Bacillus subtilis Natto Fermentation for High-Yield MK-7
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Solution Overview
Problem
Current methods for producing vitamin K2 (MK-7) are inefficient, environmentally harmful, and yield low quantities, making it difficult to meet the global demand for this essential nutrient.
Innovation Solution
A novel Bacillus subtilis natto strain, CGMCC No. 17894, is developed through mutagenesis to enhance vitamin K2 (MK-7) production, using a fermentation process with optimized media and purification techniques to achieve high yields and pure trans structure.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If chemical synthesis method is used to produce vitamin K2 (MK-7), then production capacity is improved, but environmental harm and organic solvent residues increase
Solution Approach 1:
The patent replaces chemical synthesis methods with biological fermentation using Bacillus subtilis natto strain. This substitution eliminates the need for organic solvents and harsh chemical reagents, thereby reducing environmental harm and solvent residues while maintaining production capacity through efficient microbial fermentation processes.
Solution Approach 2:
The patent optimizes fermentation parameters including temperature (30-37°C), pH (6.5-7.5), and cultivation time (16-24 hours) to maximize MK-7 yield. By adjusting these parameters, the fermentation process achieves high productivity comparable to chemical synthesis while avoiding its harmful effects.
2Ease of manufacture
If extraction from solid fermented natto is used to produce vitamin K2 (MK-7), then production method is simple, but vitamin K2 (MK-7) content is extremely low
Solution Approach 1:
The patent transforms the fermentation medium from solid to liquid state, which fundamentally changes the extraction efficiency. Liquid fermentation allows for better mass transfer and higher cell density, resulting in 200-300 mg/L MK-7 concentration - a 20-fold increase compared to solid fermented natto, while maintaining process simplicity.
Solution Approach 2:
The patent transitions from solid-phase fermentation to liquid-phase fermentation, adding the dimension of fluidity to the system. This dimensional change enables better nutrient distribution, oxygen transfer, and product extraction, dramatically increasing MK-7 yield while keeping the manufacturing process straightforward.
3Ease of manufacture
If liquid fermentation of natural Bacillus natto is used to produce vitamin K2 (MK-7), then production method is efficient, but fermentation period is long and yield is low
Solution Approach 1:
The patent applies mutagenesis treatment to Bacillus natto to create strain CGMCC No. 17894 with superior MK-7 production capabilities. This genetic modification, combined with optimized liquid fermentation parameters (temperature 30-37°C, pH 6.5-7.5, aeration rate 1.5-2.0 v/v/min), reduces fermentation time to 16-24 hours and increases yield to 200-300 mg/L, resolving the contradiction between efficiency and productivity.
4Device complexity
If conventional fermentation is used to produce vitamin K2 (MK-7), then production process is straightforward, but vitamin K2 (MK-7) yield is low
Solution Approach 1:
The patent employs mutagenesis-induced strain improvement to create CGMCC No. 17894, which naturally produces high levels of MK-7. Combined with optimized fermentation conditions (initial inoculum 5-10%, cultivation temperature 30-37°C, pH 6.5-7.5), the process achieves 200-300 mg/L yield without complicating the production workflow, maintaining straightforward operation while dramatically boosting yield.
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 process significantly increases vitamin K2 (MK-7) yield by 20 folds, achieving 200-300 mg/L within 16-24 hours, providing a sustainable and effective method for producing high-purity MK-7 for health and medical applications.
Implementation Method 1
a method for producing vitamin K2, including culturing the Bacillus subtilis natto strain CGMCC No. 17894 in a medium to produce vitamin K2 in the strain cells and the medium
Data Source
AI summary
Provided is a Bacillus subtilis natto strain ST-1008, deposited under the accession number CGMCC No. 17894. Also provided is a method for producing MK-7. The method comprises: culturing the Bacillus subtilis natto strain of CGMCC No. 17894 in a culture medium so as to produce MK-7 in the strain cell and the culture medium, and recovering and purifying MK-7 from the strain cell and the culture medium. Also provided are a MK-7 preparation, a MK-7 pure premix, and a pure MK-7.