Enzymatic Degradation of Alginate and in Vitro Immunological Activity of the Degradation Products

Abstract

Abstract: In this study, alginate was used as a substrate to explore the process parameters of enzymatic hydrolysis of alginate to prepare alginate oligosaccharides. On this basis, the in vitro immunological activity of the enzymatic hydrolysis products of alginate with different molecular weight ranges was evaluated. The results showed that the molecular weight of alginate decreased significantly after degradation by alginate lyase, and three different molecular weight ranges of degradation products were obtained through ethanol grading fractionation. Their weight-average molecular weights were 13.4 kDa, 5.73 kDa and 3.85 kDa respectively. The optimal process parameters for enzymatic preparation of alginate oligosaccharides were obtained by single factor experiment optimization: pH 7.0, alginate lyase dosage 15 U/g substrate, 24 hours. At this time, the yield of alginate oligosaccharides reached 28.05%. The in vitro immune activity of alginate and its hydrolysis products with three molecular weight ranges were evaluated on a mouse macrophage cell model. It was found that both alginate and its enzymatic hydrolysis products had certain immune enhancing activity, and the enzymatic hydrolysis products with a weight-average molecular weight of 5.73 kDa had the best immune enhancing activity, which was better than alginate oligosaccharide. Meanwhile，by adding a blocker of macrophage receptor protein TLR4 (TAK-242), it was verified that the alginate enzymatic hydrolysis products regulated macrophage immune activity by inducing the secretion of TLR4, causing a cascade reaction to increase the secretion of NO, TNF-α and IL-6. The research results can provide a theoretical basis for the high-value utilization of alginate.

Key words: alginate oligosaccharides, immunoreactivity, molecular weight, alginate enzymatic products, cytokines

CLC Number:

TS201.1

References

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