Nitrile hydratases (NHases): At the interface of academia and industry

Prasad, Shreenath; Bhalla, Tek Chand

doi:10.1016/j.biotechadv.2010.05.020

Cited by 193 publications

(120 citation statements)

References 165 publications

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“…1,2 NHases have attracted substantial interest as biocatalysts in preparative organic chemistry and are used in the large scale industrial production of acrylamide 1,[3][4][5][6] and nicotinamide. 7 X-ray crystallographic studies indicate that they are α2β2 heterotetramers with an active site consisting of three cysteine residues, two amide nitrogens, a water molecule, and either a nonheme Fe(III) ion (Fetype) or a noncorrin Co(III) ion (Co-type).…”

Section: Introductionmentioning

confidence: 99%

The iron-type nitrile hydratase activator protein is a GTPase

Gumataotao

Lankathilaka

Bennett

et al. 2017

Biochemical Journal

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Abstract:The Fe-type nitrile hydratase activator protein from Rhodococcus equi TG328-2 (ReNHase TG328-2) was successfully expressed and purified. Sequence analysis and homology modeling suggest that it is a G3E P-loop guanosine triphosphatase (GTPase) within the COG0523 subfamily. Kinetic studies revealed that the Fe-type activator protein is capable of hydrolyzing NOT THE PUBLISHED VERSION; this is the author's final, peer-reviewed manuscript. The published version may be accessed by following the link in the citation at the bottom of the page.Biochemical Journal, Vol 474, No. 2 (January 15, 2017): pg. 247-258. DOI. This article is © Portland Press Limited and permission has been granted for this version to appear in e-Publications@Marquette. Portland Press Limited does not grant permission for this article to be further copied/distributed or hosted elsewhere without the express permission from Portland Press Limited. 2GTP to GDP with a kcat value of 1.2 × 10 −3 s −1 and a Km value of 40 μM in the presence of 5 mM MgCl2 in 50 mM 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid at a pH of 8.0. The addition of divalent metal ions, such as Co(II), which binds to the ReNHase TG328-2 activator protein with a Kd of 2.9 μM, accelerated the rate of GTP hydrolysis, suggesting that GTP hydrolysis is potentially connected to the proposed metal chaperone function of the ReNHase TG328-2 activator protein. Circular dichroism data reveal a significant conformational change upon the addition of GTP, which may be linked to the interconnectivity of the cofactor binding sites, resulting in an activator protein that can be recognized and can bind to the NHase α-subunit. A combination of these data establishes, for the first time, that the ReNHase TG328-2 activator protein falls into the COG0523 subfamily of G3E P-loop GTPases, many of which play a role in metal homeostasis processes.

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Section: Introductionmentioning

confidence: 99%

The iron-type nitrile hydratase activator protein is a GTPase

Gumataotao

Lankathilaka

Bennett

et al. 2017

Biochemical Journal

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“…Likewise, some bacteria like Rhodococcus UKMP-5M have been recently applied to degrade cyanide (Nallapan et al 2013). A general metabolic pathway proposed by Prasad and Bhalla (2010) for nitrile synthesis and degradation in microorganisms can be seen in figure 2. Cyanide hydratases are primarily fungal enzymes which are considered the first responsible from cyanide conversion.…”

Section: Biotechnological Importance Of Nitrile-degrading Enzymes In mentioning

confidence: 99%

“…General metabolic pathway for nitrile synthesis and degradation in microorganisms. Prasad and Bhalla (2010).…”

Section: Conclusion and Future Perspectivesmentioning

confidence: 99%

Understanding nitrile-degrading enzymes: classification, biocatalytic nature and current applications

Rodríguez¹

2013

Rev Latinoam Biotecnol Ambient Algal

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Nitrile-degrading enzymes commonly known as nitrilase enzymes are able to metabolize nitrile-substituent compounds and they have several industrial applications, for example: in drugs synthesis. It is also common to observe their exploitation for obtaining chemical compounds with commercial interests related to cosmetics production, paints and additives. In addition, these are frequently used in the active metabolites synthesis of pesticides. Due to the catalytic nature of such proteins, it is possible to take advantage of their biotechnological potential to be applied in various scientific fields including synthetic biocatalysis and environmental remediation, since they have been successfully used for soils nitrile-wastes decontamination such as cyanide, bromoxynil and benzonitrile. On the other hand, these enzymes are considered very important intermediaries of metabolic pathways related to indolic compounds that are produced by bacteria, plants and superior fungi, acting in most cases as vegetal growth hormones. Given the fact that indole-derivative molecules play an important role in physiological responses in superior organisms, nitrilase enzymes may be considered as important part of unknown multi-enzymatic secondary metabolites pathways. In light of the above considerations, this review attempts to summarize the current status of nitrilase research and describing in detail the main characteristics of nitrile-converting enzymes with emphasis on fungal proteins, including their function and catalytic selectivity. Likewise, their relationship with plant metabolism and biotechnological importance in bioremediation processes is discussed. Keywords: nitrile-degrading enzymes, indolic compounds, indole-3-acetic acid, cyanide, environmental remediation. ResumenLas enzimas degradadoras de compuestos nitrilo conocidas comúnmente como enzimas nitrilasas, son capaces de metabolizar compuestos nitrilo-sustituyentes, y tienen diversas aplicaciones industriales como por ejemplo: en la síntesis de fármacos. Asimismo, es común observar su explotación para la obtención de compuestos químicos de interés comercial relacionados con la elaboración de cosméticos, pinturas y aditivos. Además, éstas suelen emplearse también de manera frecuente en la síntesis de metabolitos activos de plaguicidas. Debido a la naturaleza catalítica de dichas proteínas, actualmente es posible explotar su potencial biotecnológico para ser aplicado en diversos campos, incluyendo la biocatálisis sintética y remediación ambiental, ya que han sido utilizadas con éxito para la descontaminación de suelos impactados con cianuro, bromoxinil y benzonitrilo. Por otra parte, dichas enzimas son importantes intermediarias de rutas metabólicas de compuestos indólicos producidos por bacterias, plantas y hongos superiores, actuando en la mayoría de los 10.7603/s40 -01 -00 - casos como hormonas de crecimiento vegetal. Debido al hecho de que las moléculas derivadas de compuestos indólicos juegan un papel importante en las respuestas fisiológicas de organismos ...

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“…4 NHases are already used in several industrial applications such as the synthesis of specialty chemicals and pharmaceuticals including acrylamide and nicotinamide. 5,6 A key advantage of NHases is their stereoselectivity, which is of particular importance in the pharmaceutical arena. Moreover, given the exquisite reaction specificity, high turnover number, and benign environmental impact, NHases are becoming increasingly recognized as a "Green" catalyst for the degradation of environmentally harmful nitriles.…”

Section: Introductionmentioning

confidence: 99%

“…1,6,7 The metal ion is ligated by three cysteine sulfur atoms, two backbone amide nitrogens, and a water molecule. [8][9][10][11] Two of the active site cysteine residues are post-translationally modified to cysteine-sulfinic acid (Cys-SO2H) and cysteine-sulfenic acid (Cys-SOH) yielding an unusual metal coordination geometry, termed the "claw-setting".…”

Section: Introductionmentioning

confidence: 99%

Analyzing the catalytic role of active site residues in the Fe-type nitrile hydratase from Comamonas testosteroni Ni1

Martinez

Krzywda

et al. 2015

J Biol Inorg Chem

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A strictly conserved active site arginine residue (αR157) and two histidine residues (αH80 and αH81) located near the active site of the Fe-type nitrile hydratase from Comamonas testosteroni Ni1 (CtNHase), were mutated. These mutant enzymes were examined for their ability to bind iron and hydrate acrylonitrile. For the αR157A mutant, the residual activity (kcat = 10 ± 2 s −1 ) accounts for less than 1 % of the wild-type activity (kcat = 1100 ± 30 s −1 ) while the Km value is nearly unchanged at 205 ± 10 mM. On the other hand, mutation of the active site pocket αH80 and αH81 residues to alanine resulted in enzymes with kcat values of 220 ± 40 and 77 ± 13 s −1 , respectively, and Km values of 187 ± 11 and 179 ± 18 mM. The double mutant (αH80A/αH81A) was also prepared and provided an enzyme with a kcat value of 132 ± 3 s −1 and a Km value of 213 ± 61 mM. These data indicate that all three residues are catalytically important, but not essential. X-ray crystal structures of the αH80A/αH81A, αH80W/αH81W, and αR157A mutant CtNHase enzymes were solved to 2.0, 2.8, and 2.5 Å resolutions, respectively. In each mutant enzyme, hydrogen-bonding interactions crucial for the catalytic function of the αCys 104 -SOH ligand are disrupted. Disruption of these hydrogen bonding interactions likely alters the nucleophilicity of the sulfenic acid oxygen and the Lewis acidity of the active site Fe(III) ion.

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Nitrile hydratases (NHases): At the interface of academia and industry

Cited by 193 publications

References 165 publications

The iron-type nitrile hydratase activator protein is a GTPase

The iron-type nitrile hydratase activator protein is a GTPase

Understanding nitrile-degrading enzymes: classification, biocatalytic nature and current applications

Analyzing the catalytic role of active site residues in the Fe-type nitrile hydratase from Comamonas testosteroni Ni1

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