Vitamin E Biosynthesis Gene Cloning in Para Rubber Tree

Resolve Bottlenecks,
Find Innovative Solutions
Generate Solutions

Solution Overview

Problem

The lack of identified genes encoding enzymes involved in vitamin E biosynthesis in the Para rubber tree has hindered the ability to control and enhance vitamin E production in this species, which is crucial for its antioxidant properties and rubber aging resistance.

Innovation Solution

Identification and analysis of gene clusters encoding enzymes involved in vitamin E biosynthesis in Para rubber trees, followed by full-length cDNA cloning and introduction of these genes into plants to increase vitamin E content, utilizing techniques like EST analysis, RACE, and gene recombination to enhance enzyme expression.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If gene identification and cloning techniques are applied to Para rubber tree, then vitamin E biosynthesis can be controlled and enhanced, but the complexity of the research and development process increases

Engineering Contradiction:
Improvevitamin E contentVSAvoidgene identification and cloning process
Core Design Contradiction:
Quantity of substanceVSDevice complexity

Solution Approach 1:

The vitamin E biosynthesis pathway is divided into multiple enzymatic steps, and individual genes encoding these enzymes are identified and cloned separately. This segmentation allows for systematic study and manipulation of each biosynthetic step, enabling controlled enhancement of vitamin E content through targeted genetic modification.

Inventive Principle:
Principle #1Segmentation

Solution Approach 2:

Full-length cDNA clones of vitamin E biosynthesis-related genes are obtained and characterized in advance through EST analysis and RACE techniques. This preliminary action prepares the necessary genetic materials and knowledge base before actual transgenic manipulation, facilitating efficient subsequent experiments and applications.

Inventive Principle:
Principle #10Preliminary action

2Reliability

If transgenic techniques are used to increase vitamin E content in Para rubber tree, then antioxidant effects and aging resistance are enhanced, but the difficulty of genetic transformation increases

Engineering Contradiction:
Improveantioxidant effect and aging resistanceVSAvoidgenetic transformation process
Core Design Contradiction:
ReliabilityVSDevice complexity

Solution Approach 1:

The patent uses cDNA clones as intermediaries to transfer vitamin E biosynthesis gene information into Para rubber tree. These cloned genes serve as mediators that carry the genetic instructions necessary for enhanced vitamin E production, enabling controlled genetic transformation while maintaining reliability of the antioxidant effect.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

Full-length cDNA copies of the vitamin E biosynthesis genes are created through molecular cloning techniques. These genetic copies can be manipulated, replicated, and introduced into Para rubber tree cells, providing a reliable method to enhance vitamin E content without requiring direct manipulation of the original genomic DNA.

Inventive Principle:
Principle #26Copying

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

This approach leads to increased vitamin E biosynthesis and antioxidant effects, effectively enhancing the aging resistance of rubber products by increasing vitamin E content in transformed plants, particularly in the Para rubber tree latex.

Implementation Method 1

prephenate dehydratase which biosynthesizes p-hydroxyphenyl pyruvic acid, p-hydroxyphenyl pyruvic acid dioxygenase which biosynthesizes homogentisic acid, geranylgeranyl reductase which biosynthesizes phytyl diphosphate, homogentisic acid phytyl transferase which biosynthesizes 2-methyl-6-phytyl benzoquinone, 2-methyl-6-geranylgeranyl benzoquinone methyl transferase which biosynthesizes 2,3-dimethyl-5-geranylgeranyl-1,4-benzoquinone, homogentisic acid geranylgeranyl transferase which biosynthesizes 2-methyl-6-geranylgeranyl benzoquinone, tocopherol cyclase which biosynthesizes tocotrienols or tocopherols, and γ-tocopherol methyl transferase which biosynthesizes β-tocopherol or β-tocotrienols using δ-tocopherol or δ-tocotrienol and S-adenosyl-L-methionine as substrate, or α-tocopherol or α-tocotrienol using γ-tocopherol or γ-tocotrienol and S-adenosyl-L-methionine as substrate

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Data Source

PatentEP2143793B1Genes of enzymes participating in vitamin e biosynthesis in para rubber tree
Publication Date: 2013.09.25 BRIDGESTONE CORP
  • EP2143793B1 patent drawingFigure 1
  • EP2143793B1 patent drawingFigure 2
  • EP2143793B1 patent drawingFigure 3

AI summary

It is a subject of this invention to isolate a gene group of enzymes involved in the biosynthesis of vitamin E of Para rubber tree, and to determine the base sequence of each gene. According to this invention, genes encoding enzymes involved in the vitamin E biosynthesis were isolated from Para rubber tree and the base sequences of these genes were determined. Since vitamin E is an antioxidant existing in nature, it is expected that transformation of a plant by using the genes obtained in the invention would result in an increase in the vitamin E content of the plant and, in its turn, contribute to the prevention of rubber from aging.