Study On Immunoregulational Ingredients ISP Production By Cultivation Of Medicinal Mushroom Cordyceps Cicadae

Cordyceps cicadae, a caterpillar-shaped Chinese traditional medicinal mushroom, is one of the entomopathogenic fungi, which belongs to the class Ascomycetes and DongChongXiaCao group in Chinese herbs. Famous for its distinct pharmacological efficacy and commercial value, it has a lot of effective components and ISP is the most important active immunoregulational component, with the main ingredient ISP-1. The production of ISP by cultivation of medicinal mushroom C. cicadae has great significance. The separation and purification technique of ISP are established in this paper. Meanwhile, a wide and effective analysis in determining content of ISP in C. cicadae by ninhydrin colorimetric method was established. Accoording to the optimizision of carbon sources and complex nitrogen sources, the cell growth and the accumulation of ISP was studied and the ISP production had remarkably increaced.(1) The separation and purification technique of ISP with macroporous resin was researched and the influence of ethanol consistence over the effect of the separation and purification was studied. Meanwhile, the reliability was validated according to the result of HPLC with solid phase extraction (SPE) and MS. Then the separation and purification technique of cordycepin are established: after the adsorption balance, to elute with 4BV water solution and 4BV 50% ethanol by 2BV/h, then desorb with 4BV 95% ethanol by 1BV/h and collect the 95% ethanol solution at 4BV volume.concentrate to dry under decompression at 60℃and then dissolve with methanol.(2) The content of ISP-like ingredients was determined by ninhydrin colorimetric method. Reaction conditions of this method were optimized, and the validity and veracity of method were evaluated through some indexes such as linearity and range, precision, stability, and accuracy. Meanwhile, the reliability was validated according to the result of TLC. The above experimental results reveal that the method is convenient, rapid, accurate, and high sensitive, thereby leading to a wide and effective analysis in determining content of the ISP-like immunoregulational ingredient in C. cicadae.(3) Effects of various carbon sources and complex nitrogen sources were investigated to optimize ISP production by C. cicadae in shake flasks. The cell growth reached the maximum at the 8th day while the ISP production reached the maximum at the 12th day. And then sucrose and yeast were found to be the best for ISP production. Maximum ISP production of 32.25mg/l was achieved which increased by 83.56% compared with that in control medium.