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Screening of CGTase-producing Bacteria Strains and the Application of Cyclodextrins in Agricultural Production

Author: ZhangBin
Tutor: QiQingSheng
School: Shandong University
Course: Fermentation Engineering
Keywords: β-Cyclodextrin Cyclodextrin glucanotransferase Inclusion Chlopyrifos Characterization
CLC: TQ461
Type: Master's thesis
Year: 2009
Downloads: 50
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Abstract


The Cyclodextrin(CD) and its derivatives have been widely applied in medical industry,food industry,chemical industry,environment and many other areas for its unique inclusion performance.However,the application in agricultural production is still in the initial stage.In order to resolve the problem that the efficiency of screening strain is low in the production process of the CD,this study established a new method which could screen glycosyltransferase(CGTase)-producing strain from the soil quickly and accurately.In order to explore the possibility of the application of the CD in agriculture.The inclusion ofβ-CD and its derivatives and pesticide chlorpyrifos were made by many ways,the inclusion solid complex was characterizated,and the insecticide efficacy experiments was carried out.The results these studies are as follows:1.A method that can quickly select the CGTase-producing bacteria was set up. 149 extracellular amylase-producing strains have been screened from the soil The CD composition was detected using the High Performance Anion Exchange Chromatography(HPAEC) and the CGTase-producing bacteria was determined.In the study,11 CGTase-producing bacteria strains were detected and 6 strains mainly produced the CD6 and 5 strains CD7.Meanwhile,the content of the different CD including the large ring(CD＞8) was determined during the process.2.The preparation methods of the inclusion complex ofβ-CD,HP-β-CD, M-β-CD and the pesticide chlorpyifos were studied,respectively.And the best conditions were determined.The inclusion rate of the saturated aqueous solution was the best among the grinding method,ultrasonic method and saturated aqueous solution.The inclusion rate of the saturated aqueous solution was 45.8%,52%and 65%for the inclusion complex ofβ-CD,HP-β-CD,M-β-CD and the pesticide chlorpyifos,respetively.The best optimal reaction conditions of inclusion complex ofβ-CD and chlorpyifos by orthogonal was:feed ratio of 1:1(mole ratio),inclusion time was 3h,inclusion temperature is 60℃.3.The inclusion behavior ofβ-cyclodextrin and its derivatives with chlorpyrifos were quantitatively studied using UV spectrum.The results showed that the ratio of inclusion was 1:1,the stability constants of inclusion complexes were K=33.33mol/L (β-CD)、K=343.8 mol/L(HP-β-CD)和K=461.63 mol/L(Me-β-CD).It showed that the stability ofβ-CD inclusion complex with chlorpyrifos was the worst,HP-β-CD、 Me-β-CD inclusion complex were stable.The important thermodynamic parameters△G were calculated by△G =-RlnK,that were -8.69KJ/mol(β-CD)、-20.184KJ/mol(HP-β-CD)、-15.21KJ/mol(Me-β-CD).It showed that inclusion reactions were spontaneous process,and their inclusion constant decreases with increasing temperature,indicating that inclusion is an exothermic process.4.The liquid inclusion complexes ofβ-CD,HP-β-CD,M-β-CD and chlorpyrifos were characterized by UV spectroscopy,respectively.When concentrations of object chlorpyrifos was fixed,with increasing of the concentration of the main,the entire system of absorption there was no obvious shift,however, the maximum absorption peak at the absorbance gradually increased.It showed that chlorpyrifos have been recognized byβ-CD and its derivatives.5.The solid inclusion complexes ofβ-CD,HP-β-CD,M-β-CD and chlorpyrifos were characterized by infrared absorption spectroscopy and differential thermal analysis,respectively.DSC showed that three inclusion only appeared dehydration peak of main elements,original absorption peaks of Chlorpyrifos disappeared.Infrared absorption spectra certified the characteristic absorption peaks of Chlorpyrifos disappeared in three kinds of inclusion,mainly as the main molecular map.It confirmed thatβ-cyclodextrin,HP-β-CD and M-β-CD and chlorpyrifos have formed a stable 1:1 inclusion complex.6.The results of indoor insecticide toxicity of inclusion showed that:the median lethal concentration LC50 of inclusion complex ofβ-CD,HP-β-CD,M-β-CD and chlorpyrifos were 16.1μg/mL,10.8μg/mL and 12.3μg/mL,respectively.The composition of the CD especially the large ring CD were firstly detected using the HPAEC.A novel method that can detect the CD-producing ability of the CGTase during the different stages was set up.The inclusion of the CD and chlorpyrifos can increase the solubility of the chlorpyrifos and its efficacy.The reseach was the basis to the extension of the inclusion of the CD and pesticides.

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