Optimization Of Fermentation Conditions,Concentration And Characterization Of Pigment From Simplicillium Lanosoniveum

In this paper,the fermentation of pigment of Simplicillium lanosoniveum var.tianjinienss Q.L.Dong（DT06）was determined by single factor experiment and optimized by BBD.The physicochemical properties of the pigment and the structure of the pigment were studied.The main research results are as follows:The single factor experiment was carried out to determine the content of main components of the fermentation medium（carbon source,nitrogen source,K₂HPO₄,Mg SO₄·7H₂O）and the optimum operating parameters（initial pH,fluid volume,speed,additives）.The results showed that when the medium added 18 g/L glucose,0.7 g/L sodium nitrate,0.4 g/L K₂HPO₄and 0.003 g/L Mg SO₄·7H₂O,the yield of pigment was the highest.The results showed that the effects of the composition of the medium on the pigment were almost the same and positively correlated with the pigment concentration in a certain range.For the operating parameters,the suitable conditions were as follows:initial pH6,liquid volume 80 m L,rotation speed 125 r/min,VB（0.03%）,Tween 80（2 g/L）and culture time130h.The fermentation conditions optimized by BBD were glucose 18.36 g/L,Na NO₃ 0.78g/L,Mg SO₄·7H₂O 0.03 g/L,K₂HPO₄ 0.4 g/L,and additives（0.03%VB and 2 g/L Tween-80）.The incubation time was 130 h.The predicted pigment yield was 0.969 g/L,and the optimized pigment yield was 0.971 g/L,which was 1.99 times higher than that before the optimization.The separation and purification conditions of fungus DT06 pigment were studied.The pigment samples were purified and concentrated by silica gel column combined with macroporous resin.The macroporous resin D301 was selected after static preliminary screening.At the same time,the influence of different eluent pH ratio and flow rate on desorption was also investigated.The final adsorption conditions were determined as follows:pH was 5 and the volume of the upper column was 70 m L;Elution conditions for:pH of 8,5%Na OH+70%ethanol as the desorption agent to diameter ratio D/H for a quarter of elution velocity of 0.8 m L/min for the follow-up of the pigment concentration,pigment concentration increased by 2.3 times And the enrichment of purified pigment samples are the physical and chemical properties research,found that the polarity of pigment and close to water,at the same time of pH light temperature additives and metal ions are showed good stability,therefore,the pigment is a anthraquinones pigments industrial application potential.The quantitative method of pigment was established,and the method basis was provided for the content detection of pigment fermentation liquid and subsequent samples.To pigment at the same time has carried on the qualitative analysis,UV-vis analysis had the single absorption peak in the 284 nm,FTIR in 3446,1261 and 1072 cm^-1（OH）,1734 cm^-1（conjugated C=O）,and 1647 and 1456 cm^-1（anthraquinone ring）show the characteristics of anthraquinone material absorption,this is in line with the data structure of anthraquinone and at the same time through qualitative anthraquinone in chemical experiments,the results showed that the pigment with not only the anthraquinone in ultraviolet and infrared spectrum and the results are identical,The qualitative experimental data were also consistent with the anthraquinone substances,so the fungal pigment DT06 was preliminarily judged to be a new anthraquinone pigment.