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EN
Plant-esterase (EC 3.1.1.X) has received much attention because plant esterase and acetylcholinesterase (AChE) share a similar sensitivity towards organophosphorus (OP) pesticides detection with the same inhibition mechanism. To improve the analytical performance, tetraphenyl metal porphyrin, as an indicator was introduced to combine with plant-esterase. The time of reach equilibrium in PBS solution was shortened after adding plant-esterase by assaying the intensify change of the porphyrin spectrum. Meanwhile, intensify of porphyrin spectrum with plant-esterase was increased compared with that of only the porphyrin spectrum in solution. Tetraphenyl metal porphyrin, such as cobalt(II) meso-tetraphenyl porphyrin, is a mixed reversible inhibitor of plant-esterase from kinetic parameters. The combination ratio of plant-esterase and porphyrin is 2:1. On the other hand, the interaction between CoTPPCl and plant-esterase is the strongest among all tested tetraphenyl metal porphyrin. And the mixed system (CoTPPCl-plant-esterase) showed the best sensitivity towards the tested pesticide. All these results indicated that a complex system composed of tetraphenyl metal porphyrin and plant-esterase was fit for detecting pesticides. They make meaningful guidance on the further design of sensing material in monitoring pesticides.
EN
Three-phase partitioning (TPP) was used to purify plant-esterase from wheat flour. Effect of various process parameters has been evaluated and plant-esterase was purified to 11.35-fold by optimized single step TPP system (50%, (w/v) (NH4)2 SO4 saturation, 1:1 (v/v) ratio of crude extract: t-butanol at pH 4).The enzyme was found to be exclusively partitioned in the aqueous phase. Using TPP system, plant-esterase quickly purified to homogeneity with very high purity and activity. On the basis of single factor research, purification process was optimized by using response surface method, established a new type of high effcient purification plant-esterase method. To the best of our knowledge, this is the first report for purification and characterization of plant-esterase by using three phase partitioning (TPP). The results indicated that, TPP is a simple, quick, economical and very attractive process for purification of plant-esterase compared to conventional chromatographic protocols.
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