Screening Proteins to Boost 1,4-BDO Productivity

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Solution Overview

Problem

Current methods for producing 1,4-butanediol (1,4-BDO) are inefficient and costly due to rising oil prices, and existing biological pathways in Escherichia coli are limited in discovering enzymes for high-efficiency production.

Innovation Solution

A method for screening proteins involved in 1,4-BDO production by culturing microorganisms in media with varying 1,4-BDO concentrations, identifying proteins with increased expression, and selecting those positively involved in production, using Corynebacterium glutamicum and introducing specific genes like SucD, 4Hbd, Cat2, and AdhE2 to enhance productivity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If genetic mutations are induced in enzyme genes to discover high-efficiency enzymes, then enzyme efficiency may be improved, but the effectiveness of discovering related proteins or genes is limited

Engineering Contradiction:

Improveenzyme efficiencyVSAvoideffectiveness of discovering proteins or genes

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

Instead of mutating enzyme genes to discover high-efficiency enzymes (forward approach), the patent inverts the approach by screening for genes that are highly expressed under 1,4-BDO production conditions, then verifying their role in production. This reverse screening method overcomes the limitations of mutation-based approaches and successfully identifies proteins like citrate synthase and NCgl2145 that enhance 1,4-BDO productivity.

Inventive Principle:

Principle #13The other way round (Inversion)

2Productivity

If chemical production processes are used for 1,4-BDO, then production can be maintained, but production costs increase due to rising oil prices

Engineering Contradiction:

Improve1,4-BDO productionVSAvoidproduction cost

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The patent replaces chemical production processes with biological processes using microorganisms. By engineering microorganisms to overexpress specific genes (citrate synthase, NCgl2145) involved in the 1,4-BDO biosynthetic pathway, the invention achieves cost-effective production that is independent of oil price fluctuations, substituting petroleum-based chemistry with biologically-based synthesis.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Productivity

If existing biological pathways in Escherichia coli are used, then 1,4-BDO production is possible, but efficiency is limited

Engineering Contradiction:

Improve1,4-BDO production efficiencyVSAvoidpathway optimization capability

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent changes the biological system from Escherichia coli to Corynebacterium glutamicum, which provides better adaptability for 1,4-BDO production. Additionally, the invention changes the expression levels of key genes (citrate synthase, NCgl2145) by introducing plasmids with strong promoters, thereby optimizing the biosynthetic pathway parameters to achieve significantly higher production efficiency compared to existing E. coli systems.

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS9963721B2Method of screening gene for 1,4-BDO production

Publication Date: 2018.05.08 SAMSUNG ELECTRONICS CO LTD

AI summary

Provided is a screening method of discovering genes capable of increasing 1,4-BDO production on the basis of proteomics data. Over-expression of proteins screened by the method, NCgl0630 (citrate synthase) and NCgl2145 (hypothetical protein), increase 1,4-BDO productivity. The method may lead to screening of a protein associated with 1,4-BDO productivity, thereby increasing 1,4-BDO productivity, and thus, the method may be recognized as being industrially applicable.