Ochrobactrum Enzyme Cleavage of Algal Polysaccharides
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Solution Overview
Problem
Current methods for enzymatic cleavage of ulvan polysaccharides from the genus Ulva are inefficient, as they cannot effectively cleave specific sequences between rhamnose 3 sulfate and glucuronic acid or iduronic acid, leading to heterogeneous products and high costs due to rapid enzyme inactivation and the need for repeated enzyme additions.
Innovation Solution
A method using a microorganism from the genus Ochrobactrum, specifically Ochrobactrum tritici strain PEC.2, to produce an enzymatic substance that cleaves glycoside bonds between rhamnose 3 sulfate and glucuronic acid or iduronic acid through β elimination, maintaining activity without interference from degradation products, allowing for the production of homogeneous oligomers with higher productivity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ulvan lyase from marine bacterium is used to cleave aldobiuronic acid sequences, then cleavage of glycoside bonds between rhamnose 3 sulfate and glucuronic acid/iduronic acid is achieved, but enzyme activity decreases rapidly and repeated enzyme additions are required
Solution Approach 1:
The patent changes the source parameter of the enzyme from marine bacterium to fungus (Trichoderma reesei), which fundamentally alters the enzyme's stability characteristics and eliminates the need for repeated additions while maintaining cleavage activity
Solution Approach 2:
The patent replaces the unstable enzyme preparation that requires repeated additions with a stable fungal enzyme that maintains activity throughout the process, eliminating the need for continuous enzyme replenishment
2Manufacturing precision
If glucuronan lyase from Trichoderma CNCM I-3400 is used to cleave glucuronan sequences, then specific sequence cleavage is achieved, but the main aldobiuronic acid sequences cannot be cleaved
Solution Approach 1:
The patent employs a fungal enzyme preparation that exhibits multi-functionality, being able to cleave both glucuronan sequences and aldobiuronic acid sequences, thereby achieving comprehensive degradation of ulvan polysaccharide components
3Productivity
If repeated enzyme additions are made to maintain degradation, then continuous cleavage is achieved, but production costs become prohibitive
Solution Approach 1:
The patent replaces the expensive and unstable enzyme preparation requiring repeated additions with a stable fungal enzyme that maintains activity throughout the process, eliminating the need for continuous enzyme replenishment and reducing production costs
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
This approach enables the cost-effective production of oligomers with reduced molecular weight and improved homogeneity by maintaining enzymatic activity throughout the process, reducing the need for repeated enzyme additions and lowering production costs.
Implementation Method 1
said enzymatic substance is brought in contact with said polysaccharide sequences so as to cause cleavage of the glycoside bond according to a reaction of β elimination
Data Source
AI summary
The invention relates to a method for enzyme cleavage of polysaccharides comprising a first sequence [→4)-β-D-GlcpA-(1→4)-α-L-Rhap3 sulphate-(1→]n and a second sequence [→4)-α-L-IdopA-(1→4)-α-L-Rhap3 sulphate-(1→]m, the first and second sequences respectively comprising two monosaccharide units connected by an osidic bond, wherein said method is such that: said polysaccharide sequences are provided; a microorganism capable of producing an enzyme substance of the lyase class is provided; and said enzyme substance is brought into contact with said polysaccharide sequences in such a way as to bring about cleavage of the osidic bond according to a β-elimination reaction. The invention is characterized in that a microorganism belonging to the bacteria of the Ochrobactrum genus is chosen for producing said enzyme substance.