Purification and characterization of nitrilase from Fusarium solani IMI196840

Vejvoda, Vojtěch; Kubáč, David; Davidová, Alžběta; Kaplan, Ondřej; Šulc, Miroslav; Šveda, Ondřej; Chaloupková, Radka; Martı́nková, Ludmila

doi:10.1016/j.procbio.2010.03.033

Cited by 32 publications

(23 citation statements)

References 24 publications

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“…A sharp activity decrease was observed at slightly alkaline pH values, only trace activity being observed at pH 8.5. This pH optimum was not like the pH optima of other nitrilases reported in the range of 7.0 to 9.0 (Mueller et al, 2006;Vejvoda et al, 2010;Kim et al, 2009). However, it was similar to that of the nitrilase from Rhodococcus rhodochrous K22, the pH optimum of which was reported at 5.5 (Kobayashi et al, 1990).…”

Section: Effects Of Temperatures and Ph Valuescontrasting

confidence: 73%

“…Initial investigations suggested nitrilases to be specific for aromatic nitriles. But now, depending on the substrate spectrum, nitrilases are reported from different sources (Banerjee et al, 2002), which can be active on aromatic or heterocyclic nitriles (Vejvoda et al, 2010) and aliphatic nitriles (Bayer et al, 2011) as well as arylacetonitriles (Sosedov et al, 2010). Nitrilases can convert nitriles directly into the corresponding carboxylic acids and ammonia.…”

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confidence: 99%

“…Nitrilases can convert nitriles directly into the corresponding carboxylic acids and ammonia. Compared with chemical methods of producing carboxylic acids, nitrilases are widely used due to the mild reaction conditions, regioselectivity (Wang et al, 2014) and enantioselectivity (Vejvoda et al, 2010). Therefore, they are used to catalyze the production of various valuable carboxylic acids.…”

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confidence: 99%

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Characterization of a nitrilase from Arthrobacter aurescens CYC705 for synthesis of iminodiacetic acid

Cai

Zhu

et al. 2014

J. Gen. Appl. Microbiol.

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A nitrilase gene cyc705 from Arthrobacter aurescens CYC705 for synthesis of iminodiacetic acid (IDA) was cloned. This gene contained a 930 bp ORF, which encoded a polypeptide of 310 amino acids. A recombinant Escherichia coli BL21(DE3)/ pET28a-cyc705 was constructed to achieve the heterologous expression of cyc705. This recombinant nitrilase was purified to homogeneity with a molecular weight of 36.7 kDa on SDS-PAGE and mass spectrometry, and characterized to be an oligomer of 14 subunits by gel permeation chromatography. Using iminodiacetonitrile (IDAN) as the substrate, the V max , K m , k cat and k cat / K m were 9.05 U mg 1 , 43.17 mM −1 , 94.1 min −1 and 2.18 10 3 min −1 M −1 , respectively. The optimum temperature and pH were 25 C and 5.8. The suitable substrates for the purified nitrilase were short-chain aliphatic dinitriles. High concentration of IDAN could be hydrolyzed to IDA in a shorter time.Key words: Arthrobacter aurescens; characterization; iminodiacetic acid; nitrilase Introduction Nitrilase (EC 3.5.5.1), as an important industrial enzyme, belongs to the nitrilase superfamily (Brenner, 2002). Nitrilases are widely derived from various organisms, including bacteria, fungi and plants. Initial investigations suggested nitrilases to be specific for aromatic nitriles. But now, depending on the substrate spectrum, nitrilases are reported from different sources (Banerjee et al., 2002), which can be active on aromatic or heterocyclic nitriles (Vejvoda et al., 2010) and aliphatic nitriles (Bayer et al., 2011) as well as arylacetonitriles (Sosedov et al., 2010). Nitrilases can convert nitriles directly into the corresponding carboxylic acids and ammonia. Compared with chemical methods of producing carboxylic acids, nitrilases are widely used due to the mild reaction conditions, regioselectivity (Wang et al., 2014) and enantioselectivity (Vejvoda et al., 2010). Therefore, they are used to catalyze the production of various valuable carboxylic acids.IDA is an important fine chemical intermediate. It has wide applications and is mainly used in producing glyphosate herbicides, chelating agents and surfactants (Ni et al., 2009). Currently, the production of IDA mainly uses chemical methods, including the hydrocyanic acid method (Rmon, 1991), diethanolamine method (Bornscheuer and Kazlauskas, 2006), and chloroacetic acid method (Farbwerke Hoechst, 1969). These methods produce a large amount of waste water and byproducts, which are unfriendly to the environment (Duan et al., 2007). Enzymatic production of IDA has some advantages, such as the mild reaction conditions, environmental friendliness and low cost. So far, only a few papers have reported nitrilase-catalyzed production of IDA. A nitrilase-produced strain named Alcaligenes faecalis ZJB-09133 was screened by using IDAN as the carbon source, and the whole cells were used to catalyze IDAN to IDA, which showed the conversion only reached 65.3% at 8 h after optimization of the reaction conditions (Liu et al., 2011). The Acidovorax facilis nitrilase was cloned ...

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Section: Effects Of Temperatures and Ph Valuescontrasting

confidence: 73%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Characterization of a nitrilase from Arthrobacter aurescens CYC705 for synthesis of iminodiacetic acid

Cai

Zhu

et al. 2014

J. Gen. Appl. Microbiol.

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“…The molecular mass was found to be about 43 kDa. Previous studies showed similar molecular mass of nitrilase isolated from Rhodococcus rhodochrous K22 and Fusarium solani IMI196840 17,24 . The 3-cyanopyridinase of selected microorganism was partially purified to 2.3 fold from cell-free extract with a recovery of 46.5 %.…”

Section: Fig 9: the Graph Illustrates The Activity Of 3-cyanopyridinmentioning

confidence: 60%

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2017

IJPSR

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A thermostable bacterial 3-cyanopyridinase enzyme has been identified that catalyze 3-cyanopyridine to nicotinic acid and ammonia. Nicotinic acid is mainly used in food, pharmaceutical and biochemical industries. This study was undertaken to optimize the purification of 3-cyanopyridinase enzyme and characterization of its various kinetic properties. The partial purification of enzyme has been carried out in five steps that results in approximately 2.3 fold increased purification with 46.5 % of enzyme recovery. The partially purified enzyme was observed approximately 43 kDa of maximum activity at 50 °C temperature and pH 7.0. The partially purified enzyme showed the K m of 0.91 mM and maximum velocity (V max ) of 0.59 U. The half-life (t 1/2 ) of partially purified enzyme was about 60 min at optimal conditions. The enzyme has broad substrate specificity, hydrolyzing toxic compounds such as benzonitrile and propionitrile. Thus this novel enzyme has potential in bioremediation process.

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“…These cultures were grown in the laboratory using different media (Table 1) for the production of nitrilase activity following the procedures described earlier [17–19]. Nitrilase activity was assayed in 1.0 mL reaction mixture containing nitrile as substrate (1–10 mM) and 0.1 mL resting cells.…”

Section: Methodsmentioning

confidence: 99%

Mining of Microbial Genomes for the Novel Sources of Nitrilases

Sharma

Thakur

Raj

et al. 2017

BioMed Research International

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Next-generation DNA sequencing (NGS) has made it feasible to sequence large number of microbial genomes and advancements in computational biology have opened enormous opportunities to mine genome sequence data for novel genes and enzymes or their sources. In the present communication in silico mining of microbial genomes has been carried out to find novel sources of nitrilases. The sequences selected were analyzed for homology and considered for designing motifs. The manually designed motifs based on amino acid sequences of nitrilases were used to screen 2000 microbial genomes (translated to proteomes). This resulted in identification of one hundred thirty-eight putative/hypothetical sequences which could potentially code for nitrilase activity. In vitro validation of nine predicted sources of nitrilases was done for nitrile/cyanide hydrolyzing activity. Out of nine predicted nitrilases, Gluconacetobacter diazotrophicus, Sphingopyxis alaskensis, Saccharomonospora viridis, and Shimwellia blattae were specific for aliphatic nitriles, whereas nitrilases from Geodermatophilus obscurus, Nocardiopsis dassonvillei, Runella slithyformis, and Streptomyces albus possessed activity for aromatic nitriles. Flavobacterium indicum was specific towards potassium cyanide (KCN) which revealed the presence of nitrilase homolog, that is, cyanide dihydratase with no activity for either aliphatic, aromatic, or aryl nitriles. The present study reports the novel sources of nitrilases and cyanide dihydratase which were not reported hitherto by in silico or in vitro studies.

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Purification and characterization of nitrilase from Fusarium solani IMI196840

Cited by 32 publications

References 24 publications

Characterization of a nitrilase from Arthrobacter aurescens CYC705 for synthesis of iminodiacetic acid

Characterization of a nitrilase from Arthrobacter aurescens CYC705 for synthesis of iminodiacetic acid

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Mining of Microbial Genomes for the Novel Sources of Nitrilases

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