The immobilization of acetylcholinesterase （ACHE） on amino functionalized SBA-15 mesoporous sieves （NH2-SBA-15） was explored for detection of organophosphorus and carhamate pesticides. Several methods have been st...The immobilization of acetylcholinesterase （ACHE） on amino functionalized SBA-15 mesoporous sieves （NH2-SBA-15） was explored for detection of organophosphorus and carhamate pesticides. Several methods have been studied and the ＂adsorption-crosslinking＂ is found to be the best one. Firstly, AChE and bovine serum albumin （BSA） were adsorbed in turn on the surface of NH2-SBA-15 by electrostatic adsorption, and then glutaraldehyde was added for cross-linking the amino group of AChE. The NH2-SBA- 15 showed around 95% immobilization efficiency and the specific activity of immobilized AChE （ACHE- NH2-SBA-15） reached 130% relative to free AChE under optimal immobilization conditions. It was found AChE-NH2-SBA-15 retained 94.0% and 82.8% of its initial activity after 60days at 4℃ and 25℃, respectively. It remained 55.0% of its initial activity after 6 times recycling. During the detection of pesticide, the AChE-NH2-SBA-15 showed wider linear range and much lower limit of detection compared with free ACHE. The current method showed good recovery （93.8%-109.3%） and low RSD （〈5%） for both standard solution and real vegetable samples of Carbaryl and Trichlorfon. It was believed that AChE-NH2- SBA- 15 could be exploited as a fast, sensitive and low-cost biocatalyst towards the detection of pesticides residues which could be stored at room temperature for a long time.展开更多
基金supported by the National Natural Science Foundation of China (No.21303105)Shanghai Committee of Science and Technology (No.13430503400)
文摘The immobilization of acetylcholinesterase （ACHE） on amino functionalized SBA-15 mesoporous sieves （NH2-SBA-15） was explored for detection of organophosphorus and carhamate pesticides. Several methods have been studied and the ＂adsorption-crosslinking＂ is found to be the best one. Firstly, AChE and bovine serum albumin （BSA） were adsorbed in turn on the surface of NH2-SBA-15 by electrostatic adsorption, and then glutaraldehyde was added for cross-linking the amino group of AChE. The NH2-SBA- 15 showed around 95% immobilization efficiency and the specific activity of immobilized AChE （ACHE- NH2-SBA-15） reached 130% relative to free AChE under optimal immobilization conditions. It was found AChE-NH2-SBA-15 retained 94.0% and 82.8% of its initial activity after 60days at 4℃ and 25℃, respectively. It remained 55.0% of its initial activity after 6 times recycling. During the detection of pesticide, the AChE-NH2-SBA-15 showed wider linear range and much lower limit of detection compared with free ACHE. The current method showed good recovery （93.8%-109.3%） and low RSD （〈5%） for both standard solution and real vegetable samples of Carbaryl and Trichlorfon. It was believed that AChE-NH2- SBA- 15 could be exploited as a fast, sensitive and low-cost biocatalyst towards the detection of pesticides residues which could be stored at room temperature for a long time.
