| In this study, the method of screening microorganisms for the biotransformation of target compounds from medicinal plants was established, which was efficient and economic. The immobilized conditions were optimized. The fermentation conditions of biotransformation with immobilized microorganisms were optimized. The antioxidant activities of medicinal plants before and after biotransformation were determined. The immobilized microbial biotransformation reactor was applied in medicinal plants.1. The method of screening microorganisms for biotransformation of target compounds from medicinal plants was established.In this study, the potential edible strains were screened from kinds of microorganisms and evaluated by the biotransformation abilities with the content of resveratrol, genistein, genistein, arctigenin as an index, respectively. The microorganisms with biotransformation abilities were screened for the four medicinal plants including Polygonum cuspidatum, Pigeon pea root, Fructus Sophorae, Actium lappa by the first time.The best microorganisms with biotransformation Polygonum cuspidatum were Yeast CICC1912and Aspergillus niger M85. After biotransformation, the concent of resveratrol was increased to14.57mg/g compared to the untreated sample (3.04mg/g).The best microorganisms with biotransformation Pigeon pea root were Monascus anka3.782and Aspergillus oryzae3.951. After biotransformation, the concent of genistein was increased to1.24mg/g compared to the untreated sample (0.76mg/g).The best microorganisms with biotransformation Fructus Sophorae were Aspergillus oryzae Y29and Yeast JB. After biotransformation, the concent of genistein was increased to32.9mg/g compared to the untreated sample (27.8mg/g).The best microorganisms with biotransformation Actium lappa were Aspergillus niger M85and Monascus anka3.554. After biotransformation, the concent of arctigenin was increased to38mg/g compared to the untreated sample (18mg/g).2. The immobilized conditions were optimized.The sodium alginate embedding method of immobilized edible microorganisms was applied in biotransformation of four kinds of medicinal plants. The immobilized conditions were optimized and applied in the production of the target compounds. The effects of microbial concentration, concentration of sodium alginate, calcium chloride concentration, diameter of the immobilized microorganisms on immobilized conditions were evaluated. It was shown that the immobilized microorganisms can be used for more than15times from the stability experiments. In order to obtain the maximum yield production target compounds, the conditions of immobilized microorganisms were optimized. The above results would be provide a set of efficient, fast, low cost preparation technology for the immobilized biotransformation industry.The optimal immobilized conditions were shown as follows.The biotranformation conditions of Polygonum cuspidatum with immobilized Aspergillus niger and Yeast were that the concentration of sodium alginate5%, calcium chloride concentration2.5%, concentration of bacteria7.2log cfu/mL, immobilized microorganisms diameter4mm, the enzyme activity of immobilized microorganisms1.40U/g;The biotranformation conditions of Pigeon pea root with immobilized Aspergillus niger and Monacus anka conditions were that the concentration of sodium alginate6%, calcium chloride concentration3%, concentration of bacteria7.1log cfu/mL, immobilized microorganisms diameter4mm, the enzyme activity of immobilized microorganisms1.03U/g;The biotranformation conditions of Fructus Sophorae with immobilized Aspergillus oryzae and Yeast conditions were that the concentration of sodium alginate6%, calcium chloride concentration3%, concentration of bacteria7.1log cfu/mL, immobilized microorganisms diameter5mm, the enzyme activity of immobilized microorganisms0.94U/g;The biotranformation conditions of Actium lappa with immobilized Aspergillus niger and Monacus anka conditions were that the concentration of sodium alginate7%, calcium chloride concentration2.5%, concentration of bacteria7.4log cfu/mL, immobilized microorganisms diameter5mm, the enzyme activity of immobilized microorganisms1.31U/g.3. The fermentation conditions of biotransformation with immobilized microorganisms were optimized.In this study, the innovation method was found that the immobilized microorganisms technology was combined with biotransformation technology. The fermentation conditions of biotransformation for four medicinal plants with immobilized microorganisms were optimized. In order to evaluate the effect of fermentation conditions on the target compounds in medicinal plants yield, the fermentation process was investigated. After that, the response surface analysis was applied to obtain the optimum fermentation conditions.The biotranformation conditions of Polygonum cuspidatum with immobilized Aspergillus niger and Yeast were that pH6.5, temperture30℃, time2d, radio of solid to liquid1:12g/mL. Under this condition, the yield of resveratrol was33.45mg/g, which was11-fold to that of untreated sample (3.04mg/g).The biotranformation conditions of Pigeon pea root with immobilized Aspergillus niger and Monacus anka conditions were that pH6.2, temperture30℃, time2d, radio of solid to liquid1:12g/mL. Under this condition, the yield of genistein was1.87mg/g, which was2.65-fold to that of untreated sample (0.71mg/g). The biotranformation conditions of Fructus Sophorae with immobilized Aspergillus oryzae and Yeast conditions were that pH5, radio of liquid to solid25:1mL/g, time1d. Under this condition, the yield of genistein was18.95mg/g, which was34-fold to that of untreated sample (0.55mg/g).The biotranformation conditions of Actium lappa with immobilized Aspergillus niger and Monacus anka conditions were that pH6.5, temperture32℃, time2d, radio of liquid to solid15:1mL/g. Under this condition, the yield of arctigenin was41.01mg/g, which was5.97-fold to that of untreated sample (6.87mg/g).The pictures of four kinds of medicinal plants were detected by scanning electron microscopy (SEM). It showed that the surface structures of the fermented samples were destroyed by the immobilized microorganisms. It was proved that the medicinal plants could be transformed by immobilized microorganisms from the part of plant tissue morphology.4. The antioxidant activities of medicinal plants before and after biotransformation were determined.In this study, four plants including Polygonum cuspidatum, Pigeon pea root, Fructus Sophorae, Actium lappa were transformed by immobilized microorganisms. The antioxidant activities of their crude extracts were evaluated after biotransformation.The DPPH-radical scavenging test and β-Carotene-linoleic acid bleaching test were used to evaluate the antioxidant capacity of medicinal plants.The antioxidant capacity results were detected by DPPH radical scavenging test and β-Carotene-linoleic acid bleaching test. It showed that the fermentation sample of Polygonum cuspidatum with immobilized Yeast and Aspergillus niger had better antioxidant capacity than the untreated sample. Before fermentation the IC5o value concentration were0.10±0.14,0.43±0.35(mg/mL) and the IC50value concentration were0.074±0.27,0.19±0.27(mg/mL) after fermentation, respectively.The antioxidant capacity results were detected by DPPH radical scavenging test and and β-Carotene-linoleic acid bleaching test. It showed that the fermentation sample of Pigeon pea root with immobilized Monacus anka and Aspergillus niger had better antioxidant capacity than the untreated sample. Before fermentation the IC50value concentration and the clearance of super oxygen free radical were1.117±0.04(mg/mL),0.216±0.16(mg/mL) to the results of0.737±0.06(mg/mL),0.173±0.02(mg/mL) after fermentation, respectively.The antioxidant capacity results were detected by DPPH radical scavenging test and β-Carotene-linoleic acid bleaching test. It showed that the fermentation sample of Fructus Sophorae with immobilized Aspergillus oryzae and Yeast had better antioxidant capacity than the untreated sample. Before fermentation the IC50value concentration were0.34±0.08,0.41±0.24(mg/mL) and the IC50value concentration were0.31±0.13,0.35±0.08(mg/mL) after fermentation, respectively.The antioxidant capacity results were detected by DPPH radical scavenging test and β-Carotene-linoleic acid bleaching test. It showed that the fermentation sample of Actium lappa with immobilized Monacus anka and Aspergillus niger had better antioxidant capacity than the untreated sample. Before fermentation the IC50value concentration were0.43±0.16,0.64±0.33(mg/mL) and the IC50value concentration were0.27±0.05,0.43±0.24(mg/mL) after fermentation, respectively.Compared with the untreated medicinal plants, the fermentation samples with immobilized microorganisms showed better antioxidant activities.5. The immobilized biotransformation of medicinal plants were verified by pilot experiments.The immobilized microbial biotransformation reactor was applied in medicinal plants.According to the results of pilot experiments, the contents of target compounds were shown as follow.The content of resveratrol from Polygonum cuspidatum was32.78±0.34mg/g, which was10.78-fold to that of untreated sample. The antioxidant capacity results were detected by DPPH radical scavenging test and β-Carotene-linoleic acid bleaching test. It showed that the fermentation sample had better antioxidant capacity than the untreated sample.The content of genistein from Pigeon pea root was1.71±0.10mg/g, which was2.41-fold to that of untreated sample. The antioxidant capacity results were detected by DPPH radical scavenging test and (3-Carotene-linoleic acid bleaching test. It showed that the fermentation sample had better antioxidant capacity than the untreated sample.The content of arctigenin from Fructus Sophorae was17.84±0.89mg/g, which was32.44-fold to that of untreated sample. The antioxidant capacity results were detected by DPPH radical scavenging test and (3-Carotene-linoleic acid bleaching test. It showed that the fermentation sample had better antioxidant capacity than the untreated sample.The content of genistein from Actium lappa was40.19±0.37mg/g, which was5.85-fold to that of untreated sample. The antioxidant capacity results were detected by DPPH radical scavenging test and β-Carotene-linoleic acid bleaching test. It showed that the fermentation sample had better antioxidant capacity than the untreated sample.It was achieved that the continual charging and the continual production through the pilot experiment device with immobilized microorganisms. It was provided data support and a new the way for telarge-scale industrial production. |
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