The use of clade-specific PCR assays to identify novel nitrilase genes from environmental isolates

Tríona Marie Dooley-Cullinane; Catherine O'Reilly; Bilal Aslam; David P. Weiner; David O'Neill; Erica Owens; Denise O'Meara; Lee Coffey

doi:10.1002/mbo3.700

The use of clade-specific PCR assays to identify novel nitrilase genes from environmental isolates

Tríona Marie Dooley-Cullinane, Catherine O'Reilly, Bilal Aslam, David P. Weiner, David O'Neill, Erica Owens, Denise O'Meara, Lee Coffey

Department of Science

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

4 Citations (Scopus)

Abstract

Nitrilase enzymes (EC 3.5.5.1) are responsible for the direct hydration of nitriles to their corresponding carboxylic acids and ammonia. The utilization of nitrilase enzymes in biocatalysis toward bio-pharmaceuticals and industrial applications facilitates the move towards green chemistry. The body of research presented describes a novel clade-specific touchdown PCR protocol for the detection of novel nitrilase genes. The presented study identified partial sequences of 15 novel nitrilase genes across 7 genera, with partial DNA sequence homology (%) displayed across an additional 16 genera. This research will prove valuable in the screening of microorganisms for the identification of novel clade-specific nitrilase genes, with predicted enantioselective profiles as determined by their clade characterizations.

Original language	English
Article number	e00700
Journal	MicrobiologyOpen
Volume	8
Issue number	4
DOIs	https://doi.org/10.1002/mbo3.700
Publication status	Published - Apr 2019

Keywords

clade specific
degenerate primers
environmental isolates
gene detection
nitrilase
touchdown PCR

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10.1002/mbo3.700

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title = "The use of clade-specific PCR assays to identify novel nitrilase genes from environmental isolates",

abstract = "Nitrilase enzymes (EC 3.5.5.1) are responsible for the direct hydration of nitriles to their corresponding carboxylic acids and ammonia. The utilization of nitrilase enzymes in biocatalysis toward bio-pharmaceuticals and industrial applications facilitates the move towards green chemistry. The body of research presented describes a novel clade-specific touchdown PCR protocol for the detection of novel nitrilase genes. The presented study identified partial sequences of 15 novel nitrilase genes across 7 genera, with partial DNA sequence homology (%) displayed across an additional 16 genera. This research will prove valuable in the screening of microorganisms for the identification of novel clade-specific nitrilase genes, with predicted enantioselective profiles as determined by their clade characterizations.",

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AU - Dooley-Cullinane, Tríona Marie

AU - O'Reilly, Catherine

AU - Aslam, Bilal

AU - Weiner, David P.

AU - O'Neill, David

AU - Owens, Erica

AU - O'Meara, Denise

AU - Coffey, Lee

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KW - nitrilase

KW - touchdown PCR

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The use of clade-specific PCR assays to identify novel nitrilase genes from environmental isolates

Abstract

Keywords

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