Enzymatic Preparation of β-Glucan Oligosaccharides by Pichia Pastoris and Its Etransdermal Properties
In order to accurately convert β-glucan into small molecules,recombinant Pichia pastoris MF060.RG.Glu-13 was constructed to express β-1,3-glucan endonucliase.The activity of β-1,3-glucan endonucliase reached 982 U/mL in the fermentation broth obtained by fermentation.The β-glucan oligosaccharides with molecular weight of 1-2 kDa were prepared by enzymatic hydrolysis of β-glucan in fermentation broth.IR and NMR were used to preliminarily identify the structure of β-glucoseoligosaccharides,and Franz diffusion cell was used to test the ir transdermal property.The results showed that the main chain of the product β-glucooligosaccharide was connected by five glucosides by β-1,3-glycosidic bonds,and there was a D-glucose branch connected by β-1,6-glycosidic bonds on the second and third glucosides.The transdermal absorption rate of the product β-glucose-oligosaccharide(1-2 kDa)was 2.13%,which was 53.25 times higher than that of the enzymatic substrate,macromolecule β-glucan(800-1 000 kDa),of 0.04%.
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