Expression and properties of an endo-1,3-fucoidanase in Lactobacillus
Fucoidan is a marine polysaccharide with diverse physiological activities.Endo-1,3-fucoidanase is a favorable tool for the preparation of fucooligosaccharides.Nevertheless,the lack of food-grade fucoidanase with clear cleavage sites has hindered its application.This study aimed to obtain a food-grade endo-1,3-fucoidanase from GH174 family(Fun174Sb)and to investigate its cleavage site.The study demonstrated that the food-grade expression of Fun174Sb could be achieved through the Lactobacillus NICE system.Fun174Sb exhibits significant endo-acting hydrolysis activity towards fucoidan from Holothuria tubulosa(Ht-FUC),with an enzymatic activity of 1.90 U/mL.Fun174Sb showed significant activity within a temperature range of 35-50 ℃ and a pH range of 7.5-8.5,and it possessed good temperature and pH stability.Furthermore,the structure of the dominant oligosacchar-ide in the enzymatic product was analyzed using ultra performance size exclusion chromatography-mass spectrum(UPSEC-MS)and nuclear magnetic resonance(NMR).It could be inferred that Fun174Sb cleaves the α-1,3-glyc-osidic bond between Fucp2(OSO3-)and Fucp2,4(OSO3-)in Ht-FUC.The study showed that Fun174Sb expressed in Lactobacillus exhibited favorable biochemical properties and a clear mode of action.It laid the foundation for the application of food-grade fucoidanase.
fucoidanaseLactobacillussea cucumbercloning and expressionbiochemical propertiesaction mode
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