| This dissertation was divided into four chapters.In Chapter 1,the medicinal history of Nelumbinis Rhizoma(NR)was reviewed first.The whole and rhizome node of NR were the traditional medicinal parts.The fresh or dried of NR were all used up to now,which has the effects of antipyretic,help to digest,antidiarrheal,relieving restlessness,help produce saliva and slake thirst and so on.The juice of fresh NR can also release crab poison.Secondly,the research progress of chemical composition and pharmacological activity of Nelumbo were summarized.The alkaloids,flavonoids and terpenoids were rich in this genus.In addition,carbohydrates,proteins,and inorganic elements were also important components.The Nelumbo medicinal plants had pharmacological activity as follows:anti-tumor,anti-oxidation,anti-inflammatory,anti-aging,anti-HIV,liver protection and so on.Finally,a brief overview of the research ideas and methods of waste utilization in the process of NR harvesting was obtained.Response surface curve method was used to optimize process of extraction and utilization of potential resources.The research and application of microbial transformation was also summarized.It may provide reference for the effective and comprehensive utilization of NR resources.In Chapter 2,the resource components including betulinic acid,polysaccharides,oligosaccharides,monosaccharides,proteins,nucleoside amino acids and inorganic elements in different parts of NR collected from different origin were analyzed and evaluated.The results showed that the resource components of NR in different parts of NR were different.The content of betulinic acid in the node of NR was up to 0.757 ± 0.034%.The content was higher in the brittle NR than in the powdery NR.The content of polysaccharide was 5.110%?10.347%.Five kinds of monosaccharides and oligosaccharides were also determined:fructose,glucose,sucrose,raffinose and stachyose.The sucrose content was the highest,with 2.261%to 16.223%.The fructose content was 0.777%?2.985%,the glucose content was 0.589%?2.912%.The raffinose and stachyose content were 0.360%?1.700%,0.355%?2.432%respectively.The higher content of nucleosides and amino acids were ?-aminobutyric acid,L-alpha-alanine,L-threonine,L-glutamic acid and asparagine.The protein content in NR was 1.012%?4.921%.The contents of P,Na,K,Zn,Mg,Ca,Na and Al in the samples were higher than other elements.The skin of NR had the highest content of elements.Different drying methods also influence the contents of resource components.The freeze-dried method can preserve resource components better than others.Through the analysis and evaluation of the NR resource components,comprehensive development and utilization of NR could be accessed.In Chapter 3,Response surface curve method was used to optimize the extraction process of polysaccharide and betulinic acid in the rhizome node of NR.Betulinic acid was extracted by ethanol and determined by a HPLC method.The polysaccharide was by water and the phenol sulfuric acid method.On the basis of single factor experiments,ethanol concentration,extraction time,the ratio of material and solvent were selected to optimize the extract process of betulinic acid.Extraction temperature,extraction time and the ratio of material and solvent were selected to optimize the extract process of polysaccharide.When the concentration of ethanol was 100%,material liquid ratio was 1:49,the extraction time was 101.64 min,the extraction yield of BA could reach to 0.665%.When extracting temperature was 95?,the ratio of material and solvent was 1:50,extraction time was 125 min,the extraction yield of polysaccharide was 15.64%.The response surface model established was significantly(P<0.05).It could be applied in the analysis and optimize the extraction process of polysaccharides and betulinic acid in the rhizome node of NR.In Chapter 4,betulinic acid from NR was introduce sugar groups at specific positions by biotransformation.The fermentation products were isolated and purified.Their structural characterization and antitumor activities were also investigated,which may improve utilization of NR.With the transformation ability of strains screened.One kind of strain,identified as Bacillus amyloliquefaciens FJ18,was used in the biotransformation.The transformation products were analyzed using a UPLC-QTOF-MS method firstly.Then,the fermentation system was isolated and purified by silica gel column chromatography repeatedly.The product structure characteristics were analyzed by NMR and MS.Cell viability was evaluated by the methyl thiazolyl tetrazolium(MTT)assay.A novel glycosylation derivative of betulinic acid(BA)derivative,3?-hydroxy-lup-20(29)-en-28-oic acid 28-O-(4-oxy-4-oxobutanoic acid-4-O-?-D-glucopyranosyl)ester,together with a known derivative,28-O-?-D-glucopyranosyl-3?-hydroxy-lup-20(29)-en-28-oate were isolated from the fermentation system of BA.The biotransformation products showed in vitro cytotoxic activity against A375(melanoma),HeLa(cervical carcinoma),U251(glioma),SH-SY5Y(neuroblastoma)and MCF-7(breast cancer)cell lines.The in vitro cytotoxicity study revealed that the derivatives exhibit potent cytotoxic activity against cancer cell lines.Through the above research,the experience data was provided to further utilization of NR.It may helpful to promote green economic development of NR industry and generating social-economic-ecological benefits. |
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