Mutant HPPD Enzymes for Broad-Spectrum Herbicide Tolerance

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Solution Overview

Problem

Existing methods for creating plants tolerant to HPPD-inhibiting herbicides are inadequate for providing commercial levels of tolerance to different types of these herbicides, and there is a need for novel mutant HPPDs that confer resistance or tolerance to various HPPD-inhibiting herbicides across diverse crops and crop varieties.

Innovation Solution

Development of a mutant p-hydroxyphenylpyruvate dioxygenase (HPPD) enzyme with specific mutations that reduce sensitivity to HPPD-inhibiting herbicides, along with associated nucleic acids and expression systems for introducing these enzymes into plants, using CRISPR/Cas9 and Cpf1 gene editing techniques.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If wild-type HPPD enzyme is used in plants, then the plant can normally catalyze the conversion of HPP to homogentisate, but the plant is highly sensitive to HPPD-inhibiting herbicides

Engineering Contradiction:

Improveenzyme functionVSAvoidherbicide sensitivity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent applies parameter changes by introducing specific amino acid mutations at defined positions in the HPPD enzyme sequence (e.g., positions 93, 103, 141, 165, 191, 220, 226, 276, 277, 336, 337, 338, 342, 346, 370, 377, 386, 390, 392, 403, 410, 418, 419, 420, 430, or 431). These mutations alter the enzyme's molecular parameters to reduce herbicide binding affinity while preserving catalytic function, thereby achieving herbicide tolerance without losing enzyme activity.

Inventive Principle:

Principle #35Parameter changes

2Adaptability or versatility

If a given HPPD enzyme provides tolerance to some HPPD-inhibiting herbicides, then the plant has resistance to those herbicides, but the tolerance is inadequate for commercial levels and does not work for all HPPD-inhibiting herbicide types

Engineering Contradiction:

Improveherbicide tolerance rangeVSAvoidcommercial level tolerance

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The patent achieves universality by creating HPPD enzyme variants with multiple possible mutations that can confer tolerance across different types of HPPD-inhibiting herbicides (including isoxazoles, diketonitriles, triketones, and pyrazoline salts). The enzyme variants are designed to provide broad-spectrum tolerance rather than specificity to a single herbicide class, enabling commercial application across multiple herbicide types and crop scenarios.

Inventive Principle:

Principle #6Universality (Multi-functionality)

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 mutant HPPD enzymes significantly enhance plant tolerance to HPPD-inhibiting herbicides, allowing plants to maintain enzyme function despite herbicide presence, thereby improving herbicide tolerance and reducing phytotoxicity.

Implementation Method 1

P-hydroxyphenylpyruvate dioxygenases (HPPD) are enzymes which catalyze the reaction in which hydroxyphenylpyruvate (HPP) is converted into homogentisate. This reaction takes place in the presence of enzyme-bound iron and oxygen.

Methodology Applied

Scientific EffectCatalysis: Catalysis

Implementation Method 2

Methodology Applied

Scientific EffectOxidation: Oxidation

Data Source

PatentUS12428627B2Mutant p-hydroxyphenylpyruvate dioxygenase, and coding nucleic acid and use thereof

Publication Date: 2025.09.30 QINGDAO KINGAGROOT CHEM COMPOUNDS CO LTD

AI summary

The present invention relates to a mutant p-hydroxyphenylpyruvate dioxygenase (HPPD) protein or a bioactive fragment thereof and an isolated polynucleotide comprising a nucleic acid sequence encoding the protein or fragment thereof, wherein the mutant p-hydroxyphenylpyruvate dioxygenase (HPPD) protein or a bioactive fragment thereof retains or enhances the property of catalyzing the conversion of p-hydroxyphenylpyruvate (HPP) to homogentisate and is significantly less sensitive to HPPD-inhibiting herbicides than a wild-type HPPD. The present invention also relates to a nucleic acid construct, an expression vector and a host cell comprising the polynucleotide, as well as to a method for producing a plant that has the property of catalyzing the conversion of p-hydroxyphenylpyruvate (HPP) to homogentisate and significantly reduced sensitivity to HPPD-inhibiting herbicides.