Bacillus subtilis 6A-1 Cellulase Production Across pH 2 to 13

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Solution Overview

Problem

Current methods for utilizing microorganisms to degrade cellulose face challenges due to the sensitivity of fungal spores to environmental conditions and the limited applicability of bacteria requiring specific pH conditions, leading to inefficiencies in enzyme production and application.

Innovation Solution

A strain of Bacillus subtilis 6A-1 is developed, capable of producing three enzymatically active protein fractions that degrade cellulose across a wide pH range, from 2 to 13, and is effective on both native and modified cellulose forms, including carboxymethyl cellulose and acid detergent fiber, allowing for stable enzyme production and application in various processes.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If fungi are used to produce cellulase enzymes, then enzyme production efficiency is improved, but the sensitivity to environmental conditions and difficulty in harvesting spores worsens applicability

Engineering Contradiction:

Improveenzyme production efficiencyVSAvoidapplicability in viable enzyme producing products

Core Design Contradiction:

ProductivityVSAdaptability or versatility

Solution Approach 1:

The patent uses bacteria as a surrogate organism to copy the cellulase enzyme production capability of fungi. Instead of using actual fungal spores which are sensitive and difficult to harvest, the invention creates bacterial strains that replicate the enzyme production function, thereby resolving the contradiction between high enzyme production efficiency and practical applicability.

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent modifies the biological parameters by switching from fungal organisms to bacterial organisms. This parameter change allows the system to maintain high cellulase production capability while gaining the advantages of bacterial robustness, ease of cultivation, and stability across varying environmental conditions including pH and temperature ranges.

Inventive Principle:

Principle #35Parameter changes

2Ease of operation

If bacteria are used to produce cellulase enzymes, then ease of cultivation and stability are improved, but the requirement for specific narrow pH conditions limits applicability

Engineering Contradiction:

Improveease of cultivationVSAvoidpH range applicability

Core Design Contradiction:

Ease of operationVSAdaptability or versatility

Solution Approach 1:

The patent develops bacterial strains that possess universal pH tolerance, enabling them to function across a broad pH spectrum from acidic to alkaline conditions. This multi-functionality allows the same bacterial cellulase production system to be applied in diverse environments without requiring pH-specific strain selection, thereby resolving the contradiction between ease of cultivation and pH range applicability.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Productivity

If fungal spores are used as seed, then cellulase enzyme production is achieved, but the sensitivity to heat, moisture, and desiccation worsens viability and stability

Engineering Contradiction:

Improvecellulase enzyme productionVSAvoidviability of seeds under environmental conditions

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent employs bacterial cells as disposable, easily renewable seed material instead of fragile fungal spores. Bacterial cells can be rapidly cultured and regenerated under controlled conditions, providing a reliable and stable seed source that is not susceptible to the environmental sensitivities affecting fungal spores, thus resolving the contradiction between enzyme production capability and seed viability.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

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

Bacillus subtilis 6A-1 enables efficient cellulose degradation across diverse conditions, enhancing feed conversion efficiency, waste treatment, and biogas production, while maintaining enzyme activity and stability, thus overcoming previous limitations in microbial cellulose degradation.

Implementation Method 1

Bacillus subtilis 6A-1 is developed, capable of producing three enzymatically active protein fractions that degrade cellulose

Methodology Applied

Scientific EffectEnzymatic hydrolysis: Hydrolysis

Data Source

PatentUS11008534B2Bacteria and enzymes produced therefrom and methods of using same

Publication Date: 2021.05.18 AGRI KING

AI summary

A bacteria referred to here as Bacillus subtilis 6A-1 is provided, compositions thereof and processes for use of the bacteria, spores, cells, extracts and enzymes. The compositions which comprise the bacteria, spores, cells, extracts and/or enzymes are capable of degrading polysaccharides. Such compositions are capable of degrading cellulose, including plant-produced cellulose, microcrystalline cellulose and carboxymethyl cellulose. The bacteria produces at least two cellulose-degrading protein fractions. Cellulose degrading activity continues across pH2 to pH13.