Extraction And Purification Of Polysaccharide From Jujube And Its Inhibitory Effect On Tyrosinase

Ziziphus jujube is the one of the main industry in Hunan area, especially in hengyang.The comprehensive utilization and resources of the jujube are an important guarantee for sustainable and efficient development of citrus industry currently.In orde to promote the utilization of excavate germplasm resources of jujube, this experiment took jujube polysaccharitdes as comprehensive measuring index. This experiment took a lot of varieties jujube in Hunan as est materialsto analyze different varieties of jujube Polysaccharide content,Selection of candied jujube, determinating extraction, separation,purification and activity Etc.a series of studies,The main results were as follows:(1) Taking 18 kinds of jujube cultivars in Hunan local as test materials, Analysis the polysac charide contentd in different types varieties of jujube.The analysis showed that, The highest and the lowest polysaccharide content of 3.9 times in the different varieties of jujube, oMillet meters jujube and filial generation candied jujube cultivars of jujube highest polysaccharide, polysacchari de content were 9.75%,9.06%, The secondly were the female candied jujube, acid yuan jujube, medicinal jujube, Long jujube, YuQuan No.8 and southern dongzao, respectively,8.82%,6.2%, 5.89%,5.56%,5.54%,5.23%, and the light skin jujube content is the lowest, only 2.45%.(2) Determination of affect on the extraction rate of polysaccharide with 8 kinds of single factor. With the principal component analysis, the construction of various factors on extraction rate of mathematical model:F= 0.292X1+0.305X2+0.254X3+0.287X4+0.273X5+0.254X6+0.320 6X7+0.300X8, Through above experiment,Selecting the four main factors to thogonal test, the optimum extraction process parameters:ratio of material to liquid 1:40, ultrasound acoustictime 40min, ultrasonic power 700w, ultrasonic temperature of 70, under this condition, the jujube polysaccharide extraction rate is 9.38% in theory.(3) With the experimental design of response surface removal glycoprote in polysaccharide, and then regression analysis of test results.The two degree polynomial equation between the test conditions was:Y=76.31-0.59A+1.64B+0.94C+0.089D+0.087AB-2.09AC-0.97AD+3.40BC+2.04 BD-1.84CD-1.14A2-6.21B2-1.42C2-5.26D2, select the four main factors of experiment,the optim um process parameters:enzymolysis temperature of 56.09,1.48h, enzymolysis time,enzyme dosage solution of pH5.942.15mL,under this condition, the removal rate of jujube more glycoprotein the oretical value is 76.81%.(4) Jujube polysaccharide in turn by DEAE-52 cellulose and SephadexG-100 Sephadex column chromatography.The results showed that the DEAE-52 cellulose column eluent polysaccharide three components could be obtained, which were named DTA, DTB, DTC,The components of DTAand DTc have good symmetry, no noticeable overlap phenomenon and high purity, But the component DTB have high overlap degree and low purity; After further purification SephadexG-100 column, there are five good symmetry, no overlap, high purity of the polysaccharide components The DTB could be isolated the three components of polysaccharide, respectively named DTB1, DTB2,DTB3.(5) The study measured SephadexG-100 with dextran column of relative molecular weight range ofpolysaccharide group has been purified from the. The results showed that:the relativ emolecular weight range of 399086-248161 DTA, relative molecular weight range of DTB1 to 1956 89-133814, relative molecular weight range of DTB2 is 100624～37102, the relativemolecular wei ghtrange of DTB3 to 30681-23071, relative molecular weight range of DTC is 2000615044.(6) To determine monosaccharide composition of jujube polysaccharide with gas chromatogr-aph. The results showed that the monosaccharide composition of polysaccharides in the Candied jujube polysaccharide, with xylitol,glucose, galactose containing containing ratio was higher, respectively 19.4%,18.5%,24.2% of the total;The secondly were the Rhamnose, ribose, Arabinos e and mannose.respectively 7.1%,6.5%,8.1%,7.9%; The fucose, xylose content was lower, accounted for only 5.0%,3.4% of the total.(7) The study was aimed to analyze the tyrosinase activity, inhibition from polysaccharide in candied jujube.And on the kinetic study of strongest activity inhibitory parts.The results showed that the DTB3 inhibitory rate of enzymatic activity was the highest, reaching 77.94%. The DTc inhibitory rate of enzymatic activitywas 69.93%, The remaining components on the activity of tyrosinase inhibitory rate was less than 50%. Obviously, There were some differences between the components of Polysaccharide on tyrosinase activity, inhibition. DTB3 can quickly combine with the enzyme in a short time, and showed strong activity inhibitory ability, and presented a type of non competitive inhibition.When Polysaccharide components DTA, DTB1, DTB2 through sulfated modified was improved on inhibite tyrosinase activity than the original. Respectively were increased by 60.19%,41.66%,29.55%, DTC slightly increased by 0.23%. but the components of DTB3 were a slight decline, down 0.42%.