Multifunctional magnetic nanoparticle cloud assemblies for in situ capture of bacteria and isolation of microbial DNA

The Analyst ◽  
2021 ◽  
Author(s):  
Lucas Poncelet ◽  
Lidija Malic ◽  
Liviu Clime ◽  
Matthias Geissler ◽  
Keith J. Morton ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Magnetic Nanoparticle ◽  
Nanoparticle Assemblies ◽  
Bacterial Lysis ◽  
Microbial Dna

Suspended magnetic nanoparticle assemblies forming between magnetized micropillars are used as a multifunctional capture matrix for Escherichia coli O157:H7 and in-flow extraction of microbial DNA released upon bacterial lysis.

scholarly journals In situ delivery of biobutyrate by probiotic Escherichia coli for cancer therapy

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Chung-Jen Chiang ◽  
Yan-Hong Hong
Keyword(s):  
Escherichia Coli ◽  
Cancer Therapy ◽  
Rational Design ◽  
Hypoxic Condition ◽  
Human Colorectal Cancer ◽  
Apoptosis Pathway ◽  
Mitochondrial Apoptosis Pathway ◽  
Cycle Arrest ◽  
Human Colorectal Cancer Cell

AbstractButyrate has a bioactive function to reduce carcinogenesis. To achieve targeted cancer therapy, this study developed bacterial cancer therapy (BCT) with butyrate as a payload. By metabolic engineering, Escherichia coli Nissle 1917 (EcN) was reprogrammed to synthesize butyrate (referred to as biobutyrate) and designated EcN-BUT. The adopted strategy includes construction of a synthetic pathway for biobutyrate and the rational design of central metabolism to increase the production of biobutyrate at the expense of acetate. With glucose, EcN-BUT produced primarily biobutyrate under the hypoxic condition. Furthermore, human colorectal cancer cell was administrated with the produced biobutyrate. It caused the cell cycle arrest at the G1 phase and induced the mitochondrial apoptosis pathway independent of p53. In the tumor-bearing mice, the injected EcN-BUT exhibited tumor-specific colonization and significantly reduced the tumor volume by 70%. Overall, this study opens a new avenue for BCT based on biobutyrate.

scholarly journals Efficient synthesis of ponasterone A by recombinant Escherichia coli harboring the glycosyltransferase GTBP1with in situ product removal

RSC Advances ◽  
2017 ◽  
Vol 7 (37) ◽  
pp. 23027-23029 ◽  
Author(s):  
Bingfeng Li ◽  
Xuejun He ◽  
Bo Fan ◽  
Jianlin Chu ◽  
Bingfang He
Keyword(s):  
Escherichia Coli ◽  
Bacillus Pumilus ◽  
Recombinant Escherichia Coli ◽  
Efficient Synthesis ◽  
In Situ Product Removal ◽  
Product Removal ◽  
Ponasterone A

A glycosyltransferase GTBP1fromBacillus pumilusBF1 was isolated.

scholarly journals Manipulating ATP supply improves in situ CO2 recycling by reductive TCA cycle in engineered Escherichia coli

3 Biotech ◽  
2020 ◽  
Vol 10 (3) ◽  
Author(s):  
Ching-Hsun Chen ◽  
I.-Ting Tseng ◽  
Shou-Chen Lo ◽  
Zi-Rong Yu ◽  
Ju-Jiun Pang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Tca Cycle ◽  
Co2 Recycling ◽  
Engineered Escherichia Coli ◽  
Atp Supply

scholarly journals High-Temperature Fluorescent In Situ Hybridization for Detecting Escherichia coli in Seawater Samples, Using rRNA-Targeted Oligonucleotide Probes and Flow Cytometry

2005 ◽  
Vol 71 (12) ◽  
pp. 8157-8164 ◽  
Author(s):  
Ying Zhong Tang ◽  
Karina Yew Hoong Gin ◽  
Tok Hoon Lim
Keyword(s):  
Escherichia Coli ◽  
Signal Intensity ◽  
Fluorescence Signal ◽  
Standard Protocol ◽  
Oligonucleotide Probes ◽  
E Coli ◽  
Hybridization Efficiency ◽  
Hybridization Time ◽  
Seawater Samples

ABSTRACT Fluorescence in situ hybridization (FISH) is a widely used method to detect environmental microorganisms. The standard protocol is typically conducted at a temperature of 46°C and a hybridization time of 2 or 3 h, using the fluorescence signal intensity as the sole parameter to evaluate the performance of FISH. This paper reports our results for optimizing the conditions of FISH using rRNA-targeted oligonucleotide probes and flow cytometry and the application of these protocols to the detection of Escherichia coli in seawater spiked with E.coli culture. We obtained two types of optimized protocols for FISH, which showed rapid results with a hybridization time of less than 30 min, with performance equivalent to or better than the standard protocol in terms of the fluorescence signal intensity and the FISH hybridization efficiency (i.e., the percentage of hybridized cells giving satisfactory fluorescence intensity): (i) one-step FISH (hybridization is conducted at 60 to 75°C for 30 min) and (ii) two-step FISH (pretreatment in a 90°C water bath for 5 min and a hybridizing step at 50 to 55°C for 15 to 20 min). We also found that satisfactory fluorescence signal intensity does not necessarily guarantee satisfactory hybridization efficiency and the tightness of the targeted population when analyzed with a flow cytometer. We subsequently successfully applied the optimized protocols to E. coli-spiked seawater samples, i.e., obtained flow cytometric signatures where the E. coli population was well separated from other particles carrying fluorescence from nonspecific binding to probes or from autofluorescence, and had a good recovery rate of the spiked E. coli cells (90%).

Sign in / Sign up

Export Citation Format

Share Document