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 ◽  
...  
2014 ◽  
Vol 53 (8) ◽  
pp. 2130-2133 ◽  
Author(s):  
Junhe Ma ◽  
Sarah McLeod ◽  
Kathleen MacCormack ◽  
Shubha Sriram ◽  
Ning Gao ◽  
...  

2020 ◽  
Vol 44 (46) ◽  
pp. 20334-20340
Author(s):  
Han Gao ◽  
Ying Ge ◽  
Min-Hao Jiang ◽  
Cheng Chen ◽  
Le-Yun Sun ◽  
...  

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


2009 ◽  
Vol 121 (34) ◽  
pp. 6392-6395 ◽  
Author(s):  
Antonio Herrera ◽  
Encarnación Fernández-Valle ◽  
Roberto Martínez-Álvarez ◽  
Dolores Molero ◽  
Zulay D. Pardo ◽  
...  

2015 ◽  
Vol 54 (18) ◽  
pp. 5374-5377 ◽  
Author(s):  
He Wen ◽  
Yong Jin An ◽  
Wen Jun Xu ◽  
Keon Wook Kang ◽  
Sunghyouk Park

2018 ◽  
Vol 54 (16) ◽  
pp. 2000-2003 ◽  
Author(s):  
Jian Wang ◽  
Tomoya Yamamoto ◽  
Jia Bai ◽  
Sarah J. Cox ◽  
Kyle J. Korshavn ◽  
...  

Magic-angle-spinning NMR for monitoring amyloid aggregation reveals that mechanical rotation of Aβ1–40 monomers increases the rate of aggregation.


2015 ◽  
Vol 127 (18) ◽  
pp. 5464-5467 ◽  
Author(s):  
He Wen ◽  
Yong Jin An ◽  
Wen Jun Xu ◽  
Keon Wook Kang ◽  
Sunghyouk Park

Antioxidants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 288
Author(s):  
Habiba Kausar ◽  
Ghazala Ambrin ◽  
Mohammad K. Okla ◽  
Walid Soufan ◽  
Abdullah A. Al-Ghamdi ◽  
...  

(+)-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.


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