A high-throughput screening assay for distinguishing nitrile hydratases from nitrilases

Angelini, Leticia Mara Lima; Silva, Amanda Ribeiro Martins da; Rocco, Lucas de Freitas Coli; Milagre, Cíntia D. F.

doi:10.1590/s1517-838246120130851

Cited by 10 publications

(18 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Though a rapid high-throughput colorimetric method for screening nitrilase-producing microorganisms using pH sensitive indicators was developed by Banerjee et al [21], the method has the lacuna of differentiating between nitrilase and nitrile hydratase in bacteria. Screening for both the enzymes simultaneously under dye based method and Nesslarization method have not been reported so far and a first report is presented here where a simple and rapid pH sensitive indicators are used to evaluate the activity of nitrile hydrolyzing enzymes by exploiting the sensitivity of amidase to the substrate acrylonitrile [40]. Detection of ammonia released due to nitrilase activity by Nesslarization method directly determines the activity of nitrilase, an additional confirmatory method.…”

Section: Resultsmentioning

confidence: 99%

A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria

Sahu

Meghavarnam

Janakiraman

2019

Biotechnology Reports

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria

Sahu

Meghavarnam

Janakiraman

2019

Biotechnology Reports

View full text Add to dashboard Cite

show abstract

“…Angelini et al . () developed a high‐throughput screening assay to distinguish microorganisms containing nitrilases and those possessing nitrile hydratases. A Banerjee‐modified colorimetric and pH‐sensitive assay was used with an amidase inhibitor in microplates using mandelonitrile as a substrate.…”

Section: Nature's Task Force: Screening Biocatalysts From the Environmentioning

confidence: 98%

Whole cell biocatalysts: essential workers from Nature to the industry

Carvalho

2016

Microbial Biotechnology

219

151

View full text Add to dashboard Cite

SummaryMicroorganisms have been exposed to a myriad of substrates and environmental conditions throughout evolution resulting in countless metabolites and enzymatic activities. Although mankind have been using these properties for centuries, we have only recently learned to control their production, to develop new biocatalysts with high stability and productivity and to improve their yields under new operational conditions. However, microbial cells still provide the best known environment for enzymes, preventing conformational changes in the protein structure in non‐conventional medium and under harsh reaction conditions, while being able to efficiently regenerate necessary cofactors and to carry out cascades of reactions. Besides, a still unknown microbe is probably already producing a compound that will cure cancer, Alzeihmer's disease or kill the most resistant pathogen. In this review, the latest developments in screening desirable activities and improving production yields are discussed.

show abstract

“…TFNG‐AM is the product of flonicamid hydration pathway, which require the activity of NHase (Angelini et al . 2015). The NHase expression was induced in E. coli Rosetta (DE3) overexpressing Aminobacter sp.…”

Section: Resultsmentioning

confidence: 99%

Biotransformation of insecticide flonicamid by Aminobacter sp. CGMCC 1.17253 via nitrile hydratase catalysed hydration pathway

et al. 2020

View full text Add to dashboard Cite

Aims This study evaluates flonicamid biotransformation ability of Aminobacter sp. CGMCC 1.17253 and the enzyme catalytic mechanism involved. Methods and Results Flonicamid transformed by resting cells of Aminobacter sp. CGMCC 1.17253 was carried out. Aminobacter sp. CGMCC 1.17253 converts flonicamid into N‐(4‐trifluoromethylnicotinoyl) glycinamide (TFNG‐AM). Aminobacter sp. CGMCC 1.17253 transforms 31·1% of the flonicamid in a 200 mg l−1 conversion solution in 96 h. Aminobacter sp. CGMCC 1.17253 was inoculated in soil, and 72·1% of flonicamid with a concentration of 0·21 μmol g−1 was transformed in 9 days. The recombinant Escherichia coli expressing Aminobacter sp. CGMCC 1.17253 nitrile hydratase (NHase) and purified NHase were tested for the flonicamid transformation ability, both of them acquired the ability to transform flonicamid into TFNG‐AM. Conclusions Aminobacter sp. CGMCC 1.17253 transforms flonicamid into TFNG‐AM via hydration pathway mediated by cobalt‐containing NHase. Significance and Impact of the Study This is the first report that bacteria of genus Aminobacter has flonicamid‐transforming ability. This study enhances our understanding of flonicamid‐degrading mechanism. Aminobacter sp. CGMCC 1.17253 has the potential for bioremediation of flonicamid pollution.

show abstract

A high-throughput screening assay for distinguishing nitrile hydratases from nitrilases

Cited by 10 publications

References 22 publications

A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria

A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing bacteria

Whole cell biocatalysts: essential workers from Nature to the industry

Biotransformation of insecticide flonicamid by Aminobacter sp. CGMCC 1.17253 via nitrile hydratase catalysed hydration pathway

Contact Info

Product

Resources

About