| Lactarius hatsudake Tanaka belong to Eumycota, Basidiomycotina, Hymenomycetes,Agaicales, Russulaceae, Lactarius. Lactarius hatsudake Tanaka is mycorrhizal fungi ofChinese pine, Mason pine etc, and also a precious of wild edible mushroom. Lactariushatsudake Tanaka doesn’t only has rich nutritious, and has antimicrobial activity to a varietyof bacteria and fungi, is an important resource of the study of natural antimicrobial activesubstances. In this paper we use E. coli as the indicator bacteria, inhibition zone diameter offermentation broth of E. coli as index, using quadratic general rotational combination designmethod, to optimize the Lactarius hatsudake Tanaka fermentation substrate and environmentalconditions. and on these basis we have preliminary studed the antimicrobial activity andcomposition of Lactarius hatsudake Tanaka fermentation liquor, with a view to provide somereferences to the development of fermentation liquid of Lactarius hatsudake Tanaka. Themain results are as follows:1. Screened dominant strains SH-3.2. Through the establishment of mathematical model, the optimizing results offermentation substrate of Lactarius hatsudake Tanaka ware: Glucose was48.30g/L, maltosewas45.29g/L, peptone was4.69g/L, yeast extract was4.12g/L, KH2PO41was g/L, MgSO4was0.5g/L, the theoretical inhibition zone diameter was20.1mm. The optimizing results ofenvironment of Lactarius hatsudake Tanaka ware: Rotate speed was145r/min, initial pHwas7.0, liquid volume was108mL, temperature was25℃, the theoretical inhibition zonediameter was27.8mm.3. Lactarius hatsudake Tanaka fermentation broth had good thermal stability; Theantimicrobial activity of Lactarius hatsudake Tanaka fermentation broth staied stable whenthe pH was5.0~7.0; Lactarius hatsudake Tanaka fermentation broth was not sensitive to UVirradiation; It could be placed up to130days at25℃, and the antimicrobial activity weren’taffected. Low concentrations of H2O2and Na2SO3had less impact on the antibacterial activityof fermentation broth, high concentrations of H2O2and Na2SO3had much impact on theantibacterial activity of fermentation broth.4. Lactarius hatsudake Tanaka fermentation broth had the most strong inhibition effectof Bacillus cereus (F2), Bacillus subtilis (E3), Salmonella Lignieres (S1). Fermentation broth had strong inhibition effect of Escherichia coli (E), Pasteurella, Bacillus licheniformis (F1),Salmonella Lignieres (S2). Fermentation broth had weak inhibition effect of Aspergillusflavus, Rhizopus stoloifer, Salmonella Lignieres (S8), Shiyang pig aureus. Fermentation brothhad the weakest inhibition effect of Aspergillus oryzae, Mucor, Trichoderma viride,Penicillium citrinum.5. The test of polarity of Lactarius hatsudake Tanaka shows that the active ingredient isfat-soluble substances, the antimicrobial effect was better than the bacterial filtrate solution.Petroleum ether extracts had the most strong inhibition effect of Salmonella Lignieres (S1),and then Bacillus licheniformis (F1). Ethyl acetate extracts had the most strong inhibitioneffect of Pasteurella and Escherichia coli (E), then Salmonella Lignieres (S1). N-butanolextract had strong inhibition effect of Salmonella Lignieres (S8), ether extract inhibited thepathogen were weak.6. Fermentation liquid of Lactarius hatsudake Tanaka was separated by silica gelcolumn chromatography, finally,2antimicrobial activity components ware received (S4andS12), After TLC, the results of S4and S12showed that they had a larger conjugated system(Benzene ring, a conjugated double bond, etc.). In the test we didn’t get monomer compoundwith antimicrobial activity. |
