Research On Mutation Breeding Of Microwave On Monascus Purpureus And Liquid Fermentation For Orange Pigments Production At High Level

The use of monascus pigments as food addictive has a long history. In the admixture of monascus pigments, orange pigments are the best fresh pigments. Recently, orange pigment was the major antibiotic substance of monascus pigments, also had health effection such as enhancing immunity, resisting fatigue and antilipemic. So, mutation breeding on Monascus purpureus for orange pigments production at high level, increase of the orange pigments yield and looking for the application scopes has a great significance both of the oretic and praxis.This paper focused on mutation breeding Monascus purpureus for orange pigments production at high level using microwave apparatus with cooling water circulating mutate, liquid medium and cultural conditions were optimized for increase of the orange pigments yield, silica gel column chromatography was used to separate monascus pigments. And then research on bacteriostasis and application of monascus pigments in food. The results as follows:1. Microwave irradiation with the condition of circulating cooling water used to mutate breeding of Monascus pursuers. According the effect of microwave power and irradiation time to fatality and mutation rate of Monascus purpureus defined the best the best mutation conditions were the microwave power of 500W and irradiation length of 70s. The positive mutation rate reached 53%.7 stable Mutants of orange pigments production at high level were found. The color value of orange pigment of optimal mutant W5S7 reached 16.38U/mL 59.62% higher than the parent strain..2. Liquid fermenting mutant W5S7, the single factors of medium composition was optimized. From these results the fermentation process was optimized by response surface method, as follows:The optimized medium composition contained: yeast extract 0.57%, NaNO30.29%, corn flour 6.66%, soymeal 0.5%, ZnSO40.02%, MgSO40.1%, KH2PO4 0.5%. According the orthogonal experiment, the optimal fermentation condition was obtained:100 mL fermentation fluid in 500 mL flasks; rotation speed of 150r/min; fermentation time of 14 d; fermentation fluid initial pH of 4.5. Liquid fermenting in optimized medium on optimal fermentation condition, the color value of orange pigment of optimal Mutant W5S7 reached 17.48U/mL.3. Separation of monascus pigments with 60-80 silica gel column chromatography, defined the mixture of n-hexane and ethylacetate (volume ratio of 5:1) for isolating yellow components, ethylacetate for isolating orange components and the mixture of ethylacetate and methanol (volume ratio of 5:1) for isolating red components. The best sample was 2 mL×0.02g/mL, the best elution agents velocity was 3 mL/min. Separating with silica gel column chromatography, we could know the proportion of each pigment component in monascus pigments:yellow component 4.05%, orange component 3.04%, red component 89.61%.4. Bacteriostatic test of monascus pigments showed: The inhibitory effect of monascus pigments to Bacillus subtilis and E.coli is strong, to yeast is weak,and no inhibitory effect to Staphylococcus albus or mycete. Marinated in 25.6mg/mL monascus pigment solution, cooked marbledmeat could be extended the shelf 2d and spicy cabbage be extended the shelf 4d. Comparing the antibacterial activity of three components separated from pigment, the results showed that the antibacterial activity orange components is strongest in the three components. There was little effect of temperature to bacteria restraint of monascus pigments, but sunlight could continue weaken the bacteriostatic activity of monascus pigments with irradiation time prolonging. Marinated in 4.0mg/mL orange pigment solution, chicken could be extended the shelf 4d.