Recombinant Bacillus subtilis Strain for Riboflavin Yield Enhancement
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current industrial production of riboflavin by Bacillus subtilis strains falls short of meeting industrial demands, with maximum yields not sufficient for commercial needs.
Innovation Solution
Development of a recombinant B. subtilis strain, B. subtilis RF1-6, through gene modification and mutagenesis, incorporating zwf, ywlf, and ribBA genes with a strong promoter P43, and knocking down pgi and purR genes to enhance metabolic flux and yield, resulting in a microbial inoculant for increased riboflavin production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional B. subtilis strains are used for riboflavin production, then the production process is simple and stable, but the riboflavin yield is insufficient (maximum 15.7 g/L)
Solution Approach 1:
The strain construction is divided into modular steps: starting with a high-yield parent strain (RF1), performing targeted gene modifications (inserting zwf, ywlf, ribBA genes; knocking down pgi and purR genes), and creating a systematic mutant series (RF1-6, RF1-6ZYRS) that segments the complexity of metabolic engineering into manageable, incremental improvements
Solution Approach 2:
The patent systematically changes genetic parameters by modifying specific gene expressions - inserting genes (zwf, ywlf, ribBA) to enhance riboflavin synthesis pathways and knocking down genes (pgi, purR) to redirect metabolic flux - thereby achieving a yield increase from 15.7 g/L to 25.2 g/L through controlled genetic parameter modification
2Productivity
If gene modification and mutagenesis are performed to increase riboflavin yield, then the riboflavin yield increases (22.8% improvement), but the strain construction complexity increases
Solution Approach 1:
The patent performs preliminary actions by first identifying and modifying key genes (zwf, ywlf, ribBA, pgi, purR) that control riboflavin synthesis pathways, establishing a foundation of genetic knowledge and experimental protocols that simplifies subsequent strain construction and optimization
Solution Approach 2:
The patent employs feedback mechanisms through systematic screening of mutant strains (RF1-6 series) to identify the optimal combination of gene modifications that maximize riboflavin yield while managing construction complexity, using performance data to guide further genetic engineering efforts
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 recombinant strain achieves a 22.8% increase in riboflavin yield compared to the parent strain and a maximum yield of 25.2 g/L when engineered as B. subtilis RF1-6ZYRS, representing a 69.58% increase over the initial strain, effectively addressing the shortfall in industrial production.
Implementation Method 1
The present disclosure belongs to the technical field of microbial fermentation, and particularly relates to a Bacillus subtilis strain, a recombinant B. subtilis strain and use thereof
Data Source
AI summary
The present disclosure provides a Bacillus subtilis strain, a recombinant B. subtilis strain and use thereof, belongs to the technical field of microbial fermentation. B. subtilis strain RF1-6 provided by the present disclosure is a mutant strain with the highest riboflavin yield screened by gene modification and mutagenesis using a high riboflavin-producing strain RF1 as a starting strain, deposited at China Center for Type Culture Collection (CCTCC) under accession number M 2022565. Riboflavin yield can be increased by 22.8% compared with that of the high riboflavin-producing strain RF1.