Study On The Isolation,Purification And Biological Activity Of Alcohol-Solube Pigment From Paecilomyces Sinensis Mycelial

Paecilomyces sinensis is a kind of medicinal fungus that contain a variety of active natural ingredients. The objective of this research is to examine the natural pigment produced by P. sinens through liquid fermentation, including the optimal fermentation conditions, the purification process of pigment, pigment structure, stability and biological activity.Flavonoids pigment was identified throughthe hydrochloric acid-magnesium powder reaction of pigment samples. The examination of the neutral lead acetate reaction exhibited no occurance of precipitation or air bubble, which indicated that the pigment samples did not contain the phenol hydroxyl groups. The color was not tanned in the ferric chloride chromogenic reaction, demonstrating the same result.The optimal culture medium for the process was determined through single factor experiment as follows:glucose 2%, peptone 1%, potassium dihydrogen 0.3%,phosphate magnesium sulfate 0.15%. The response surface experiment confirmed the best speed, temperature and best pH set as 160 r. min-1,25℃ and natural pH. After the evaporate spinning of alcohol solution, obtained from dissolution of dried mycelium of P. sines in 80% ethanol, the absorbance value of the solution was 0.397 under 229 nm condition, illustrating that the alcohol soluble pigment content was 3.96 mg/L.The monomer was obtained by AB-8 macroporous adsorption resin, silica gel column separation, purification and HPLC analysis from pigment samples. The examination of infrared spectroscopy, Ms spectrometry and Nuclear Magnetic Resonance （NMR） spectroscopy and comprehensive analysis methods exhibited that the molecular weight of the flavonoid pigments was 425, the formula was C₂₃H₂₇O₇ and it contained flavonoid glycosides key, a methyl substituents and a methoxyl group, which was considered to be the first extraction of this pigment.The stability experiments exhibited that the pigment is stable to a certain concentration of sucrose and citric acid and the change of temperature and light. It is high tolerance to oxidant and reducing agent. However, iron, aluminum and metal ions had great effects on the stability of the pigment. The examination of oxidation tolerance and bacteriostatic experiment displayed that the pigment scaveng free radicals effectively, which had very significant anti-bacterial effect as well as mold bacteriostatic effect.The flavonoid pigments was extracted from the P. sinens mycelium, expanded the sources of flavonoid compounds, while providing people with a new sources of flavonoids, which can alleviate the use of natural resources and protect the natural resources conflicts.