Purification and Characterization of Nitphym, a Robust Thermostable Nitrilase From Paraburkholderia phymatum - Archive ouverte HAL Access content directly
Journal Articles Frontiers in Bioengineering and Biotechnology Year : 2021

Purification and Characterization of Nitphym, a Robust Thermostable Nitrilase From Paraburkholderia phymatum

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Abstract

Despite the success of some nitrilases in industrial applications, there is a constant demand to broaden the catalog of these hydrolases, especially robust ones with high operational stability. By using the criteria of thermoresistance to screen a collection of candidate enzymes heterologously expressed in Escherichia coli , the enzyme Nit phym from the mesophilic organism Paraburkholderia phymatum was selected and further characterized. Its quick and efficient purification by heat treatment is of major interest for large-scale applications. The purified nitrilase displayed a high thermostability with 90% of remaining activity after 2 days at 30°C and a half-life of 18 h at 60°C, together with a broad pH range of 5.5–8.5. Its high resistance to various miscible cosolvents and tolerance to high substrate loadings enabled the quantitative conversion of 65.5 g⋅L –1 of 3-phenylpropionitrile into 3-phenylpropionic acid at 50°C in 8 h at low enzyme loadings of 0.5 g⋅L –1 , with an isolated yield of 90%. This study highlights that thermophilic organisms are not the only source of industrially relevant thermostable enzymes and extends the scope of efficient nitrilases for the hydrolysis of a wide range of nitriles, especially trans -cinnamonitrile, terephthalonitrile, cyanopyridines, and 3-phenylpropionitrile.

Dates and versions

cea-03664095 , version 1 (10-05-2022)

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Thomas Bessonnet, Aline Mariage, Jean-Louis Petit, Virginie Pellouin, Adrien Debard, et al.. Purification and Characterization of Nitphym, a Robust Thermostable Nitrilase From Paraburkholderia phymatum. Frontiers in Bioengineering and Biotechnology, 2021, 9, ⟨10.3389/fbioe.2021.686362⟩. ⟨cea-03664095⟩
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