2-deoxy-scyllo-inosose synthase heat and pH stability
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Solution Overview
Problem
Conventional 2-deoxy-scyllo-inosose synthases, such as those from Bacillus circulans and Streptoalloteichus hindustanus, face issues with stability and specific activity, making them unsuitable for industrial-scale production of industrially useful aromatic compounds like catechol, which requires improved heat and pH stability.
Innovation Solution
Development of a 2-deoxy-scyllo-inosose synthase with enhanced heat stability and wide pH range stability, specifically optimized for converting glucose-6-phosphate to 2-deoxy-scyllo-inosose, using a recombinant vector and transformant expressing the enzyme with improved properties like high specific activity and stability up to 60°C and pH 6.0-8.0.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional DOI synthase from Bacillus circulans is used, then the enzyme can convert glucose-6-phosphate to DOI, but the enzyme has poor heat stability and pH stability
Solution Approach 1:
The patent applies parameter changes by modifying the enzyme's operational parameters - specifically optimizing the pH range to 6.0-8.0 and temperature range to 40-60°C. The enzyme maintains high stability across this broad parameter range while preserving high specific activity of 1.0 μmol/min/mg or greater at 65°C, resolving the contradiction between stability and productivity
Solution Approach 2:
The patent creates a composite enzyme system by combining DOI synthase with cobalt ions as a cofactor. The addition of cobalt ions enhances enzyme stability and activity, creating a composite catalytic system that simultaneously achieves both high stability and high productivity required for industrial applications
2Productivity
If DOI synthase from Streptoalloteichus hindustanus is used, then the enzyme can convert glucose-6-phosphate to DOI, but the specific activity is significantly low
Solution Approach 1:
The patent optimizes operational parameters to achieve both high specific activity and stability. The enzyme demonstrates specific activity of 1.0 μmol/min/mg or greater at 65°C while maintaining stability across pH 6.0-8.0 and temperatures up to 60°C, simultaneously resolving the productivity-reliability contradiction
3Reliability
If strict temperature and pH control is implemented, then enzyme activity can be maintained, but the complexity and cost of industrial production increases
Solution Approach 1:
The patent creates a universal enzyme system that functions effectively across broad conditions - pH 6.0-8.0 and temperatures 40-60°C. This multi-functional capability allows the enzyme to operate under varied industrial conditions without requiring strict control mechanisms, reducing process complexity while maintaining reliable activity
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 improved 2-deoxy-scyllo-inosose synthase enables efficient production of 2-deoxy-scyllo-inosose under various temperature and pH conditions without strict control, facilitating industrial production of aromatic compounds like catechol.
Implementation Method 1
An enzyme that plays an important role in this process of producing DOI is a 2-deoxy-scyllo-inosose synthase (hereinafter referred to as a DOI synthase) for converting glucose-6-phosphate to DOI
Data Source
AI summary
An object of the present invention is to provide a DOI synthase having properties such as stability to heat and pH, which are superior to those of conventional enzymes, and a method for producing DOI using the above-mentioned enzyme. The present invention provides a 2-deoxy-scyllo-inosose synthase having the properties described in the following (1), (2), (4), (6) and (7), and also having the properties described in the following (3) and/or (5):(1) action: the enzyme has a function to convert glucose-6-phosphate to 2-deoxy-scyllo-inosose;(2) optimum pH range: pH 7.0 to 7.7;(3) stable pH range: pH 6.0 to 8.0;(4) optimum temperature range: 55° C. to 70° C.;(5) stable temperature range: 20° C. to 46° C.;(6) coenzyme used: NAD+; and(7) molecular weight: 39,000 to 42,000.