Research On Constructing Genetic Bacteria For The Biodegradation Of Chlorinated Organic Compounds

The epoxy propane wastewater contains high concentration of chlorinated organic compounds,such as chloropropanol,propylene glycol,dichloroisopropyl ether and chloroacetone etc,which is the rub of the biodegradation treatment.Taking use of epoxy propane wastewater as a starting point,the main research contents include three aspects:the biodegradation mechanism of organic chloride,the experiment for identifying the activity of the key enzyme,the biodegradation study of 1,3-dicloro-2-propanol structural resembling chloropropanol in the field of biology and the cloning and expression research of the key enzyme of the goal gene in the biodegradation pathway by molecular biology,which aims at constructing genetic bacteria for the efficient biodegradation of chlorinated organic compounds.Firstly,it is the study of the biodegradation mechanism of organic chloride.By using bioinformatics in the field of biology,research the biodegradation mechanism of 1,3-dicloro-2-propanol.The relevant information and the sequence information of DNA of halohydrin epoxidase A and halohydrin epoxidase B,which is the key enzymes in the process of the biodegradation of 1,3-Dichloro-2-propanol,are acquired.And the homology analysis of halohydrin epoxidase A and halohydrin epoxidase B is made.The result shows that relevant micro-biotin that can synthesize those key enzymes is widely spread in the nature.And the typical relevant micro-biotin is as follows:CoryneBacterium sp.,Arthrobacter sp.,Burkholderia sp. etc.Secondly,Select Halobacterium sp.J7,E.coli DHSαand bacteriological WH-ZJ,study their adaptability mechanism for propylene oxide,the result shows that the major factor which limits the growth of bacterial is that the bacterial can not degrade the organic matter in epoxypropane waste water.Select Corynebacterium (Corynebacterium sp.) CICC 10189 for the test strains,through the experiment for identifying the activity of the key enzyme,it proves that Corynebacterium CICC 10189 can not efficiently use 1,3-dichloro-2-propanol as a carbon source,but the bacteria have the gene which can synthesize the enzyme(halohydrin epoxidase A or halohydrin epoxidase B) for degradating the 1,3-dichloro-2-propanol.Finally,It is he biodegradation study of 1,3-dicloro-2-propanol structural in the field of biology and the cloning and expression research of the key enzyme of goal gene in the biodegradation pathway by molecular biology.700 bp of the gene fragments from Corynebacterium sp.CICC 10189 was amplified by PCR and ligated into vector PHX303.The recombined plasmid was constructed and transferred into E. coli DH5αat appropriate temperature.Screening four positive bacteria,the double-enzyme digestion and PCR amplification identification results exist contradictory,sequencing and analysis show that the cloned gene sequence has high similarity with the non-potential target genes in Burkholderia reported in GenBank. In the further study of cloning hheA and hheB,the potential aimed gene fragment is not amplified.Untill now the whole trial is end,the paper analyzes the results and made recommendations in the end.