Branched Cyclodextrin Purification via Selective Enzymatic Hydrolysis
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Solution Overview
Problem
Current methods for preparing branched cyclodextrin suffer from low yield and purity due to difficulties in separating branched cyclodextrin from cyclodextrin, which has similar molecular weights, making purification challenging.
Innovation Solution
A method involving the use of cyclodextrin glycosyltransferase (CGTase) and a saccharifying enzyme to treat a mixture containing branched cyclodextrin, controlling CGTase activity to 0.1-0.4 U/g of the mixture by dry weight, allowing for the selective hydrolysis of cyclodextrin into glucose, thereby facilitating separation and purification of branched cyclodextrin.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If pullulanase is used to prepare branched cyclodextrin from cyclodextrin and maltose/maltotriose, then branched cyclodextrin can be synthesized, but the remaining cyclodextrin in the mixture is relatively large and difficult to separate due to similar molecular weights
Solution Approach 1:
The patent extracts and removes the unwanted cyclodextrin component from the reaction mixture through selective enzymatic hydrolysis. By using CGTase to specifically hydrolyze cyclodextrin into glucose while leaving branched cyclodextrin intact, the method effectively separates the desired product from the unwanted byproduct, solving the purification problem caused by similar molecular weights.
Solution Approach 2:
The patent changes the molecular weight parameter of the unwanted cyclodextrin by enzymatically converting it into glucose, which has a significantly different molecular weight. This parameter change enables effective separation through techniques like membrane filtration or chromatography, resolving the separation difficulty caused by similar molecular weights between cyclodextrin and branched cyclodextrin.
2Ease of manufacture
If CGTase is used to act on starch or starch derivatives to prepare branched cyclodextrin, then the process can be simplified, but the yield and purity are low
Solution Approach 1:
The patent performs a preliminary action by first synthesizing branched cyclodextrin using pullulanase, then subsequently removing the unwanted cyclodextrin through CGTase treatment. This two-step approach ensures high initial synthesis efficiency while enabling effective purification, thereby achieving both high yield and high purity that cannot be obtained by using CGTase alone on starch.
3Productivity
If high-concentration cyclodextrin is used as substrate with pullulanase, then branched cyclodextrin can be produced efficiently, but the large amount of remaining cyclodextrin complicates separation and purification
Solution Approach 1:
The patent introduces CGTase as an intermediary enzyme that selectively acts on the unwanted cyclodextrin to convert it into glucose. This intermediary action transforms the separation problem from separating two similar molecules (cyclodextrin and branched cyclodextrin) into separating glucose from branched cyclodextrin, which is much easier and reduces purification complexity.
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 method enhances the yield and purity of branched cyclodextrin by exploiting the molecular weight difference between branched cyclodextrin and glucose, making subsequent separation and purification more effective.
Implementation Method 1
the activity of CGTase is controlled and the coupling effect of the CGTase is used to make the CGTase and the saccharifying enzyme according to 0.1-0.4 U/g of the mixture by dry weight act on the above mixture
Implementation Method 2
selective hydrolysis of cyclodextrin into glucose
Implementation Method 3
the large molecular weight difference between the branched cyclodextrin and the glucose is favorable for subsequent separation and purification
Data Source
AI summary
The disclosure herein relates to a method for preparing branched cyclodextrin and application thereof, and belongs to the technical field of synthesis of branched cyclodextrin. The method comprises the following steps: (1) dissolving maltodextrin in a phosphate buffer solution, and adding CGTase for reacting; (2) reducing enzyme activity by a physical method; and (3) adding a saccharifying enzyme to the reaction system of step (2), and performing high-temperature enzyme deactivation to obtain the branched cyclodextrin. The method has mild reaction conditions, and at the same time, cyclodextrin and unreacted substrates can be hydrolyzed via the weak coupling activity of CGTase, thereby realizing effective separation of the branched cyclodextrin.