Isolation Of Antimicrobial Material From Pine Needle And Research Of Its Biological Activity

This study with double pine needles as raw materials, in the form with water, ethanol extraction of crude extract, leaves and fresh leaves and pine needles, in comparison with the bacteriostatic effect of screening bacteriostatic effect of solidifying, namely fresh pine needle leaves for subsequent tests.First of all, the single factor experiment, pine needles the determination of the optimal extraction conditions of antibacterial material is using the response surface optimization test.Then crude extract by dextran gel chromatography for purification, determine its protein concentration.According to Tricine sds-page electrophoresis- test, get the relative molecular mass.Finally the pine needles antibacterial material heat resistance, acid-proof alkaline, fight ion, and the roles of uv irradiation, determine the pine needle against the minimum bacteriostasis concentration of content.The results show:1, The ethanol extraction is the best way to pine needle antibacterial extraction effect.Single factor experiment, in order to determine the three optimal levels of each factor, made to measure the size of the bacteriostatic circle diameter.Response surface after the test as the three factors three levels, get the pine needles antibacterial the best extraction conditions, namely the material liquid ratio of 1:20, ethanol concentration was 75%, the extracting time of 5 h.2, Pine needles, purification of antibacterial materialGlucan gel Sephadex G- 25 chromatography separation, get a pure substance.Protein concentration is 0.87 mg/m L.3, Pine needle antibacterial material identificationTricine sds-page electrophoresis- test, the results showed that: a single band in 4.03 KD place, prove that it has reached the electrophoresis net.The relative molecular mass of 2.583 KD.4, The biological activity of pine needle antibacterial(1) Yhermal stability is one of the biological activities of pine needle antibacterial material.In 60 ℃, 100 ℃ hot water bath after 30 min, bacteriostatic activity of difference is not significant.But in 121 ℃ hot water bath 30 min bacteriostatic activity disappeared.(2) The pine needles antibacterial has the acid and alkaline.Placed in the acidic(p H < 6.0) and alkaline(p H > 8.0) environment, bacteriostatic activity basically remain unchanged.(3) Resistance to ion interaction and pine needles antibacterial biological activities.When the pine needle antibacterial content showed antimicrobial activity, is in the salt ion concentration less than 5%, and when the pine needle antibacterial salt ion concentration reaches 7% 9%, bacteriostatic activity largely disappeared.(4) The pine needles antibacterial content at the same time also has the role of resistance to ultraviolet radiation.After 5 h under uv irradiation, antibacterial activity, antibacterial activity will weaken after the disappeared, is when the irradiation time at 6 h above.(5) The pine needle antibacterial will showed inhibition to part of the growth of fungi, including: e. coli(Escherichia coli), Staphylococcus aureus, Staphylococcus aureus), Salmonella enteritidis(Salmonella enteritidis), brewer’s yeast(Saccharomyces cerevisiae Hansen).But for Penicillium(Penicillium notatum) and Aspergillus Niger, Aspergillus Niger) this kind of fungi is not show the bacteriostatic effect.(6) Purification of pine needle antibacterial in e. coli, the minimum bacteriostasis concentration is 0.386 mg/m L, staphylococcus aureus was 0.571 mg/m L, of salmonella enteritidis, beer yeast was about 0.164 mg/m L.Has no bacteriostasis penicillium and aspergillus Niger.