

Methodology Applied
Scientific EffectGlycosyl transfer: Chemical Bonding

Data Source

PatentUS12188055B2Engineered glucosyltransferases
Publication Date: 2025.01.07 NUTRITION & BIOSCIENCES USA 4 INC

AI summary

Disclosed herein are glucosyltransferases with modified amino acid sequences. Such engineered enzymes exhibit improved alpha-glucan product yields and/or lower leucrose yields, for example. Further disclosed are reactions and methods in which engineered glucosyltransferases are used to produce alpha-glucan.