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

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• 作者：Tríona Marie Dooley-Cullinane ; Catherine O’Reilly ; Lee Coffey
• 关键词：Aldoxime dehydratase ; Aldoxime–nitrile pathway ; Amidase ; High ; throughput ; Nitrile hydratase ; Real ; time PCR
• 刊名：Antonie van Leeuwenhoek
• 出版年：2017
• 出版时间：February 2017
• 年：2017
• 卷：110
• 期：2
• 页码：271-279
• 全文大小：
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Microbiology; Medical Microbiology; Plant Sciences; Soil Science & Conservation;
• 出版者：Springer International Publishing
• ISSN：1572-9699
• 卷排序：110

文摘

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.