Real-time PCR detection of aldoxime dehydratase genes in nitrile-degrading microorganisms

Tríona Marie Dooley-Cullinane; Catherine O’Reilly; Lee Coffey

doi:10.1007/s10482-016-0786-2

Real-time PCR detection of aldoxime dehydratase genes in nitrile-degrading microorganisms

Tríona Marie Dooley-Cullinane, Catherine O’Reilly, Lee Coffey

Department of Science

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

Aldoxime dehydratase catalyses the conversion of aldoximes to their corresponding nitriles. Utilization of the aldoxime–nitrile metabolising enzyme pathway can facilitate the move towards a greener chemistry. In this work, a real-time PCR assay was developed for the detection of aldoxime dehydratase genes in aldoxime/nitrile metabolising microorganisms which have been purified from environmental sources. A conventional PCR assay was also designed allowing gene confirmation via sequencing. Aldoxime dehydratase genes were identified in 30 microorganisms across 11 genera including some not previously shown to harbour the gene. The assay displayed a limit of detection of 1 pg/μL DNA or 7 CFU/reaction. This real-time PCR assay should prove valuable in the high-throughput screening of micro-organisms for novel aldoxime dehydratase genes towards pharmaceutical and industrial applications.

Original language	English
Pages (from-to)	271-279
Number of pages	9
Journal	Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology
Volume	110
Issue number	2
DOIs	https://doi.org/10.1007/s10482-016-0786-2
Publication status	Published - 01 Feb 2017

Keywords

Aldoxime dehydratase
Aldoxime–nitrile pathway
Amidase
High-throughput
Nitrile hydratase
Real-time PCR

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10.1007/s10482-016-0786-2

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abstract = "Aldoxime dehydratase catalyses the conversion of aldoximes to their corresponding nitriles. Utilization of the aldoxime–nitrile metabolising enzyme pathway can facilitate the move towards a greener chemistry. In this work, a real-time PCR assay was developed for the detection of aldoxime dehydratase genes in aldoxime/nitrile metabolising microorganisms which have been purified from environmental sources. A conventional PCR assay was also designed allowing gene confirmation via sequencing. Aldoxime dehydratase genes were identified in 30 microorganisms across 11 genera including some not previously shown to harbour the gene. The assay displayed a limit of detection of 1 pg/μL DNA or 7 CFU/reaction. This real-time PCR assay should prove valuable in the high-throughput screening of micro-organisms for novel aldoxime dehydratase genes towards pharmaceutical and industrial applications.",

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AU - Coffey, Lee

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KW - High-throughput

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Real-time PCR detection of aldoxime dehydratase genes in nitrile-degrading microorganisms

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Keywords

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