古尼拟青霉胞内多糖的提取分离工艺及条件

The technical route and condition of polysaccharides extraction and isolation from Paecilomyces gunnii

通过对古尼拟青霉胞内多糖(简称PGPS)的提取分离工艺及条件的研究,结果表明,正交试验优化后的PGPS提取条件:一份脱脂菌粉,加10倍菌粉量的蒸馏水,70℃条件下浸提2h,浸提3次,合并浸提液并浓缩至其总体积的1/6,再加至质量分数85%的乙醇,醇析12h。大孔吸附树脂脱色效果优于H2O2。PGPS半精制品上DEAE-SephadexA-25层析柱和SephadexG-200层析柱获得4个主要组分。PGPS中多糖含量为13 9%,其中含有结合蛋白的多糖组分。

In this paper,the general methods of extraction and isolation of intrapolysaccharide (PGPS) from strain 5-19 of Paecilomyceps gunnii were investigated.The best technical route was gained by orthogonal test,the powder of dry mycelium was washed and degreased with aether for 12h.The residue,collected by centrifugation and dried at room temperature,was extracted (3 times) with distilled water at 70℃ for 2h(the ratio of residue and distilled water was 1∶10 every time),and then centrifuged at 3 000 r.p.m.for 15min.The supernatants were collected and concentrated to 1/6 total vol.Ethanol was added to concentrate liquor.The final concentration of ethanol was 85%.The sample was then precipitated at 4℃ for 12h and centrifuged at 3 000 r.p.m.for 15min again.In the end,the precipitator portion was washed 3 times with 95% ethanol and dried in vacuo.It is better to depigment by marcoporous adsorbent resin column chromatography than H2O2 oxidation.Partial fine PGPS was further isolated and purified by column chromatography on DEAE-Sephadex A-25 and Sephadex G-200,and was obtained four fractions.The polysaccharides content of PGPS was 139%,and at least PGPS included a pure polysaccharide fraction that was proteinbound polysaccharide.