M23A Protease Production via Bacillus Extracellular Processing
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Solution Overview
Problem
M23A subfamily proteases cannot be efficiently produced in mature form using heterologous expression systems, such as Escherichia coli, due to lack of self-processing, limiting their industrial applications.
Innovation Solution
Culturing bacteria of the genus Bacillus with a polynucleotide encoding a proprotein of the M23A family protease to produce a mature form extracellularly, utilizing the host's extracellular proteases for processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If heterologous expression systems such as Escherichia coli are used to produce M23A subfamily proteases, then the production process can be simplified and controlled, but the proteases cannot be converted into mature forms due to lack of self-processing
Solution Approach 1:
The patent introduces a heterologous protease (subtilisin or papain) as an intermediary to process the proprotein form of M23A subfamily proteases into mature forms. The heterologous protease acts as a mediator that compensates for the lack of self-processing capability, enabling mature protein production in heterologous expression systems.
2Manufacturing precision
If natural M23A subfamily protease-producing strains are cultured to obtain mature enzymes, then self-processing capability is maintained, but productivity is low and production cost is high
Solution Approach 1:
The patent uses bacteria of the genus Bacillus as a universal host that can perform multiple functions: expressing the M23A subfamily protease gene, secreting the proprotein extracellularly, and providing heterologous proteases for mature form conversion. This multi-functional host system combines the advantages of both natural strains and heterologous expression systems.
Solution Approach 2:
The patent introduces the gene encoding M23A subfamily protease into Bacillus bacteria, creating a recombinant strain that copies the essential functions of natural producing strains while adding the capability for efficient heterologous expression and extracellular secretion, thereby improving productivity.
3Productivity
If proprotein is expressed in heterologous hosts, then the gene can be efficiently produced, but the proprotein cannot be converted into mature protein form
Solution Approach 1:
The patent introduces heterologous proteases (subtilisin or papain) as intermediary enzymes that specifically process the proprotein form into mature protein form. These intermediary proteases compensate for the lack of self-processing capability, enabling efficient conversion while maintaining high expression productivity.
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 efficient collection of mature M23A family proteases with improved productivity and industrial applicability, overcoming previous limitations in production methods.
Implementation Method 1
utilizing the host's extracellular proteases for processing
Data Source
AI summary
Provided is a method for efficiently producing an M23A family protease. The method for producing an M23A family protease includes culturing bacteria of the genus Bacillus having a polynucleotide encoding a proprotein of the M23A family protease introduced thereinto to produce a mature form of the M23A family protease extracellularly from the bacteria of the genus Bacillus.