Study On The Isolation,Purification And Activity Of Poilsaccharide From Euglena Gracilis

Euglena gracilis is the name of the ancient protozoan eyeworm.Eyecells contain chloroplasts and can perform photosynthesis,so they have both animal and plant characteristics.Euglena gracilis has no cell wall and can be easy to be digested.Euglena gracilis is also rich in nutrients,such as protein,polysaccharides,vitamins,mineral nutrients and so on.Euglena gracilis also has the function of removing harmful cholesterol and heavy metals,and solving constipation and constipation.Polysaccharides are a class of sugars with anti-cancer,anti-viral,anti-aging,hypoglycemic,lipid-lowering,and bacteriostatic functions.However,there are few reports on euglena gracilis polyccharides?EGPS?.Euglena gracilis polysaccharide was added to this kind of cosmetics,but its extraction process have not been studied.This paper uses euglena gracilis as raw materials,and uses ultrasonic-assisted enzymolysis to optimize the extraction process of euglena polysaccharides.The cavitation of ultrasound is used to make cells easy to break and help polysaccharides to dissolve.Enzyme engineering technology can decompose under milder conditions.These two methods have the characteristics of high efficiency,energy saving,etc.,and have been widely used in the extraction of active ingredients of plants and animals.The euglena gracilis polysaccharide was divided into three molecular weight segments by ultrafiltration technology,and its antioxidant activity was analyzed.The euglena gracilis polysaccharide with molecular weight segment with better antioxidant activity was subjected to DEAE-cellulose column and G-100 glucan gel.Chromatography was used for separation and purification,and its purity identification and antioxidant activity analysis were performed.The purified euglena gracilis polysaccharides with better antioxidant activity were subjected to molecular weight determination,monosaccharide composition,and other physiological activity analysis.The purpose of this study is to obtain euglena gracilis polysaccharides with the best antioxidant activity.Firstly,based on the evaluation of the polysaccharide extraction rate,four factors including ultrasonic power,ultrasonic time,enzymatic hydrolysis time,and enzyme addition amount were used to extract ultrasound-assisted enzymatic extraction of the effect of euglena gracilis polysaccharide yield.The results showed that the optimal extraction process was ultrasonic power of 360 W,ultrasonic time of 19 min,neutral protease addition amount of 3.7%,and enzymolysis time of 3.5h.Under these conditions,the polysaccharide yield was 3.68%.The euglena gracilis polysaccharide was fractionated by ultrafiltration method to obtain euglena gracilis polysaccharides with 3 different molecular weight segments?EGPS-1?EGPS-2?EGPS-3?.The in vitro anti-oxidation experiments of euglena polysaccharides with 3different molecular weight segments were performed.The antioxidant activity analysis showed that EGPS-3 had the strongest hydroxyl radical scavenging ability,superoxide anion scavenging ability,Fe²⁺chelating ability,and reducing power,and its IC₅₀was the strongest.At 0.23 mg/m L,10 mg/m L,and 0.07 mg/m L,respectively,EGPS-1 had the strongest lipid peroxidation inhibitory ability,with IC₅₀ of 28.27 mg/m L.There was no DPPH free radical scavenging ability of Euglena polysaccharides in each molecular weight range.EGPS-3 was separated and purified by DEAE-cellulose column and G-100 glucan gel chromatography to obtain three purified polysaccharides EGPS-3A1,EGPS-3A2,and EGPS-3C1.The purity identification results of ultraviolet scanning and the showed that the purity of the three purified polysaccharides was high;the analysis of infrared spectra showed that each component had a typical absorption characteristic of sugars;the results of antioxidant experiments showed that each component had Certain antioxidant activity,EGPS-3A1 has higher antioxidant activity,its molecular weight is1.368×10⁵Da,monosaccharide composition includes:mannose,rhamnose,glucuronic acid,galacturonic acid,glucose,xylose galactose,arabic;The ratio of the amount of sugar and fucose is 0.072:0.43:0.049:0.01:0.26:0.62:0.53:0.18:1.01.Physiological activity experiments show that EGPS-3A1 has a strong inhibitory activity against Staphylococcus aureus and E.coli,and the inhibitory activity against E.coli is greater than that of Staphylococcus aureus,and it is also predicted to have blood sugar lowering,blood lipid lowering,and cholesterol lowering.effect.This study explores the extraction process of euglena gracilis polysaccharides and the molecular weight,monosaccharide composition and other physiological activities of euglena gracilis polysaccharides with better antioxidant activity.It provides a basis for the further deveiopment and utilization of Euglena gracilis polysaccharides.