Thermophilic Bacillus Sucrose Fermentation via Enzymatic Modification

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Solution Overview

Problem

Moderately thermophilic Bacillus strains used in industrial fermentations often lack the capability to utilize sucrose as a carbon source, limiting their fermentation efficiency and production potential.

Innovation Solution

Genetic modification of these strains by introducing specific genes and polypeptides involved in sucrose catabolism, such as sucrose-specific phosphotransferase and sucrose-6-phosphate hydrolase, to enable sucrose utilization, using transformation methods like electroporation or chromosomal integration.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Temperature

If moderately thermophilic Bacillus strains are used for industrial fermentation, then fermentation temperature can be raised above 50°C with advantages of higher product purity and faster reactions, but these strains lack the capability to utilize sucrose as a carbon source

Engineering Contradiction:

Improvefermentation temperatureVSAvoidsucrose utilization capability

Core Design Contradiction:

TemperatureVSAdaptability or versatility

Solution Approach 1:

The patent introduces foreign genes (scrA and scrB) encoding sucrose-specific phosphotransferase and sucrose-6-phosphate hydrolase enzymes into Bacillus strains, fundamentally changing the metabolic parameters of the organism to enable sucrose catabolism. This genetic modification allows the bacteria to acquire new functional capabilities without altering the optimal fermentation temperature parameters.

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent uses plasmid vectors as intermediary carriers to transfer sucrose utilization genes into the Bacillus strain genome. These plasmids serve as mediators that facilitate the introduction of external genetic material, enabling the host organism to acquire sucrose metabolism capabilities while maintaining its thermophilic fermentation properties.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Productivity

If Bacillus coagulans strain DSM 1 is used, then it shows poor fermentation capability on sucrose with only scarce growth and acid formation, but genetic modification can improve this capability

Engineering Contradiction:

Improvesucrose fermentation productivityVSAvoidfermentation reliability on sucrose

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent enables the Bacillus strain to produce its own sucrose-catabolizing enzymes (sucrose-specific phosphotransferase and sucrose-6-phosphate hydrolase) through genetic modification. The modified strain becomes self-sufficient in utilizing sucrose as a carbon source, converting it efficiently into lactic acid and other fermentation products without requiring external enzyme supplementation.

Inventive Principle:

Principle #25Self-service

3Ease of manufacture

If sucrose is used as a low-cost carbon source in fermentation media, then production costs can be reduced, but not all moderately thermophilic Bacillus strains possess the capability to utilise sucrose

Engineering Contradiction:

Improveproduction costVSAvoidsubstrate utilization range

Core Design Contradiction:

Ease of manufactureVSAdaptability or versatility

Solution Approach 1:

The patent confers multi-functionality to the Bacillus strain by enabling it to utilize multiple carbon sources effectively. The genetically modified strain can now ferment both traditional sugars (glucose, fructose) and sucrose, making it a universal fermenter that can process various inexpensive carbohydrate substrates available in industrial settings.

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

Enables the modified strains to efficiently ferment sucrose, enhancing their industrial fermentation capabilities and production potential by allowing growth on sucrose-containing substrates, thereby improving product yield and process efficiency.

Implementation Method 1

Fermentation of moderately thermophilic bacilli on sucrose

Methodology Applied

Scientific EffectFermentation: Fermentation

Implementation Method 2

Genetic modification of these strains by introducing specific genes and polypeptides involved in sucrose catabolism, such as sucrose-specific phosphotransferase and sucrose-6-phosphate hydrolase

Methodology Applied

Scientific EffectEnzymatic catabolism: Enzyme

Data Source

PatentUS8663954B2Fermentation of moderately thermophilic bacilli on sucrose

Publication Date: 2014.03.04 PURAC BIOCHEM BV

AI summary

A method for the construction of a moderately thermophilic Bacillus strain capable of utilizing sucrose as a carbon source includes the transformation of a parent moderately thermophilic Bacillus strain not capable of utilizing sucrose as a carbon source with a polynucleotide comprising a DNA sequence that encodes a polypeptide having sucrose-specific phosphotransferase activity and having i) an amino acid sequence of SEQ ID NO:1 or ii) an amino acid sequence with an identity of at least 70% to the sequence of SEQ ID NO:1 and/or comprising a DNA sequence that encodes a polypeptide having sucrose-6-phosphate hydrolase activity and having iii) an amino acid sequence of SEQ ID NO:2 or iv) an amino acid sequence with an identity of at least 70% to the sequence of SEQ ID NO:2.