| dc.contributor.advisor | Warsiki, Endang | |
| dc.contributor.advisor | Rahayuningsih, Mulyorini | |
| dc.contributor.author | Maulidin, Ilham | |
| dc.date.accessioned | 2020-08-24T05:37:49Z | |
| dc.date.available | 2020-08-24T05:37:49Z | |
| dc.date.issued | 2020 | |
| dc.identifier.uri | http://repository.ipb.ac.id/handle/123456789/103567 | |
| dc.description.abstract | Paraoxon (diethyl (4-nitrophenyl) phosphate) was chosen as a model compound of organophosphate insecticide because of their high toxicity level toward human health, animals and environment. Phosphotriesterase (PTE) as one of the potential organophosphorus hydrolase enzymes has a special ability in degrading the paraoxon in various concentration. This research aims to produce the PTE derived from Sphingobium fuliginis and evaluate its ability of PTE in hydrolyzing the paraoxon. The preparation of microbial PTE production was successfully obtained by cultivating the inoculum of Sphingobium fuliginis in LB-medium. The purified PTE was obtained by the purification process using DEAE-Toyopearl Column equilibrated by 20mM of PPB (pH=7.2). PTE enzyme assay was observed by spectrophotometric UV-vis with measured absorbance is 410nm and it was examined on two different types of the buffer which is NaHCO3 buffer 100mM (pH=8.5) and Glycine-NaOH buffer 50mM (pH=9.5). The expression of PTE was confirmed by SDS-PAGE analysis. The measurement for the time-course of pH decreased in the presence and the absence of PTE was evaluated by using the pH-ISFET sensor. The best result showed that LB-medium was chosen as the most suitable medium for Sphingobium fuliginis cultivation due to its perfect nutrients content. The molecular mass of PTE subunits protein was well-expressed in 35kDa. Moreover, the amount of paraoxon concentration and the PTE’s performance such as PTE’s specific activity, productivity, sensitivity and the higher amount of total protein of PTE (mg) was performed well in 100mM NaHCO3 buffer pH 8.5. PTE’s Km value obtained on 8.106 µM and its maximum rate velocity (Vmax) showed in 2.497 unit paraoxon mg-1 protein. Lastly, the time course of pH decreased dramatically when the PTE applied to the system. It showed a rapid detection for less than 2-5 seconds the hydrolyzation of paraoxon has been already finished. So that, PTE has been confirmed able to recognize the paraoxon in various concentration in rapid detection. | id |
| dc.description.sponsorship | The author would like to express his sincere appreciation to Dr. Endang Warsiki, STP MSi and Dr. Ir Mulyorini Rahayuningsih, MSi for the support and patience in supervising the author to conduct the research. The honorable appreciation also go to Prof Suprihatin, as the examiner lecturer on the final assignment of examination for his valuable advice and correction so that this document can be improved. The author also would like to thank Professor Wakako Tsugawa for welcoming me to the Tokyo University of Agriculture and Technology, Koganeishi Tokyo, Japan, as a co-supervisor and it is my great appreciation for all their kindness effort. Thank you also, author, would like to say for Professor Koji Sode for correction and improvement given to this work. The award and regard go to all of staffs and students at the Ikebukuro Tsugawa Asano Laboratory, Division of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology who helped and guided me during the research and data collection. Especially, the author thanks for Maya Fitriana to give me advice for thesis writing and all of the staffs of the STEP @TUAT program from The Organization of the Advancement of Education and Global Learning (EAGLE), Tokyo University of Agriculture and Technology who help and take care during this program. | id |
| dc.language.iso | en | id |
| dc.publisher | IPB University | id |
| dc.title | Organophosphate Insecticide Sensing Using
Phosphotriesterase (PTE) Enzyme Derived from Sphingobium fuliginis ATCC
27551 Based-on pH-ISFET Sensor Measuremen | id |
| dc.subject.keyword | Organophosphate insecticide, Paraoxon, pH-ISFET, Phosphotriesterase, Sphingobium fuliginis ATCC 27551 | id |
| dc.subject.keyword | Organophosphate insecticide | |
| dc.subject.keyword | Paraoxon | |
| dc.subject.keyword | pH-ISFET | |
| dc.subject.keyword | Phosphotriesterase | |
| dc.subject.keyword | Sphingobium fuliginis ATCC 27551 | |
