Studies On Extraction, Properties,Identifications And Optimization Of Fermentation Conditions Of The Pigment From(Leccinum Extremiorientale(L. Vass.)Singer)

There was a world interest in process development for the production of safe pigment from natural sources. It was desirable to produce natural pigment from microorganisms for their high productivity. Leccinum extremiorientale(L.Vass.)Singer was a kind of being domesticated wild edible and medicinal fungus. It was the firet time to study on extraction, properties, identifications and optimization of culture conditions of the pigment from Leccinum extremiorientale.In the experiment, four means of extraction were compared in extracting effect, and six means of solvent were determined to be the optimal, The results showed that the best extraction method was ultrasonic assist, the best solvent was ethanol, the maximum wavelength of the pigment was 410nm, Based on single-factor studies, orthogonal design was used to optimize the extraction technology of Leccinum extremiorientale pigments in mycelia by ultrasonic assist, the most important factor was the liquid-solid ratio, the optimum conditions were determined as:90% ethanol, liquid-solid ratio 75mL/g, power 400W, temperature 60℃, ultrasonic time 50 minutes.GC-MS analysis showed that the moity of pigment of Leccinum extremiorientale were steroids such as ergosterol, the chromogenic compound was Ergosterol-4,6,8(14),22-tetraen-3-one, that meant the pigment is a compound with good value of physiological and remedium activity.The pigment was stable to alkaline environment and low temperature(<60℃), it was relatively stable to most metal ions, it was not sensitive to outdoor light, indoor light or UV-ray and oxidizer. The low concertration of reducing agent and preservation was advantage to preserve the pigment.The fermentation of pigment production with Leccinum extremiorientale in shake flask was studied.The optimal carbon source and nitrogen source for pigment production with Leccinum extremiorientale were obtained by monofactorial experiment, which were maltose and corn steep respectively,the optimal temperature was 27℃,the optimal culture time was 144h,the optimal liquid volume was 80mL/250mL;the fermentation time and liquid volume were found to have major effects on pigment production through Plackett-Burman experimental design; the steepest ascent experiment and response surface analysis were used to estimate optimum fermentation conditions:maltose 20(g/L),corn steep 5(g/L),KH2PO4 3(g/L),MgSO4·7H2O 1.5(g/L),VB1 0.01(g/L),temperature 27℃,culture time 141.85h,liquid volume 87.70mL/250mL,and the maximum content of pigment under optimum conditions was improved 11.37-fold compared to the original conditions.