title: Rapid and specific detection of herbicides using a self-assembled photosynthetic reaction center from purple bacterium on an SPR chip.
abstract: In this study, a direct detection system for herbicides inhibiting photosynthetic electron transfer was developed using the photosynthetic reaction center (RC) from the purple bacterium, Rhodobacter sphaeroides, and surface plasmon resonance (SPR) apparatus. The heavy-subunit-histidine-tagged RCs (HHisRCs) were immobilized on an SPR sensor chip via nickel chelation chemistry as a binder for one of the triazine herbicides, atrazine. Immediately after injection of atrazine solution on the HHisRCs-immobilized chip, the SPR responses increased and reached plateaus within 1 min. The SPR signals were proportional to the sample concentrations of atrazine in the range 1-100 microg/ml. To evaluate the binding specificity to atrazine, chlorinated aromatic herbicides, DCMU and MCPP, were investigated using the HHisRCs-immobilized chip. An RC inhibitor, DCMU, could also be detected with a higher detection limit of 20 microg/ml than atrazine (1 microg/ml). MCPP showed no signals because its inhibition mechanism against plants is different from that of atrazine and DCMU. These results indicated that the sensor chip immobilized RCs could be used for the specific detection of photosynthetic inhibitors.
authors.affiliation: Tissue Biosensor Team, Tissue Engineering Research Center, National Institute of Advanced Industrial Science and Technology, 1-1-1 Higashi, 305-8562, Tsukuba, Japan. chikashi-nakamura@aist.go.jp
authors.forename: Chikashi
authors.initials: C
authors.lastname: Nakamura
authors.forename: Miki
authors.initials: M
authors.lastname: Hasegawa
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Nakamura
authors.forename: Jun
authors.initials: J
authors.lastname: Miyake
mesh_terms: D008456:2-Methyl-4-chlorophenoxyacetic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000145:classification; D001280:Atrazine / Q000032:analysis / Q000145:classification; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004237:Diuron / Q000032:analysis / Q000145:classification; D004800:Enzymes, Immobilized / Q000737:chemistry; D006540:Herbicides / Q000032:analysis* / Q000145:classification*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry*; D011485:Protein Binding; D012242:Rhodobacter sphaeroides / Q000737:chemistry / Q000201:enzymology; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
chemical_list: D004800:Enzymes, Immobilized; D006540:Herbicides; D045322:Photosynthetic Reaction Center Complex Proteins; C008340:mecoprop; D004237:Diuron; D008456:2-Methyl-4-chlorophenoxyacetic Acid; D001280:Atrazine