Study On The Synthesis Of O-Benzoquinone And Catechol Compounds By Enzymatic Catalysis

With rapid development of economic and bad charge of environment,people are paying much attention to protecting environment.The application of o-benzoquinone and catechol compounds is very extensive,such as medicine,environment,chemicals,sensors,materials,especially in polymer polymerization,grafting,modification and functionalization,used as a anti-biofouling coating for the hull.However people care much about the pollution from the production and application process.This kind of problem was mainly solved by chemical method in the field of industry.In recent years,tyrosinase has been found to have dual catalytic activities for the synthesis of monophenols and catechols.It can not only directly compose o-benzoquinone compounds but also catechol compounds.Therefore,exploring a large number of efficient tyrosinases to compose o-benzoquinone and catechol compounds by catalytic method has broad prospects,the hotspots of research field at present.Based on the tyrosinase information collected in NCBI database,this subject selects the tyrosinase gene of Dadun Armillaria,using gene project technology to structure it in prokaryotic expression vector,then leading it into E.coli pGro7/BL21(DE3)to finish expression.As a result,Ao tyr with high purity was purified by Ni-metal affinity chromatography.After comparing the effects of different catalytic systems and conditions on the efficiency of enzymatic reaction,products of different phenolic compounds catalyzed by Ao tyr were proved by high performance liquid chromatography and quadrupole flight mass spectrometry,so the feasibility of this reaction in the treatment of phenolic wastewater is verified.At last,the subject researches the Ao tyr can catalyze p-TBC(synthetic pair ofp-TBP)under the circumstance of ascorbic acid existing,the same characterization.So the feasibility of application of the method in catechin compounds is verified.The research concludes:Prepared Ao tyr has catalytic activity,in the aqueous micelle system with 5mmol/L SDS,under the condition of 25℃ pH 5.0,the catalytic effectivity of Ao tyr without pre-treatment is the highest;Characterization result shows phenol,2-F-4-methylphenol,2,4-dimethylphenol,3,4-dimethylphenol and 2,3,5-trimethylphenol can be catalyzed into corresponding o-benzoquinone;1 mmol/L ascorbic acid can inhibit the formation of o-benzoquinone and improve the yield of p-TBC.