Study on Extraction of Biological Polyphenols from Bark Based on Enzyme Catalysis Technology

Author:Ma Zuo Yue

Supervisor:gu ji you shi junyou


Degree Year:2019





Due to the presence of large amounts of lumber and pulp industry processing residues,the concept of green polymer has been widely attentioned,more and more researchers focus on the development of new polymer based on bark extract.This paper used the enzyme,alkali and UV/H2O2 process fir and poplar bark extract polyphenols extract comparison,using chromatography determination of bark extract some polyphenol compounds in chemical composition,to observe the effects of different extraction method on extraction yield.The research and effect of enzyme,alkali and UV/H2O2 treatment on the yield and content of extracts were studied.The UV/H2O2 process showed the highest yield,to extract as much as 54%of the dry bark weight,found that compared with enzyme and alkali treatment,the amount of lignin/tannin,enzyme treatment is favorable for heald fiber recycling.By studying the enzymatic reactions of different lignin degradation enzymes,the potential of lignin to effectively degrade lignin in green process can be shown.Three different fungal strains were selected from ten different lignin production strains.In submerged fermentation after 10 days,in the partial separation of fungal strains enzyme was observed in the largest production,research on the separation of the fungus strains producing lignin enzymes,mainly manganese peroxidase(pod),and the enzymatic oxidation of various usually MnP substrates,including lignin phenol.In addition,the degradation products of MnP were determined by the rapid protein liquid chromatography,and the molecular weight change in the degradation process was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis.Choice is studied through 18 s ribosomal DNA sequence analysis used in production of lignin enzyme fungus species,choose two primer set to the main part of the amplification of 18 s rDNA,the results show that the method based on 18 s rDNA is unknown for the identification of lignin enzyme production potential fungus species,the separation of fungal species mainly produces manganese peroxidase(MnP),it will be all kinds of typical oxide substrate and lignin of MnP polyphenols.The temperature and pH of the medium were optimized to maximize the production of manganese dioxide.The time series study showed that,under optimal conditions,the strain culture was developed 10 days later,and the maximum yield of the lignin decomposition enzyme MnP(64 IUL-1),LiP(26.4 IUL-1)and lactase(5.44 IUL’’)was achieved respectively.In addition,the biological decolorization rate of reactive brilliant blue(RBBR)dyes was 94 percent after 10 days of cultivation.Eight different strains of fungi were tested during the fermentation period for the production of lacquer enzymes.Compared with other fungal strains studied,the lateral ear species were found to be good producers.Klebsiella pneumoniae mainly in the initial pH 5.5 and 30℃ in the production of medium under the condition of laccase production.The bark was depolymerized in a petri dish,and after four weeks the polyphenol/polyaromatic compound was identified,which was induced by every bark in the late stage with 50mg/100ml.During the training period,honey was used as a substitute for natural medium in the medium,and the enzyme activity was about 1.5 times higher than that in the culture medium without honey.These samples were analyzed by GC-MS/MS.The purified lacquer enzyme was purified by anion exchange and 10%ABTS were used as substrate.The maximum velocity(Vmax),mie’s constant(Km)and turnover number(Kcat)were 76.9 mu Mmin-1,909 mu M and 739min-1 respectively.In addition,the lacquer enzyme is used to transform the bark life into high value bark biopolymer/bio-chemical application.The py-gc-ms analysis of bark showed the different effects of fungal activity on the bark composition.The differences observed in GC-MS spectra of aromatic compounds are due to the use of two different species of bark.Using high performance liquid chromatography(HPLC)method to inverse model separation of polyphenols,in a petri dish is studied in the bark of single flower honey and impregnated with honey materials instead of natural mediators,and studied the source of fungus laccase influence on bark sources of polyphenols.