Pectate Lyase Thermostability via Amino Acid Substitution
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current thermostable pectate lyases lose activity at temperatures above 70 degrees Celsius, limiting their application in industrial processes that require higher temperatures.
Innovation Solution
Introduction of specific amino acid substitutions, such as replacing lysine at position 235 with threonine, serine, asparagine, or cysteine, and optionally altering the amino acid at position 231 to leucine, enhances the thermostability of pectate lyases, allowing them to maintain activity at elevated temperatures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If thermostable pectate lyase is used in industrial processes, then enzyme stability at higher temperatures is improved, but enzyme activity is lost at temperatures above 70 degrees Celsius
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence of the pectate lyase enzyme through site-directed mutagenesis. Specific amino acid residues at positions 235 and 231 are altered to change the enzyme's thermal stability parameters while maintaining its catalytic function, allowing the enzyme to remain active at temperatures above 70°C
Solution Approach 2:
The patent creates modified copies of the original pectate lyase enzyme by introducing specific amino acid substitutions. These copied enzyme variants retain the core structure and function of the wild-type enzyme while incorporating improvements in thermostability through the copied and modified sequence information
Data Source
Figure 1
Figure 2
Figure 3
AI summary
The invention is in the field of protein chemistry, in particular in the field of enzymology. It provides pectinases, i.e. polypeptides with pectin-degrading properties. In particular the invention provides polypeptides with pectate lyase activity (EC 4.2.2.2). Enzymes according to the invention have improved properties, such as improved thermostability and decreased calcium dependence.