Bacillus subtilis CS1802 Whole-Cell Lipase Synthesis

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current enzymatic synthesis methods for vitamin A palmitate rely on expensive immobilized enzymes like Novozymes 435, and there is a lack of lipase-producing strains derived from natural fermented foods for organic phase whole-cell transformation, which limits cost-effectiveness and diversity in microbial sources.

Innovation Solution

Identification and preservation of Bacillus subtilis CS1802, a lipase-producing strain isolated from opossum shrimp paste, used in a whole-cell transformation method for enzymatic synthesis of vitamin A palmitate, involving inoculation in a beef extract peptone medium, followed by fermentation in an organic phase system with a vitamin A to palmitic acid ratio of 1:1 in n-hexane, achieving a yield of 15.35 mg/mL with a transformation rate of 76.75%.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If commercial immobilized enzymes like Novozymes 435 are used for enzymatic synthesis of vitamin A palmitate, then the synthesis efficiency and product quality are improved, but the production cost increases significantly

Engineering Contradiction:

Improvesynthesis efficiencyVSAvoidproduction cost

Core Design Contradiction:

ProductivityVSEase of manufacture

Solution Approach 1:

The patent uses whole cells of Bacillus subtilis CS1802 as a disposable biocatalyst system instead of expensive commercial immobilized enzymes. The whole cells perform the lipase-catalyzed synthesis of vitamin A palmitate and can be discarded after use, eliminating the need for expensive enzyme procurement while maintaining synthesis efficiency

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The Bacillus subtilis CS1802 strain naturally produces lipase enzyme within its cellular structure, eliminating the need to separately procure and immobilize commercial enzymes. The cell itself serves as both the enzyme source and the reaction vessel, reducing overall production costs while maintaining catalytic activity

Inventive Principle:

Principle #25Self-service

2Productivity

If traditional chemical synthesis methods are used for vitamin A palmitate, then the production process is simple and fast, but environmental pollution and equipment corrosion occur

Engineering Contradiction:

Improveproduction speedVSAvoidenvironmental pollution

Core Design Contradiction:

ProductivityVSObject-generated harmful factors

Solution Approach 1:

The patent replaces chemical synthesis mechanisms with biological enzymatic mechanisms. The lipase enzyme within Bacillus subtilis CS1802 cells catalyzes the esterification reaction between vitamin A and palmitic acid, substituting harsh chemical reagents and conditions with a mild, environmentally friendly biological system that maintains high production speed

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Adaptability or versatility

If lipase-producing strains are screened from diverse natural sources, then the microbial diversity and enzyme performance are improved, but the screening complexity and time increase

Engineering Contradiction:

Improvemicrobial diversityVSAvoidscreening complexity

Core Design Contradiction:

Adaptability or versatilityVSDevice complexity

Solution Approach 1:

The patent uses opossum shrimp paste as an intermediary medium that naturally contains diverse microorganisms including lipase-producing strains. This pre-enriched sample serves as a convenient starting point for screening, reducing the complexity of having to screen from raw environmental samples while still providing access to diverse microbial sources

Inventive Principle:

Principle #24Intermediary (Mediator)

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

The use of Bacillus subtilis CS1802 in the whole-cell transformation method provides a cost-effective and efficient production of vitamin A palmitate, overcoming the high cost of commercial enzymes and expanding microbial diversity in food industry applications.

Implementation Method 1

The use of Bacillus subtilis CS1802 in the whole-cell transformation method provides a cost-effective and efficient production of vitamin A palmitate

Methodology Applied

Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

inoculating the strain cultured at the step (1) into a fermentation medium for shake fermentation

Methodology Applied

Scientific EffectFermentation: Fermentation

Data Source

PatentUS12180528B2Strain for producing lipase and application thereof

Publication Date: 2024.12.31 CHANGSHU INSTITUTE OF TECHNOLOGY

AI summary

The present invention relates to a lipase-producing strain and application thereof. The strain is classified and named Bacillus subtilis CS1802, with a preservation number of CCTCC NO: M2018262. The strain can be used to produce vitamin A palmitate through whole-cell transformation of vitamin A and palmitic acid. The Bacillus subtilis CS1802 of the present invention is derived from traditional natural fermented food and is a microorganism generally recognized as safe. The strain can be easily cultured and preserved. The highest content of vitamin A palmitate obtained through whole-cell transformation of vitamin A and palmitic acid is 15.35 mg/L. The highest transformation efficiency is 76.75%. The strain provides a new path for enzymatic synthesis of vitamin A palmitate and has important application prospects.