Purification and characterization of a nitrilase from Fusarium solani O1

Vejvoda, Vojtěch; Kaplan, Ondřej; Bezouška, Karel; Pompach, Petr; Šulc, Miroslav; Cantarella, Maria; Benada, Oldřich; Uhnáková, Bronislava; Rinágelová, Anna; Lutz-Wahl, Sabine; Fischer, Lutz; Křen, Vladimı́r; Martı́nková, Ludmila

doi:10.1016/j.molcatb.2007.09.006

Cited by 54 publications

(35 citation statements)

References 23 publications

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“…2a) and pH 8.0 (Fig. 2b), respectively, which were similar to those from Alcaligenes faecalis JM3 [9], Alcaligenes faecalis ATCC 8750 [10], Bradyrhizobium japonicum USDA110 [26], Aspergillus niger K10 [27] and Fusarium solani O1 [28], as listed in Table 1. Although their optimal temperatures were the same with the nitrilase from Alcaligenes sp.…”

Section: Temperature and Ph Optimasupporting

confidence: 75%

Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(−)-mandelic acid production

Zhang

et al. 2010

Bioprocess Biosyst Eng

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A nitrilase gene from Alcaligenes sp. ECU0401 was cloned and overexpressed in Escherichia coli BL21 (DE3) in a soluble form. The encoded protein with a His₆-tag was purified to nearly homogeneity as revealed by SDS-PAGE with a molecular weight of approximately 38.5 kDa, and the holoenzyme was estimated to be composed of 10 subunits of identical size by size exclusion chromatography. The V(max) and K(m) parameters were determined to be 27.9 μmol min⁻¹ mg⁻¹ protein and 21.8 mM, respectively, with mandelonitrile as the substrate. The purified enzyme was highly thermostable with a half life of 155 h at 30 °C and 94 h at 40 °C. Racemic mandelonitrile (50 mM) could be enantioselectively hydrolyzed to (R)-(-)-mandelic acid by the purified nitrilase with an enantiomeric excess of 97%. The extreme stability, high activity and enantioselectivity of this nitrilase provide a solid base for its practical application in the production of (R)-(-)-mandelic acid.

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Section: Temperature and Ph Optimasupporting

confidence: 75%

Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(−)-mandelic acid production

Zhang

et al. 2010

Bioprocess Biosyst Eng

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“…The strong inhibition of the enzymes by these ions is similar to that observed for nitrilase of F. solani (Vejvoda et al, 2008). To date, it is not clear why nitrilases behave differently in the presence of different ions.…”

Section: Discussionsupporting

confidence: 69%

Purification and characterization of a cyanide-degrading nitrilase fromTrichoderma harzianum VSL291

Ricaã'o-Rodriguez¹,

Lepe²

2015

Turk J Biol

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An intracellular nitrilase (Nit1) with cyanide-degrading activity was isolated from Trichoderma harzianum VSL291, cultivated on benzonitrile as the sole carbon source. Nit1 was purified to homogeneity by ion exchange and gel filtration chromatography with a recovery of 7.15% and a fold of 22.5. The molecular weight was estimated to be 47.7 kDa and the purified enzyme was sequenced with a system of liquid chromatography and mass spectrometry (LC-MS). The enzyme consists of 436 amino acids with a predicted molecular weight of 47.088 kDa. The sequence revealed conserved domains for a nitrilase super family such as putative active and binding sites and a Glu-Lys-Cys catalytic triad. Nit1 exhibited maximum activity (19.6 U mg -1 ) at 40 °C and a pH of 7.5. Nit1 had a strong inhibition in the presence of Al 3 +, Cu 2+ , Zn 2+ , and Ag + ions and was able to degrade KCN completely at 0.02 mmol/L, 0.05 mmol/L, and 0.1 mmol/L in 15 min, 40 min, and 45 min, respectively. The effect on KCN (0.02 mmol/L) degradation was tested in the presence of Cu2+ and Ag+ ions (0.025 mmol/L to 1.0 mmol/L) and the enzymatic activity was not affected significantly at 0.025 mmol/L, 0.075 mmol/L, and 0.125 mmol/L concentrations. However, when both ions were combined, the activity of the enzyme decreased significantly.

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“…Fusarium sp. was reported to be a ''factory" producing different kinds of enzymes, such as cellulose, nitrilase as well as ligninolytic enzymes (Fernaud et al, 2006;Vejvoda et al, 2008). Investigation of the metabolic pathways has demonstrated that the fungal species should transform the PAHs into the quinone substances, and the lignin-degrading system (LDS) enzymes (including LiP, MnP and laccase) were involved in this transformation (Haritash and Kaushik, 2009).…”

Section: Production Of Ligninolysis-related Extracellular Enzymes In mentioning

confidence: 99%

Biodegradation of anthracene and benz[a]anthracene by two Fusarium solani strains isolated from mangrove sediments

Luo

Vrijmoed

2010

Bioresource Technology

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Purification and characterization of a nitrilase from Fusarium solani O1

Cited by 54 publications

References 23 publications

Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(−)-mandelic acid production

Cloning and biochemical properties of a highly thermostable and enantioselective nitrilase from Alcaligenes sp. ECU0401 and its potential for (R)-(−)-mandelic acid production

Purification and characterization of a cyanide-degrading nitrilase fromTrichoderma harzianum VSL291

Biodegradation of anthracene and benz[a]anthracene by two Fusarium solani strains isolated from mangrove sediments

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