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Profiles of volatile indole emitted by Escherichia coli based on CDI-MS.

ABSTRACT: Escherichia coli is an important pathogen of nosocomial infection in clinical research, Thus, exploring new methods for the rapid detection of this pathogen is urgent. We reported the early release of molecular volatile indole vapour of E. coli cultures and blood cultures analyzed by direct atmospheric corona discharge ionization mass spectrometry (CDI-MS). The concentration of indole in E. coli cultures remarkably increases during the early log and lag phases of bacterial growth, thereby enabling early detection. Technical replicates were cultivated for 3 days for reference diagnosis using current conventional bacteraemia detection. A reference MS screen of common microbes from other genera confirmed that the peaks at m/z 116 signal corresponded to indole were specifically present in E. coli. Our results indicated that volatile indole based on CDI-MS without the need for any sample pretreatment is highly suitable for the reliable and cost-efficient differentiation of E. coli, especially for bacteraemia in humans.

SUBMITTER: Zhong Q

PROVIDER: S-EPMC6739388 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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Profiles of volatile indole emitted by Escherichia coli based on CDI-MS.

Zhong Qiaoshi Q Cheng Feng F Liang Juchao J Wang Xiaozhong X Chen Yanhui Y Fang Xueyao X Hu Longhua L Hang Yaping Y

Scientific reports 20190911 1

Escherichia coli is an important pathogen of nosocomial infection in clinical research, Thus, exploring new methods for the rapid detection of this pathogen is urgent. We reported the early release of molecular volatile indole vapour of E. coli cultures and blood cultures analyzed by direct atmospheric corona discharge ionization mass spectrometry (CDI-MS). The concentration of indole in E. coli cultures remarkably increases during the early log and lag phases of bacterial growth, thereby enabli ...[more]

PMID: 31511564

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Project description:Indole pyruvic acid (IPA) is a versatile platform intermediate and building block for a number of high-value products in the pharmaceutical and food industries. It also has a wide range of applications, such as drugs for the nervous system, cosmetics, and luminophores. Chemical synthesis of IPA is a complicated and costly process. Moreover, through the biosynthesis route employing L-amino acid oxidase, the byproduct hydrogen peroxide leads the degradation of IPA. TdiD, identified as a specific tryptophan aminotransferase, could be an alternative solution for efficient IPA biosynthesis.Escherichia coli strain W3110, which demonstrates basic production when supplied with tryptophan, was engineered for IPA biosynthesis. Several strategies were implemented to improve IPA production. First, through incorporating the codon-optimized tdiD into W3110, IPA levels increased from 41.54 ± 1.26 to 52.54 ± 2.08 mg/L. Second, after verifying the benefit of an increased phenylpyruvate pool, a YL03 strain was constructed based on a previously reported mutant strain of W3110 with a plasmid carrying aroF fbr and pheA fbr to further improve IPA production. The recombinant YL03 strain accumulated IPA at 158.85 ± 5.36 mg/L, which was 3.82-fold higher than that of the wild-type W3110 strain. Third, optimization of tdiD co expression was carried out by replacing the Trc promoter with a series of constitutively active promoters along with increasing the plasmid copy numbers. The highest IPA production was observed in YL08, which achieved 236.42 ± 17.66 mg/L and represented a greater than 5-fold increase as compared to W3110. Finally, the effects of deletion and overexpression of tnaA on IPA biosynthesis were evaluated. The removal of tnaA led to slightly reduced IPA levels, whereas the overexpression of tnaA resulted in a considerable decline in production.This study illustrates the feasibility of IPA biosynthesis in E. coli through tdiD. An efficient IPA producing strain, YL08, was developed, which provides a new possibility for biosynthesis of IPA. Although the final production was limited, this study demonstrates a convenient method of IPA synthesis.

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Profiles of volatile indole emitted by Escherichia coli based on CDI-MS.

Publications

Profiles of volatile indole emitted by Escherichia coli based on CDI-MS.

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