Study On Production Of New Polysaccharide By Bacillus Amyloliquefaciens With Inulin As Carbon Source

Objective: Polysaccharides have a wide range of physiological activities,and the search for new active polysaccharides is of great significance in the pharmaceutical field.In this paper,using inulin as the only carbon source,liquid fermentation Bacillus amyloliquefaciens and controlling fermentation conditions,three new polysaccharides obtained structural analysis and preliminary activity of one of them.Methods: Using inulin as the only carbon source,liquid fermentation Bacillus amyloliquefaciens.the purified polysaccharides were extracted and isolated from the fermentation fluid,and the structural characterization was studied to determine their biological activity.Fermentation conditions: at 28 ℃,Bacillus amylstarch was cultured with 72 h by Culture with inulin medium.Separation and purification: Fermentation Solution-Centrifugal-Over 0.22 μm Filter Disinfection → 70% Ethanol Depression → Large Porous Resin Decolor → sevage Reagent Deprotein → Over deae-52 column → Over sephadex g200 Gel Column →Dialysis-Frozen Dry →HPLC Inspection.Structural characterization study: HPSEC determination of molecular weight,HPLC-MS determination of monysaccharide composition,methylation experiment,and determination of monosaccharide connection method by NMR method.Bioactivity determination: determination of α-glucosinase inhibitory activity by spectrophotometry;hydroxyfree radical clearance experiment;determination of polysaccharide proliferation activity on raw264.7 cells;and polysaccharide inhibitory activity on ct26 cells.Results: Using inulin as the only carbon source,liquid fermentation Bacillus amyloliquefaciens produced 1.15 g/L.Three components are obtained after coarse polysaccharide separation and purification,named PPBFI-1,PPBFI-2 and PPBFI-3.PPBFI-2 and PPBFI-3,both of them were inactive,and the PPBFI-1 was studied in detail.PPBFI-1: 86.90% for hydroxyfree radical clearance at 8mg/ml(p< 0.01);It has a certain inhibitory effect on ct26 cancer cells,The inhibition rate was 15.44% at 1.6mg/ml(p< 0.01);Significant inhibition of α-glycosidase,36.90% at 1.6mg/ml(p< 0.01);PPBFI-1(0.4mg/ml)can significantly stimulate the raw264.7proliferation of macrophages,24 h proliferation rate of over 300%(p< 0.01).A structural characterization of the PPBFI-1 component is made,and the monosaccharide composition of the PPBFI-1 is: Fucose,Amminogalactose hydrochloride,Rhamnose,Arabic sugar,Glucosamine hydrochloride,Galactose glucose,Galacuronic acid,Glucuronic acid,and moor ratio is: 0.011:0.068: 0.133: 0.158: 0.031: 0.255: 0.030: 0.184: 0.130.PPBFI-1’s sugar main chain is:→ 6-β-D-galactose polycan-1 → 6-β-D-galactose polycan-1 → 6-β-D-galactose glycan-1 →2-α-L-rat risan-1 → 4-α-D-galactaldehyde acid-1 → connection.Conclusion: The way to ferment Bacillus amyloliquefaciens with polysaccharide of Jerusalem artichoke as carbon source to obtain polysaccharide with novel structure was discussed.The obtained PPBFI-1 has potential application value.