Study On The Bisphenol A Degradation Gene And Enzyme Of Pseudomonas Sp.W2

Nowadays, endocrine disruptor chemicals are endangering human health. Of all EDCs, Bisphenol A is very important one. With Bisphenol A used in great quantity, it is detected in all kinds of waters all over the world. It is very necessary to control of bisphenol A pollution. Some pure cultures of bacterial able to degrade bisphenol A have been reported, but the study about degradation gene and enzyme was not found.We previously identified a Pseudomonas sp. W2 including a plasmid pW2, which rapidly metabolized bisphenol A. pW2 was responsible for degradation of bisphenol A. Based on this, we studied on the bisphenol A degradation gene and enzyme of Pseudomonas sp.W2.1. Construction and analysis of library of plasmid pW2. Plasmid pW2 was isolated and purified. A gene library of plasmid pW2 was constructed and sequenced. By PCR and primer walking, gaps were closed. By the BLAST series of programs provided by the National Center for Biotechnology Information (http://www.ncbi.nih.gov), the information on the Plasmid pW2 was studied.2. Characteristic and founction of the bisphenol A degradation gene. All ORFs without homology sequence and were longer than 450bp were ligated into pET-32a(+) in E. coli BL21(DE3) cells. The clone including gene that encodes bisphenol A degradation enzyme was found. Bisphenol A degradation activity was analysed by HPLC.3. Purification and analysis of bisphenol A degradation enzyme. By ultrasonic and ammonium sulfate salting-out, we got the crude enzyme. Then we purified the crude enzyme by chromatogram.