Study On The Antibacterial Enhancement Active Constituents From Sour Jujube And Their Biological Activities

Sour jujube(Ziziphus jujuba Mill.var.spinosa(Bunge)Hu ex H.F.Chow)belongs to family Rhamnaceae.It has many biological activities,such as calming and hypnotic,improving learning and memory,strong heart,antitumor,antibacterial,antioxidation,antidepressant,hypolipidemic,protecting myocardium,enhancing humoral immunity and cell immunity.Sour jujube is an important part of China's important wild fruit trees and medicinal plant resources,and has great potential for development and utilization.In this paper,sour jujube fruit was used as the raw material to extract,separate and purify the chemical composition of the antibacterial enhancement activity,composition analysis of the obtained products,the broad-spectrum antibacterial enhancement activity analysis from the obtained products.The obtained products were tested on the bacterial biofilm and motility,and that own the potential of a food preservative.This study could provide theoretical basis for the further development and utilization of the sour jujube.The test results are as follows:1.Ultrasonic assisting different polar solvents: petroleum ether,chloroform,ethyl acetate,80% methanol and hot water(polarity from small to large)from 10 kg sour jujube extract effective constituent of different polar range,respectively obtained petroleum ether extract 73.28 g,chloroform extract 278.16 g,ethyl acetate extract 7.79 g,80% methanol extract 1717.04 g,hot water extract 356.83 g.The extraction rates were 0.733%,2.782%,0.078%,17.170%,3.568%,respectively.2.The bacteriostasis effect of each extract was measured by filter paper method.The results showed that chloroform extract had good bacteriostasis effect on pseudomonas aeruginosa,Acinetobacter baumannii,Staphylococcus aureus,Bacillus subtilis,Escherichia coli,Monilia albican,Salmonella typhi,Shigella flexneri,Pichia pastoris,but the other extracts had no bacteriostasis effect.3.Evaluation of antibacterial enhancement activity in vitro.the results showed that the chloroform extract could significantly reduce the minimum inhibitory concentration(MIC)of ampicillin(Amp)to pseudomonas aeruginosa,and showed better antibacterial enhancement activity in vitro.while petroleum ether extract,ethyl acetate extract,80% methanol extract and hot water extract did not have antibacterial enhancement activity.4.The best process for isolation and purification of sour jujuba chloroform extract is through the silica gel column chromatography with antibacterial enhancement activity tracer is as follows: the first column chromatography uses petroleum ether,petroleum ether:chloroform 1:1,We get the active component Fr.3 through petroleum ether:ethyl acetate 1:1.Then the Fr.3 is separated and purified by second column chromatography,and the extraction are eluted in turn through the petroleum ether and petroleum ether,ethyl acetate 10:1.Because of loss of activity,column chromatography can not be used to separate Fr.2a.Therefore,Fr.2a is identified as a refined product of antibacterial enhancement activity.5.Fr.2a components were analyzed by GC-MS,its main components: 49.59% 1,3-two chloro propanol,5.49% 1,1-dichloromethyl ether,0.96% hexachloroethane,7.81% 1,2,3,3-tetrachloroprop-1-ene,1.33% lauric acid,1.34% myristic acid,0.87% palmitoleic acid,7.37% palmitic acid,9.75% dibutyl-o-phthalate,2.02% trans-13-octadecenoic acid,1.88% Oleamide,3.06% ?-amyrin,0.93% ?-Acorenol,6.20% lupeol,1.42% ursolic aldehyde and so on.6.The study of the broad-spectrum antibacterial activity to the refined products showed that: When Fr.2a was combined with gentamicin(Gm),kanamycin(Km),tobramycin(Tob),Amp,tetracycline(Tc),carbenicillin(Cb)and streptomycin(St),have synergistic antibacterial effect on pseudomonas aeruginosa;When Fr.2a was combined with Gm,Km,Tob,Amp,and St,have synergistic antibacterial effect on Staphylococcus aureus;Fr.2a was combined with Gm,Km,Tob and Tc,have synergistic antibacterial effect on Bacillus subtilis;Fr.2a is combined with Gm,Km,Tob,Amp,Tc,chloramphenicol(Cm),have synergistic antibacterial effect on Escherichia coli,Monilia albican,Salmonella typhi and Shigella flexneri.7.Fr.2a concentration promoted the formation of biofilm in a dose dependent and without affecting the growth of pseudomonas aeruginosa and Monilia albican,while different concentrations of Fr.2a had no significant effect on the formation of the biofilm of Acinetobacter baumannii.8.1/4 MIC Fr.2a can inhibit motility of pseudomonas aeruginosa,Acinetobacter baumannii,Escherichia coli,Salmonella typhi and Shigella flexneri.9.The application of Fr.2a in food preservation shows that the addition of 0.5% Fr.2a to milk can prevent the decrease of milk pH and significantly inhibit the growth of microorganism in milk.Therefore,Fr.2a can be used as a potential food additives in the food industry.