液态发酵法生产云芝胞内糖肽

Production of Coriolus versicolor Polysaccharopeptide by Submerged Fermentation

通过筛选比较 6株云芝菌种 ,获得液态发酵生产云芝糖肽较佳菌株CV S和CV H ,经优化摇瓶发酵 ,将菌株CV S放大至 3 0L罐发酵 ,2 8℃发酵 4d ,菌丝体干重达 2 4 9g/L ,菌丝体破碎后 ,超滤浓缩 ,醇沉干燥后 ,云芝胞内糖肽产率达 2 2 3 g/L ,糖肽总糖为 5 5 5 8% ,肽 3 3 5 3 % ,经HPLC检测其分子质量高于 1 0ku的有 2个组分 ,分别为 742 2和 2 5 2ku ,所提取的产品 ,其出峰时间与标准品基本一致 ,总糖、单糖、肽含量符合国家药品标准WS—XG 0 2 1 2 0 0 2的要求。

By comparison of six different Coriolus versicolor strains, strain CV-H was selected for polysaccharopeptide production by submerged fermentation. After optimization of shake-flask culture, polysaccharopeptide production was scale-up to 30 liter fermentor. Maximal dry mycelium weight was 24.9?g/L after cultivation for 4 days at 28℃. Intracellular polysaccharopeptide was extracted by crashing mycelium, ultra-filtration concentration, ethanol precipitation and dehydration. The yield of polysaccharopeptide was 2.23?g/L, total sugar content was 55.58%, and peptide content was 33.53%. High Performance Liquid Chromatography(HPLC, Ultrahudrgel TM Linear 7.8×300?mm columns) was used in the fraction and molecular weight(M W) estimation of polysaccharopeptide. Two components with molecular weight more than 10?ku were found and their molecular weights are 742.2?ku and 25.2?ku respectively. HPLC retention time of extracted polysaccharopeptide was consistent with that of standard. Total sugar content, reducing sugar content and peptide content of extracted polysaccharopeptide were in accordance with China medicine standard WX-XG-021-2002.