A D-allulose-3-epimerase immobilized enzyme preparation and its immobilization method

By using the encapsulation-crosslinking method of sodium alginate and calcium chloride containing cobalt chloride, and the adsorption technology of weakly basic anion exchange resin D301P, the stability problem of D-allulose-3-epimerase was solved, achieving high enzyme activity and low-cost industrial production.

CN116355888BActive Publication Date: 2026-07-17SHANDONG BAILONG CHUANGYUAN BIO TECH CO LTD
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
SHANDONG BAILONG CHUANGYUAN BIO TECH CO LTD
Filing Date
2021-12-28
Publication Date
2026-07-17

AI Technical Summary

Technical Problem

Existing D-allulose-3-epimerases lack good stability and are easily affected by pH, temperature and organic matter, making them unusable for repeated use and resulting in high costs for industrial applications.

Method used

A sodium alginate solution containing cobalt chloride was used as a carrier for immobilized cells, and calcium chloride was used as a cross-linking agent. D-alulose-3-epomerase was immobilized using an embedding-cross-linking method, and then adsorbed using a weakly basic anion exchange resin D301P to form an immobilized enzyme preparation of D-alulose-3-epomerase.

Benefits of technology

It improves the stability and activity of enzymes, increases the optimal reaction temperature of immobilized enzyme preparations by 10°C, increases the conversion rate from 30% to 35%, and maintains an enzyme activity recovery rate of over 65% after 10 reuses, thereby reducing industrial production costs.

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Abstract

This invention relates to a method for immobilizing D-allulose-3-epimerase. The method uses a sodium alginate solution containing cobalt chloride as a carrier for immobilized cells and a calcium chloride solution as a cross-linking agent, immobilizing the cells using an embedding-cross-linking method to obtain D-allulose-3-epimerase immobilized cells. The immobilized cells are then adsorbed onto a resin to obtain the D-allulose-3-epimerase immobilized enzyme preparation. The D-allulose-3-epimerase immobilized enzyme preparation prepared by this invention has an optimal enzyme reaction temperature 10°C higher than that of the free enzyme preparation, and the conversion rate of the immobilized enzyme preparation is increased from 30% to 35%. After being reused 10 times, the enzyme activity recovery rate can still be stably maintained at more than 65% of the initial enzyme activity. It can be repeatedly recycled in the industrial production of D-allulose, greatly reducing costs.
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