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無乳鏈球菌莢膜多糖的粗提及多糖含量測(cè)定條件優(yōu)化

發(fā)布時(shí)間:2018-11-05 09:19
【摘要】:為提取無乳鏈球菌莢膜多糖,并對(duì)苯酚-硫酸法測(cè)定多糖含量條件進(jìn)行優(yōu)化。離心收集發(fā)酵上清液,超濾濃縮,經(jīng)800mL/L乙醇沉淀提取無乳鏈球菌莢膜多糖;采用單因素試驗(yàn)對(duì)苯酚硫酸法測(cè)定條件進(jìn)行優(yōu)化,并對(duì)測(cè)定方法的可靠性進(jìn)行驗(yàn)證。結(jié)果表明,從1L發(fā)酵液中提取到含量為38.51%的莢膜多糖0.36g;測(cè)定的最佳條件為:2mL樣品液中,加入1 mL 50 mL/L的苯酚,5 mL濃硫酸,混勻后80℃作用20min,在1.5h內(nèi)測(cè)定485nm處的光密度。在該條件下,平均加樣回收率為99.96%,相對(duì)標(biāo)準(zhǔn)偏差為0.40%。說明從無乳鏈球菌發(fā)酵上清液中成功提取到莢膜多糖,建立了苯酚硫酸法測(cè)定莢膜多糖的最優(yōu)條件。
[Abstract]:In order to extract the capsular polysaccharides from Streptococcus lactis and optimize the conditions for the determination of polysaccharides by phenol-sulfuric acid method. The supernatant of fermentation was collected by centrifugation and concentrated by ultrafiltration. The capsule polysaccharide of Streptococcus acuminatum was extracted by 800mL/L ethanol precipitation. The conditions of phenol sulfuric acid method were optimized by single factor test and the reliability of the method was verified. The results showed that the content of capsule polysaccharides was 38.51% and 0.36 g from 1L fermentation broth. The optimum conditions were as follows: 1 mL 50 mL/L phenol and 5 mL concentrated sulfuric acid were added to 2mL sample solution, then mixed at 80 鈩,

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