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Isolation, Characterization and PM Degrading Enzyme Production of Organophosphorus Pesticide Degrading Fungi

Author: JieShunChang
Tutor: CaiHuiNong
School: Jimei University
Course: Microbiology
Keywords: Organophosphorus pesticides Microbial degradation Bronk Candida Aspergillus oryzae Yellow-blue -like bacteria Isolation and identification of Enzyme production characteristics
CLC: X172
Type: Master's thesis
Year: 2011
Downloads: 86
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Abstract


The extensive use of organophosphorus pesticides has caused serious environmental problems, and potential harm to human health. The removal of organophosphorus pesticide residues is particularly important organophosphorus pesticide degrading microorganisms produce enzymes that degrade the elimination of organophosphorus pesticide pollution, safe, and most promising. Degrading bacteria isolated from organophosphorus pesticide-contaminated soil in three organophosphorus pesticides: JMUPMD-1-2 JMUPMD,-3 JMUPMD, the use of morphological observation, physiological and biochemical experiments and rDNA ITS sequence analysis of the three bacteria were identified . Organophosphorus pesticide hydrolase production JMUPMD-3 strains optimized shake flask cultures, 7 L tank fermentation, the fermentation kinetics of strain analysis based on the optimization results. The JMUPMD-1 rDNA ITS sequence homology with the Bronk fake silk yeast (Candida blankii), 99%, and the morphological characteristics and physiological and biochemical characteristics consistent with Bronk Candida, and therefore identification Bronk Candida. JMUPMD-2 rDNA ITS sequence of Aspergillus oryzae (Aspergillus oryzae) 99% homology, colony morphology and morphology are consistent with the characteristics of Aspergillus oryzae, identified as Aspergillus oryzae. The JMUPMD-3 rDNA ITS sequences with the yellow-blue-like bacteria (Talaromyces flavus) 99% homology, the morphological characteristics of the yellow and blue like the bacteria basically in line with, identified as the yellow-blue-like bacteria (asexual generation as by Penicillium vermiculatum Dangeard) the. The three bacteria produce methyl parathion (Parathion-methyl, PM) degrading enzymes are intracellular enzyme, the T. flavus JMUPMD-producing enzyme strongest. Optimized shake flask media and culture conditions: sucrose 19.0 g / L, yeast extract 3.2 g / L, MgSO4 · 7H2O 0.5 g / L, KCl 0.5 g / L of FeSO4 · 7H2O 0.018 g / L, Triton the X- 100 2% (v / v), the methyl parathion emulsifiable 0.2% (v / v), and the fermentation liquid initial pH 8.0, culture temperature 24 ° C. 7 L tank fermentation with an initial medium the T. flavus JMUPMD-3 PM degrading enzyme activity up to 46.1 U / L, using optimized fermentation media and culture conditions, PM degrading enzyme activity of 127.1 U / L, than the first batch of 183%. 7 L tank fermentation on the basis of the second batch, the abolition of Triton X-100 Add and to maintain fermentation process medium pH is 6.0 biomass increased to 3.2 g / L, the highest level of enzyme activity was flat, but the stable phase delay. Kinetic analysis showed that the strains of the logarithmic phase of 24 h? 56 h, PM degrading enzyme synthesis and cell growth phase coupling, PM degradation related to the enzymatic synthesis and PM consumption, while the consumption of PM from enzymes to produce the impact. PM degrading enzymes to dichlorvos the good degradation may belong to the broad-spectrum organophosphorus pesticide degrading enzyme. Separated into three organophosphorus pesticide degrading fungus: of Candida blankii Aspergillus oryzae, Talaromyces flavus, the of Candida blankii and Talaromyces flavus few reports on the degradation of organophosphorus pesticides to increase the resources of the new organophosphorus pesticide degrading fungus. The of T. flavus JMUPMD-3 production PM degrading enzymes were the shake flask Optimization and 7 L tank fermentation first test, and laid the foundation for degrading enzyme activity in order to further improve the strain produced PM.

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CLC: > Environmental science, safety science > The basic theory for the Environment and Science > Environmental Biology > Environmental Microbiology
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