The superiority of the third-generation catalyst, Rhodococcus rhodochrous J1 nitrile hydratase, for industrial production of acrylamide

Nagasawa, Tôru; Shimizu, Hitoshi; Yamada, Hideaki

doi:10.1007/bf00170364

Cited by 163 publications

(98 citation statements)

References 31 publications

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“…Among ten metal ions tested, Ag + (Ag NO 3 ) caused complete inhibition, while Pb2 + (Pb(CH 3 COO) 2 ) and Hg 2+ (HgCl 2 ) inhibited the NHase to extent of 95 and 89% respectively. This meant that cysteine is involved at the active site of the enzyme and these results supported the earlier fi ndings [14].…”

Section: Effect Of Metal Ions and Inhibitorsmentioning

confidence: 99%

“…Nitrile hydratase has emerged as enzyme of choice in chemical industries practicing white biotechnology for the organic synthesis. Currently, acrylamide, nicotinamide and 3-cyanovaleramide are being produced at industrial scale using the immobilized resting cells of NHase producing organism [2]. We have earlier reported a hyperactive propionitrile-induced NHase of R. rhodochrous PA-34 which have been used to synthesise acrylamide (600 g/l), butyramide (585 g/l) and nicotinamide (855 g/l) [3][4][5].…”

Section: Introductionmentioning

confidence: 99%

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Purification of a hyperactive nitrile hydratase from resting cells of Rhodococcus rhodochrous PA-34

2009

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A propionitrile-induced nitrile hydratase (NHase), a promising biocatalyst for synthesis of organic amides has been purifi ed from cell-free extract of Rhodococcus rhodochrous PA-34. About 11-fold purifi cation of NHase was achieved with 52% yield. The SDS-PAGE of the purifi ed enzyme revealed that it consisted of two subunits of 25.04 kD and 30.6 kD. However, the molecular weight of holoenzyme was speculated to be 86 kD by native-PAGE. This NHase exhibited maximum activity at pH 8.0 and temperature 40°C. Half-life was 2 h at 40°C and 0.5 h at 50°C. The Km and Vmax were 167 mM and 250 μmole/min/mg using 25 mM 3-cyanopyridine as substrate. AgNO 3 , Pb(CH 3 COO) 2 and HgCl 2 inhibited the NHase to extent of 89-100%.

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Section: Effect Of Metal Ions and Inhibitorsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Purification of a hyperactive nitrile hydratase from resting cells of Rhodococcus rhodochrous PA-34

2009

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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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“…Surfacial nitrile groups of acrylic fibers were hydrolyzed by the R. rhodochrous enzymes to a maximum of only 16%, although acrylonitrile is known to be a very good substrate for both nitrilases and nitrile hydratases from these organism (15,25). However, the enzyme reaction on PAN is restricted by several factors related to the properties of the polymer.…”

Section: Discussionmentioning

confidence: 99%

“…Although acrylonitrile seems to be one of most suitable substrates for both nitrile hydratase (25) and nitrilase from R. rhodochrous (15), there are no reports in the literature on the enzymatic hydrolysis of PANs.…”

mentioning

confidence: 99%

Nitrile Hydratase and Amidase from Rhodococcus rhodochrous Hydrolyze Acrylic Fibers and Granular Polyacrylonitriles

Tauber

Cavaco‐Paulo

Robra

et al. 2000

Appl Environ Microbiol

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Rhodococcus rhodochrous NCIMB 11216 produced nitrile hydratase (320 nkat mg of protein ؊1) and amidase activity (38.4 nkat mg of protein ؊1 ) when grown on a medium containing propionitrile. These enzymes were able to hydrolyze nitrile groups of both granular polyacrylonitriles (PAN) and acrylic fibers. Nitrile groups of PAN40 (molecular mass, 40 kDa) and PAN190 (molecular mass, 190 kDa) were converted into the corresponding carbonic acids to 1.8 and 1.0%, respectively. In contrast, surfacial nitrile groups of acrylic fibers were only converted to the corresponding amides. X-ray photoelectron spectroscopy analysis showed that 16% of the surfacial nitrile groups were hydrolyzed by the R. rhodochrous enzymes. Due to the enzymatic modification, the acrylic fibers became more hydrophilic and thus, adsorption of dyes was enhanced. This was indicated by a 15% increase in the staining level (K/S value) for C.I. Basic Blue 9.

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The superiority of the third-generation catalyst, Rhodococcus rhodochrous J1 nitrile hydratase, for industrial production of acrylamide

Cited by 163 publications

References 31 publications

Purification of a hyperactive nitrile hydratase from resting cells of Rhodococcus rhodochrous PA-34

Purification of a hyperactive nitrile hydratase from resting cells of Rhodococcus rhodochrous PA-34

The iron-type nitrile hydratase activator protein is a GTPase

Nitrile Hydratase and Amidase from Rhodococcus rhodochrous Hydrolyze Acrylic Fibers and Granular Polyacrylonitriles

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