Transposon Vector for Corynebacterium Lysine Production

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Solution Overview

Problem

Conventional methods for enhancing lysine production in Corynebacterium sp. face limitations in inserting multiple genes without affecting microorganism growth, particularly due to restricted gene insertion sites and potential destruction of rDNA copies.

Innovation Solution

A vector for transformation using a transposon gene with a multicloning site is developed, allowing for the insertion of multiple copies of target genes such as aspartate kinase, dihydrodipicolinate synthase, and dihydropicolinate reductase, along with the additional insertion of fructokinase, without affecting microorganism growth, using secondary crossover for efficient gene integration.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If multiple target genes are inserted into Corynebacterium sp. to enhance lysine production, then lysine productivity is improved, but the growth of microorganism is affected due to destruction of rDNA copies

Engineering Contradiction:

Improvelysine productivityVSAvoidmicroorganism growth stability

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The transposon vector acts as an intermediary tool that enables gene insertion into non-rDNA regions of the Corynebacterium genome. The vector contains a transposon sequence that can randomly integrate target genes (lysC, asd, dapA, dapB) into safe genomic locations that do not disrupt rDNA copies, thereby maintaining microorganism growth stability while achieving enhanced lysine production capability

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention shifts the gene insertion strategy from the limited one-dimensional approach of targeting specific rDNA regions to a multi-dimensional approach using transposon-mediated random integration across the entire genome. This allows simultaneous insertion of multiple target genes into various safe locations, expanding the available genomic real estate for gene amplification without compromising essential genetic functions

Inventive Principle:

Principle #17Another dimension (Dimensionality change)

2Productivity

If conventional gene insertion methods are used to increase gene copies, then lysine biosynthesis is enhanced, but the number of insertable genes is limited due to restricted insertion sites

Engineering Contradiction:

Improvelysine biosynthesisVSAvoidgene insertion flexibility

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The transposon vector system provides universal applicability for inserting any target gene of interest into the Corynebacterium genome. The vector can accommodate multiple different gene combinations (lysC, asd, dapA, dapB) and can be applied to various Corynebacterium strains, making it a versatile tool for metabolic engineering beyond the limitations of site-specific recombination methods

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Adaptability or versatility

If rDNA region is used for gene insertion to overcome insertion site limitations, then multiple genes can be inserted, but growth of microorganism is affected

Engineering Contradiction:

Improvegene insertion capacityVSAvoidmicroorganism growth

Core Design Contradiction:

Adaptability or versatilityVSReliability

Solution Approach 1:

The invention converts the potential harm of random transposon insertion into a benefit by utilizing the transposon's random integration capability to insert genes into safe genomic locations. The system transforms the uncontrolled nature of transposon movement from a disadvantage into an advantage, as it naturally avoids disrupting essential rDNA regions while still achieving effective gene amplification for lysine production

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

Data Source

PatentUS8323933B2Vector for transformation using transposons, microorganisms transformed by the vector, and method for producing L-lysine using the same

Publication Date: 2012.12.04 CJ CHEILJEDANG CORP

AI summary

The present invention relates to a vector for transformation using transposon genes, microorganisms transformed by the vector, and a method for producing L-lysine using the microorganisms.