1.
Designing an Artificial Pathway for the Biosynthesis of a Novel Phenazine N-Oxide in Pseudomonas chlororaphis HT66.
Guo, S, Liu, R, Wang, W, Hu, H, Li, Z, Zhang, X
ACS synthetic biology. 2020;(4):883-892
Abstract
Aromatic N-oxides are valuable due to their versatile chemical, pharmaceutical, and agricultural applications. Natural phenazine N-oxides possess potent biological activities and can be applied in many ways; however, few N-oxides have been identified. Herein, we developed a microbial system to synthesize phenazine N-oxides via an artificial pathway. First, the N-monooxygenase NaphzNO1 was predicted and screened in Nocardiopsis sp. 13-12-13 through a product comparison and gene sequencing. Subsequently, according to similarities in the chemical structures of substrates, an artificial pathway for the synthesis of a phenazine N-oxide in Pseudomonas chlororaphis HT66 was designed and established using three heterologous enzymes, a monooxygenase (PhzS) from P. aeruginosa PAO1, a monooxygenase (PhzO) from P. chlororaphis GP72, and the N-monooxygenase NaphzNO1. A novel phenazine derivative, 1-hydroxyphenazine N'10-oxide, was obtained in an engineered strain, P. chlororaphis HT66-SN. The phenazine N-monooxygenase NaphzNO1 was identified by metabolically engineering the phenazine-producing platform P. chlororaphis HT66. Moreover, the function of NaphzNO1, which can catalyze the conversion of 1-hydroxyphenazine but not that of 2-hydroxyphenazine, was confirmed in vitro. Additionally, 1-hydroxyphenazine N'10-oxide demonstrated substantial cytotoxic activity against two human cancer cell lines, MCF-7 and HT-29. Furthermore, the highest microbial production of 1-hydroxyphenazine N'10-oxide to date was achieved at 143.4 mg/L in the metabolically engineered strain P3-SN. These findings demonstrate that P. chlororaphis HT66 has the potential to be engineered as a platform for phenazine-modifying gene identification and derivative production. The present study also provides a promising alternative for the sustainable synthesis of aromatic N-oxides with unique chemical structures by N-monooxygenase.
2.
Experimental study of Love-wave immunosensors based on ZnO/LiTaO3 structures.
Zhou, FM, Li, Z, Fan, L, Zhang, SY, Shui, XJ
Ultrasonics. 2010;(3):411-5
Abstract
Experimental study of Love-mode immunosensors based on structures of ZnO/36 degrees YX-LiTaO3 is presented, in which the ZnO films with c-axis (002) orientation have been successfully grown on the 36 degrees YX-LiTaO3 substrates by RF magnetron sputtering technique. Then the Love-mode immunosensors based on the ZnO/36 degrees YX-LiTaO3 structures and monitoring antibody-antigen immunoreactions in aqueous solutions in real time are fabricated. The experimental results show that the optimal thickness of ZnO layers is about 1.20 microm in the structures deposited on 36 degrees YX-LiTaO3 substrates, which is much less than that of SiO2 overlayers about 6 microm. The antibody-antigen immunoreaction experiments also show that the frequency shifts of the sensors with 1.33 microm ZnO films are proportional to the concentration of antigen in solution as the concentration range less than 100 microg/ml.