Bacillus Protein Production via YmaH Overexpression

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Solution Overview

Problem

There is a need for improved methods to enhance the expression and production of proteins, such as proteases, in Bacillus species, as current technologies do not effectively optimize protein production in these microorganisms.

Innovation Solution

Genetically modified Bacillus host cells are developed to overexpress the YmaH protein by using polynucleotide constructs and expression vectors that include SigA and SigH promoters, allowing for increased production of proteins like proteases.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If conventional genetic engineering methods are used in Bacillus species, then some protein production improvement is achieved, but production efficiency and expression levels remain insufficient

Engineering Contradiction:

Improveprotein production efficiencyVSAvoidexpression optimization

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies parameter changes by modifying promoter strength (using strong SigA and SigH promoters), optimizing ribosome binding sites, and adjusting gene copy numbers to enhance protein expression levels and production efficiency in Bacillus species

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses molecular chaperones and signaling molecules as intermediaries to facilitate proper protein folding, stabilize expression, and optimize the transcription-translation process, thereby improving overall protein production reliability

Inventive Principle:

Principle #24Intermediary (Mediator)

2Quantity of substance

If protein expression is increased in Bacillus host cells, then production volume improves, but expression timing and regulation become less optimized

Engineering Contradiction:

Improveprotein production volumeVSAvoidexpression timing optimization

Core Design Contradiction:

Quantity of substanceVSLoss of time

Solution Approach 1:

The patent employs periodic action through inducible promoter systems that allow controlled timing of protein expression at specific growth phases, enabling optimized expression schedules that maximize production volume while maintaining temporal regulation

Inventive Principle:

Principle #19Periodic action

Solution Approach 2:

The patent applies preliminary action by pre-inducing expression at optimal growth stages and using signaling molecules that prepare the cellular machinery for efficient protein synthesis before the actual production phase

Inventive Principle:

Principle #10Preliminary action

Data Source

PatentUS8512982B2Enhanced protein production in Bacillus

Publication Date: 2013.08.20 DANISCO US INC

AI summary

The present invention relates to cells that have been genetically manipulated to have an altered capacity to express and/or produce proteins of interest. In particular, the present invention relates to modified host cells of Gram-positive microorganisms, such as Bacillus species that are capable of overexpressing ymaH. The invention encompasses polynucleotide constructs and expression vectors containing polynucleotide sequences that encode YmaH, and the modified host cells comprising them. In particular, the present invention relates to compositions and methods of overexpressing YmaH for enhancing the expression and production of proteins of interest (e.g., proteases) in Bacillus species.