Real-time monitoring and inhibition of the activity of carbapenemase in live bacterial cells: application to screening of β-lactamase inhibitors

2020 ◽  
Vol 44 (46) ◽  
pp. 20334-20340
Author(s):  
Han Gao ◽  
Ying Ge ◽  
Min-Hao Jiang ◽  
Cheng Chen ◽  
Le-Yun Sun ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Real Time ◽  
Bacterial Cells ◽  
Real Time Monitoring ◽  
Live Bacterial Cells

Antibiotic resistance mediated by β-lactamases including metallo-β-lactamases (MβLs) has become an emerging threat.

Real-Time Monitoring of New Delhi Metallo-β-Lactamase Activity in Living Bacterial Cells by1H NMR Spectroscopy

2014 ◽  
Vol 126 (8) ◽  
pp. 2162-2165 ◽  
Author(s):  
Junhe Ma ◽  
Sarah McLeod ◽  
Kathleen MacCormack ◽  
Shubha Sriram ◽  
Ning Gao ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Real Time ◽  
New Delhi ◽  
Bacterial Cells ◽  
Real Time Monitoring

scholarly journals Metabolic Flux Analysis of Catechin Biosynthesis Pathways Using Nanosensor

Antioxidants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 288
Author(s):  
Habiba Kausar ◽  
Ghazala Ambrin ◽  
Mohammad K. Okla ◽  
Walid Soufan ◽  
Abdullah A. Al-Ghamdi ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Real Time ◽  
Metabolic Flux Analysis ◽  
Metabolic Flux ◽  
Regulatory Element ◽  
Flux Analysis ◽  
Bacterial Cells ◽  
Biosynthesis Pathway ◽  
Real Time Monitoring ◽  
E Coli

(+)-Catechin is an important antioxidant of green tea (Camelia sinensis (L.) O. Kuntze). Catechin is known for its positive role in anticancerous activity, extracellular matrix degradation, cell death regulation, diabetes, and other related disorders. As a result of enormous interest in and great demand for catechin, its biosynthesis using metabolic engineering has become the subject of concentrated research with the aim of enhancing (+)-catechin production. Metabolic flux is an essential concept in the practice of metabolic engineering as it helps in the identification of the regulatory element of a biosynthetic pathway. In the present study, an attempt was made to analyze the metabolic flux of the (+)-catechin biosynthesis pathway in order to decipher the regulatory element of this pathway. Firstly, a genetically encoded fluorescence resonance energy transfer (FRET)-based nanosensor (FLIP-Cat, fluorescence indicator protein for (+)-catechin) was developed for real-time monitoring of (+)-catechin flux. In vitro characterization of the purified protein of the nanosensor showed that the nanosensor was pH stable and (+)-catechin specific. Its calculated Kd was 139 µM. The nanosensor also performed real-time monitoring of (+)-catechin in bacterial cells. In the second step of this study, an entire (+)-catechin biosynthesis pathway was constructed and expressed in E. coli in two sets of plasmid constructs: pET26b-PT7-rbs-PAL-PT7-rbs-4CL-PT7-rbs-CHS-PT7-rbs-CHI and pET26b-T7-rbs-F3H-PT7-rbs- DFR-PT7-rbs-LCR. The E. coli harboring the FLIP-Cat was transformed with these plasmid constructs. The metabolic flux analysis of (+)-catechin was carried out using the FLIP-Cat. The FLIP-Cat successfully monitored the flux of catechin after adding tyrosine, 4-coumaric acid, 4-coumaroyl CoA, naringenin chalcone, naringenin, dihydroquercetin, and leucocyanidin, individually, with the bacterial cells expressing the nanosensor as well as the genes of the (+)-catechin biosynthesis pathway. Dihydroflavonol reductase (DFR) was identified as the main regulatory element of the (+)-catechin biosynthesis pathway. Information about this regulatory element of the (+)-catechin biosynthesis pathway can be used for manipulating the (+)-catechin biosynthesis pathway using a metabolic engineering approach to enhance production of (+)-catechin.

scholarly journals Real-Time Monitoring of New Delhi Metallo-β-Lactamase Activity in Living Bacterial Cells by1H NMR Spectroscopy

2014 ◽  
Vol 53 (8) ◽  
pp. 2130-2133 ◽  
Author(s):  
Junhe Ma ◽  
Sarah McLeod ◽  
Kathleen MacCormack ◽  
Shubha Sriram ◽  
Ning Gao ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Real Time ◽  
New Delhi ◽  
Bacterial Cells ◽  
Real Time Monitoring

1618: Real-Time Monitoring of Nitric Oxide and Blood Flow During Ischemia-Reperfusion in the Rat Testis

2006 ◽  
Vol 175 (4S) ◽  
pp. 521-521
Author(s):  
Motoaki Saito ◽  
Tomoharu Kono ◽  
Yukako Kinoshita ◽  
Itaru Satoh ◽  
Keisuke Satoh
Keyword(s):  
Nitric Oxide ◽  
Blood Flow ◽  
Real Time ◽  
Ischemia Reperfusion ◽  
Real Time Monitoring ◽  
Rat Testis

GaN heteroepitaxy by remote plasma MOCVD : Real time monitoring by laser reflectance interferometry

2001 ◽  
Vol 11 (PR3) ◽  
pp. Pr3-1175-Pr3-1182 ◽  
Author(s):  
M. Losurdo ◽  
A. Grimaldi ◽  
M. Giangregorio ◽  
P. Capezzuto ◽  
G. Bruno
Keyword(s):  
Real Time ◽  
Real Time Monitoring ◽  
Remote Plasma ◽  
Laser Reflectance
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